Rationale, formative research, and protocol for Calma, Conversa, y Cría: A pilot mindful parenting intervention with Latina women.

Background: Latina mothers' stress is associated with their children's health behaviors and risk for obesity; however, existing pediatric health promotion programs have not focused on maternal stress reduction. Methods: Herein we describe a study design that will examine the acceptability and feasibility of Calma, Conversa, y Cría (CCC) a 6-week mindful parenting intervention designed to reduce stress. We present the results of qualitative research with Latina mothers and experts in Latinx health and mindfulness who provided culturally-relevant feedback on existing mindful parenting strategies to inform the development of CCC. Fifty Latina mothers of children ages 3-11 years will be randomly assigned to CCC or an enhanced usual care health education intervention. Acceptability will be assessed through participant satisfaction surveys and exit interviews. Feasibility will be determined through detailed tracking of recruitment, retention, and attendance rates. A signal regarding any group differences in maternal stress, health-related parenting practices, child diet, child physical activity, and child quality of life will be explored. Discussion: The development of interventions that can reduce maternal stress and risk for obesity in Latinx children is critical to significantly reduce negative health impacts in this underserved population. Our approach includes the identification of effective cultural adaptations that should improve the feasibility and acceptability of mindful parenting strategies in Latinx families, ideally reducing maternal stress and improving parenting behaviors related to child health. If successful, CCC will be examined in a larger efficacy trial involving the measurement of objective biomarkers of children's chronic disease risk.

Resiliencia y violencia en el noviazgo: un estudio explicativo-predictivo e instrumental con adolescentes colombianos

Introducción: los estudios sobre resiliencia y violencia psicológica en parejas adolescentes en Colombia son escasos, pues se ha explorado con mayor frecuencia la violencia en parejas adultas; además, la disposición de óptimos instrumentos de medida de la violencia psicológica en el cortejo adolescente es limitada. Objetivos: analizar la influencia de la resiliencia en la violencia psicológica en parejas adolescentes; para ello, inicialmente se validó el Cuestionario de Violencia Psicológica en el Cortejo (PDV-Q). Método: el estudio es explicativo-predictivo y de tipo instrumental. La muestra incidental estuvo conformada por 700 adolescentes colombianos, con edades entre 15 a 20 años (M = 18.40; DT = 1.56; 29.3 % hombres). Se reconoció la confiabilidad, la validez de contenido y de constructo con análisis factoriales; posteriormente, se analizaron los modelos de ecuaciones estructurales (SEM). Resultados: se reconocieron evidencias adecuadas de validez y valores óptimos de confiabilidad del PDV-Q. El modelo SEM sugiere influencias inversas de la competencia personal sobre el rol de agresión y victimización; así como una influencia directa de la aceptación de uno mismo y la vida sobre la victimización. Conclusiones: los bajos niveles de aceptación de sí mismo y de la vida podrían ser un factor de riesgo de victimización; mientras que un alto desarrollo de competencias personales sería un factor de protección ante la victimización y la agresión en el cortejo adolescente. El PDV-Q posee óptimas propiedades psicométricas y es adecuado para medir la violencia psicológica en parejas de adolescentes colombianos.

Introduction: Studies on resilience and psychological violence in adolescent couples in Colombia are scarce, since violence in adult couples has been explored more frequently; in addition, the availability of optimal instruments for measuring psychological violence in adolescent courtship is limited. Objectives: Analyze the influence of resilience on psychological violence in adolescent couples; for this, the Psychological Violence Questionnaire in Partners (PVD-Q) was initially validated. Method: The study is explanatory-predictive and instrumental. The incidental sample consisted of 700 Colombian adolescents, aged 15 to 20 years (M = 18.40; SD = 1.56; 29.3 % men). Reliability, content and construct validity were recognized with factor analysis; Subsequently, structural equation models (SEM) were analyzed. Results: Adequate evidence of validity and optimal reliability values of the PVD-Q were recognized. The SEM model suggests inverse influences of personal competence on the role of aggression and victimization; as well as a direct influence of the acceptance of oneself and life on victimization. Conclusions: Low levels of acceptance of oneself and of life could be a risk factor for victimization; while a high development of personal skills would be a protective factor against victimization and aggression in adolescent courtship. The PVD-Q has optimal psychometric properties and is suitable for measuring psychological violence in couples of Colombian adolescents.

Physiological and transcriptome analyses of Kluyveromyces marxianus reveal adaptive traits in stress response.

Kluyveromyces marxianus is a non-conventional yeast with outstanding physiological characteristics and a high potential for lignocellulosic ethanol production. However, achieving high ethanol productivity requires overcoming several biotechnological challenges due to the cellular inhibition caused by the inhibitors present in the medium. In this work, K. marxianus SLP1 was adapted to increase its tolerance to a mix of inhibitory compounds using the adaptive laboratory evolution strategy to study the adaptation and stress response mechanisms used by this non-Saccharomyces yeast. The fermentative and physiological parameters demonstrated that the adapted K. marxianus P8 had a better response against the synergistic effects of multiple inhibitors because it reduced the lag phase from 12 to 4 h, increasing the biomass by 40% and improving the volumetric ethanol productivity 16-fold than the parental K. marxianus SLP1. To reveal the effect of adaptation process in P8, transcriptome analysis was carried out; the result showed that the basal gene expression in P8 changed, suggesting the biological capability of K. marxianus to activate the adaptative prediction mechanism. Similarly, we carried out physiologic and transcriptome analyses to reveal the mechanisms involved in the stress response triggered by furfural, the most potent inhibitor in K. marxianus. Stress response studies demonstrated that P8 had a better physiologic response than SLP1, since key genes related to furfural transformation (ALD4 and ALD6) and stress response (STL1) were upregulated. Our study demonstrates the rapid adaptability of K. marxianus to stressful environments, making this yeast a promising candidate to produce lignocellulosic ethanol. KEY POINTS: • K. marxianus was adapted to increase its tolerance to a mix of inhibitory compounds • The basal gene expression of K. marxianus changed after the adaptation process • Adapted K. marxianus showed a better physiological response to stress by inhibitors • Transcriptome analyses revealed key genes involved in the stress response.

Identification of genes related to hydrolysis and assimilation of Agave fructans in Candida apicola NRRL Y-50540 and Torulaspora delbrueckii NRRL Y-50541 by denovo transcriptome analysis.

Fructans are the main sugar in agave pine used by yeasts during mezcal fermentation processes, from which Candida apicola NRRL Y-50540 and Torulaspora delbrueckii NRRL Y-50541 were isolated. De novo transcriptome analysis was carried out to identify genes involved in the hydrolysis and assimilation of Agave fructans (AF). We identified a transcript annotated as SUC2, which is related to ß-fructofuranosidase activity, and several differential expressed genes involved in the transcriptional regulation of SUC2 such as: MIG1, MTH1, SNF1, SNF5, REG1, SSN6, SIP1, SIP2, SIP5, GPR1, RAS2, and PKA. Some of these genes were specifically expressed in some of the yeasts according to their fructans assimilation metabolism. Different hexose transporters that could be related to the assimilation of fructose and glucose were found in both the transcriptomes. Our findings provide a better understanding of AF assimilation in these yeasts and provide resources for further metabolic engineering and biotechnology applications.

Diploid genome assembly of Kluyveromyces marxianus NRRL Y-50883 (SLP1).

The yeast Kluyveromyces marxianus SLP1 has the potential for application in biotechnological processes because it can metabolize several sugars and produce high-value metabolites. K. marxianus SLP1 is a thermotolerant yeast isolated from the mezcal process, and it is tolerant to several cell growth inhibitors such as saponins, furan aldehydes, weak acids, and phenolics compounds. The genomic differences between dairy and nondairy strains related to K. marxianus variability are a focus of research attention, particularly the pathways leading this species toward polyploidy. We report the diploid genome assembly of K. marxianus SLP1 nonlactide strain into 32 contigs to reach a size of ∼12 Mb (N50 = 1.3 Mb) and a ∼39% GC content. Genome size is consistent with the k-mer frequency results. Genome annotation by Funannotate estimated 5000 genes in haplotype A and 4910 in haplotype B. The enriched annotated genes by ontology show differences between alleles in biological processes and cellular component. The analysis of variants related to DMKU3 and between haplotypes shows changes in LAC12 and INU1, which we hypothesize can impact carbon source performance. This report presents the first polyploid K. marxianus strain recovered from nonlactic fermenting medium.

Conocimiento y aplicación de la política de seguridad del paciente por el personal de enfermería, para la prevención y reporte del evento adverso en un hospital de segundo nivel en Cundinamarca / Knowledge and application of the patient safety policy by the nursing staff, for the prevention and reporting of medical errors in a second level hospital in Cundinamarca

Objetivo Determinar el efecto de una intervención educativa sobre el conocimiento y la aplicación de la política de seguridad del paciente por el personal de enfermería, para la prevención y reporte del evento adverso en un Hospital de segundo nivel en Cundinamarca. Metodología Estudio cuantitativo, cuasi experimental con diseño pretest ­ postest de grupo único. 40 participantes dentro del personal de Enfermería de los diferentes servicios del Hospital; esta investigación tuvo en cuenta las consideraciones éticas para la Investigación en salud. Se elaboró un pre y pos-test a partir del reporte estadístico de eventos adversos de mayor frecuencia y la intervención educativa, con ello se aplicó una prueba U de Mann-Whitney apareada. Resultados Con este estudio se puede afirmar que la existencia de EA a nivel asistencial es causada por las acciones inseguras en la atención, ejecutadas por los profesionales de salud en medio del cumplimiento de la alta carga laboral. Con el apareamiento del pre y pos-test no se obtuvieron diferencias significativas, pero se infiere que la implementación de estrategias educativas son un importante soporte en la realización de los diferentes procedimientos para la prevención del evento adverso. Conclusión Se afirma que el cuidado de enfermería lleva implícito en su quehacer, la prevención de los eventos adversos, estos deben articularse a su vez con una disponibilidad de recursos Institucionales (humanos, de infraestructura y materiales, entre otros) que fortalezcan la política de seguridad del paciente, disminuyan el riesgo de incidentes y promuevan un cuidado de calidad y una atención segura.

Objective To determine the effect of an educational intervention on the knowledge and application of the patient safety policy by the nursing staff, for the prevention and reporting of medical errors in a second level Hospital in Cundinamarca. Methodology Quantitative, quasi-experimental study with pretest-posttest single group design. 40 participants within the nursing staff of the different services of the Hospital; this research considered the ethical considerations for health research. A pretest and posttest were elaborated based on the statistical report of the most frequent Medical Errors (ME) and the educational intervention, and a paired U Mann-Whitney test was applied. Results With this study it can be affirmed that the existence of ME's at the assistance level is caused by unsafe actions in care, executed by health professionals in the midst of the high workload. With the pairing of the pre- and post-test, no significant differences were obtained, but it can be inferred that the implementation of educational strategies is an important support in the performance of the different procedures for the prevention of ME. Conclusion It is affirmed that nursing care has implicit in their work, the prevention of medical errors, these must be articulated in turn with the availability of institutional resources (human, infrastructure, and materials, among others) that strengthen the patient safety policy, reduce the risk of incidents, and promote quality care and safe care.

Enzymatic synthesis of phlorizin fructosides.

Phlorizin is a low soluble dihydrochalcone with relevant pharmacological properties. In this study, enzymatic fructosylation was approached to enhance the water solubility of phlorizin, and consequently its bioavailability. Three enzymes were assayed for phlorizin fructosylation in aqueous reactions using sucrose as fructosyl donor. Levansucrase (EC 2.4.1.10) from Gluconacetobacter diazotrophicus (Gd_LsdA) was 6.5-fold more efficient than invertase (EC 3.2.1.26) from Rhodotorula mucilaginosa (Rh_Inv), while sucrose:sucrose 1-fructosyltransferase (EC 2.4.1.99) from Schedonorus arundinaceus (Sa_1-SST) failed to modify the non-sugar acceptor. Gd_LsdA synthesized series of phlorizin mono- di- and tri-fructosides with maximal conversion efficiency of 73 %. The three most abundant products were identified by ESI-MS and NMR analysis as ß-D-fructofuranosyl-(2→6)-phlorizin (P1a), phlorizin-4'-O-ß-D-fructofuranosyl-(2→6)-D-fructofuranoside (P2c) and phlorizin-4-O-monofructofuranoside (P1b), respectively. Purified P1a was 16 times (30.57 g L-1 at 25 °C) more soluble in water than natural phlorizin (1.93 g L-1 at 25 °C) and exhibited 44.56 % free radical scavenging activity. Gd_LsdA is an attractive candidate enzyme for the scaled synthesis of phlorizin fructosides in the absence of co-solvent.

Synthesis of b(1 ? 3) and b(1 ? 6) galactooligosaccharides from lactose and whey using a recombinant b-galactosidase from Pantoea anthophila

BACKGROUND: Milk whey, a byproduct of the dairy industry has a negative environmental impact, can be used as a raw material for added-value compounds such as galactooligosaccharides (GOS) synthesis by bgalactosidases. RESULTS: B-gal42 from Pantoea anthophila strain isolated from tejuino belonging to the glycosyl hydrolase family GH42, was overexpressed in Escherichia coli and used for GOS synthesis from lactose or milk whey. Crude cell-free enzyme extracts exhibited high stability; they were employed for GOS synthesis reactions. In reactions with 400 g/L lactose, the maximum GOS yield was 40% (w/w) measured by HPAEC-PAD, corresponding to 86% of conversion. This enzyme had a strong predilection to form GOS with b(1 ? 6) and b (1 ? 3) galactosyl linkages. Comparing GOS synthesis between milk whey and pure lactose, both of them at 300 g/L, these two substrates gave rise to a yield of 38% (60% of lactose conversion) with the same product profile determined by HPAEC-PAD. CONCLUSIONS: B-gal42 can be used on whey (a cheap lactose source) to produce added value products such as galactooligosaccharides.

One-pot bi-enzymatic cascade synthesis of puerarin polyfructosides.

Enzymatic glycosylation is an efficient way to increase the water solubility and the bioavailability of flavonoids. Levansucrases from Bacillus subtilis (Bs_SacB), Gluconacetobacter diazotrophicus (Gd_LsdA), Leuconostoc mesenteroides (Lm_LevS) and Zymomonas mobilis (Zm_LevU) were screened for puerarin (daidzein-8-C-glucoside) fructosylation. Gd_LsdA transferred the fructosyl unit of sucrose onto the glucosyl unit of the acceptor forming ß-d-fructofuranosyl-(2→6)-puerarin (P1a), while Bs_SacB, Lm_LevS and Zm_LevU synthesized puerarin-4'-O-ß-D-fructofuranoside (P1b) and traces of P1a. The Gd_LsdA product P1a was purified and assayed as precursor for the synthesis of puerarin polyfructosides (PPFs). Bs_SacB elongated P1a more competently forming a linear series of water-soluble PPFs reaching at least 21 fructosyl units, as characterized by HPLC-UV-MS, HPSEC and MALDI-TOF-MS. Simultaneous or sequential Gd_LsdA/Bs_SacB reactions yielded PPFs directly from puerarin with the acceptor conversion ranging 82-92 %. The bi-enzymatic cascade synthesis of PPFs in the same reactor avoided the isolation of the intermediate product P1a and it is appropriate for use at industrial scale.

2-Phenylethanol and 2-phenylethylacetate production by nonconventional yeasts using tequila vinasses as a substrate.

Vinasses from the tequila industry are wastewaters with highly elevated organic loads. Therefore, to obtain value-added products by yeast fermentations, such as 2-phenylethanol (2-PE) and 2-phenylethylacetate (2-PEA), could be interesting for industrial applications from tequila vinasses. In this study, four yeasts species (Wickerhamomyces anomalus, Candida glabrata, Candida utilis, and Candida parapsilosis) were evaluated with two different chemically defined media and tequila vinasses. Differences in the aroma compounds production were observed depending on the medium and yeast species used. In tequila vinasses, the highest concentration (65 mg/L) of 2-PEA was reached by C. glabrata, the inhibitory compounds decreased biomass production and synthesis of 2-PEA, and biochemical and chemical oxygen demands were reduced by more than 50 %. Tequila vinasses were suitable for the production of 2-phenylethylacetate by the shikimate pathway. A metabolic network was developed to obtain a guideline to improve 2-PE and 2-PEA production using flux balance analysis (FBA).

Biodegradation of NSAIDs and their effect on the activity of ligninolytic enzymes from Pleurotus djamor.

In this work, the white-rot fungus Pleurotus djamor was used for the first time to determine the degradation kinetics of the nonsteroidal anti-inflammatory drugs diclofenac, naproxen and, ketoprofen, either individually or in mixtures, in submerged cultures. Removal of 93% individual diclofenac and 99% diclofenac in mixtures with naproxen and ketoprofen at 6 h of incubation with the fungus was achieved. The elimination levels of naproxen and ketoprofen individually were 90% and 87%, respectively, after 48 h of incubation. However, the removal levels of these compounds in mixtures were 85% and 83%, respectively. On the other hand, during the degradation kinetics analysis, the enzymatic activities of laccases, manganese peroxidases, and lignin peroxidases were evaluated, yielding values of 3700, 270 and 31 U/L, respectively. Additionally, it was demonstrated that during degradation of diclofenac or the three drugs mixed in the submerged cultures, the enzymatic activity of extracellular laccases expressed by P. djamor increased by 200% and 300%, respectively. The activity of manganese peroxides increased by 126% with diclofenac and 138% when the mixture of drugs was added to the cultures. On the other hand, lignin peroxidase only increased activity by 123% with the drug mixture.

Cellular and Synaptic Dysfunctions in Parkinson's Disease: Stepping out of the Striatum.

The basal ganglia (BG) are a collection of interconnected subcortical nuclei that participate in a great variety of functions, ranging from motor programming and execution to procedural learning, cognition, and emotions. This network is also the region primarily affected by the degeneration of midbrain dopaminergic neurons localized in the substantia nigra pars compacta (SNc). This degeneration causes cellular and synaptic dysfunctions in the BG network, which are responsible for the appearance of the motor symptoms of Parkinson's disease. Dopamine (DA) modulation and the consequences of its loss on the striatal microcircuit have been extensively studied, and because of the discrete nature of DA innervation of other BG nuclei, its action outside the striatum has been considered negligible. However, there is a growing body of evidence supporting functional extrastriatal DA modulation of both cellular excitability and synaptic transmission. In this review, the functional relevance of DA modulation outside the striatum in both normal and pathological conditions will be discussed.

Multicenter prospective observational long-term follow-up study of endocardial cardiac resynchronization therapy using the Jurdham procedure.

BACKGROUND: Endocardial cardiac resynchronization therapy (eCRT) avoids the limitations and failures of coronary sinus (CS) resynchronization. However, data regarding long-term outcomes are lacking. OBJECTIVE: The purpose of this study was to report the long-term outcome of eCRT performed using the Jurdham procedure in a real-world setting. METHODS: eCRT was performed in patients who failed a CS implant or failed to respond to cardiac resynchronization therapy (CRT), or in selected patients requiring lifelong oral anticoagulation (OAC). Left ventricular ejection fraction (LVEF), New York Heart Association functional class (NYHA FC), and left ventricular stimulation parameters were assessed during long-term follow-up (FU). RESULTS: From August 2009 to March 2018, the Jurdham procedure was performed in 88 patients at 15 centers in 8 countries, with FU of 32.88 ± 61.52 months (range 0-88 months; 196 patient-years). NYHA FC improved from 2.9 preimplant to 1.3 during FU. LVEF increased <10 percentage points from baseline in 7% of patients, between 10 and 20 percentage points in 11% of patients, and >20 percentage points in 82% of patients. All-cause mortality at 60 months was 30.5%. Three transient ischemic attacks (1.53 per 100 patient-years) and 6 strokes (3.06 per 100 patient-years) occurred. Of the 6 patients with stroke, 4 (66%) had almost complete recovery. CONCLUSION: eCRT using the Jurdham procedure is an effective and safe technique in anticoagulated patients. This approach may be an attractive option for patients with failed CS implants or nonresponders to CS CRT. In addition, it might be a reasonable approach as a first option for treatment of patients requiring lifelong OAC.

Cell surface display of proteins on filamentous fungi.

Protein display approaches have been useful to endow the cell surface of yeasts with new catalytic activities so that they can act as enhanced whole-cell biocatalysts. Despite their biotechnological potential, protein display technologies remain poorly developed for filamentous fungi. The lignocellulolytic character of some of them coupled to the cell surface biosynthesis of valuable molecules by a single or a cascade of several displayed enzymes is an appealing prospect. Cell surface protein display consists in the co-translational fusion of a functional protein (passenger) to an anchor one, usually a cell-wall-resident protein. The abundance, spacing, and local environment of the displayed enzymes-determined by the relationship of the anchor protein with the structure and dynamics of the engineered cell wall-are factors that influence the performance of display-based biocatalysts. The development of protein display strategies in filamentous fungi could be based on the field advances in yeasts; however, the unique composition, structure, and biology of filamentous fungi cell walls require the customization of the approach to those microorganisms. In this prospective review, the cellular bases, the design principles, and the available tools to foster the development of cell surface protein display technologies in filamentous fungi are discussed.

Fructosylation of phenolic compounds by levansucrase from Gluconacetobacter diazotrophicus.

Fructosylation can significantly improve the solubility, stability and bioactivity of phenolic compounds, increasing their health benefits. Levansucrase from Gluconacetobacter diazotrophicus (LsdA, EC 2.4.1.10) was found to transfer the fructosyl unit of sucrose to different classes of phenolic compounds. Among the various acceptors tested, the isoflavone puerarin and the phenol coniferyl alcohol were the most efficiently fructosylated compounds, with conversion rates of 93% and 25.1%, respectively. In both cases, mono-, di-, and trifructosides were synthesized at a ratio of 37:14:1 and 32:8:1, respectively. Structural characterization of the puerarin mono-fructoside revealed that the enzyme transferred the fructosyl moiety of sucrose to the O6-position of the glucosyl unit of puerarin. The water solubility of fructosyl-ß-(2→6)-puerarin was increased 23-fold, up to 16.2 g L-1, while its antioxidant capacity was only decreased 1.25-fold compared with that of puerarin.

Volatile compounds flavoring obtained from Brazilian and Mexican spirit wastes by yeasts.

Vinasse is a waste obtained from the production of beverages, such as tequila and cachaça. The presence of acids, alcohols, sugars, minerals, amino acids, peptides, and nitrogen salts make vinasse a hazardous liquid waste to the environment, affecting the fauna, flora, and microbiota of rivers and lagoons. This study used biological treatment concomitant to volatile compound production. The yeasts used in the study were Saccharomyces cerevisiae (CCMA 0187 and CCMA 0188), Candida parapsilosis (CCMA 0544), and Pichia anomala (CCMA 0193). A higher percentage reduction in chemical and biochemical oxygen demand was observed in the tequila vinasse than in the cachaça vinasse. However, a higher production of volatile compounds was observed in the cachaça vinasse. C. parapsilosis CCMA 0544 produced the highest concentration of 2-phenylethanol (162 mg L-1). These results indicated that the environmental damage of vinasse can be reduced by treating vinasse with yeasts, and this treatment produces aroma compounds. This biological treatment has high economic potential, especially for the tequila industry.

Bacillus subtilis with endocellulase and exocellulase activities isolated in the thermophilic phase from composting with coffee residues / Bacillus subtilis con actividades de endocelulasa y exocelulasa aislado en la fase termofílica del compostaje con residuos de café

The goal of this study was to isolate, select and characterize bacteria with cellulolytic activity from two different coffee residue composting piles, one of which had an internal temperature of 57 -#9702;C and pH 5.5 and the other, a temperature of 61 -#9702;C, and pH 9.3. Culture media were manipulated with carboxymethylcellulose and crystalline cellulose as sole carbon sources. The enzyme activity was assessed by hydrolysis halo formation, reducing sugar production and zymograms. Three out of twenty isolated strains showed higher enzymatic activity and were identified as Bacillus subtilis according to their morphological, physiological, biochemical characteristics and based on the sequence analysis of 16S rDNA regions. The enzymatic extracts of the three selected strains showed exocellulase and endocellulase maximum activity of 0.254 and 0.519 U/ml, respectively; the activity of these enzymes was maintained even in acid pH (4.8) and basic (9.3) and at temperatures of up to 60°C. The enzymatic activities observed in this study are within the highest reported for cellulose produced by bacteria of the genus Bacillus. Endocellulase activity was shown in the zymograms from 24 h until 144 h of incubation. Furthermore, the pH effect on the endocellulase activity is reported for the first time by zymograms. The findings in this study entail the possibility to use these enzymes in the procurement of fermentable substrates for the production of energy from the large amount of residues generated by the coffee agroindustry.

El objetivo de este estudio fue aislar, seleccionary caracterizar bacterias con actividad celulolítica a partir de 2 diferentes pilas de compostaje de residuos de café, una con temperatura interna de 57°C y pH 5,5; la otra con temperatura interna de 61 °C y pH 9,3. Se utilizaron medios de cultivo con carboximetilcelulosa y celulosa cristalina como únicas fuentes de carbono. La actividad enzimàtica fue evaluada por formación de halos de hidrólisis, producción de azúcares reductores y zimogramas. De 20 cepas aisladas, 3 presentaron mayor actividad enzimàtica y fueron identificadas como Bacillus subtilis sobre la base de sus características morfológicas, fisiológicas y bioquímicas y del análisis de las secuencias de la región 16S del ADNr. Los extractos enzimáticos de las 3 cepas seleccionadas presentaron actividad de exocelulasa y de endocelulasa, con máximos de 0,254 y 0,519 U/ml, respectivamente; la actividad de estas enzimas se mantuvo incluso a pH ácido (4,8) o básico (9,3) y a temperaturas de hasta 60 °C. Las actividades enzimáticas halladas en este estudio se ubican dentro de las más altas reportadas para celulasas producidas por bacterias del género Bacillus. En los zimogramas se demostró actividad de endocelulasa desde las 24h hasta las 144h de incubación. Asimismo, se reporta por primera vez el efecto del pH sobre la actividad de endocelulasa observado por zimogramas. Los resultados de este estudio abren la posibilidad de hacer uso de estas enzimas en la obtención de sustratos fermentables para la producción de energía a partir de los residuos generados en grandes cantidades por la agroindustria del café.

Scale-up and evaluation of hydrothermal pretreatment in isothermal and non-isothermal regimen for bioethanol production using agave bagasse.

The production of tequila in Mexico generates a large amount of agave bagasse per year. However, this biomass can be considered as a potential source for biofuel production. In this study, it is described how the hydrothermal pretreatment was scaled in a bench scale, considering the severity index as a strategy. The best condition was at 180 °C in isothermal regime for 20 min with 65.87% of cellulose content and high concentration of xylooligosaccharides (15.31 g/L). This condition was scaled up (using severity factor: [logR0] = 4.11), in order to obtain a rich pretreated solid in cellulose to perform the enzymatic hydrolysis, obtaining saccharification yields of 98.5 and 99.5% at high-solids loading (10 and 15%, respectively). The pre-saccharification and fermentation strategy was used in the bioethanol production at 10 and 15% of total pretreated solids, obtaining 38.39 and 55.02 g/L of ethanol concentration, corresponding to 90.84% and 87.56% of ethanol yield, respectively.

Bacillus subtilis with endocellulase and exocellulase activities isolated in the thermophilic phase from composting with coffee residues.

The goal of this study was to isolate, select and characterize bacteria with cellulolytic activity from two different coffee residue composting piles, one of which had an internal temperature of 57°C and pH 5.5 and the other, a temperature of 61°C, and pH 9.3. Culture media were manipulated with carboxymethylcellulose and crystalline cellulose as sole carbon sources. The enzyme activity was assessed by hydrolysis halo formation, reducing sugar production and zymograms. Three out of twenty isolated strains showed higher enzymatic activity and were identified as Bacillus subtilis according to their morphological, physiological, biochemical characteristics and based on the sequence analysis of 16S rDNA regions. The enzymatic extracts of the three selected strains showed exocellulase and endocellulase maximum activity of 0.254 and 0.519 U/ml, respectively; the activity of these enzymes was maintained even in acid pH (4.8) and basic (9.3) and at temperatures of up to 60°C. The enzymatic activities observed in this study are within the highest reported for cellulose produced by bacteria of the genus Bacillus. Endocellulase activity was shown in the zymograms from 24h until 144h of incubation. Furthermore, the pH effect on the endocellulase activity is reported for the first time by zymograms. The findings in this study entail the possibility to use these enzymes in the procurement of fermentable substrates for the production of energy from the large amount of residues generated by the coffee agroindustry.

High heterogeneity, mixed infections and new genotypes in human congenital toxoplasmosis cases in the mega-metropolis of Central Mexico.

Mexico presents high prevalence of Toxoplasma gondii infection, including the congenital form, but there are few data about the genetic diversity of the parasite, so we attempted parasite isolation and genotyping in nine mother/children pairs with congenital toxoplasmosis (CT), living in the Valley of Mexico, who were part of a 30 cases cohort that started 12 years ago. They were recruited through research projects which included pre- and postnatal screening of congenital infections or directly CT, and cases referred to INP for management because they had clinical abnormalities. Genotyping was performed by PCR-RFLP of SAG1, SAG2, SAG3, BTUB GRA6, c22-8, c29-2, L358, PK1 and Apico markers, followed by sequencing. Sixty seven percent of samples were typed for the SAG3 locus, 39% for Apico and 33% for BTUB, while Alt. SAG2, GRA6 and c29-2 types could be labelled in less cases. Type I alleles predominated, followed by II and III. We isolated the first strain obtained from humans in Mexico and found three genotypes not previously found in the world. The presence of ToxoDB#10 clonal type was documented in one pair, as well as mixed infections in five mothers. No relation of genotype or parasite load with clinical signs was found. In conclusion, we encountered great genetic diversity and mixed T. gondii infections among mother/children pairs with congenital toxoplasmosis in the mega-metropolis of the Valley of Mexico.