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The New Culture of Water (NCW) is a catalyst for sustainable transformation between political agreements and the unsustainability of the real world, thus requiring an integral participation for an efficient water management where the academy will try to make a substantial contribution. Therefore, this study aimed to design, implement and evaluate four competencies: Knowledge, Attitude, Perception and Practices (KAPP) of a formal program of Education for Sustainability (EfS) on the New Water Culture (NWC), addressing issues such as prevention of water quality, conservation and efficient use at home. To carry out the evaluation, a semi-structured survey of 47 items was conducted in three stages: at the beginning (diagnostic), during the development of the (EfS) (formative) and at the end (summative). A total of 103 nursing students participated in the (EfS), divided into two groups A and B, with the addition of group C as a control group. The results showed that group A had the best results, in general. Statistical tests to calculate the mean, standard deviation (SD) and a validation with the nonparametric Wilcoxon test showing a significant change in the participants of group A, which indicated an important impact of the (EfS). This study also provides an overview of the community situation that can help trigger early warnings to identify weaknesses in water management.

Botrytis cinerea causes postharvest fruit rot of an infinity of crops, the infective capacity is due to its physiological diversity that shown, even inside the same crop. For its control, the use of antagonistic microoganisms is emerging as a sustainable option. In the present work, 40 Botrytis isolates from three vineyards were characterized by their ability to infect grape fruit (Thomson Seedless), the results showed that all produced lesions diameters from 6.5 to 22.2 mm. Ten of these isolates that presented differences in terms of their virulence, were subject to in vitro antagonism test, using the yeasts Metschnikowia sp. NB9 and FLL17 (Kodamaea sp. FLL17 and the bacteria FR4B12 Bacillus sp. R4B12 from must and flower and fruit, respectively. The results showed that, on average, FRB412 had the highest inhibitory activity on the growth of Botrytis strains, exhibiting mycelial growth inhibition percentages from 51 to 81 %, followed by FLL17 (21 to 53 %) and NB9 (15 to 51 %). In conclusion, the three study strains have different ranges of biocontrol on Botrytis, whose application could reduce gray rot in grapes.

A pre-adaptation approach to osmotic stress was used to improve the colonisation and biocontrol capacity of Yarrowia lipolytica M18/3b on the apple under pre- and postharvest conditions. For this, ...

The low quality of the apple produced in Queretaro, Mexico, impedes its efficient fresh marketing. One possible solution to this problem is to produce champagne-type sparkling cider. Therefore, the objective of this work was to select strains of Saccharomyces yeasts isolated from apple varieties of the region, based on desirable traits to make sparkling ciders. To select the yeasts, we induced the spontaneous fermentation of the must of 14 varieties, from which 135 strains (102 Saccharomyces spp.) were isolated and three (MM7, 436.4 and RY5) were selected, all with killer phenotype, β-glucosidase activity, tolerance to 8 % ethanol and 50 mg∙L-1 of SO2, a different degree of flocculation and with the ability to produce more than 3 atm of pressure in the bottle. With MM7, a base cider of good quality was obtained (glucose < 2 g∙L-1, pH 3.57, total titratable acidity of 4.03 g∙L-1 and volatile acidity of 0.27 g∙L-1). For the second fermentation, the base ciders made with MM7 were bottled, 15 g∙L-1 of sucrose were added and then they were inoculated separately with MM7, 436.4, RY5 or K1-V1116. By day 21, they all reached about 6 atm of pressure. Sensorially, ciders made with MM7 stood out in effervescence speed, and those made with RY5 in taste acceptability. The amplification and sequencing of the ITS1/ITS4 domain identified MM7 and 436.4 as S. cerevisiae, and RY5 as S. paradoxus. The selected yeasts showed desirable traits for the production of quality sparkling ciders typical of the region.

Background Two edible coating (EC) emulsions based on potato starch (F6 and F10) alone or formulated with sodium benzoate (SB, 2% w/w) (F6/SB and F10/SB) were evaluated to maintain postharvest quality of cold-stored 'Fino' lemons and control sour rot on lemons artificially inoculated with Geotrichum citri-aurantii. Previous research showed the potential of these ECs to improve the storability of 'Orri' mandarins and reduce citrus green and blue molds caused by Penicillum digitatum and Penicillium italicum, respectively. Results The coatings F6/SB and F10/SB significantly reduced sour rot incidence and severity compared to uncoated control samples on lemons incubated at 28 °C for 4 and 7 days. The F6/SB coating reduced weight loss and gas exchange compared to uncoated fruit after 2 and 4 weeks of storage at 12 °C plus a shelf life of 1 week at 20 °C, without adversely affecting the lemon physicochemical quality. Conclusion Overall, the F6/SB coating formulation, composed of pregelatinized potato starch, glyceryl monostearate, glycerol, emulsifiers and SB, with a total solid content of 5.5%, showed the best results in reducing citrus sour rot and maintaining the postharvest quality of cold-stored 'Fino' lemons. Therefore, it showed potential as a new cost-effective postharvest treatment suitable to be included in integrated disease management programs for citrus international markets with zero tolerance to chemical residues. © 2021 Society of Chemical Industry.

Starch is a biodegradable polymeric carbohydrate that can easily form films and coatings and can readily be obtained from some food industry by-products and wastes, which may contribute to the circular bioeconomy. In this work, we studied the potential of two edible coating emulsions based on pregelatinized potato starch (PPS) and glyceryl monostearate (GMS) alone (F6 and F10) or formulated with the food additive sodium benzoate (SB, 2%) (F6/SB and F10/SB) to control sour rot, an important citrus postharvest disease caused by the fungus Geotrichum citri-aurantii, and maintain postharvest quality of cold-stored ‘Orri’ mandarins. The PPS-GMS coating application was compared to dipping in water (uncoated controls) and dipping in a 2% SB (w/v) aqueous solution. The results showed that the coating F10/SB was the most promising treatment to control sour rot on mandarins, with reductions in disease incidence with respect to the uncoated control samples of 94, 69, and 55% after 2, 4, and 6 weeks of storage at 5 °C, respectively. Coatings formulated without SB were ineffective. Regarding fruit quality, the coating F10 was the most effective to reduce weight loss, maintain firmness, and provide gloss on mandarins stored at 5 °C for up to 6 weeks followed by a shelf-life period of 1 week at 20 °C. The addition of SB to the PPS-GMS coatings adversely affected these coating properties, but the coating F10/SB still reduced weight loss compared to uncoated controls without negatively affecting the fruit physicochemical (juice titratable acidity, soluble solids content, and volatiles content) and sensory quality (overall flavor, off-flavors, external aspect). Overall, the coating F10/SB showed the greatest potential for commercial use as an efficient and environmentally friendly alternative to conventional fungicides and waxes for sour rot control and quality preservation of cold-stored mandarins.

Esta obra contiene siete aportaciones de académicos que examinan la dimensión territorial de variantes del turismo de intereses especiales. En el desarrollo de estas investigaciones se privilegió examinar, desde una perspectiva geográfica, la interrelación naturaleza-sociedad materializada en el acomodo del turismo en diferentes lugares de México, Costa Rica y Paraguay. Las estrategias metodológicas conducentes fueron distintas: algunas optaron por el acopio de información proporcionada por las comunidades involucradas; otras privilegiaron los recorridos y trabajo en campo, hechos en forma intensiva; también se recurrió a novedosas maneras de compilar referencias valiosas por medio de las actuales tecnologías en información y comunicación. Al ser este un libro sobre Geografía del turismo, la producción cartográfica original ocupa un sitio privilegiado desde el que se proclaman los principales hallazgos de investigación de cada capítulo. Es intención de los autores que los estudios aquí contenidos resulten, en un futuro, significativos para esfuerzos académicos similares.

Se presenta el resultado de un proceso de investigación desarrollada en dos Escuelas Secundarias Generales del estado de Durango, México; con el objetivo de identificar la percepción que tienen los estudiantes acerca de los procesos de atención a la multiplicidad de diferencias dentro de sus escuelas de pertenencia. En el proceso de desarrollo de la investigación se hacen hallazgos en el campo de la conceptualización en lo relativo: a) la forma de abordar laatención a la multiplicidad de diferencias, término que surge de una asunción crítica, histórica, semiótica y epistémica en la que se incorpora la idea de lo individual como elemento de enriquecimiento y aprendizaje; b) la perspectiva crítica y acrítica con la que los estudiantes se representan la inclusión y c) la certeza metodológica de abordar los proceso de investigación en este campo desde una perspectiva de complementariedad metodológica que dé cuenta de los elementos generales y particulares que dan forma a las prácticas educativas y sociales en las propias escuelas. Se concluye que los estudiantes de secundaria cuyas edades oscilan entre los 12 y 15 años de edad, perciben en forma escasamente crítica los procesos de atención a la multiplicidad de diferencias en sus instituciones; se advierte un desconocimiento de la educación y convivencia como derecho básico, que permita explorar los factores incidentes en el éxito o fracaso de la implementación de políticas inclusivas. Se detecta un campo de oportunidad en la exploración de la representación social de laInclusión Educativainstitucionalizada por el sistema educativo nacional y las posibilidades de institucionalización, que los propios colectivos escolares pueden incorporar en sus prácticas de manera que sea posible conceptualizar de una manera crítica y transformadora las prácticas educativas, de tal forma que a partir de la asunción de nuevas formas de acción no segregadoras se transite desde un modelo de inclusiónreformista institucionalizantehacia un modelo que pugne por unatransformación institucionalizada.

Se presenta el resultado de un proceso de investigación desarrollada en dos Escuelas Secundarias Generales del estado de Durango, México; con el objetivo de identificar la percepción que tienen los estudiantes acerca de los procesos de atención a la multiplicidad de diferencias dentro de sus escuelas de pertenencia. En el proceso de desarrollo de la investigación se hacen hallazgos en el campo de la conceptualización en lo relativo: a) la forma de abordar la atención a la multiplicidad de diferencias, término que surge de una asunción crítica, histórica, semiótica y epistémica en la que se incorpora la idea de lo individual como elemento de enriquecimiento y aprendizaje; b) la perspectiva crítica y acrítica con la que los estudiantes se representan la inclusión y c) la certeza metodológica de abordar los proceso de investigación en este campo desde una perspectiva de complementariedad metodológica que dé cuenta de los elementos generales y particulares que dan forma a las prácticas educativas y sociales en las propias escuelas. Se concluye que los estudiantes de secundaria cuyas edades oscilan entre los 12 y 15 años de edad, perciben en forma escasamente crítica los procesos de atención a la multiplicidad de diferencias en sus instituciones; se advierte un desconocimiento de la educación y convivencia como derecho básico, que permita explorar los factores incidentes en el éxito o fracaso de la implementación de políticas inclusivas. Se detecta un campo de oportunidad en la exploración de la representación social de la Inclusión Educativa institucionalizada por el sistema educativo nacional y las posibilidades de institucionalización, que los propios colectivos escolares pueden incorporar en sus prácticas de manera que sea posible conceptualizar de una manera crítica y transformadora las prácticas educativas, de tal forma que a partir de la asunción de nuevas formas de acción no segregadoras se transite desde un modelo de inclusión reformista institucionalizante hacia un modelo que pugne por una transformación institucionalizada.

Antifungal composite edible coatings (ECs) formulated with pregelatinized potato starch (PPS, 1.0-2.0 % w/w) as biopolymer, glyceryl monostearate (GMS, 0.5-1.5 %, w/w) as hydrophobe, glycerol (Gly, 0.5-1.5 %, w/w) as plasticizer, and sodium benzoate (SB, 2 % w/w) as antifungal agent were optimized using the Box–Behnken response surface methodology to extend the postharvest life of Orri’ mandarins. The second order polynomial models satisfactorily fitted the experimental data, with high values of the coefficient of determination for the different variables (R2>0.91). The individual linear effect of GMS concentration was significant in all the responses evaluated, whereas PPS only affected emulsion viscosity, fruit tacking, and weight loss of coated mandarins. Gly only affected acetaldehyde content in the juice of coated mandarins when interacted with PPS and in the quadratic effect. The optimum concentrations of PPS, GMS, and Gly for the starch-based EC based on maximum fruit quality and required emulsion properties were predicted to be 2.0, 0.5 and 1.0 % (w/w), respectively. The optimal EC reduced weight loss of mandarins and created a modified atmosphere within the fruit without negatively affecting the overall acceptability of the fruit. On the other hand, the optimized antifungal EC containing SB significantly reduced postharvest green and blue molds and sour rot on mandarins artificially inoculated with the pathogens Penicillium digitatum, Penicillium italicum and Geotrichum citri-aurantii, respectively. Therefore, the optimized antifungal EC showed potential to control the main postharvest diseases and maintain the overall quality of ‘Orri’ mandarins and could be a suitable alternative to commercial citrus waxes formulated with conventional chemical fungicides.

Background The Mexican arm of the Camino Verde trial of community mobilisation for dengue prevention covered three coastal regions of Guerrero state: Acapulco, Costa Grande and Costa Chica. A baseline cross-sectional survey provided data for community mobilisation and for adapting the intervention design to concrete conditions in the intervention areas. Methods Trained field teams constructed community profiles in randomly selected clusters, based on observation and key informant interviews. In each household they carried out an entomological inspection of water containers, collected information on socio-demographic variables and cases of dengue illness among household members in the last year, and gathered paired saliva samples from children aged 3–9 years, which were subjected to ELISA testing to detect recent dengue infection. We examined associations with dengue illness and recent dengue infection in bivariate and then multivariate analysis. Results In 70/90 clusters, key informants were unable to identify any organized community groups. Some 1.9% (1029/55,723) of the household population reported dengue illness in the past year, with a higher rate in Acapulco region. Among children 3–9 years old, 6.1% (392/6382) had serological evidence of recent dengue infection. In all three regions, household use of anti-mosquito products, household heads working, and households having less than 5 members were associated with self-reported dengue illness. In Acapulco region, people aged less than 25 years, those with a more educated household head and those from urban sites were also more likely to report dengue illness, while in Costa Chica and Costa Grande, females were more likely to report dengue illness. Among children aged 3–9 years, those aged 3–4 years and those living in Acapulco were more likely to have evidence of recent dengue infection. Conclusions The evidence from the baseline survey provided important support for the design and implementation of the trial intervention. The weakness of community leadership and the relatively low rates of self-reported dengue illness were challenges that the Mexican intervention team had to overcome. The higher dengue illness occurrence among women in Costa Grande and Costa Chica may help explain why women participated more than men in activities during the Camino Verde trial.

Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) for postharvest diseases of citrus and pome fruits. However, for registration purposes and to implement their use as effective control strategy, it is necessary to study the traceability and survival of BCAs in their target application sites. The main objective of this work was to evaluate the persistence and quantify the population of CPA-2 after its postharvest application on orange cultivar 'Valencia Late' by molecular techniques. After application, the persistence of CPA-2 was evaluated by sampling the packing line and storage chambers, as well as on clothing of the workers by conventional PCR. The results showed that the maximum persistence of CPA-2 was lower than 3 days in surfaces of packing line. Furthermore, CPA-2 did not survive more than 1 day on working clothes, while in the environment or on different storage chamber surfaces it was not detected. In addition, the CPA-2 populations were quantified by quantitative PCR (qPCR) combined with a DNA intercalating reagent, propidium monoazide dye (qPCR-PMA) to quantify the CPA-2 viable cells on fruit surface. The qPCR-PMA method was compared with qPCR and dilution plating method. Results showed that CPA-2 populations quantified by qPCR-PMA were significantly different compared with those obtained by qPCR during the time-course of the assay; however, no significant differences were observed between qPCR-PMA and dilution plating. In conclusion, the persistence of CPA-2 was low at different sampling areas, suggesting that it cannot grow and survive on the surfaced sampled. Furthermore, qPCR-PMA method can be a quick and specific tool to monitor the viable population of CPA-2 on fruit surface.

Background: Oxidative stress is the predominant pathogenic mechanism of ischaemia-reperfusion (IR) injury. The noble gas argon has been shown to alleviate oxidative stress-related myocardial and cerebral injury. The risk of lung IR injury is increased in some major surgeries, reducing clinical outcome. However, no study has examined the lung-protective efficacy of argon preconditioning. The present study investigated the protective effects of argon preconditioning on airway epithelial cells exposed to hydrogen peroxide (H2O2) to induce oxidative stress. Methods: A549 airway epithelial cells were treated with a cytotoxic concentration of H2O2 after exposure to standard air or 30 or 50% argon/21% oxygen/5% carbon dioxide/rest nitrogen for 30, 45 or 180 min. Cells were stained with annexin V/propidium iodide, and apoptosis was evaluated by fluorescence-activated cell sorting. Protective signalling pathways activated by argon exposure were identified by Western blot analysis for phosphorylated candidate molecules of the mitogen-activated protein kinase and protein kinase B (Akt) pathways. Results: Preconditioning with 50% argon for 30, 45 and 180 min and 30% argon for 180 min caused significant protection of A549 cells against H2O2-induced apoptosis, with increases in cellular viability of 5-47% (p < 0.0001). A small adverse effect was also observed, which presented as a 12-15% increase in cellular necrosis in argon-treated groups. Argon exposure resulted in early activation of c-Jun N-terminal kinase (JNK) and p38, peaking 10- 30 min after the start of preconditioning, and delayed activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway, peaking after 60-90 min. Conclusions: Argon preconditioning protects airway epithelial cells from H2O2-induced apoptotic cell death. Argon activates the JNK, p38, and ERK1/2 pathways, but not the Akt pathway. The cytoprotective properties of argon suggest possible prophylactic applications in surgery-related IR injury of the lungs.

BACKGROUND The noble gas argon induces cardioprotection in a rabbit model of myocardial ischemia and reperfusion. However, no studies in human primary cells or subjects have been performed so far. We used human cardiac myocyte-like progenitor cells (HCMs) to investigate the protective effect on the cellular level. METHODS HCMs were pretreated with 30% or 50% argon before oxygen-glucose deprivation (OGD) and reperfusion. We evaluated apoptotic states by flow cytometry and the activation of mitogen-activated protein kinase (MAPKs) members extracellular signal-regulated kinase (ERK), c-jun N-terminal kinase (JNK), p38 MAPkinase, and protein kinase B (Akt) by Westernblot analysis and by activity assays of downstream transcription factors. Specific inhibitors were used to proof a significant participation of these pathways in the protection by argon. Beneficial effects were further assessed by TdT-mediated dUTP-biotin nick end labeling (TUNEL) assay, lactate dehydrogenase (LDH), mitochondrial deoxyribonucleic acid (mtDNA), and cytokine release. RESULTS Pretreatment with 30% or 50% argon for 90 min before OGD resulted in a significant protection of HCMs against apoptosis. This effect was reversed by the application of MAPK and Akt inhibitors during argon exposure. Argon 30% reduced the release of LDH by 33% and mtDNA by 45%. The release of interleukin 1β was reduced by 44% after OGD and more than 90% during reperfusion. CONCLUSIONS Pretreatment with argon protects HCMs from apoptosis under ischemic conditions via activation of Akt, Erk, and biphasic regulation of JNK. Argon gas is cheap and easily administrable, and might be a novel therapy to reduce myocardial ischemia-reperfusion injury.

Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) against the major postharvest pathogens present on pome and citrus fruits. Dehydration, such as freeze-drying, spray-drying and fluidized bed drying is one of the best ways to formulate BCAs. In this work, the survival of CPA-2 cells after formulation was determined by dilution plating and molecular methods as qPCR alone and combined with a sample pretreatment with a propidium monoazide dye (PMA-qPCR) and they were used to calculate treatment concentrations in efficacy trials on postharvest oranges. Furthermore, no significant differences in CPA-2 survival were observed as determined by dilution plating and PMA-qPCR after both the freeze drying and fluidized bed drying processes; however, an interesting significant difference was observed in the spray dried product comparing all quantitative methods. A difference of 0.48 and 2.17 log10 CFU or cells g/dw was observed among PMA-qPCR with qPCR and dilution plating, respectively. According to our study, dilution plating was shown to be an unreliable tool for monitoring the survival of CPA-2 after spray drying. In contrast, the combination of PMA and qPCR enabled a quick and unequivocal methodology to enumerate viable and VBNC CPA-2 cells under stress-dried conditions. Efficacy trials showed that, after 3 days, spray drying formulation rehydrated with 10% non-fat skimmed milk (NFSM) was as effective as fresh cells to control Penicillium digitatum in oranges.

The aim of this study was to evaluate the environmental fate and behaviour of the biocontrol product based on Pantoea agglomerans CPA-2 cells after its release into the field. Three different methods, dilution plating, quantitative real-time PCR (qPCR) and propidium monoazide combined with qPCR (PMA-qPCR) were used to track the CPA-2 population. Populations quantified by qPCR were statistically different compared with those by the PMA-qPCR and dilution plating methods. In addition, the spray dispersion of the CPA-2 treatment was evaluated using water-sensitive papers. A lack of dispersion of the CPA-2 treatment was observed at distances of 2.5 ± 0.5 m. The presence and persistence of CPA-2 in the environment were monitored in different areas, by dilution plating and confirmed by conventional PCR. The results showed that CPA-2 can survive but not proliferate on weed and leaves. The persistence on inert surfaces, such as motorised backpack sprayer, gloves and working clothes, was less than 7 days. In conclusion, PMA-qPCR was a potential tool for fast and specific monitoring of viable populations of CPA-2 and it gave valuable information on population behaviour. Moreover, the results demonstrate that CPA-2 would not present a risk for the environment because of its low establishment, survival and dispersion.

Pantoea agglomerans strain CPA-2 is an effective biocontrol agent (BCA) for postharvest diseases of citrus and pome fruit. To implement their use as a control strategy is necessary to study the traceability of BCAs in the environment during application, for registration issues. In this study, the presence and persistence of CPA-2 was monitored in the packing line, storage chambers and on working clothes by conventional PCR. After postharvest application, the presence of CPA-2 was not detectable in the environment and storage chambers, whereas on working clothes and the packing line its persistence was less than 1 and 3 days, respectively. Additionally, the CPA-2 population was quantified on oranges stored at two different temperatures (20 °C and 4 °C) by quantitative PCR (qPCR), sample pretreatment with a propidium monizade dye (PMA–qPCR) and the dilution plating method. At the initial time of the assay, no differences were observed in CPA-2 populations quantified by qPCR, PMA–qPCR, and dilution plating, at both storage temperatures. However, CPA-2 populations quantified by PMA–qPCR were significantly different compared with those obtained by qPCR during the time-course of the assay; no significant differences were observed between PMA–qPCR and dilution plating. In conclusion, the persistence of P. agglomerans CPA-2 at different sampling areas after postharvest application was low. Furthermore, PMA–qPCR gave valuable information on viable population behavior and the presence of residual DNA from dead cells. In general, these studies help to understand the persistence of antagonists when applied under postharvest conditions and will lead to optimization of time and mode of application.

BACKGROUND Perioperative oxygen (O2) therapy can cause hyperoxia. Extreme hyperoxia can injure the cardiovascular system and remote organs. OBJECTIVE Our primary objective was to test the hypothesis that exposure to moderate hyperoxia will induce injury to human umbilical vein endothelial cells (HUVECs), a model for studying the vascular endothelium under controlled conditions. DESIGN In-vitro cell culture study. SETTING Department of Anaesthesia, General Intensive Care and Pain Management, Medical University of Vienna, Austria. Study period from the beginning of October 2013 to the end of July 2014. CELLS HUVECs were isolated from fresh umbilical cords. INTERVENTIONS HUVECs were exposed to constant hyperoxia (40% O2), cyclic hyperoxia/anoxia (40%/0% O2, average 20% O2), constant normoxia (21% O2) and constant anoxia (0% O2) using a cell culture bioreactor. MAIN OUTCOME MEASURES Cell growth, viability and release of IL-6, IL-8 and macrophage migration inhibitory factor were assessed at baseline and after 6, 12, 24 and 48 h of treatment. A phosphokinase array was performed after 60 min of treatment to identify activated cellular signalling pathways. RESULTS Constant hyperoxia and cyclic hyperoxia/anoxia impeded cell growth, reduced viability, triggered a proinflammatory response, proven by IL-6, IL-8 and migration inhibitory factor release, and induced apoptosis and necrosis. The inflammatory and cytotoxicity responses were highest in the constant hyperoxia group. Phosphokinase arrays revealed that different O2 concentrations activated distinct sets of cytoprotective and cell death-associated kinases, including mitogen-activated protein kinases, Src kinases, p53, Akt, mitogen-activated and stress-activated kinase, Lyn, Lck, p70S6, signal transducers and activators of transcription 5b and 6, glycogen synthase kinase 3a/b and 5' AMP-activated protein kinases 1/2. CONCLUSION Continuous moderate hyperoxia and cyclic moderate hyperoxia/anoxia-induced endothelial inflammation, apoptosis and necrosis. Given the large surface area of the vascular endothelium, moderately elevated O2 levels may contribute to cardiovascular inflammation and injury. TRIAL REGISTRATION This in-vitro study was not registered in a database.

The aim of this study was to determine the effects of maturity stage, ripening time, year, and several quality parameters of the fruit (total soluble solids, acidity, pH, firmness, and color) on the susceptibility to Penicillium expansum of apple genotypes established in Queretaro, Mexico. The accessions consisted in cultivars, creoles and low chilling hybrids obtained in Mexico. The development of the disease was determined by measuring the lesion diameter of decay (severity) on fruits inoculated with 104 spores mL−1 and incubated for 8 days at 26 °C. Data were analyzed for significant difference and for significant correlation by analysis of variance and principal component analysis (PCA), respectively. In 2011, commercial maturity apples exhibited a greater tolerance to P. expansum than did over-mature apples (lesion diameters of 31.3 and 34.9 mm, respectively); however, the ripening time of genotypes did not influence the development of the pathogen in either year of study (21.8, 23.2, and 25.5 mm in diameter for early-, middle-, and late-ripening genotypes, respectively, in 2012). Differences in the susceptibility of genotypes were observed in both harvest years; the most outstanding were ‘436’, ‘442-9’ and ‘441’ (17.4, 17.5 and 17.6 mm, respectively) in contrast to ‘Golden Delicious’ (34.2 mm) in 2012; nevertheless, no correlation was observed in genotypes behavior between the two harvest years (r = 0.11NS). Excluding total soluble solids in 2012, PCA analysis did not reveal any correlation between pathogen severity and fruit quality parameters, suggesting the possibility that other mechanisms are involved in the fruit-pathogen interaction. This study demonstrated for the first time that ripening time does not affect the susceptibility to P. expansum in apple fruits.

Dilution plating is the quantification method commonly used to estimate the population level of postharvest biocontrol agents, but this method does not permit a distinction among introduced and indigenous strains. Recently, molecular techniques based on DNA amplification such as quantitative real-time PCR (qPCR) have been successfully applied for their high strain-specific detection level. However, the ability of qPCR to distinguish viable and nonviable cells is limited. A promising strategy to avoid this issue relies on the use of nucleic acid intercalating dyes, such as propidium monoazide (PMA), as a sample pretreatment prior to the qPCR. The objective of this study was to optimize a protocol based on PMA pre-treatment samples combined with qPCR to distinguish and quantify viable cells of the biocontrol agent P. agglomerans CPA-2 applied as a postharvest treatment on orange. The efficiency of PMA-qPCR method under the established conditions (30μM PMA for 20min of incubation followed by 30min of LED light exposure) was evaluated on an orange matrix. Results showed no difference in CFU or cells counts of viable cells between PMA-qPCR and dilution plating. Samples of orange matrix inoculated with a mixture of viable/dead cells showed 5.59log10 CFU/ml by dilution plating, 8.25log10 cells/ml by qPCR, and 5.93log10 cells/ml by PMA-qPCR. Furthermore, samples inoculated with heat-killed cells were not detected by dilution plating and PMA-qPCR, while by qPCR was of 8.16log10 cells/ml. The difference in quantification cycles (Cq) among qPCR and PMA-qPCR was approximately 16cycles, which means a reduction of 65,536 fold of the dead cells detected. In conclusion, PMA-qPCR method is a suitable tool for quantify viable CPA-2 cells, which could be useful to estimate the ability of this antagonist to colonize the orange surface.

The registration of biological control agents requires the development of monitoring systems to detect and quantify the agent in the environment. Pantoea agglomerans CPA-2 is an effective biocontrol agent for postharvest diseases of citrus and pome fruits. The monitoring of CPA-2 in postharvest semi-commercial trials was evaluated by Rodac impression plates and the colonies isolated were confirmed by conventional PCR using the SCAR primers PAGA1 and PAGB1. Samples were taken from different surfaces that had contact with CPA-2, the surrounding environment and working clothes worn by handlers. Moreover, population dynamics of the strain CPA-2 were determined on apple surfaces using both the classical plating technique and real-time quantitative PCR (qPCR). A qPCR assay using a 3'-minor groove-binding (MGB) probe was developed for the specific detection and quantification of P. agglomerans strain CPA-2. Based on the nucleotide sequence of a SCAR fragment of CPA-2, one primer set and TaqMan MGB probe were designed. The primers SP2-F/SP2-R and the TaqMan MGB probe showed a specific detection of strain CPA-2 on apple surfaces, which was verified tested against purified DNA from 17 strains of P. agglomerans, 4 related Pantoea species, and 21 bacterial strains from other genera isolated from whole and also freshly-cut fruit and vegetables. The detection level was approximately 10(3) cells per reaction, and the standard curve was linear within a range of 5log units. Results from semi-commercial trials showed that CPA-2 had a low impact. The maximum persistence of P. agglomerans CPA-2 was not longer than 5days in plastic boxes stored at 0°C. Significant differences in CPA-2 population level dynamics were observed in results obtained by qPCR and dilution plating. These differences may indicate the presence of non-degraded DNA from non-viable cells. In conclusion, qPCR is a novel potential tool to quickly and specifically monitor recent surface colonisation by CPA-2 populations on apple surfaces during large-scale experiments that could ensure efficient and successful treatments.

Native lactic acid bacteria (LAB) are capable of growing during winemaking, thereby strongly affecting wine quality. The species of LAB present in musts, wines during malolactic fermentation (MLF), and barrels/filters were investigated in wineries from the emerging wine region of Queretaro, México using multiplex PCR and culture. The resistance to wine-like conditions (WLC): ethanol (10, 12, and 13%), SO2 (30 mg⋅l-1), and low pH (3.5) of native LAB strains was also studied. Five species were detected within 61 samples obtained: Oenococcus oeni, Lactobacillus plantarum, Pediococcus parvulus, Lactobacillus hilgardi, and Lactobacillus brevis. Four species (excepting L. brevis) were found in must; O. oeni and P. parvulus were ubiquitous in wine and L. plantarum and L. brevis were mainly present at the initial stage of MLF, while L. hilgardii was mostly detected at the advanced stage. Furthermore, some species detected in barrel/filter, prove them to be hazardous reservoirs. From 822 LAB isolates, only 119 resisted WLC with 10% ethanol; the number of strains able to grow in WLC with 13% ethanol decreased approximately by 50%, O. oeni being the most versatile species with 65% of resistant isolates, while Lactobacillus spp. and P. parvulus were the most strongly affected, especially those recovered from barrel/filter, with less than 10% of resistant isolates. This study evidences the presence of local strains able to be used as starter cultures, and also enabled the assessment of the risks derived from the presence of spoilage LAB strains resistant to WLC.

Objective To investigate biomarkers of cytogenotoxicity in women exposed to mercury in a mining area. Mercury has been associated with cytogenotoxicity in human and animal models but has not been considered carcinogenic in humans, even though genotoxic effects following exposure to inorganic mercury compounds have been observed. Methods A cross-sectional study and micronucleus assay in uroepithelial cells were performed in 104 women (12e84 years of age). First urine void samples were taken to determine creatinine-adjusted mercury levels in urine (HgUCr), to isolate cells and to quantify cytogenetic damage. Results The geometric average level for HgUCr was 4.9 mg/g (range, 0.4e85). In the generalised linear model, after controlling for other co-variables, we observed that for each 10 mg/g increase in HgUCr, the OR of developing a genotoxic effect was 2.37 (95% CI 1.79 to 2.84), while for cytotoxic damage in uroepithelial cells the OR was 1.34 (95% CI 1.10 to 1.79). These results suggest a possible association between cytogenotoxicity and HgUCr. Conclusion Living in a mining area with exposure to inorganic mercury and having higher mercury levels in urine increased the risk of developing uroepithelial cytogenotoxicity.

OBJECTIVES/SPECIFIC AIMS: To diversify the biomedical research workforce by training postdoctoral scholars and junior faculty from 6 Minority Serving Institutions (MSIs) on practical research skills such as Critical and Creative Thinking, Formulating the Problem, Asking the Right Question, Grant Writing, and Team Science METHODS/STUDY POPULATION: In collaboration with our partners, we identified 11 topics where trainees lack research funding. Next, we identified instructors for these topics. We converted the topics to online module with modules ranging from 2 to 8 weeks. In working with an online education expert, we developed innovative online training using Moodle as the content management system. Scholars complete readings, videos, self-assessments and participate in discussion board each week. In addition, we have weekly synchronous sessions for each module. All scholars are required to take the grant writing module and 8 other modules. After each module, trainees complete a brief survey to evaluate the module. The leaders at the MSI participated in an intensive face-to-face training session on how to be a career coach so that they could be career coaches for the LEADS Scholars at their home institutions. RESULTS/ANTICIPATED RESULTS: In the first year, we selected 13 LEADS Scholars. All but 3 scholars elected to take every module. The 3 scholars did not enroll in the Peer Reviewing module. Results of the brief survey at the end of each module indicate that the scholars value each of the modules and rate them very highly. When 1 scholar wanted to leave the program, we decided to have a conference call with all of the LEADS Scholars to determine what was working and what was not working with the program. All scholars recognized the value of LEADS. Some scholars felt that the weekly synchronous session was too demanding as they have competing demands on their time. We consulted with the leadership at the MSI and decided to modify the requirements of the program such that every synchronous call was not required for successful completion of the module and to earn a badge. Scholars need to have at least 9 badges to earn a certificate. In addition to the training, we decided that scholars would also benefit from mock reviews of their grants. This will help them submit successful grants. We learned that the best way to serve the needs of the scholars is to work iteratively with the scholars and leadership to develop a successful program that most effectively meets their needs of the scholars and helps them launch a successful career. DISCUSSION/SIGNIFICANCE OF IMPACT: Postdoctoral scholars and junior faculty from MSI need practical research training to help launch their research career. We suspect that this is true of many institutions and plan to develop these modules so that they can be widely disseminated to other institutions.

Blood lead levels (BLL) in children are associated with lead in soil and represent a major public health problem; however, there are few reports of lead contamination related to mining waste sites in Mexico. Therefore, we conducted a cross-sectional study in the State of Guerrero to identify the association between proximity of residence to mining site waste and BLL in children. The impact of the different variables related to BLL were analyzed with logistic regression. Geometric mean BLL was 13.6 μg/dL, 15.9 μg/dL in communities proximal to waste sites and 5.5 μg/dL in a distant control community. Children living in communities near the mining waste have higher BLL that children living in communities far away from the waste. Our results are similar to studies in other countries and indicate that mining waste sites remain an active source of BLL contamination that affect children’s health.

To determine the risk factors associated with recurrent retinal detachment in patients vitrectomized.Case-control study nested in a cohort. We included patients with a diagnosis of rhegmatogenous retinal detachment treated with vitrectomy for one year, with a six months follow-up. We monitored the presence of incident event in the emergency department, and the examination room. Statistical analysis was performed by descriptive statistics (median, minimum and maximum) and inferential statistics (chi-squared test, Cox regression, Kaplan-Meier).A population of 94 patients with a median age of 55 years (21-83) was analyzed. The silicone was the most widely used type of tamponade in 62 patients (66 %) and recurrent retinal detachment occurred in 19 cases (20 %). In the relation between the non-relapse group and the relapse group, the statistically significant variable was the use of silicone (p = 0.05, OR = 0.03, 0.013-0.98), wich had a lower probability of recurrent detachment compared with the use of sulfur hexafluoride (SF6 gas) (p = 0.012).The use of silicone compared with the use of SF6 gas has shown to be a protective factor for recurrent retinal detachment; however, the choice of method of tamponade should be individualized for each patient.determinar los factores de riesgo quirúrgico relacionados con el desprendimiento de retina recidivante en pacientes que habían sido tratados con vitrectomía. MÉTODOS: estudio de casos y controles anidado en una cohorte recolectada durante un año. Se incluyeron pacientes con diagnóstico de desprendimiento regmatógeno de retina tratados con vitrectomía, con un seguimiento de seis meses. Se monitorizó la presencia del evento incidente en el servicio de urgencias y en la consulta. El análisis de los datos se realizó mediante estadística descriptiva (medianas, mínimos y máximos) e inferencial (chi-squared, regresión de Cox y método de Kaplan-Meier).se analizaron 94 pacientes con mediana de edad de 55 años (rango de 21 a 83 años). La silicona fue el material de taponamiento utilizado en 62 pacientes (66 %) y la recidiva se presentó en 19 (20 %). En la relación entre el grupo sin recidiva y el grupo con recidiva, la variable estadísticamente significativa fue el uso de silicona (p = 0.05, OR = 0.03, 0.013-0.98), el cual tuvo menor probabilidad de redesprendimiento comparado con el del hexafluoruro de azufre (p = 0.012).el uso de silicona comparado con el del gas SF6 mostró ser un factor protector para el desarrollo de redesprendimiento de retina, sin embargo, la decisión de que elemento emplear para el taponamiento debe individualizarse.