Genome Comparisons between Botrytis fabae and the Closely Related Gray Mold Fungus Botrytis cinerea Reveal Possible Explanations for Their Contrasting Host Ranges.

While Botrytis cinerea causes gray mold on many plants, its close relative, Botrytis fabae, is host-specifically infecting predominantly faba bean plants. To explore the basis for its narrow host range, a gapless genome sequence of B. fabae strain G12 (BfabG12) was generated. The BfabG12 genome encompasses 45.0 Mb, with 16 chromosomal telomere-to-telomere contigs that show high synteny and sequence similarity to the corresponding B. cinerea B05.10 (BcB0510) chromosomes. Compared to BcB0510, it is 6% larger, due to many AT-rich regions containing remnants of transposable elements, but encodes fewer genes (11,420 vs. 11,707), due to losses of chromosomal segments with up to 20 genes. The coding capacity of BfabG12 is further reduced by nearly 400 genes that had been inactivated by mutations leading to truncations compared to their BcB0510 orthologues. Several species-specific gene clusters for secondary metabolite biosynthesis with stage-specific expression were identified. Comparison of the proteins secreted during infection revealed high similarities, including 17 phytotoxic proteins that were detected in both species. Our data indicate that evolution of the host-specific B. fabae occurred from an ancestral pathogen with wide host range similar to B. cinerea and was accompanied by losses and degeneration of genes, thereby reducing its pathogenic flexibility.

New Benzodihydrofuran Derivatives Alter the Amyloid ß Peptide Aggregation: Strategies To Develop New Anti-Alzheimer Drugs.

Alzheimer's disease is a neurodegenerative disorder that is the leading cause of dementia in elderly patients. Amyloid-ß peptide (1-42 oligomers) has been identified as a neurotoxic factor, triggering many neuropathologic events. In this study, 15 chalcones were synthesized employing the Claisen-Schmidt condensation reaction, starting from a compound derived from fomannoxine, a natural benzodihydrofuran whose neuroprotective activity has been proven and reported, and methyl aromatic ketones with diverse patterns of halogenated substitution. As a result, chalcones were obtained, with good to excellent reaction yields from 50 to 98%. Cytotoxicity of the compounds was assessed, and their cytoprotective effect against the toxicity associated with Aß was evaluated on PC-12 cells. Out of the 15 chalcones obtained, only the 4-bromo substituted was cytotoxic at most tested concentrations. Three synthesized chalcones showed a cytoprotective effect against Aß toxicity (over 37%). The 2,4,5-trifluoro substituted chalcone was the most promising series since it showed a cytoprotective impact with more than 60 ± 5% of recovery of cellular viability; however, 3-fluoro substituted compound also exhibited important values of recovery (50 ± 6%). The fluorine substitution pattern was shown to be more effective for cytoprotective activity. Specifically, substitution with fluorine in the 3,5-positions turned out to be particularly effective for cytoprotection. Furthermore, fluorinated compounds inhibited the aggregation rate of Aß, suggesting a dual effect that can be the starting point of new molecules with therapeutic potential.

Ferrocene-based nitroheterocyclic sulfonylhydrazones: design, synthesis, characterization and trypanocidal properties.

A series of new ferrocenyl nitroheterocyclic sulfonylhydrazones (1a-4a and 1b-2b) were prepared by the reaction between formyl (R = H) or acetyl (R = CH3) nitroheterocyclic precursors [4/5-NO2(C5H2XCOR), where X = O, S)] and ferrocenyl tosyl hydrazine [(η5-C5H5)Fe(η5-C5H4SO2-NH-NH2)]. All compounds were characterized by conventional spectroscopic techniques. In the solid state, the molecular structures of compounds 1a, 2b, and 3a were determined by single-crystal X-ray diffraction. The compounds showed an E-configuration around the C=N moiety. Evaluation of trypanocidal activity, measured in vitro against the Trypanosoma cruzi and Trypanosoma brucei strains, indicated that all organometallic tosyl hydrazones displayed activity against both parasite species with a higher level of potency toward T. brucei than T. cruzi. Moreover, the biological evaluation showed that the 5-nitroheterocyclic derivatives were more efficient trypanocidal agents than their 4-nitroheterocyclic counterparts.

Heterobimetallic complexes containing organometallic acylhydrazone ligands as potential inhibitors of human carbonic anhydrases.

In the search of new bioorganometallic compounds as potential inhibitors of human (h) carbonic anhydrases (hCAs, EC 4.2.1.1), heterobinuclear ruthenium(II) complexes based on organometallic-acylhydrazones have been obtained. The complexes (1a-b, 2a-b) were prepared by reaction between the corresponding organometallic-acylhydrazone of the general formula [{(η5-C5H4)CH=N-NH-C(O)-C6H4-4-SO2NH2}]MLn or [{(η5-C5H4)CH=N-NH-C(O)-CH2CH2-NH-C6H4-4-SO2NH2}]MLn (where MLn = Re(CO)3; FeCp) and [Ru(p-cymene)Cl2]2. All compounds were characterized by conventional spectroscopic techniques and cyclic voltammetry. Biological evaluation as CA inhibitors (CAIs) was carried out and showed derivatives 1a, 2a and 2b to behave as selective inhibition against the tumors associate isoforms hCA IX and XII making them interesting candidates for preclinical evaluation in various hypoxic tumors in which the two enzymes are overexpressed.

Deciphering the Monilinia fructicola Genome to Discover Effector Genes Possibly Involved in Virulence.

Brown rot is the most economically important fungal disease of stone fruits and is primarily caused by Monilinia laxa and Monlinia fructicola. Both species co-occur in European orchards although M. fructicola is considered to cause the most severe yield losses in stone fruit. This study aimed to generate a high-quality genome of M. fructicola and to exploit it to identify genes that may contribute to pathogen virulence. PacBio sequencing technology was used to assemble the genome of M. fructicola. Manual structural curation of gene models, supported by RNA-Seq, and functional annotation of the proteome yielded 10,086 trustworthy gene models. The genome was examined for the presence of genes that encode secreted proteins and more specifically effector proteins. A set of 134 putative effectors was defined. Several effector genes were cloned into Agrobacterium tumefaciens for transient expression in Nicotiana benthamiana plants, and some of them triggered necrotic lesions. Studying effectors and their biological properties will help to better understand the interaction between M. fructicola and its stone fruit host plants.

Dynamics in Secondary Metabolite Gene Clusters in Otherwise Highly Syntenic and Stable Genomes in the Fungal Genus Botrytis.

Fungi of the genus Botrytis infect >1,400 plant species and cause losses in many crops. Besides the broad host range pathogen Botrytis cinerea, most other species are restricted to a single host. Long-read technology was used to sequence genomes of eight Botrytis species, mostly pathogenic on Allium species, and the related onion white rot fungus, Sclerotium cepivorum. Most assemblies contained <100 contigs, with the Botrytis aclada genome assembled in 16 gapless chromosomes. The core genome and pan-genome of 16 Botrytis species were defined and the secretome, effector, and secondary metabolite repertoires analyzed. Among those genes, none is shared among all Allium pathogens and absent from non-Allium pathogens. The genome of each of the Allium pathogens contains 8-39 predicted effector genes that are unique for that single species, none stood out as potential determinant for host specificity. Chromosome configurations of common ancestors of the genus Botrytis and family Sclerotiniaceae were reconstructed. The genomes of B. cinerea and B. aclada were highly syntenic with only 19 rearrangements between them. Genomes of Allium pathogens were compared with ten other Botrytis species (nonpathogenic on Allium) and with 25 Leotiomycetes for their repertoire of secondary metabolite gene clusters. The pattern was complex, with several clusters displaying patchy distribution. Two clusters involved in the synthesis of phytotoxic metabolites are at distinct genomic locations in different Botrytis species. We provide evidence that the clusters for botcinic acid production in B. cinerea and Botrytis sinoallii were acquired by horizontal transfer from taxa within the same genus.

Variabilidad de la frecuencia cardíaca como indicador de carga interna en mujeres no deportistas: estudio piloto / Heart rate variability as an indicator of internal load in non-athlete women: pilot study

La variabilidad de la frecuencia cardiaca (VFC) es una herramienta no invasiva que permite evaluar la modulación simpática y parasimpática y se ha propuesto como un método válido para valorar la respuesta individual a una carga de trabajo y, por tanto, la carga de entrenamiento. El objetivo es utilizar la RMSSD-Slope (La pendiente de la raíz cuadrada de la media de las diferencias de la suma de los cuadrados entre intervalos RR adyacentes) para analizar la recuperación tras dos intensidades diferentes en tapiz rodante en mujeres no deportistas, como medida de carga interna (CI) y su posible relación con la carga externa (CE).Participaron 9 mujeres sanas, físicamente activas. Se realizaron dos test, separados entre sí por 48-72 h. El primero fue una prueba máxima en tapiz rodante, en el que se determinó la velocidad aeróbica máxima (VAM). En la segunda sesión, se realizó una prueba al 80% de la VAM. En cada una de las sesiones se hizo un seguimiento la escala de Borg y de la VFC (reposo, ejercicio y recuperación) para su posterior análisis con la RMSSD-Slope.El valor de la RMSSD-Slope en la prueba del 80% de intensidad fue de 0,97 (±0,78), y en la Prueba Máxima fue 0,84 (±0,36). Ambas pruebas presentan una R2 con la escala de Borg (0,62 y 0,62) respectivamente. En el caso de la R2 entre la CE y la RMSSD-Slope fue de 0,04 y 0,14 respectivamente. La pendiente de recuperación de la RMSSD es una buena herramienta de valoración de CI en mujeres físicamente activas pero no deportistas

Heart rate variability (HRV ) is a non-invasive tool capable to evaluate the sympathetic and parasympathetic modulation and it has been proposed as a valid method to assess the individual response to a workload and, therefore, the training load. The objective is to use the RMSSD-Slope (square root of the mean of the differences of the sum of the squares between adjacent RR intervals) to analyze the recovery after two different treadmill intensities in non-athletic women, as an internal training load (ITL) measure and its possible relation with the external training load (ETL) 9 healthy, physically active women participated in the study. Two tests were performed, separated from each other for 48-72h. The first was a maximum treadmill test, in which the maximal aerobic speed (MAS) was determined. In the second session, an 80% test of the MAS was carried out. In each of the sessions, Borg scale and HRV was monitored (rest, exercise and recovery) for further analysis with the RMSSD-Slope. The RMSSD-Slope value in the 80% intensity test was 0.97 (± 0.78), and in the Maximum Test it was 0.84 (± 0.36). Both tests show an R2 with Borg scale of 0.62 and 0.62 respectively. In the case of the R2 between the ETL and the RMSSD-Slope it was 0.04 and 0.14 respectively. The recovery slope of the RMSSD is a good ITL assessment tool in physically active women but not athletes

Basal and post-exercise heart rate variability correlates with training load in endurance athletes / La variabilidad de la frecuencia cardíaca basal y posterior al ejercicio correlaciona con la carga de entrenamiento en atletas de resistencia / A variabilidade da frequência cardíaca basai e pós-exercício está correlacionada com a carga de treinamento em atletas de resistência

OBJECTIVE: Heart rate variability has been proposed as a valid method to examine the individual response to training load in endurance athletes. Thanks to this tool, the relationship between basal and post-exercise Heart rate variability measurements can be analyzed during a microcycle (one week) using straight values or their coefficients of variation. METHOD: Ten amateur endurance athletes (n = 5 men, n = 5 women) were monitored during a 7-day microcycle that included three road-cycling sessions, two running sessions and two trail-running sessions. The RR series were measured for 5 minutes upon wake up and after training, in a seating position, using a chest strap. RESULTS: Basal and post-exercise Heart rate variability measurements showed high correlation when weekly mean values were used, very similar to when coefficients of variation values were used. In women, the root mean square of successive differences (RMSSD) was: r= 0.73; RMSSD coefficients of variation (RMSSDcv) was: r= 0.66; natural logarithm (Ln) RMSSD: r= 0.68; LnRMSSDcv: r= 0.79; and in men it RMSSD was: r= 0.78; RMSSDcv: r= -0.62; LnRMSSD: r= 0.75; LnRMSSDcv: r= -0.73). CONCLUSION: the relationship between these two measurements could be useful to program the training loads of the following microcycle

OBJETIVO: La variabilidad de la frecuencia cardíaca se ha propuesto como un método válido para examinar la respuesta individual a la carga de entrenamiento en atletas de resistencia. El objetivo de este estudio fue analizar la relación entre las mediciones basales y post ejercicio durante un microciclo (una semana) utilizando valores directos o sus coeficientes de variación. MÉTODO: Se monitorizó a diez atletas aficionados de resistencia durante un microciclo de 7 días, que incluyó tres sesiones de ciclismo de ruta, dos sesiones de carrera y dos sesiones de trail running. Las series RR se midieron durante 5 minutos al despertar y después del entrenamiento, en posición sentado, utilizando una banda torácica. RESULTADOS: Las mediciones de variabilidad de la frecuencia cardíaca basales y post ejercicio mostraron una alta correlación cuando se usaron valores medios semanales, como cuando se usaron valores de los coeficientes de variación. En mujeres, la raíz cuadrada de la media de las diferencias de la suma de los cuadrados entre intervalos RR adyacentes (RMSSD) fue: r = 0.73; el coeficiente de variación (cv) de la RMSSD fue RMSSDcv: r = 0.66; el logaritmo natural (Ln) de la RMSSD (LnRMSSD) fue: r = 0.68; LnRMSSDcv: r = 0.79; y en los hombres fueron RMSSD: r = 0.78; RMSSDcv: r = -0.62; LnRMSSD: r = 0.75; LnRMSSDcv: r = -0.73. CONCLUSIONES: La relación entre estas dos mediciones podría ser útil para el programa de entrenamiento del microciclo posterior

OBJETIVO: A variação da frequência cardíaca tem sido utilizada como método de análise de respostas individuais a carga de treino em atletas de endurance. Graças a esta ferramenta, a relação entre a variação da frequência cardíaca basal e pós-exercício pode ser analisada durante um microciclo (uma semana) usando valores diretos ou seus coeficientes de variação. MÉTODO: Dez atletas de endurance amadores (n = 5 homens, n = 5 mulheres) foram monitorados durante microciclos de 7 dias que incluíram 3 sessões de ciclismo em estrada, duas sessões de corrida e duas sessões de corrida em trilha. As series de RR foram medidas durante 5 minutos depois de acordar e depois do exercício, em posição sentada, com recurso a cardiofrenquencímetros toráxicos. RESULTADOS: Os resultados mostram uma correlação alta entre as medidas de variação de frequência cardíaca basal e pós-exercício quando utilizados valores médios semanais, muito semelhantes aos resultados quando utilizados coeficientes de variação. Nas mulheres, a raiz quadrada media das diferenças sucessivas (RMSSD) foi: r= 0.73; coeficientes de variação RMSSD, (RMSSDcv) foi: r= 0.66; logaritmo natural (Ln) RMSSD: r = 0.68; LnRMSSDcv: r= 0.79; e nos homens, raiz quadrada media das diferenças sucessivas r= 0.78; RMSSDcv: r= -0.62; LnRMSSD: r= 0.75; LnRMSSDcv: r= -0.73). CONCLUSÃO: a relação entre estas duas medidas poderia ser útil para prescrição de cargas de treino dos microciclos seguintes

Synthesis and Cytotoxic Analysis of Novel Myrtenyl Grafted Pseudo-Peptides Revealed Potential Candidates for Anticancer Therapy.

Myrtenal is a natural monoterpene isolated from essential oils of several plants and their derivates have shown to have several biological properties including cytotoxicity. The cytotoxic activity of these derivates are being investigated for their antitumor effect leading to the development of potential anticancer agents. In this study, novels Myrtenyl grafted pseudo-peptides were designed, synthesized and functionally characterized as possible therapeutic agents for cancer treatment. Thirteen novel Myrtenyl grafted pseudo-peptides were prepared in high atom economy and efficiency by a classic Ugi-4CR and sequential post-modification. Their structures were confirmed by NMR, and ESI-MS, and its cytotoxic activity was evaluated in three cancer cell lines and primary CD4+ T cells at different proliferative cycles. Our results revealed that some of these compounds showed significant cytotoxicity against human gastric, breast and colon adenocarcinoma cells lines, but not against human dermal fibroblast cell line. Moreover, from the thirteen novel myrtenyl synthesized the compound (1R,5S)-N-{[1-(3-chlorophenyl)-1H-1,2,3-triazol-4-yl]methyl}-N-[2-(cyclohexylamino)-2-oxoethyl]-6,6-dimethylbicyclo[3.1.1]hept-2-ene-2-carboxamide (3b) proved to be the best candidate in terms of acceptable EC50, and Emax values in cancer cell lines and at inducing cytotoxicity in CD4+ T cells undergoing active proliferation, without affecting non-proliferating T cells. Overall, the synthesis and characterization of our Myrtenyl derivates revealed novel potential anticancer candidates with selective cytotoxic activity.

Heart rate variability to assess the effect of sleep deprivation in mountain troops of the chilean army: a pilot study / Variabilidad de la frecuencia cardíaca para evaluar el efecto de la privación del sueño en tropas de montaña del ejército chileno: un estudio piloto

BACKGROUND: Our objective was to identify the effect of sleep deprivation on a stress test simulating a military march, via changes in heart rate variability (HRV) in special mountain troops. Eight subjects from special mountain troops carried out a simulated march test on a treadmill. The incremental march test had 7 stages of 3 minute duration at a constant velocity of 5 km/h and slopes of 1, 3, 5, 7, 8, 9 and 10 %. To assess the HRV, two heartbeat records were taken over 5 minutes in dorsal decubitus position before and after the march test; the first session took place without sleep deprivation, and the following day with sleep deprivation. RESULTS: The main finding of this study is that the physiological stress imposed by the simulated treadmill march is the same with and without sleep deprivation. There were no significant differences between pre and post HRV data in any of the situations, but effect size was moderate or large (d=0.2 was considered as the Smallest Worthwhile Change). indicating a highly relevant response. However, after comparing with and without sleep deprivation tests no changes were found (non-significant and non-relevant). CONCLUSIONS: The stress test performed, did not present differences in physical and physiological responses while being deprived of sleep over 24 hours. A simple test is proposed to evaluate the effect of sleep deprivation as a stressor agent. A treadmill test at a constant speed with increasing slopes would be performed and repeated the following day after 24 hours of sleep deprivation

INTRODUCCIÓN: Nuestro objetivo fue identificar el efecto de la falta de sueño en una prueba de esfuerzo que simula una marcha militar, a través de cambios en la variabilidad de la frecuencia cardiaca (VFC) en tropas especiales de montaña. Ocho sujetos de tropas especiales de montaña realizaron una prueba de marcha simulada en una cinta de correr. La prueba de marcha incremental tuvo 7 etapas de 3 minutos de duración a una velocidad constante de 5 km/h y pendientes de 1, 3, 5, 7, 8, 9 y 10%. Para evaluar la VFC, se tomaron los registros de latidos latido del corazón durante 5 minutos en posición de decúbito dorsal antes y después de la prueba de marcha; la primera sesión tuvo lugar sin privación de sueño y al día siguiente con privación de sueño. RESULTADOS: El principal hallazgo de este estudio es que el estrés fisiológico impuesto por la marcha simulada de la cinta rodante es el mismo con y sin privación del sueño. No hubo diferencias significativas entre los datos de VFC anteriores y posteriores en ninguna de las situaciones, pero el tamaño del efecto fue moderado o grande (d = 0.2 se consideró como umbral de cambio pequeño). Indica una respuesta altamente relevante. Sin embargo, después de comparar con y sin las pruebas de privación de sueño, no se encontraron cambios (no significativos y no relevantes). CONCLUSIONES: La prueba de esfuerzo realizada no presentó diferencias en las respuestas físicas y fisiológicas al estar privada de sueño durante 24 horas. Se propone una prueba simple para evaluar el efecto de la falta de sueño como agente estresante. Se realizaría una prueba de la cinta rodante a una velocidad constante con pendientes crecientes y se repetiría al día siguiente después de 24 horas de falta de sueño

Cambios hormonales en soldados aclimatados durante una marcha en gran altitud con esquí de montaña / Hormonal changes in acclimatized soldiers during a march at a high altitude with mountain skis

Background: The aim of the present study is to identify the physiological impact of acute exposure to high altitudes on special acclimatized troops of the Chilean Army. Twenty-nine soldiers carried out a nocturnal winter march on mountain skis at an initial altitude of 2,800 m and up to 3,640 m. Two separate blood measurements were taken. The first one was taken the day before the march (Pre-sample) and the second one just after returning to the base camp (Post-sample). All subjects had been acclimatized prior to the study. For hypothesis comparison purposes, the normality of the distribution was tested using the Shapiro-Wilk test. To determine if there were significant differences between the Pre and Post tests, a paired-samples Student t-test was applied for the variables with a normal distribution, and the Wilcoxon test was applied for the variables without a normal distribution. In all cases, a level of significance of 95% (p<0.05) was taken into consideration. Results: Exposure of acclimatized troops to altitudes of 2,800 m to 3,640 m has an impact on the endocrine parameters and on the reduction of cortisol (p<0.01), total testosterone (p<0.0001), free testosterone (p<0.0001) and the free testosterone-cortisol ratio (p<0.01). Likewise, an increase in total leukocytes (p<0.0001), neutrophils (p<0.0001), monocytes (p<0.0001) and basophils (p<0.001), as well as a decrease of eosinophils (p<0.0001) and lymphocytes (p<0.01), was observed. No hematological changes were detected. Conclusions: Endocrine changes were observed during high-altitude winter marches on mountain skis carried out by accli-matized Special Operation Troops, resulting in decreased cortisol and free and total testosterone levels. A stress condition due to the high altitudes also affected the anabolic/catabolic environment, which manifested as a significant decrease in the free testosterone/cortisol ratio. No hematological changes were identified. Marked changes were observed in some white cell series

Introducción: El objetivo del presente estudio es identificar el impacto fisiológico (con especial atención a los parámetros endocrinos y hematológicos) de la exposición aguda a gran altitud (GA) en tropas especiales aclimatadas del Ejército de Chile. Veintinueve soldados llevaron a cabo una marcha nocturna con esquí de montaña invernal a una GA de 2.800 m. hasta 3.640 m. Se tomaron dos muestras de sangre. La primera muestra fue tomada el día antes de la marcha (Pretest) y la segunda muestra justo después al regresar al campamento base Post test (a los 2.800 m). Todos los sujetos se encontraban aclimatados antes del estudio. Para cada análisis se testeo la normalidad de las distribuciones empleando el test de Shapiro-Wilk. Se calculó el promedio y la desviación estándar para cada medición. Para determinar si existían diferencias significativas entre el pre y post test se aplicó la prueba de t-Student pareada para las variables con distribución normal y el test de Wilcoxon para las variables que no tenían distribución normal. En todos los casos se consideró un nivel de confianza de 95% (valor p < 0,05). Resultados: La exposición de las tropas aclimatadas a GA tiene un impacto en los parámetros endocrinos y en la reducción de cortisol (p <0,01), testosterona total (p <0,0001), testosterona libre (p <0,0001) y el ratio testosterona libre-cortisol (p <0.01). Asimismo, se observaron un aumento de leucocitos (p <0,0001), neutrófilos (p <0,0001), monocitos (p <0,0001) y basófilos (p <0,001), así como una decrease de eosinófilos (p <0,0001) y linfocitos (p < 0.01). No se observaron cambios en la serie roja. Conclusiones: La marcha invernal nocturna con esquí de montaña en GA para tropas de operaciones especiales aclimatadas presento cambios endocrinos con disminución del cortisol, testosterona libre y total. Una condición de estrés por la marcha en GA también afectó al ambiente anabólico/catabólico, lo que se ve reflejado en una disminución significativa en el cociente testosterona libre/cortisol. No se observaron cambios hematológicos. Se observaron cambios significativos en algunas células de la serie blanca

Comparative genomics of plant pathogenic Botrytis species with distinct host specificity.

BACKGROUND: Fungi of the genus Botrytis (presently containing ~ 35 species) are able to infect more than 1400 different plant species and cause losses in a wide range of crops of economic importance. The best studied species is B. cinerea, which has a broad host range and is one of the best studied necrotrophic plant pathogenic fungi. Most other Botrytis spp. have a narrow host range and have been studied in less detail. To characterize genomic variation among different representatives of Botrytis spp., we sequenced and annotated the draft genomes of nine Botrytis species: B. calthae, B. convoluta, B. elliptica, B. galanthina, B. hyacinthi, B. narcissicola, B. paeoniae, B. porri and B. tulipae. RESULTS: Bioinformatics and comparative genomics tools were applied to determine a core of 7668 shared protein families in all Botrytis species, which grouped them in two distinct phylogenetic clades. The secretome of all nine Botrytis spp. was similar in number (ranging from 716 to 784 predicted proteins). A detailed analysis of the molecular functions of the secretome revealed that shared activities were highly similar. Orthologs to effectors functionally studied in B. cinerea were also present in the other Botrytis species. A complex pattern of presence/absence of secondary metabolite biosynthetic key enzymes was observed. CONCLUSIONS: Comparative genomics of Botrytis show that overall, species share the main signatures and protein families in the secreted proteins, and of known effectors. Our study provides leads to study host range determinants in the genus Botrytis and provides a stepping stone to elucidate the roles of effector candidates in the infection process of these species.

Spatio-temporal patterns of genetic variation in Arbacia lixula, a thermophilous sea urchin in expansion in the Mediterranean.

The genetic structure of 13 populations of the amphiatlantic sea urchin Arbacia lixula, as well as temporal genetic changes in three of these localities, were assessed using ten hypervariable microsatellite loci. This thermophilous sea urchin is an important engineer species triggering the formation of barren grounds through its grazing activity. Its abundance seems to be increasing in most parts of the Mediterranean, probably favoured by warming conditions. Significant genetic differentiation was found both spatially and temporally. The main break corresponded to the separation of western Atlantic populations from those in eastern Atlantic and the Mediterranean Sea. A less marked, but significant differentiation was also found between Macaronesia (eastern Atlantic) and the Mediterranean. In the latter area, a signal of differentiation between the transitional area (Alboran Sea) and the rest of the Mediterranean was detected. However, no genetic structure is found within the Mediterranean (excluding Alboran) across the Siculo-Tunisian Strait, resulting from either enough gene flow to homogenize distance areas or/and a recent evolutionary history marked by demographic expansion in this basin. Genetic temporal variation at the Alboran Sea is as important as spatial variation, suggesting that temporal changes in hydrological features can affect the genetic composition of the populations. A picture of genetic homogeneity in the Mediterranean emerges, implying that the potential expansion of this keystone species will not be limited by intraspecific genetic features and/or potential impact of postulated barriers to gene flow in the region.

The obligate alkalophilic soda-lake fungus Sodiomyces alkalinus has shifted to a protein diet.

Sodiomyces alkalinus is one of the very few alkalophilic fungi, adapted to grow optimally at high pH. It is widely distributed at the plant-deprived edges of extremely alkaline lakes and locally abundant. We sequenced the genome of S. alkalinus and reconstructed evolution of catabolic enzymes, using a phylogenomic comparison. We found that the genome of S. alkalinus is larger, but its predicted proteome is smaller and heavily depleted of both plant-degrading enzymes and proteinases, when compared to its closest plant-pathogenic relatives. Interestingly, despite overall losses, S. alkalinus has retained many proteinases families and acquired bacterial cell wall-degrading enzymes, some of them via horizontal gene transfer from bacteria. This fungus has very potent proteolytic activity at high pH values, but slowly induced low activity of cellulases and hemicellulases. Our experimental and in silico data suggest that plant biomass, a common food source for most fungi, is not a preferred substrate for S. alkalinus in its natural environment. We conclude that the fungus has abandoned the ancestral plant-based diet and has become specialized in a more protein-rich food, abundantly available in soda lakes in the form of prokaryotes and small crustaceans.

Encapsulation of curcumin using supercritical antisolvent (SAS) technology to improve its stability and solubility in water.

This study aims to develop a formulation of a turmeric-based dye extract using supercritical antisolvent (SAS) technology and different encapsulating polymers to improve the aqueous stability and solubility of curcumin. The dye formulation obtained by SAS consists of a mixture of Eudragit® L100 and Pluronic® 127 using tween 20 as a surfactant The characterization and quantification of curcumin extracts and the encapsulation product were conducted using high performance liquid chromatography (HPLC). The properties of the dye were determined using SEM, DSC, X-ray diffraction, particle size, zeta potential, and oxygen radical absorbance capacity (ORAC) to measure antioxidant capacity. The dye formulation contained a curcumin content of 4.45 µg/mL. The average diameter of the amorphous particles was 5667.4 nm, and the zeta potential was 11.21 mV. The largest aqueous stability and solubility was observed at pH 4. From color comparison with a solution of tartrazine 200 µg/mL solution of dye formulation based on curcuminoids is equivalent to an approximated 30 µg/mL tartrazine solution.

Impact of Hypothermic Stress During Special Operations Training of Chilean Military Forces.

Introduction: The Chilean Army considers processes that can optimize physical capacities for responding to the impact of situations and given stressors. The study of the effect of hypothermia as a stressor agent (HSA) and its relationship with cardiovascular, hematological, anthropometric, endocrine, and immunological parameters has not been fully addressed experimentally in military populations. Objective: To identify the endocrine, hematological, cardiovascular, and immunological changes caused by HSA and to associate these variables with body composition and physical fitness in the military special operation courses of the Chilean Army. Materials and Methods: Forty-two male subjects were exposed to remain in cold water (10.6 °C) in the context of regular military operations training, the longest time of exposure was determined by individual volitional limits. The measurements were taken in pre-hypothermia conditions, then 2 d later under acute hypothermia condition, and finally during the course period of lesser physical and psychological stressors where the baseline measurements were taken. The statistical analysis consisted of testing normality of the distribution through the Shapiro-Wilk test, assessing the equality of variances through the Levene test, and variance analysis by applying the ANOVA test (analysis of variance). The Bonferroni test was used for multiple comparison correction and the Pearson test for correlations between two variables. The level of significance was of p < 0.05. Results: The main finding of this study is that HSA has a significant impact at the cardiovascular level and produces an increment in the cell population of the immune and hematologic systems. Significant hormonal changes were observed: ACTH (r = 0.50, p < 0.002), cortisol (r = 0.32, p < 0.03), free testosterone (r = 0.13, p < 0.002), total testosterone r = 0.31, p < 0.002), and anthropometrics (r = -0.51, p < 0.05). However, there is no significant correlation between physical fitness and HAS. Conclusions: All subjects experienced hypothermia stress elicited by immersion in cold water. This was evidenced by the decrease in core temperature as well as cardiovascular, endocrine, anthropometric, and immunological changes. Individual differences exist between subjects and their resistance to hypothermia in cold water. These differences are not explained by the physical fitness profile but rather respond to a greater body adiposity index and minor changes in the adrenocorticotropic hormone and cortisol hormone. An acute hypothermia stress condition also affects the anabolic/catabolic environment. Finally, HSA produces an increase in the cell population of the immune system. The authors believe that this study allows to standardize HSA exposure times during regular military operations training by identifying the physiological impacts under this extreme environment. At present, the availability of intra-abdominal temperature measurement apparatus with capsule thermometers raises the interest of corroborating the findings of the current study through the use of such measuring devices. Likewise, an interesting line of research for the future would be to compare the HSA against a psychological evaluation with the purpose of identifying the stress management mechanisms among subjects of these characteristics and include heart rate variability measurements as an indicator of sympathetic stress.

Extraction and identification of endopeptidases in convection dried papaya and pineapple residues: A methodological approach for application to higher scale.

The use of agro-industrial waste for application in the obtention of products with high added value has become a trend in recent years, especially in tropical countries whose main economic sector is agricultural exports. In the present study, an applicable method to food industry of extracting proteolytic enzymes from dryed papaya and pineapple residues by convection was developed. Different to other scientific reports the heat treatment at 40 °C of waste residues, to reach 20% moisture, allowed an increase in total soluble protein content and did not alter the proteolytic activity of the extracts when phosphate buffer pH 7.0 was used as solvent. In the residues evaluated as candidates for the extraction of endopeptidases, we observed that green dried papaya peel and dried pineapple core, had higher activity values (914.34 ±â€¯25.47 U/mg and 2152.36 ±â€¯75.99 U/mg, respectively). These results, combined with one-dimensional electrophoresis and protein identification methods by MALDI TOF-TOF, showed the presence of signal peptides characteristic of papain, bromelain and other endopeptidases previously reported in extracts of fresh papaya and pineapple residues. These findings show that the drying of the residues by convection does not alter neither the activity nor the structure of the proteolytic enzymes. Finally, it is confirmed that the use of 20% ammonium sulfate as a precipitating agent allows to reach an efficiency of 74% in different work scales the use of purification and identification protocols in a more adaptable way, making them the most promising waste in Colombia, due to its potential for the production of bromelain on a larger scale.

Experimental evolution to increase the efficacy of the entomopathogenic fungus Beauveria bassiana against malaria mosquitoes: Effects on mycelial growth and virulence.

Entomopathogenic fungi such as Beauveria bassiana are currently considered as a potential control agent for malaria mosquitoes. The success of such strategies depends among others on the efficacy of the fungus to kill its hosts. As B. bassiana can use various resources for growth and reproduction, increasing the dependency on mosquitoes as a nutritional source may be instrumental for reaching this goal. Passage of entomopathogenic fungi through an insect host has been shown to increase its virulence. We evaluated the virulence, fungal outgrowth, mycelial growth rate, and sporulation rate of two B. bassiana isolates (Bb1520 and Bb8028) that underwent 10 consecutive selection cycles through malaria mosquitoes (Anopheles coluzzii) using an experimental evolution approach. This cycling resulted in an altered capacity of evolved B. Bassiana lineages to grow on different substrates while maintaining the ability to kill insects. Notably, however, there were no significant changes in virulence or speed of outgrowth when comparing the evolved lineages against their unevolved ancestors. These results suggest that fungal growth and sporulation evolved through successive and exclusive use of an insect host as a nutritional resource. We discuss the results in light of biocontrol and provide suggestions to increase fungal virulence.

Comparative genomics of Beauveria bassiana: uncovering signatures of virulence against mosquitoes.

BACKGROUND: Entomopathogenic fungi such as Beauveria bassiana are promising biological agents for control of malaria mosquitoes. Indeed, infection with B. bassiana reduces the lifespan of mosquitoes in the laboratory and in the field. Natural isolates of B. bassiana show up to 10-fold differences in virulence between the most and the least virulent isolate. In this study, we sequenced the genomes of five isolates representing the extremes of low/high virulence and three RNA libraries, and applied a genome comparison approach to uncover genetic mechanisms underpinning virulence. RESULTS: A high-quality, near-complete genome assembly was achieved for the highly virulent isolate Bb8028, which was compared to the assemblies of the four other isolates. Whole genome analysis showed a high level of genetic diversity between the five isolates (2.85-16.8 SNPs/kb), which grouped into two distinct phylogenetic clusters. Mating type gene analysis revealed the presence of either the MAT1-1-1 or the MAT1-2-1 gene. Moreover, a putative new MAT gene (MAT1-2-8) was detected in the MAT1-2 locus. Comparative genome analysis revealed that Bb8028 contains 163 genes exclusive for this isolate. These unique genes have a tendency to cluster in the genome and to be often located near the telomeres. Among the genes unique to Bb8028 are a Non-Ribosomal Peptide Synthetase (NRPS) secondary metabolite gene cluster, a polyketide synthase (PKS) gene, and five genes with homology to bacterial toxins. A survey of candidate virulence genes for B. bassiana is presented. CONCLUSIONS: Our results indicate several genes and molecular processes that may underpin virulence towards mosquitoes. Thus, the genome sequences of five isolates of B. bassiana provide a better understanding of the natural variation in virulence and will offer a major resource for future research on this important biological control agent.