Light and focused ion beam microscopy workflow for resin-embedded tissues

Although the automated image acquisition with the focused ion beam scanning electron microscope (FIB-SEM) provides volume reconstructions, volume analysis of large samples remains challenging. Here, we present a workflow that combines a modified sample protocol of the classical transmission electron microscope with FIB-SEM volume imaging. The proposed workflow enables efficient 3D structural surveys of rabbit ovaries collected at consecutive developmental stages. The precise trimming of the region of interest adds the time dimension to the volume, constructing a virtual 4D electron microscopy. We found filopodia-like processes emitted by oocyte cysts allowing contact between oocytes not previously observed

PARÁSITOS POTENCIALMENTE ZOONÓTICOS HALLADOS EN SEIS GRANJAS PORCÍCOLAS DE CUNDINAMARCA, COLOMBIA

In Colombia, the departments of Cundinamarca, Huila and Tolima, among others, represent 19.16% of the total domestic production of pig farms. Gastrointestinal parasites (PGI) limit the productive potential of infected animals, affecting parameters such as average daily gain and food conversion ratio. The aim of the study was to determinate the prevalence of PGI with a zoonotic potential in backyard farms and semi-technical production. Farmyard pigs were sampled in three semi-technified farms with full production cycle (breeding, fattening and lift) in the municipalities of Vega, Chia, Tibirita and in four backyard pig farms (fattening) in the towns of Chía and Santandersito. The farms were chosen for convenience. Three samplings were performed serially at intervals of 15 days and analyzed with a total of 119 samples, 94 farms and 25 semi-technified backyard farms. Stool samples in "pool" per pen, were analyzed by direct analysis, qualitative flotation technique and staining with modified Ziehl-Neelsen. In semi-technified farms, Balantidium coliMalmsten, 1857 cysts (40.5%), Giardiaspp. (26.9%) cysts, Balantidium colitrophozoites (3.5%), Eimeriaspp. (3.8%), eggs of Anoplocephalidaeeggs (1.8%), hepatic Fasciolaeggs (2.0%), Ascaris suumGoeze, 1782 eggs (0.6%), cysts of Entamoeba(1.5%) and larvae of Strongyloidesspp. (0.3%) were detected. In backyard farms we found cysts of Entamoebaspp. (42.5%), cysts of Giardiaspp. (30%), Blastocystis spp (37.5%) and Taenia soliumLinnaeus, 1758 eggs (2.5%). The presence of Giardia, Entamoebaand Blastocystissuggests the possible transmission of parasite populations between pigs and humans thus increasing the transmission of parasites zoonotic potential.En Colombia, en los departamentos de Cundinamarca, Huila y Tolima se encuentra el 19,16% de granjas porcícolas del total de la producción nacional y como en otras regiones del mundo, la producción se ve afectada por parásitos gastrointestinales (PGI), que limitan el potencial productivo de los animales infestados, afectando parámetros como la ganancia diaria de peso y el índice de conversión alimenticias. El objetivo del estudio fue determinar la prevalencia de PGI .con potencial zoonótico en granjas traspatio y de producción semitecnificada. El estudio fue de tipo descriptivo transversal. Se muestrearon cerdos en corral (no pastoreo) de tres granjas semitecnificadas con ciclo productivo completo (cría, levante y ceba) en los municipios de la Vega, Chía, Tibirita y cuatro granjas de cerdos traspatio (ceba) en los municipios de Chía y Santandersito, elegidas por conveniencia. Fueron realizados tres muestreos seriados con intervalos de 15 días y se analizaron un total de 119 muestras, 94 de granjas semitecnificadas y 25 en granjas traspatio. Las muestras de materia fecal, fueron analizadas mediante: análisis directo, técnica de flotación cualitativa y tinción con Ziehl-Neelsen modificado. En granjas semitecnificadas se observó: Quistes de Balantidium coliMalmsten, 1857 (40,5%), Quistes de Giardiaspp. (26,9%), Trofozoitos de B. coli(3,5%), Ooquistes de Eimeriaspp. (3,8%), Huevos compatibles con familia Anoplocephalidae (1,8%), Huevos de Fasciola hepatica(2,0%), Huevos de Ascaris suum(0,6%), Quistes de Entamoebaspp. (1,5%) y Larvas de Strongyloidesspp. (0,3%). En las granjas traspatio se encontró: Quistes de Entamoebaspp. (42,5%), Quistes de Giardiaspp. (30%), Blastocystisspp (37,5%) y huevos de Taenia soliumLinnaeus, 1758 (2,5%). La presencia de Giardia, Entamoebay Blastocystissugiere la posible rotación de poblaciones parasitarias entre cerdos y humanos, ampliando la transmisión de parásitos con potencial zoonótico entre las dos especies

An SiO Toroid and Wide-angle Outflow associated with the Massive Protostar W75N(B)-VLA2

We have carried out ALMA observations of the massive star-forming region W75N(B), which contains the massive protostars VLA1, VLA2, and VLA3. Particularly, VLA2 is an enigmatic protostar associated with a wind-driven H$_2$O maser shell, which has evolved from an almost isotropic outflow to a collimated one in just 20 years. The shell expansion seemed to be halted by an obstacle located to the northeast of VLA2. Here we present our findings from observing the 1.3 mm continuum and H$_2$CO and SiO emission lines. Within a region of $\sim 30"$ ($\sim 39,000$ au) diameter, we have detected 40 compact mm-continuum sources, three of them coinciding with VLA1, VLA2, and VLA3. While the H$_2$CO emission is mainly distributed in a fragmented structure around the three massive protostars, but without any of the main H$_2$CO clumps spatially coinciding with them, the SiO is highly concentrated on VLA2, indicating the presence of very strong shocks generated near this protostar. The SiO emission is clearly resolved into an elongated structure ($\sim 0.6"\times0.3"$; $\sim 780$ au$\times$390 au) perpendicular to the major axis of the wind-driven maser shell. The structure and kinematics of the SiO emission are consistent with a toroid and a wide-angle outflow surrounding a central mass of $\sim 10$ M$_{\odot}$, thus supporting previous theoretical predictions regarding the evolution of the outflow. Additionally, we have identified the expected location and estimated the gas density of the obstacle that is hindering the expansion of the maser shell.Comment: To be published in The Astrophysical Journal Letters. Sixteen pages, seven figures. Updated metadat

Minimal impacts of invasive Scaevola taccada on Scaevola plumieri via pollinator competition in Puerto Rico

IntroductionScaevola taccada and Scaevola plumieri co-occur on shorelines of the Caribbean. Scaevola taccada is introduced in this habitat and directly competes with native dune vegetation, including S. plumieri, a species listed as locally endangered and threatened in Caribbean locations. This study addresses whether the invasive S. taccada also impacts the native S. plumieri indirectly by competing for pollinators and represents the first comparative study of insect visitation between these species.MethodsInsect visitation rates were measured at sites where species co-occur and where only the native occurs. Where species cooccur, insect visitors were captured, identified and analyzed for the pollen they carry. Pollen found on open-pollinated flowers was analyzed to assess pollen movement between the two species. We also compared floral nectar from each species by measuring volume, sugar content, and presence and proportions of amine group containing constituents (AGCCs).ResultsOur results demonstrate that both species share insect visitors providing the context for possible pollinator competition, yet significant differences in visitation frequency were not found. We found evidence of asymmetrical heterospecific pollen deposition in the native species, suggesting a possible reproductive impact. Insect visitation rates for the native were not significantly different between invaded and uninvaded sites, suggesting that the invasive S. taccada does not limit pollinator visits to S. plumieri. Comparisons of nectar rewards from the invasive and the native reveal similar volumes and sugar concentrations, but significant differences in some amine group containing constituents that may enhance pollinator attraction.ConclusionOur analysis finds no evidence for pollination competition and therefore S. taccada’s main impacts on S. plumieri are through competitive displacement and possibly through reproductive impacts as a consequence of heterospecific pollen deposition

Design of the nutritional therapy for overweight and obese Spanish adolescents conducted by registered dieticians: the EVASYON study

Background: Dietary treatment for obese adolescents should aim to ensure adequate growth and development, by reducing excessive fat mass accumulation, avoiding loss of lean body mass, improving well-being and selfesteem and preventing cyclical weight regain. The aim of this article is to describe the dietary intervention design and the methods used to evaluate nutritional knowledge and behavior in the EVASYON study (Development, implementation and evaluation of the efficacy of a therapeutic programme for overweight/obese adolescents). Methods/design: EVASYON is a multi-centre study conducted in 5 Spanish hospital settings (Granada, Madrid, Pamplona, Santander and Zaragoza), where 204 overweight/obese Spanish adolescents were treated in groups of 9 to 11 subjects over 20 visits. The study was implemented in two stages: an intensive, calorie-restricted period for the first 9 weeks, and an extensive body-weight follow-up period for the last 11 months. A moderate energy intake restriction was applied in the intensive period according to the degree of obesity, on the basis of a balanced diet supplying 50-55% of daily energy as carbohydrates; 30-35% as fats and 10-15% as proteins. In the intensive period, adolescents were prescribed both a fixed full-day meal plan for the first three weeks and a full day meal plan with different food-choices for 6 weeks. Later, adolescents received a flexible meal plan based on food exchanges for the follow-up period until the end of the trial. Data on food intake, dietary and meal-related habits and behavior were collected by means of dietary questionnaires. To analyse nutritional knowledge, adolescents were examined regarding nutrient concepts and food items for a healthy diet with the appropriate tools. Participants were given nutritional information with complementary teaching material, which was available on the EVASYON website (www.estudioevasyon.com). Discussion: The dietary intervention of the EVASYON programme with a moderate calorie restriction for a limi - ted period of time could be a good strategy in treating overweight and obese adolescents and that will be tested further. Moreover, combining fixed plan with free-choice menus may help adolescents and their families to make right decisions for every day meals

Psychometric properties of the Clarke questionnaire for hypoglycemia awareness in the Spanish population with type 2 diabetes

Objectives: The Clarke questionnaire, validated in Spanish language, assesses hypoglycemia awareness in patients with type 1 diabetes. This study aimed to analyze its psychometric properties in patients with type 2 diabetes (T2DM). Methods: This was a questionnaire validation study. Patients with T2DM and treated with insulin, sulfonylureas or glinides were consecutively recruited from six endocrinology consultations and six primary care centers. The internal structure of the 8-item Clarke questionnaire was analyzed by exploratory (training sample) and confirmatory (testing sample) factor analysis; the internal consistency using Omega’s McDonald coefficient; and goodness of fit with comparative fit index (CFI, cutoff >0.9), Goodness of Fit Index (GFI, cutoff >0.9), and root mean-square error of approximation (RMSEA, cutoff <0.09), as well as unidimensionality indicators. Results: The 265 participants (56.8% men) had a mean age of 67.8 years. Confirmatory factor analysis for one dimension obtained poor indicators: fit test (p < 0.001); CFI = 0.748; RMSEA = 0.122 and SRMR = 0.134. Exploratory factor analysis showed 2 or 3 dimensions with poor adjustment indicators. Omega’s McDonald was 0.739. Conclusions: The Spanish version of the Clarke questionnaire was not valid or reliable for assessing hypoglycemia awareness in people with T2DM in Spanish population

In silico Analyses of Immune System Protein Interactome Network, Single-Cell RNA Sequencing of Human Tissues, and Artificial Neural Networks Reveal Potential Therapeutic Targets for Drug Repurposing Against COVID-19

Background: There is pressing urgency to identify therapeutic targets and drugs that allow treating COVID-19 patients effectively.Methods: We performed in silico analyses of immune system protein interactome network, single-cell RNA sequencing of human tissues, and artificial neural networks to reveal potential therapeutic targets for drug repurposing against COVID-19.Results: We screened 1,584 high-confidence immune system proteins in ACE2 and TMPRSS2 co-expressing cells, finding 25 potential therapeutic targets significantly overexpressed in nasal goblet secretory cells, lung type II pneumocytes, and ileal absorptive enterocytes of patients with several immunopathologies. Then, we performed fully connected deep neural networks to find the best multitask classification model to predict the activity of 10,672 drugs, obtaining several approved drugs, compounds under investigation, and experimental compounds with the highest area under the receiver operating characteristics.Conclusion: After being effectively analyzed in clinical trials, these drugs can be considered for treatment of severe COVID-19 patients. Scripts can be downloaded at

Defining the clonal dynamics leading to mouse skin tumour initiation.

The changes in cell dynamics after oncogenic mutation that lead to the development of tumours are currently unknown. Here, using skin epidermis as a model, we assessed the effect of oncogenic hedgehog signalling in distinct cell populations and their capacity to induce basal cell carcinoma, the most frequent cancer in humans. We found that only stem cells, and not progenitors, initiated tumour formation upon oncogenic hedgehog signalling. This difference was due to the hierarchical organization of tumour growth in oncogene-targeted stem cells, characterized by an increase in symmetric self-renewing divisions and a higher p53-dependent resistance to apoptosis, leading to rapid clonal expansion and progression into invasive tumours. Our work reveals that the capacity of oncogene-targeted cells to induce tumour formation is dependent not only on their long-term survival and expansion, but also on the specific clonal dynamics of the cancer cell of origin.C.B. is an investigator of WELBIO. A.S-D. and JC.L. are supported by a fellowship of the FNRS and FRIA respectively. B.D.S. and E.H. are supported by the Wellcome Trust (grant number 098357/Z/12/Z and 110326/Z/15/Z). EH is supported by a fellowship from Trinity College, Cambridge. This work was supported by the FNRS, the IUAP program, the Fondation contre le Cancer, the ULB fondation, the foundation Bettencourt Schueller, the foundation Baillet Latour, a consolidator grant of the European Research Council.This is the author accepted manuscript. The final version is available from Nature Publishing Group via http://dx.doi.org/10.1038/nature1906

Use of water-Cherenkov detectors to detect Gamma-Ray-Bursts at the Large Aperture GRB Observatory (LAGO)

The Large Aperture GRB Observatory (LAGO) project aims at the detection of high energy photons from Gamma Ray Bursts (GRB) using the single particle technique in ground-based water-Cherenkov detectors (WCD). To reach a reasonable sensitivity, high altitude mountain sites have been selected in Mexico (Sierra Negra, 4550 m a.s.l.), Bolivia (Chacaltaya, 5300 m a.s.l.) and Venezuela (Me´ rida, 4765 m a.s.l.). We report on detector calibration and operation at high altitude, search for bursts in 4 months of preliminary data, as well as search for signal at ground level when satellites report a burst.Fil: Allard, D.. Université Paris Diderot - Paris 7; FranciaFil: Allekotte, Ingomar. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Alvarez, C.. Facultad de Ciencias Fısico-Matematicas; MéxicoFil: Asorey, Hernán Gonzalo. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Barros, H.. Universidad Simon Bolivar; VenezuelaFil: Bertou, Xavier Pierre Louis. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Burgoa, O.. Instituto de Investigaciones Fisicas; BoliviaFil: Gomez Berisso, Mariano. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Martinez, O.. Facultad de Ciencias Fısico-Matematicas; MéxicoFil: Miranda Loza, P.. Instituto de Investigaciones Fısicas; BoliviaFil: Murrieta, T.. Facultad de Ciencias Fısico-Matematicas; MéxicoFil: Perez, G.. Facultad de Ciencias Fısico-Matematicas; MéxicoFil: Rivera, H.. Instituto de Investigaciones Fısicas; BoliviaFil: Rovero, Adrian Carlos. Consejo Nacional de Investigaciónes Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Saavedra, O.. Istituto Nazionale di Fisica Nucleare; ItaliaFil: Salazar, H.. Facultad de Ciencias Fısico-Matematicas ; MéxicoFil: Tello, J. C.. Universidad Simon Bolıvar; VenezuelaFil: Ticona Peralda, R.. Instituto de Investigaciones Fısicas; BoliviaFil: Velarde, A.. Instituto de Investigaciones Fısicas; BoliviaFil: Villaseñor, L.. Universidad de Michoacan; MéxicoFil: Areso, Omar Antonio. Consejo Nacional de Investigaciónes Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Arnaldi, Luis Horacio. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Dasso, Sergio Ricardo. Consejo Nacional de Investigaciónes Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Gonzalez, M.. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Gulisano, Adriana Maria. Consejo Nacional de Investigaciónes Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Astronomía y Física del Espacio. - Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Astronomía y Física del Espacio; ArgentinaFil: Martin, R.. Ministerio de Relaciones Exteriores, Comercio Interno y Culto. Dirección Nacional del Antártico. Instituto Antártico Argentino; ArgentinaFil: Masías Meza, Jimmy Joel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Física de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Física de Buenos Aires; ArgentinaFil: Sidelnik, Iván Pedro. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; ArgentinaFil: Alvarez, W.. Universidad de San Carlos; GuatemalaFil: The LAGO Collaboration

Telomere Length as a Biomarker for Adiposity Changes after a Multidisciplinary Intervention in Overweight/Obese Adolescents: The EVASYON Study

[Context] Telomeres are biomarkers of biological aging. Shorter telomeres have been associated with increased adiposity in adults. However, this relationship remains unclear in children and adolescents. [Objective] To evaluate the association between telomere length (TL) and adiposity markers in overweight/obese adolescents after an intensive program. We hypothesize that greater TL at baseline would predict a better response to a weight loss treatment. Design, Setting, Patients and Intervention The EVASYON is a multidisciplinary treatment program for adolescents with overweight and obesity that is aimed at applying the intervention to all possibly involved areas of the individual, such as dietary habits, physical activity and cognitive and psychological profiles. Seventy-four participants (36 males, 38 females, 12–16 yr) were enrolled in the intervention program: 2 months of an energy-restricted diet and a follow-up period (6 months). [Main Outcome] TL was measured by quantitative real-time polymerase chain reaction at baseline and after 2 months; meanwhile, anthropometric variables were also assessed after 6 months of follow-up. [Results] TL lengthened in participants during the intensive period (+1.9±1.0, p<0.001) being greater in overweight/obese adolescents with the shortest telomeres at baseline (r = −0.962, p<0.001). Multivariable linear regression analysis showed that higher baseline TL significantly predicted a higher decrease in body weight (B = −1.53, p = 0.005; B = −2.25, p = 0.047) and in standard deviation score for body mass index (BMI-SDS) (B = −0.22, p = 0.010; B = −0.47, p = 0.005) after the intensive and extensive period treatment respectively, in boys. [Conclusion] Our study shows that a weight loss intervention is accompanied by a significant increase in TL in overweight/obese adolescents. Moreover, we suggest that initial longer TL could be a potential predictor for a better weight loss response.Research relating to this work was funded by grants from the Health Research Fund from the Carlos III Health Institute from Ministry of Health and Consumption, Fondo de Investigación Sanitaria (FIS; PI051579, PI051080) for the EVASYON project; Línea Especial, Nutrición y Obesidad (University of Navarra); Spanish Ministry of Science and Innovation (MICINN) [SAF2010-20367]; Carlos III Health Institute [Centro de Investigación Biomédica en Red (CIBER) project, CB06/03/1017], and RETICS network. The scholarship to S. García-Calzón from the FPU ‘Formación de Profesorado Universitario’ from the Spanish Ministry is fully acknowledged