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KEY POINTS Least shrews were studied to explore the relationship between metabolic function, mitochondrial morphology and protein content in different tissues. Liver and kidney mitochondrial content and enzymatic activity approaches the heart indicating similar metabolic demand among tissues that contribute to basal and maximum metabolism. This allows examination of mitochondrial structure and composition in tissues with similar maximum metabolic demands. Mitochondrial networks only occur in striated muscle. In contrast, the liver and kidney maintain individual mitochondria with limited reticulation. Muscle mitochondrial reticulation is the result of dense ATPase activity and cell-spanning myofibrils which require networking for adequate metabolic support. In contrast, liver and kidney ATPase activity is localized to the endoplasmic reticulum and basolateral membrane respectively, generating a locally balanced energy conversion and utilization Mitochondrial morphology is not driven by maximum metabolic demand, but by the cytosolic distribution of energy utilizing systems set by the functions of the tissue. ABSTRACT Mitochondrial adaptations are fundamental to differentiated function and energetic homeostasis in mammalian cells. But the mechanisms that underlie these relationships remain poorly understood. Here, we investigated organ-specific mitochondrial morphology, connectivity and protein composition in a model of extreme mammalian metabolism, the Least shrew (Cryptotis parva). This was achieved through a combination of high-resolution 3D focused-ion-beam EM imaging and tandem-mass-tag MS proteomics. We demonstrate that liver and kidney mitochondrial content are equivalent to the heart permitting assessment of mitochondrial adaptations in different organs with similar metabolic demand. Muscle mitochondrial networks (cardiac and skeletal) are extensive, with a high incidence of nanotunnels - which collectively support the metabolism of large muscle cells. Mitochondrial networks were not detected in the liver and kidney as individual mitochondria are localized with sites of ATP consumption. This configuration is not observed in striated muscle, likely due to a homogenous ATPase distribution and the structural requirements of contraction. These results demonstrate distinct, fundamental mitochondrial structural adaptations for similar metabolic demand that are dependent on the topology of energy utilization process in a mammalian model of extreme metabolism. Abstract figure. This study investigates the role of mitochondrial morphology and protein composition in setting the extreme metabolic rates of one of the smallest extant mammals - the North American least shrew (Cryptotis parva). To do this, mitochondrial characteristics from liver, kidney, skeletal muscle and heart tissues were compared as these tissues are major contributors to basal and maximum metabolic states. Liver and kidney mitochondrial volume density and protein content approach levels observed in the heart - indicating that these former tissues are major contributors to the high basal metabolic rates of small mammals. Despite this high mitochondrial content, the liver and kidney do not exhibit mitochondrial networking - structures that are proposed to conduct mitochondrial proton motive force at the scale of the cell. Shrew skeletal muscle and cardiac mitochondrial network organization is consistent with networks observed in larger mammals while also exhibiting increased connectivity at the nm-scale. Instead of forming networks, kidney and liver mitochondria are directly associated with sites of ATP utilization. These results identify conditions that dictate the formation of mitochondrial networks and processes that drive mammalian allometric scaling of metabolic rates. This article is protected by copyright. All rights reserved.

Knowing the potential of natural resources associated with mining activity is critical to decision-making especially at the exploration stage, this allows organizations to assess whether the area of interest is viable, to continue with post-exploration processes of mining deposits. So, it is important to get new exploration processes that cover very wide surfaces, be low-cost and reveal high reliability, one of these alternatives is the use of satellite imagery based on the principles of spectral responses. Therefore, The objective of the research was to search for a technique that considers the quantitative criteria to relate and identify the objects by using the spectral response values whose spectral response patterns are different, in an area whose cutting law for exploitation is feasible (zones of epithermal alteration), with the spectral response values of compounds associated with alteration zones of the United States Geological Survey (USGS), using two statistical criteria, correlation analysis and quadratic mean error, which have revealed good performance in identifying objects from the comparison of spectral responses.

Background Appropriate thrombus-device interaction is critical for recanalization. Histology can serve as a proxy for mechanical properties, and thus inform technique selection. Objective To investigate the value of histologic characterization, we conducted a systematic review and meta-analysis on the relationship between thrombus histology and recanalization, technique, etiology, procedural efficiency, and imaging findings. Methods In this meta-analysis, we identified studies published between March 2010 and March 2020 reporting findings related to the histologic composition of thrombi in large vessel occlusion stroke. Studies with at least 10 patients who underwent mechanical thrombectomy using stent retriever or aspiration were considered. Only studies in which retrieved thrombi were histologically processed were included. Patient-level data were requested when data could not be directly extracted. The primary outcome assessed was the relationship between thrombus histology and angiographic outcome. Results A total of 22 studies encompassing 1623 patients met inclusion criteria. Clots associated with good angiographic outcome had higher red blood cell (RBC) content (mean difference [MD] 9.60%, 95% CI 3.85-15.34, P = .008). Thrombi retrieved by aspiration had less fibrin (MD -11.39, 95% CI -22.50 to -0.27, P = .046) than stent-retrieved thrombi. Fibrin/platelet-rich clots were associated with longer procedure times (MD 13.20, 95% CI 1.30-25.10, P = .037). Hyperdense artery sign was associated with higher RBC content (MD 14.17%, 95% CI 3.07-25.27, P = .027). No relationship was found between composition and etiology. Conclusion RBC-rich thrombi were associated with better recanalization outcomes and shorter procedure times, suggesting that preinterventional compositional characterization may yield important prognostic and therapeutic guidance.

Cavia tschudii inhabits coastal and Andean wetlands where it is important prey for medium carnivores, but its habitat selection and its role in the wetlands are unknown. In order to reduce this lack of knowledge, we evaluated changes in the abundance for two seasons and the habitat selection of C. tschudii in a wetland on the central coast of Peru. Additionally, we report information on their movement distances and provide comments on their predators. We carried out six evaluations during the autumn and spring of 2019, with nine grids in three plant communities: grassland, bulrush community, and cattail community. Each captured individual was marked with a numbered ear tag. We compared the relative abundance between seasons using the Mann-Whitney U test, and calculated the relative abundance per evaluated month and carried out regressions to model its behavior. The use/availability of habitat was evaluated with the Chi-square test together with Bonferroni confidence intervals to show habitat selection. Finally, we estimate the mean maximum distance moved (MMDM) of recaptured individuals. The results show significant differences between the seasons, with higher relative abundance in autumn. The relative abundance showed a peak in April, from which the values decrease. Likewise, we found significant differences in habitat selection, showing positive selection for the cattail community, neutral selection for the bulrush community, and negative selection for the grassland. The MMDM was 36.5 ± 15.7 m. We report a decrease in the relative abundance of C. tschudii , possibly related to seasonal changes in habitat quality or to the presence of predators. The peak of abundance in mid-autumn and the apparent decrease in the population until late spring resembles the annual dynamics reported for C. aperea . It is also confirmed that C. tschudii , like other Cavia , select environments with greater plant coverage, possibly as an anti-predation strategy. We report dogs predating C. tschudii and provide a list of other potential predators. This study increases the information on C. tschudii in coastal wetlands and gives a first approach to the necessary knowledge for its management and conservation within these fragile ecosystems.

Background The biophysics of an organism span multiple scales from subcellular to organismal and include processes characterized by spatial properties, such as the diffusion of molecules, cell migration, and flow of intravenous fluids. Mathematical biology seeks to explain biophysical processes in mathematical terms at, and across, all relevant spatial and temporal scales, through the generation of representative models. While non-spatial, ordinary differential equation (ODE) models are often used and readily calibrated to experimental data, they do not explicitly represent the spatial and stochastic features of a biological system, limiting their insights and applications. However, spatial models describing biological systems with spatial information are mathematically complex and computationally expensive, which limits the ability to calibrate and deploy them and highlights the need for simpler methods able to model the spatial features of biological systems. Results In this work, we develop a formal method for deriving cell-based, spatial, multicellular models from ODE models of population dynamics in biological systems, and vice versa. We provide examples of generating spatiotemporal, multicellular models from ODE models of viral infection and immune response. In these models, the determinants of agreement of spatial and non-spatial models are the degree of spatial heterogeneity in viral production and rates of extracellular viral diffusion and decay. We show how ODE model parameters can implicitly represent spatial parameters, and cell-based spatial models can generate uncertain predictions through sensitivity to stochastic cellular events, which is not a feature of ODE models. Using our method, we can test ODE models in a multicellular, spatial context and translate information to and from non-spatial and spatial models, which help to employ spatiotemporal multicellular models using calibrated ODE model parameters. We additionally investigate objects and processes implicitly represented by ODE model terms and parameters and improve the reproducibility of spatial, stochastic models. Conclusion We developed and demonstrate a method for generating spatiotemporal, multicellular models from non-spatial population dynamics models of multicellular systems. We envision employing our method to generate new ODE model terms from spatiotemporal and multicellular models, recast popular ODE models on a cellular basis, and generate better models for critical applications where spatial and stochastic features affect outcomes.

The effects of the use of solvents (96% ethanol and PEG 400), solute ratios: solvent (1: 4 and 1: 6 w/w), extraction temperatures (50 and 60 °C) and leaching techniques (conventional and assisted by ultrasound) on the concentration and time of extraction of bixin from Achiote (Bixa Orellana L.) were evaluated. The extraction kinetics were constructed, and their orders were determined. The experimental values were adjusted through the mathematical models of Peleg and Azuara. ANOVA was used based on a 23 design, blocking type of technology. The kinetics showed two stages of extraction: rapid and slow. The reaction orders were decimal numbers, which indicates the complexity of the extraction mechanism. The highest orders of reaction (2.97 and 2.075) were presented with extraction with ultrasound-assisted and ethanol as solvent, at 50 and 60 °C and solute solvent ratio 1: 4 and 1: 6, respectively. The concentration of Bixin was increased by approximately 30% by increasing the temperature. The Azuara model presented the highest adjustment with a standard error

16 páginas.- 5 figuras.- 3 tablas.- 67 referencias, This study was conducted to evaluate the effect of compost produced with agricultural residues and oat-based biochar, iron oxide and halloysite nanoparticles as additives of the process of composting on soil chemical properties, nutrient status and growth of ryegrass Lolium perenne L. For this, a 90-day mesocosm experiment was carried out under greenhouse conditions. Bare soil and a basal fertilization treatment were compared to soils amended with nonadditive compost (NA compost), compost supplied with oat-based biochar (Bioch compost), iron oxide nanoparticles (Fe compost), and halloysite nanoparticles (Ha compost). Compost supplied with nanoparticles and biochar combined were also considered. The incorporation of compost with or without additives increased the content of total C and N in soil, with N diminishing (total and mineral forms) and C/N modifications after 90 days. The addition of compost and co-composted treatments also increased the total contents of main nutrients such as Ca, K, P and S. Furthermore, the supply of additives into composting did not increase the concentration of trace toxic elements. At the end of the experiment, plant biomass increased by the addition of the different organic amendments, with the highest shoot biomass in soils amended with compost supplied with nanoparticles. These results suggest that the addition of compost based on agricultural residues with additives such as halloysite or biochar improves chemical properties and nutritional status of soil that favor and increase plant growth of Lollium perenne stablished in soils from the Mediterranean Region., This research was funded by Agencia Nacional de Desarrollo (ANID), through the program FONDECYT by the postdoctoral project N◦ 3170677 and FONDECYT Initiation project N◦ 11201107

Dihaploid production from elite tetraploid cultivars is key to both traditional and novel breeding approaches that seek to simplify potato genetics. For this purpose, efficient and widely compatible haploid inducers (HIs) are needed. We compared PL-4, a new HI developed at the International Potato Center, to known HIs IvP101 and IvP35. By pollination of elite tetraploid breeding lines, we showed that PL-4 performed significantly better and had a homogeneous response regardless of the genetic background of the pistillate parents, on the most important efficiency traits—number of dihaploids per 100 fruits and haploid induction rate. Moreover, PL-4 exhibited a reduced proportion of hybrid seeds, a convenient trait for efficient screening. In this context, we recommend PL-4 as an enhanced HI for the potato breeding community.

Coral snakes of the genus Micrurus have been recorded as prey of rapacious birds, some mammals and other snakes, but so far not as prey of fishes. The black spotted piranha, Pygocentrus cariba, is an opportunistic predator that feeds mainly on fishes, occasionally on other vertebrates, but has so far not been reported to prey on snakes. Herein, based on morphological and molecular data (mitochondrial 16S fragment), we report the first case of predation of P. cariba on the coral snake Micrurus filiformis. Additionally, we briefly discuss the significance of this new finding for the diet of P. cariba and comment on the systematics and distribution of M. filiformis.

Functional analysis of the Mtl1 protein in Saccharomyces cerevisiae has revealed that this transmembrane sensor endows yeast cells with resistance to oxidative stress through a signaling mechanism called the cell wall integrity pathway (CWI). We observed upregulation of multiple heat shock proteins (HSPs), proteins associated with the formation of stress granules, and the phosphatase subunit of trehalose 6-phosphate synthase which suggests that mtl1Δ strains undergo intrinsic activation of a non-lethal heat stress response. Furthermore, quantitative global proteomic analysis conducted on TMT-labeled proteins combined with metabolome analysis revealed that mtl1Δ strains exhibit decreased levels of metabolites of carboxylic acid metabolism, decreased expression of anabolic enzymes and increased expression of catabolic enzymes involved in the metabolism of amino acids, with enhanced expression of mitochondrial respirasome proteins. These observations support the idea that Mtl1 protein controls the suppression of a non-lethal heat stress response under normal conditions while it plays an important role in metabolic regulatory mechanisms linked to TORC1 signaling that are required to maintain cellular homeostasis and optimal mitochondrial function.

18 Päg. Departamento de Medio Ambiente y Agronomía​, The structure and functioning of temperate forests are shifting due to changes in climate. Foreseeing the trajectory of such changes is critical to implementing adequate management practices and defining long-term strategies. This study investigated future shifts in temperate forest species composition and abundance expected to occur due to climate change. It also identified the ecological mechanisms underpinning such changes. Using an altitudinal gradient in the Romanian Carpathian temperate forests encompassing several vegetation types, we explored forest change using the Landis-II landscape model coupled with the PnET ecophysiological process model. We specifically assessed the change in biomass, forest production, species composition and natural disturbance impacts under three climate change scenarios, namely, RCP 2.6, 4.5 and 8.5. The results show that, over the short term (15 years), biomass across all forest types in the altitudinal gradient will increase, and species composition will remain unaltered. In contrast, over the medium and long terms (after 2040), changes in species composition will accelerate, with some species spreading (e.g., Abies alba Mill.) and others declining (e.g., Fagus sylvatica L.), particularly under the most extreme climate change scenario. Some forest types (e.g., Picea abies (L.) karst forests) in the Southern Carpathians will notably increase their standing biomass due to climate change, compared to other types, such as Quercus forests. Our findings suggest that climate change will alter the forest composition and species abundance, with some forests being particularly vulnerable to climate change, e.g., F. sylvatica forests. As far as productivity and forest composition changes are concerned, management practices should accommodate the new conditions in order to mitigate climate change impacts., This work was supported financially by EO-ROFORMON project, ID P_37_651/SMIS 105058 and PN 19070101

Leishmania RNA virus (LRV) is a double-stranded RNA virus belonging to the Totiviridae family detected as cytoplasmic inclusions in some strains of the human parasite Leishmania spp. Experimental evidence supports the hypothesis that human coinfection with Leishmania spp.–LRV triggers an exacerbated immune response in the host that can be responsible for the observed complicated outcomes in cutaneous leishmaniasis (CL), such as mucosal leishmaniasis (ML) and treatment failure of CL. However, the reported frequencies of LRV associated with complicated outcomes in patient’s series are highly variable, diminishing the relevance on the virus presence in the pathogenesis of the disease. To assess whether or not the inconsistent information about the frequency of LRV associated with CL complicated outcomes could be related to the virus detection approach, the present study evaluated the LRV presence in clinical samples using a diagnostic algorithm according to the type of the sample. In 36 samples with diagnosis of complicated forms of CL (15 of ML and 21 of CL antimony treatment failure) and six samples with non–Leishmania spp. infection, the LRV presence was assessed by RT-PCR, RT-qPCR, and nested RT-PCR. Viral load was estimated in parasite clinical isolates. By combining the methods, LRV1 presence was confirmed in 45% (9/20) of isolates and 37.5% (6/16) of the incisional biopsies. Remarkably, in some cases (4/8), LRV1 was undetectable in the isolates but present in their respective biopsies, and less frequently, the opposite was observed (1/8), suggesting the possibility of loss of parasites harboring LRV1 during the in vitro growth.

Aims The purpose of this study was to assess the value of genetic testing in addition to a comprehensive clinical evaluation, as part of the diagnostic work-up of elite and/or amateur Italian athletes referred for suspicion of inherited cardiac disease, following a pre-participation screening programme. Methods Between January 2009–December 2018, of 5892 consecutive participants, 61 athletes were investigated: 30 elite and 31 amateur athletes. Elite and amateur athletes were selected, on the basis of clinical suspicion for inherited cardiac disease, from two experienced centres for a comprehensive cardiovascular evaluation. Furthermore, the elite and amateur athletes were investigated for variants at DNA level up to 138 genes suspected to bear predisposition for possible cardiac arrest or even sudden cardiac death. Results Of these 61 selected subjects, six (10%) had diagnosis made possible by a deeper clinical evaluation, while genetic testing allowed a definite diagnosis in eight (13%). The presence of >3 clinical markers (i.e. family history, electrocardiogram and/or echocardiographic abnormalities, exercise-induced ventricular arrhythmias) was associated with a higher probability of positive genetic diagnosis (75%), compared with the presence of two or one clinical markers (14.2%, 8.1%, respectively, p-value = 0.004). Conclusion A combined clinical and genetic evaluation, based on the subtle evidence of clinical markers for inherited disease, was able to identify an inherited cardiac disease in about one-quarter of the examined athletes.

Dominant yeast species in Salsiccia Sarda, a traditional fermented sausage produced in Sardinia (Italy), were evaluated through the monitoring three typical production processes. Six different species were identified by molecular techniques, but Debaryomyces (D.) hansenii proved to be dominant. A D. hansenii strain was selected according to its technological features and used in three experimental sausage productions at farm scale with the aim to evaluate its antifungal effect. In all cases, two batches were inoculated with a previously selected autochthonous starter cultures (Lactobacillus plantarum and Staphylococcus xylosus), whereas two batches were left to spontaneous fermentation. D. hansenii was inoculated on the sausages surface by brushing after the sausages drying, by immersion in a yeast suspension after the stuffing, or, alternatively, casings were dipped in a yeast suspension before the dough stuffing. Microbial counts in the sausages core did not appear to be affected by D. hansenii application, while outcomes obtained for casings appeared soundly diversified. Brushing on the sausages surface at the onset of fermentation proved to be the best approach to treat sausages. Yeast inoculation exerted a noteworthy anti-mould effect, independently of the mode of application and, on the other hand, did not affect the overall quality and typical features of the product.

Rhizophora mangle es una especie abundante en el manglar de Sontecomapan. Sus individuos presentan el parásito Coccotrypes rhizophorae, perforador de corteza en los propágulos y corteza de R. mangle. Se determinó la abundancia de larvas, juveniles y adultos de C. rhizophorae en propágulos antes de ser dispersados en época de lluvias. El estudio se hizo en el sitio Ramsar 1342 “Manglares y Humedales de la Laguna de Sontecomapan”. A cada propágulo recolectado se le midió la longitud y diámetro medio del hipocótilo, y se determinó la abundancia del parásito en fase de larva, juvenil y adulto. Se encontró diferencia significativa en la abundancia de larvas y adultos de C. rhizophorae en propágulos predispersión en época de lluvias, siendo ausente la fase juvenil. No se encontró tendencia positiva al correlacionar la abundancia de C. rhizophorae en sus tres fases de desarrollo con la longitud y diámetro de los propágulos. Los resultados sugieren que los propágulos predispersión en la época de lluvias son más vulnerables para establecerse y desarrollarse al presentar mayor abundancia de C. rhizophorae.

Resumen Introducción: Los arroyos asociados a bofedales se hallan en ambientes donde las condiciones son extremas con temperaturas muy variables, baja presión atmosférica e intensa radiación solar. Sin embargo, existen ensamblajes de macroinvertebrados asociados a estos ambientes, cuya información es escasa. A escala global y regional, las características de los macroinvertebrados cambian con el aumento de la latitud y altitud (disminución de la riqueza y variación de la composición). A nivel local, también se puede agregar el efecto de las aguas ácidas y de elevada conductividad proveniente de manantiales que se unen a los arroyos, que generan heterogeneidad en la calidad de agua y consecuentemente en las características de los macroinvertebrados. Objetivo: Comparar la diversidad, composición y abundancia de macroinvertebrados y las características fisicoquímicas del agua de arroyos asociados a dos bofedales altoandinos, ubicados en el departamento de Ayacucho, Perú. Metodología: Se estudiaron once estaciones ubicados en arroyos de dos bofedales: 7 en Guitarrachayocc (G) de 43 ha (n = 28) y 4 en Pichccahuasi (P) de 28 ha (n = 8). Se empleó una red de Surber (0.3 mm de malla, área de muestreo de 1 200 cm2), para realizar muestreos mensuales de octubre de 2016 a abril de 2017. Análisis de componentes principales (PCA) y análisis de varianza multivariante no paramétrico (NP-MANOVA) se aplicaron para comparar las características del agua. Además, se utilizó un análisis de coordenadas principales (PCoA) y NP-MANOVA para comparar el ensamblaje. La prueba no paramétrica de Kruskal-Wallis (α = 0.05) se utilizó para comparar las abundancias absolutas de los taxa principales. Resultados: Las características del agua fueron heterogéneas, con pH de 3.2 (± 0.4) a 7.6 (± 0.1), conductividad de 168.9 (± 91) µS/cm a 1 117 (± 159.3) µS/cm; diferencias que fueron significativas (p < 0.05) al comparar las estaciones de muestreo, donde el agua de G6 presenta los mínimos valores de pH y los máximos de conductividad, al igual que cloruros. Se recolectaron un total de 8 126 individuos, distribuidos en 26 taxa de 20 familias, 11 órdenes y cinco clases. Los insectos fueron el grupo más diverso y abundante. A pesar a la cercanía de los arroyos de los bofedales, presentaron diferencias: en G, se registró 25 taxa, con dominancia de Macrelmis sp., Dicrotendipes sp., Claudioperla sp., Meridialaris sp. y en P, 9, con dominancia de Pedrowygomya sp., Hyalella sp. y Dicrotendipes sp. La densidad fue de 475 ind./m2 y de 31 ind./m2 para el bofedal G y P respectivamente, estos valores fueron estadísticamente diferentes (p < 0.05). El PCoA y el PCA confirmados por NP-MANOVA mostraron que G6 es diferente (p < 0.05) dentro de G porque recibe el aporte de agua de un manantial. Conclusiones: La riqueza, composición y densidad de los macroinvertebrados son diferentes en los arroyos según el bofedal al cual se halla asociado. La contribución de los manantiales a los arroyos genera cambios drásticos en la calidad del agua y los macroinvertebrados, determinando la formación de ensamblajes con diferente riqueza y estructura. Abstract Introduction: Streams associated with high Andean wetlands are found in environments where conditions are extreme, highly variable temperature, low atmospheric pressure, intense solar radiation. The macroinvertebrate assemblages have been poorly studied compared to streams located at lower altitudes. On a global and regional scale, the characteristics of macroinvertebrates change with the increase in latitude and altitude (decreasing in richness and variation in composition). At the local level, the effect of acidic waters with high conductivity from springs that join streams generate heterogeneity in water quality and consequently in the characteristics of macroinvertebrate structure. Objective: To compare the diversity, composition and abundance of macroinvertebrates and the physicochemical characteristics of stream water in two high Andean wetlands in Peru. Methodology: Eleven stations located in streams of two high Andean wetlands were studied: 7 in Guitarrachayocc (G) of 43 ha (n = 28) and 4 in Pichccahuasi (P) of 28 ha (n = 8). A Surber sampler (0.3 mm mesh, sampling area of 1 200 cm2) was used to carry out monthly samplings from October 2016 to April 2017. Principal Component Analysis (PCA) and Non-Parametric-Multivariate-Analysis-Of-Variance (NP-MANOVA) were applied to compare water characteristics. The macroinvertebrate assemblages from the two streams were compared by using a Principal coordinate analysis (PCoA), NP-MANOVA, and a non-parametric Kruskal-Wallis test (α = 0.05). Results: The characteristics of the water were heterogeneous, with significant differences in pH which ranged from 3.2 (± 0.4) to 7.6 (± 0.1) and conductivity ranging from 168.9 (± 91.0) µS / cm to 1 117.0 (± 159.3) µS / cm. In particular, G6 presented the lowest pH values and the highest electrical conductivity, as well as chlorides. A total of 8 126 individuals were collected, distributed in 26 taxa from 20 families, 11 orders and five classes. Insects were the most diverse and abundant group. Differences were found between the streams of the highland wetlands, despite their proximity: in G, 25 taxa were registered, with a dominance of Macrelmis sp., Dicrotendipes sp., Claudioperla sp. and Meridialaris sp. and in P, 9, with a dominance of Pedrowygomya sp., Hyalella sp. and Dicrotendipes sp. The density was 475 ind./m2 and 31 ind./m2 for G and P respectively, these values were statistically different (p < 0.05). The PCoA and PCA confirmed by NP-MANOVA showed that G6 was different (p < 0.05) within G because it receives water contribution from a spring. Conclusions: The richness, composition and density of macroinvertebrates were different in streams depending on the bofedal with which it is associated. The contribution of the springs to the streams generates drastic changes in the quality of the water and the macroinvertebrates, determining the formation of assemblages with different richness and structure.

Context: Tetra Pak® is a common material used for the production of food containers. Currently, those containers are recycled by physical separation of the cellulose fibers through a hydro pulped process, but sometimes separating the individual components is not economically viable. This work evaluates an alternative process to obtain composites from recycled Tetra Pak®. Methodology: Tetra Pak® used containers were collected and cut into small pieces at the laboratory. Then, the containers were hot-pressed to obtain sheets in a manual hydraulic press by using different configurations. Samples were cut, and their tensile strength was evaluated (ASTM D3039). Failure analysis of samples was carried out by FE-SEM to identify issues related to processing and to understand the differences in mechanical properties. Results: The results showed that the lowest tensile strength was 9.5 MPa (type I sample) and the highest tensile strength was 37.4 MPa (type III sample). Conclusions: The results of mechanical tests show that this material can be used for non-structural purposes in the building industry. Failure analysis shows that fiber pull-out and delamination are the most important failure mechanisms in type I samples. For type III sample, failure was produced by a sequence of intralaminar fractures. Financing: Instituto Tecnológico Metropolitano de Medellín Resumen Contexto: Tetra Pak® es un material común que se utiliza para recipientes de alimentos. Actualmente, esos contenedores se reciclan mediante la separación física de las fibras de celulosa a través de un proceso de hidropulpeado, pero a veces no es económicamente viable separar los componentes individuales. En este trabajo, se evaluó un proceso alternativo para obtener materiales compuestos de Tetra Pak® reciclado. Metodología: Inicialmente, los contenedores de Tetra Pak® usados se recolectaron y cortaron en trozos pequeños en el laboratorio. Después, los contenedores se prensaron en caliente para obtener laminados en una prensa hidráulica manual utilizando diferentes configuraciones. Después de obtener los materiales compuestos, se cortaron las muestras y se evaluó su resistencia a la tracción (ASTM D3039). El análisis de fallas de las muestras fue realizado por FE-SEM para identificar problemas relacionados con el procesamiento y para comprender las diferencias en las propiedades mecánicas. Resultados: Los resultados mostraron que la menor resistencia a la tracción era de 9,5 MPa (muestras de tipo I). La mayor resistencia a la tracción fue de 37,4 MPa para muestras de tipo III. Conclusiones: Los resultados de las pruebas mecánicas permitieron concluir que el material se puede utilizar para fines no estructurales en la industria de la construcción. El análisis de fallas mostró que el desprendimiento de la fibra es el mecanismo más importante en las muestras de tipo I. Para las muestras de tipo III, la fractura se produjo por una secuencia de fallas interlaminares. Financiamiento: Instituto Tecnológico Metropolitano de Medellín

Antifungal drug discovery and design is very challenging because of the considerable similarities in genetic features and metabolic pathways between fungi and humans. However, cell wall composition represents a notable point of divergence. Therefore, a research strategy was designed to improve our understanding of the mechanisms for maintaining fungal cell wall integrity, and to identify potential targets for new drugs that modulate the underlying protein-protein interactions in Saccharomyces cerevisiae. This study defines roles for Wsc2p and Wsc3p and their interacting protein partners in the cell wall integrity signaling and cell survival mechanisms that respond to treatments with fluconazole and hydrogen peroxide. By combined genetic and biochemical approaches, we report the discovery of 12 novel protein interactors of Wsc2p and Wsc3p. Of these, Wsc2p interacting partners Gtt1p and Yck2p, have opposing roles in the resistance and sensitivity to fluconazole treatments respectively. The interaction of Wsc2p with Ras2p was confirmed by iMYTH and IP-MS approaches and is shown to play a dominant role in response to oxidative stress induced by hydrogen peroxide. Consistent with an earlier study, Ras2p was also identified as an interacting partner of Wsc1p and Mid2p cell wall integrity signaling proteins. Collectively, this study expands the interaction networks of the mechanosensory proteins of the Cell Wall Integrity pathway.

Carbon storage is one of the most important ecosystem services provided by kelp beds. Laminarialean kelps are widely harvested along the Warm Temperate South-eastern Pacific coast, a marine province shared by Chile and Peru. Carbon storage assessments of kelps in Peru are lacking. From a blue economy and sustainable management perspectives, information on the carbon storage of kelps is important. We conduct the first carbon storage assessment of Lessonia trabeculata in Peru and contribute to the development of biomass estimation models in order to monitor kelps using the least destructive methodologies. We chose three commonly harvested sites in San Juan de Marcona to haphazardly extract Lessonia trabeculata sporophytes using transects. Sporophyte height (m), wet biomass (kg), maximum (D) and minimum (d) holdfast disk widths (cm) were measured in the field. In the laboratory, C content was measured to calculate the best-fitting coefficients for future estimations by using allometric equations. Individual biomass was best estimated from sporophyte height through a rational model, while holdfast area (ellipse in cm2) was a good proxy of biomass with a sinusoidal model. The southernmost, least accessible, and most exposed site (Elefante) had significantly higher values of stored carbon. We estimated a carbon standing stock (from sporophytes only) of 2044 t C in these kelp beds. Nevertheless, additional and more detailed measurements will likely produce more accurate estimates both in time and space. We provide allometric equations for future carbon assessments. Our results highlight the importance carbon assessment for kelp management and blue carbon estimates and the need to develop science-based marine planning strategies.

Neutrophils release their intracellular content, DNA included, into the bloodstream to form neutrophil extracellular traps (NETs) that confine and kill circulating pathogens. The mechanosensitive adhesive blood protein, von Willebrand Factor (vWF), interacts with the extracellular DNA of NETs to potentially immobilize them during inflammatory and coagulatory conditions. Here, we elucidate the previously unknown molecular mechanism governing the DNA–vWF interaction by integrating atomistic, coarse-grained, and Brownian dynamics simulations, with thermophoresis, gel electrophoresis, fluorescence correlation spectroscopy (FCS), and microfluidic experiments. We demonstrate that, independently of its nucleotide sequence, double-stranded DNA binds to a specific helix of the vWF A1 domain, via three arginines. This interaction is attenuated by increasing the ionic strength. Our FCS and microfluidic measurements also highlight the key role shear-stress has in enabling this interaction. Our simulations attribute the previously-observed platelet-recruitment reduction and heparin-size modulation, upon establishment of DNA–vWF interactions, to indirect steric hindrance and partial overlap of the binding sites, respectively. Overall, we suggest electrostatics—guiding DNA to a specific protein binding site—as the main driving force defining DNA–vWF recognition. The molecular picture of a key shear-mediated DNA–protein interaction is provided here and it constitutes the basis for understanding NETs-mediated immune and hemostatic responses.

This study was carried out to evaluate the effects of foliar sprays containing boron (B) nano-fertilizer (NF) on the growth and physiology of lettuce (Lactuca sativa) and zucchini (Cucurbita pepo). Plants were grown under greenhouse conditions for 60 days on a modified Hoagland solution with the presence and absence of boron (+B or –B). A synthesized B–NF foliar spray and a commercial B foliar fertilizer (Bortrac™ 150, BT) was applied at a concentration of 30 mg B L−1 at 10-d intervals throughout the experiment. The B–NF treatment increased the growth of lettuce 2.7- and 1.9-fold for shoots and roots, respectively, with an average production of lettuce biomass by ~58%. Similarly, the NF increased the growth of zucchini by 18 and 66% compared with Control–B (the absence of B), and 13 and 36% compared with BT, both for shoots and roots, respectively. Nevertheless, NF + B mostly decreased lettuce growth with symptoms of B toxicity in leaves. In lettuce, addition of B did not affect concentrations of phenols; however, in zucchini, Control–B induced a higher production of phenolic compounds possibly related to B deficiency. The B addition in lettuce reduced the DPPH activity by 32 and 21% in NF and BT, respectively, compared to Control-B. These responses were similar in zucchini; however, the effect of B was product of its presence in mineral solution rather than due the foliar product applied. This suggests that a NF-based delivery system for B may be highly effective at boosting plant productivity on B-limited soils.

Riparian ecosystems are among the most degraded worldwide as they are subject to a range of human-mediated disturbances at different scales. As riparian vegetation plays a key role in maintaining waterway and landscape health, restoration often focuses on promoting riparian vegetation re-establishment. The role of below-ground processes and agents in the process of revegetation is often overlooked even though agents like mycorrhizal fungi enhance seedling establishment and plant growth. In this study we aimed to understand whether colonisation by local mycorrhizal fungi can be advantageous to the early phases of plant establishment during revegetation, whether revegetation technique influences the extent of this potential colonisation, and has the potential to enhance revegetation outcomes. We examined mycorrhizal colonisation rate and species composition of root fungal communities in direct seeded and tube-stock plants and compared them with local native vegetation in a retarding basin in Victoria, Australia, then modelled the relationship between root fungal community attributes and plant size. Results indicate that revegetation technique influenced the colonisation rate, diversity and composition of root fungal communities. Moreover, greater fungal diversity and greater similarity of fungal communities to the mycobiota of native vegetation were positively related with growth of seeded plants but showed negative or little relationship to size of tube-stock plants. High colonisation rates of plant species studied, and relationships between fungal community characteristics and plant growth of seeded plants provide the first evidence of the potential importance of mycorrhizal associations for these plant species in a revegetation context.

This is the species account about the Banded Tree-Anole (Anolis transversalis), a chapter in the Reptiles of Ecuador book. This account summarizes novel as well as historical information (recognition, natural history, distribution, conservation, etymology, and relevant literature) about the species and provides maps and images to facilitate identification of this reptile in the field.

Sourdough is an old example of a natural starter composed of a mixture of flour, water, and metabolites and is produced by naturally occurring lactic acid bacteria and yeasts that influence bread aroma. In this work, four types of sourdough were used to prepare bread: one sourdough with yeast beer and three with bacteria and yeasts. The physicochemical parameters (pH, moisture, water activity, and organic acids) of the bread and sourdoughs were assessed. Lactic, acetic, and succinic acids were found in considerable amounts in sourdoughs and the corresponding breads. The fermentation quotient (molar ratio between lactic and acetic acid) ranged from 0.39 to 3.4 in sourdoughs. Lactic acid was prevalent in all types of bread and showed the highest value in bread made from sourdough with a 1.5 bacteria/yeast ratio (8722.24 mg/kg). Moreover, volatile organic compounds were identified in bread samples. Alcohols, aldehydes, and acetic acid were mainly found. The alcohol concentration ranged from 140.88 to 401.20 ng/g. Aldehydes ranged from 185.01 to 454.95 ng/g, and acetic acid ranged from 91.40 to 173.81 ng/g. Bread prepared from sourdough with a 1.5 bacteria/yeast ratio showed a considerable amount of alcohols and acetic acid.

Realistic mappings of genes to morphology are inherently multivariate on both sides of the equation. The importance of coordinated gene effects on morphological phenotypes is clear from the intertwining of gene actions in signaling pathways, gene regulatory networks, and developmental processes underlying the development of shape and size. Yet, current approaches tend to focus on identifying and localizing the effects of individual genes and rarely leverage the information content of high-dimensional phenotypes. Here, we explicitly model the joint effects of biologically coherent collections of genes on a multivariate trait – craniofacial shape – in a sample of n = 1145 mice from the Diversity Outbred (DO) experimental line. We use biological process Gene Ontology (GO) annotations to select skeletal and facial development gene sets and solve for the axis of shape variation that maximally covaries with gene set marker variation. We use our process-centered, multivariate genotype-phenotype (process MGP) approach to determine the overall contributions to craniofacial variation of genes involved in relevant processes and how variation in different processes corresponds to multivariate axes of shape variation. Further, we compare the directions of effect in phenotype space of mutations to the primary axis of shape variation associated with broader pathways within which they are thought to function. Finally, we leverage the relationship between mutational and pathway-level effects to predict phenotypic effects beyond craniofacial shape in specific mutants. We also introduce an online application that provides users the means to customize their own process-centered craniofacial shape analyses in the DO. The process-centered approach is generally applicable to any continuously varying phenotype and thus has wide-reaching implications for complex trait genetics.

Cocaine use disorders (CUD) are devastating for affected individuals and impose a significant burden on society, but there are currently no FDA-approved therapies. The development of novel and effective treatments has been hindered by substantial gaps in our knowledge about the etiology of these disorders. The risk for developing a CUD is influenced by genetics, the environment and complex interactions between the two. Identifying specific genes and environmental risk factors that increase CUD risk would provide an avenue for the development of novel treatments.Rodent models of addiction-relevant behaviors have been a valuable tool for studying the genetics of response to drugs of abuse. Traditional genetic mapping using genetically and phenotypically divergent inbred mice has been successful in identifying numerous chromosomal regions that influence addiction-relevant behaviors, but these strategies rarely result in identification of the causal gene or genetic variant. To overcome this challenge, reduced complexity crosses (RCC) between closely related inbred mouse substrains have been proposed as a method for rapidly identifying and validating functional variants. The RCC approach is dependent on identifying phenotypic differences between substrains. To date, however, the study of addiction-relevant behaviors has been limited to very few sets of substrains, mostly comprising the C57BL/6 lineage.The present study expands upon the current literature to assess cocaine-induced locomotor activation in 20 inbred mouse substrains representing six inbred strain lineages (A/J, BALB/c, FVB/N, C3H/He, DBA/2 and NOD) that were either bred in-house or supplied directly by a commercial vendor. To our knowledge, we are the first to identify significant differences in cocaine-induced locomotor response in several of these inbred substrains. The identification of substrain differences allows for the initiation of RCC populations to more rapidly identify specific genetic variants associated with acute cocaine response. The observation of behavioral profiles that differ between mice generated in-house and those that are vendor-supplied also presents an opportunity to investigate the influence of environmental factors on cocaine-induced locomotor activity.

SummaryThe spleen and lymph node represent important hubs for both innate and adaptive immunity1,2. Herein, we map immune, endothelial, and neuronal cell networks within these tissues from “normal”/non-diseased organ donors, collected through the NIH Human BioMolecular Atlas Program (HuBMAP)3, using highly multiplexed CODEX (CO-Detection by indEXing) imaging and 3D light sheet microscopy of cleared tissues. Building on prior reports4–6, we observed the lymph node subcapsular sinus expressing podoplanin, smooth muscle actin, and LYVE1. In the spleen, LYVE1 was expressed by littoral cells lining venous sinusoids, whereas podoplanin was restricted to arteries and trabeculae. 3D visualization of perivascular innervation revealed a subset of axonal processes expressing choline acetyl transferase in both tissues, in contrast with prior literature on human spleen7. We further report our novel observations regarding the distinct localization of GAP43 and β3-tubulin within the vascular anatomy of both lymph node and spleen, with Coronin-1A+ cells forming a dense cluster around β3-tubulin positive GAP43 low/negative segments of large vessels in spleen. These data provide an unprecedented 2D and 3D visualization of cellular networks within secondary lymphoid tissues, laying the groundwork for future disease-specific and system-wide studies of neural regulation of immunity in human lymphatics.

The brocket deer (Genus Mazama) is a highly diverse cervid group distributed from Mexico to Argentina, with a downward population trend. However, literature on the basic reproductive biology of the genus is scarce. This work aimed to study biometric, histological and stereological aspects of the testes of Dwarf Red Brocket (Mazama rufina). Testes from free-ranging adult brockets (n = 3) were retrieved from necropsies. Testes were histologically processed. From histological images, several stereological parameters were estimated, and seminiferous epithelium cycle morphology was described. Testes volumes were between 8.2 and 18.4 ml and weights from 8.3 to 19.4 g. Gonadosomatic index (% paired-testes weight to body weight) went from 0.17 to 0.64. The tubular cross-sectional diameter was 179.8 ± 2.8 µm. Estimated volume densities for parenchyma and interstitium were 78.8% and 21.2% respectively. There were (in millions/ml) 96.0 ± 13.1 germ cells and 37.7 ± 6.0 somatic cells. Specific cell densities were (all expressed in millions/ml) as follows: spermatogonia 13.1 ± 4.2; primary spermatocytes 43.1 ± 5.0; round spermatids 36.8 ± 8.0 (lower density near the caudal pole, p

Neurodegenerative diseases exhibit extensive genetic heterogeneity and complex etiology with varying onset and severity. To deduce the mechanism leading to retinal degeneration, we adopted a temporal multi-omics approach and examined molecular and cellular events before the onset of photoreceptor cell death in the widely-used Pde6brd1/rd1 (rd1) mouse model. Transcriptome profiling of neonatal and developing rods revealed early downregulation of genes associated with anabolic pathways and energy metabolism. Quantitative proteomics of rd1 retina showed early changes in calcium signaling and oxidative phosphorylation, with specific partial bypass of complex I electron transfer, which precede the onset of cell death. Concurrently, we detected alterations in central carbon metabolism, including dysregulation of components associated with glycolysis, pentose phosphate and purine biosynthesis. Ex vivo assays of oxygen consumption and transmission electron microscopy validated early and progressive mitochondrial stress and abnormalities in mitochondrial structure and function of rd1 rods. These data uncover mitochondrial over-activation and related metabolic alterations as early determinants of pathology and implicate dysregulation of calcium signaling as the initiator of higher mitochondrial stress, which then transitions to mitochondrial damage and photoreceptor cell death in retinal degeneration. Our studies support the “one hit model” arguing against the cumulative damage hypothesis but suggest that cell death in neurodegenerative disease is initiated by specific rather than a random event.

Traditional packaging made from non-renewable compounds such as petroleum has a negative impact on the environment, which has led to a growing body of research on biodegradable packaging. This paper details the characterization of a biocomposite which was made from cassava bran, coffee husk, and water (BCCW). After this, a layer of polylactic acid (PLA) was added, mixed with nisaplin 1000UI, and 8, 10, 12% concentrations of oregano (Origanum vulgare L) essential oil (OVEO) in order to create an active biocomposite (AB) which could be used as an antimicrobial material in the packaging of pork. In vitro inhibitory effects were evaluated against Listeria monocytogenes, Escherichia coli and Salmonella typhimurium. After obtaining the active biocomposite, the characterization was performed by means of Infrared Spectroscopy (FT-IR), Water Vapor Permeability (WVP), a Mechanical Property analysis, Differential Scanning Calorimetry (DSC), and Scanning Electron Microscopy (SEM). Using the concentration with the highest inhibition against pathogenic bacteria, antimicrobial tests were carried out after the application of the AB in pork meat for 0, 1, 3, 5, and 10 days. Inhibition tests against pathogenic bacteria showed that nisin had a greater effect on Gram positive bacteria, while OVEO had an effect on both Gram positive and Gram negative bacteria. FT-IR analysis allowed for the characterization of the molecular interactions of OVEO and nisin in the matrix compounds of the different biocomposites by identifying and interpreting their respective vibration bands. According to the statistical analysis, the different concentrations of the biocomposites affect WVP (p 0.05). The DSC test on the ABs showed two melting peaks and an increase in the glass transition temperature (Tg) of the PLA, a finding that indicates greater thermal stability in the ABs. SEM images revealed that the nisaplin and OVEO particles were uniformly distributed in the PLA polymer matrix on the surface of the ABs. The ABs showed a significant reduction in Listeria monocytogenes in pork as of day 3 and in the other pathogens as of the 10th day. The biocomposite containing OVEO and nisaplin, showed good potential to be used to food packaging.

Mitochondrial networks provide coordinated energy distribution throughout muscle cells. However, pathways specifying mitochondrial network-type separately from contractile fiber-type remain unclear. Here, we show that natural energetic demands placed on Drosophila melanogaster muscles yield native cell-types among which contractile and mitochondrial network-types are regulated independently. Proteomic analyses of indirect flight, jump, and leg muscles together with muscles misexpressing known fiber-type specification factor salm identified transcription factors H15 and cut as potential mitochondrial network regulators. We demonstrate H15 operates downstream of salm regulating flight muscle contractile and mitochondrial network-type. Conversely, H15 regulates mitochondrial network configuration but not contractile type in jump and leg muscles. Further, we find that cut regulates salm expression in flight muscles and mitochondrial network configuration in leg muscles. These data indicate cell type-specific regulation of muscle mitochondrial network organization separately from contractile type, mitochondrial content, and mitochondrial size through an evolutionarily conserved pathway involving cut, salm, and H15.

We describe a colorful and distinctive new species of tanager from the lower slopes of the Andes of southeastern Peru and western Bolivia. The species was first noted from southeastern Peru in 2000, but little of its natural history was uncovered until the 2011 discovery of a breeding population in deciduous forest in an intermontane valley, the Machariapo valley, in Bolivia. This species appears to be an intratropical migrant, breeding in deciduous forest during the rainy season (November–March) and spending the dry season dispersed along the lower slopes of the Andes, apparently favoring Guadua bamboo-dominated habitats in both seasons. Phylogenetic evidence suggests this tanager is embedded within a clade of thraupids that includes Ramphocelus, Coryphospingus, Loriotus, Tachyphonus, and related genera in the subfamily Tachyphoninae. Within this subfamily, the new species falls in a clade with two monotypic genera, Eucometis penicillata (Gray-headed Tanager) and Trichothraupis melanops (Black-goggled Tanager). There is strong support for a sister relationship between the new tanager and T. melanops, but because all three species in this clade are highly distinctive phenotypically, we propose erecting a new genus and species name for the new tanager. LAY SUMMARY We discovered a new species of tanager, from the foothills of the Andes mountains in Peru and Bolivia, which we name the Inti Tanager. The name “Inti” is the Quechua (a language indigenous to the region) word that means “sun,” referring to the bright yellow color of the bird. Unlike most new bird species discovered recently, which typically differ only subtly from their closest relatives, this species is so different in appearance and genetics that we also described a new genus just for it. The tanager family, to which the new species belongs, is one of the largest bird families in the world (with about 377 species), restricted almost entirely to the American tropics and including many beautifully colorful species. We uncovered a surprising piece of natural history about the Inti Tanager: it migrates from its breeding grounds in a little-explored valley in Bolivia to “winter” along the lower slopes of the Andes in Peru. Such migration within the tropics is rare within the tanagers. That this unique, bright yellow bird could have gone unnoticed by ornithologists until recently attests to the remoteness of the areas where it lives and to the importance of continued biological surveys off-the-grid in South America.

Respiratory viruses present major public health challenges, as evidenced by the 1918 Spanish Flu, the 1957 H2N2, 1968 H3N2, and 2009 H1N1 influenza pandemics, and the ongoing severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Severe RNA virus respiratory infections often correlate with high viral load and excessive inflammation. Understanding the dynamics of the innate immune response and its manifestations at the cell and tissue levels is vital to understanding the mechanisms of immunopathology and to developing strain-independent treatments. Here, we present a novel spatialized multicellular computational model of RNA virus infection and the type-I interferon-mediated antiviral response that it induces within lung epithelial cells. The model is built using the CompuCell3D multicellular simulation environment and is parameterized using data from influenza virus-infected cell cultures. Consistent with experimental observations, it exhibits either linear radial growth of viral plaques or arrested plaque growth depending on the local concentration of type I interferons. The model suggests that modifying the activity of signaling molecules in the JAK/STAT pathway or altering the ratio of the diffusion lengths of interferon and virus in the cell culture could lead to plaque growth arrest. The dependence of plaque growth arrest on diffusion lengths highlights the importance of developing validated spatial models of cytokine signaling and the need for in vitro measurement of these diffusion coefficients. Sensitivity analyses under conditions leading to continuous or arrested plaque growth found that plaque growth is more sensitive to variations of most parameters and more likely to have identifiable model parameters when conditions lead to plaque arrest. This result suggests that cytokine assay measurements may be most informative under conditions leading to arrested plaque growth. The model is easy to extend to include SARS-CoV-2-specific mechanisms or to use as a component in models linking epithelial cell signaling to systemic immune models., Author summary Respiratory lung infections form lesions in the lungs, whose number and size correlate with the severity of illness. In some severe cases, respiratory disease triggers a severe inflammatory condition known as a cytokine storm. In order to link molecular signaling at the site of infection to its impact on the overall interferon response and the occurrence of severe inflammation, we created a computational model of the early stages of infection that simulates lung cells infected with RNA viruses, such as those responsible for COVID-19 and flu, to help explore how the disease forms viral plaques, an in vitro analog to lesion growth in the lung. Consistent with experimental observations, our model suggests that the treatment of cells with type-I interferons, which are currently being evaluated for the treatment of COVID-19, may have a protective effect. We found that enhancing certain aspects of the inflammatory response, such as the JAK/STAT pathway, may be able to arrest viral plaque growth, suggesting molecules involved in this pathway as possible drug candidates. Analysis of the model also shows that to better estimate the parameter values needed to model interferon signaling and viral replication, experiments should be performed under conditions that inhibit viral growth, e.g., by pretreating cells with interferon. We present a computational framework built using the CompuCell3D multicellular simulation environment that lays the groundwork for constructing larger models of respiratory-infection-induced immune responses and that can inform experiments to improve our fundamental understanding of the mechanisms regulating the immune response.

In order to evaluate dominance/implantation of starter cultures for wine fermentation, both commercial starters and wild strains were monitored during the fermentation of Greco di Tufo (GR) and Aglianico of Taurasi (AGL) musts. Preliminary characterization of commercial strains was carried out by several molecular markers. Five fermentations&mdash, four starter-inoculated and one spontaneous&mdash, were carried out in duplicates by using grapes from GR and AGL. Trials were monitored, and yeast cultures were isolated within the dominant microflora. Comparison of Interdelta patterns allowed to assess the real occurrence of both starters and indigenous strains. A high genetic diversity within S. cerevisiae strains was detected. In starter-led fermentations (except for few cases), in addition to the starter strains, indigenous S. cerevisiae biotypes were found, as well. Native strains isolated from replicates of the same fermentation showed different genetic profiles. Spontaneous fermentations were conducted, during the first 5 days, by non-Saccharomyces yeasts and, afterwards, by a high number (16 in the AGL and 20 in the GR) of S. cerevisiae biotypes. Indigenous biotypes isolated by GR revealed a high variability in oenological features and, in several cases, showed better performances than those recorded for commercial strains. The study further highlighted the low dominance of some commercial starter cultures. Moreover, autochthonous yeast strains proved to be sometimes more aggressive in terms of fermentation vigor in GR must, likely because better adapted to ecological and technological conditions occurring during winemaking. Finally, the use of such strains for production of autochthonous &ldquo, pied de cuve&rdquo, may be a useful strategy for lowering production cost of winemaking.

Chestnut is a nut cultivated in a variety of growing conditions and climates, being globally popular and valued for its sensory, nutritional and healthy properties. European sweet chestnut (Castanea sativa Mill.) is mainly grown in the temperate regions of continental Europe and has represented one of the most important and sustainable food resources of rural areas for many centuries. Chestnut fruit is highly appreciated and extensively consumed throughout Europe, America and Asia, because is considered a high nutritional value food, extremely versatile and safe. Recently, there is an increasing demand of both the fresh and transformed fruit e.g. chestnut flour for its gluten-free characteristic and low fat content compared with other nuts for the development of glutenfree for celiac disease patients, non-celiac gluten sensitivity people and consumers who avoid gluten for lifestyle reasons and of health-related food products. The market for edible chestnuts has considerable potential for increase in production and demand given growing consumer interest in alternative and healthy foods (Gold et al., 2004). Almost 20% of the total production is used to make chestnut flour, dried chestnuts and the marrons glacé sweet.

Probiotics are considered as the twenty-first century panpharmacon due to their competent remedial power to cure from gastrointestinal dysbiosis, systematic metabolic diseases, and genetic impairments up to complicated neurodegenerative disorders. They paved the way for an innovative managing of various severe diseases through palatable food products. The probiotics’ role as a “bio-therapy” increased their significance in food and medicine due to many competitive advantages over traditional treatment therapies. Their prophylactic and therapeutic potential has been assessed through hundreds of preclinical and clinical studies. In addition, the food industry employs probiotics as functional and nutraceutical ingredients to enhance the added value of food product in terms of increased health benefits. However, regardless of promising health-boosting effects, the probiotics’ efficacy still needs an in-depth understanding of systematic mechanisms and factors supporting the healthy actions.

Flour from grass pea, a legume that is adapted to arid conditions containing high levels of proteins, was characterized according to microstructure, some functional properties and digestibility. Microstructural results showed that grass pea starch granules appeared surrounded by an integral matrix with heterogeneous sizes from 6 to 30 µm. Thermal properties displayed a single endothermic transition corresponding to starch gelatinization transition. The digestibility of starch was 79.6%, expressed as the ratio of non-resistant starch to the total amount of resistant and non-resistant starch. The flour was also relatively rich in phenolic substances possessing antioxidant properties as demonstrated by the 2-diphenyl-1-picrylhydrazyl radical method. Adult and elderly in vitro digestion demonstrated that proteins were easily digested. These findings suggest that this legume is suitable for feeding of a large spectrum of population, being endowed with attractive properties that make it potential enough as functional food.

The combined effect of ozone gaseous treatment and passive refrigeration system on microbiological, biochemical, and sensory changes of fresh cod (Merluccius merluccius), red shrimp (Aristeus antennatus), scald fish (Arnoglossus laterna) and musky octopus (Eledone moschata) was evaluated by monitoring quality during 12 days of storage. Except for hake, ozonation exerted a significant bactericidal effect on psychrotrophs, H2S-producing bacteria, Aeromonas and Brochothrix spp. TVB-N and TMA-N experienced the same trend in the four fish types with higher increases in scald fish. The remarkable contents recorded in musky octopus for both parameters appeared to be due to autolytic activity. Except for red shrimp, the level of fat hydrolysis rarely appeared to be different within the ozonated and control samples. According to sensory assessment, ozonation may have a varying impact depending on the fish product: the treatment extended the scald fish shelf-life up to one week, while in red shrimp no difference occurred for the first week. Based on the outcomes, the combined approach developed here has proven to be an effective technology for fish shelf-life extension, with the best influence on microbial control in scald fish and musky octopus. At any rate, conditions for treatment need to be fine-tuned to the specific fish product.

Prevention of COVID-19 on a global scale will require the continued development of high-volume, low-cost platforms for the manufacturing of vaccines to supply ongoing demand. Vaccine candidates based on recombinant protein subunits remain important because they can be manufactured at low costs in existing large-scale production facilities that use microbial hosts like Komagataella phaffii (Pichia pastoris). Here, we report an improved and scalable manufacturing approach for the SARS-CoV-2 spike protein receptor-binding domain (RBD); this protein is a key antigen for several reported vaccine candidates. We genetically engineered a manufacturing strain of K. phaffii to obviate the requirement for methanol induction of the recombinant gene. Methanol-free production improved the secreted titer of the RBD protein by >5X by alleviating protein folding stress. Removal of methanol from the production process enabled to scale up to a 1200 L pre-existing production facility. This engineered strain is now used to produce an RBD-based vaccine antigen that is currently in clinical trials and could be used to produce other variants of RBD as needed for future vaccines.

Global containment of COVID-19 still requires accessible and affordable vaccines for low- and middle-income countries (LMICs).1 Recently approved vaccines provide needed interventions, albeit at prices that may limit their global access.2 Subunit vaccines based on recombinant proteins are suited for large-volume microbial manufacturing to yield billions of doses annually, minimizing their manufacturing costs.3 These types of vaccines are well-established, proven interventions with multiple safe and efficacious commercial examples.4–6 Many vaccine candidates of this type for SARS-CoV-2 rely on sequences containing the receptor-binding domain (RBD), which mediates viral entry to cells via ACE2.7,8 Here we report an engineered sequence variant of RBD that exhibits high-yield manufacturability, high-affinity binding to ACE2, and enhanced immunogenicity after a single dose in mice compared to the Wuhan-Hu-1 variant used in current vaccines. Antibodies raised against the engineered protein exhibited heterotypic binding to the RBD from two recently reported SARS-CoV-2 variants of concern (501Y.V1/V2). Presentation of the engineered RBD on a designed virus-like particle (VLP) also reduced weight loss in hamsters upon viral challenge.

Phytonutrients are plant foods that contain many natural bioactive compounds, called phytochemicals, which show specific biological activities. These phytonutrients and their phytochemicals may play an important role in health care maintaining normal organism functions (as preventives) and fighting against diseases (as therapeutics). Phytonutrients’ components are the primary metabolites (i.e., proteins, carbohydrates, and lipids) and phytochemicals or secondary metabolites (i.e., phenolics, alkaloids, organosulfides, and terpenes). For years, several phytonutrients and their phytochemicals have demonstrated specific pharmacological and therapeutic effects in human health such as anticancer, antioxidant, antiviral, anti-inflammatory, antibacterial, antifungal, and immune response. This review summarizes the effects of the most studied or the most popular phytonutrients (i.e., turmeric, garlic, cinnamon, graviola, and oregano) and any reported contraindications. This article also presents the calculated physicochemical properties of the main phytochemicals in the selected phytonutrients using Lipinski’s, Veber’s, and Ghose’s rules. Based on our revisions for this article, all these phytonutrients have consistently shown great potential as preventives and therapeutics on many diseases in vitro, in vivo, and clinical studies.

Aponte, Ameris, Gyaltshen, Yangtsho, Burns, John A., Heiss, Aaron A., Kim, Eunsoo, Warring, Sally D. (2021): The Bacterial Diversity Lurking in Protist Cell Cultures. American Museum Novitates 2021 (3975): 1-16, DOI: 10.1206/3975.1, URL: https://bioone.org/journals/american-museum-novitates/volume-2021/issue-3975/3975.1/The-Bacterial-Diversity-Lurking-in-Protist-Cell-Cultures/10.1206/3975.1.full

Nowadays, one of the major global health concerns is coronavirus disease 2019 (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Even though numerous treatments and vaccines to combat this virus are currently under development, the detailed molecular mechanisms underlying the pathogenesis of this disease are yet to be elucidated to design future therapeutic tools against SARS-CoV-2 variants. MicroRNAs (miRNAs) are small (20-24 nucleotides), non-coding RNA molecules that regulate post-transcriptional gene expression. Recently, it has been demonstrated that both host and viral-encoded miRNAs are crucial for the successful infection of SARS-CoV-2. For instance, dysregulation of miRNAs that modulate multiple genes expressed in COVID-19 patients with comorbidities (e.g., type 2 diabetes, lung adenocarcinoma, and cerebrovascular disorders) could affect the severity of the disease. Thus, altered expression levels of circulating miRNAs might be helpful to diagnose this illness and forecast whether a COVID-19 patient could develop a severe state of the disease. Besides, researchers have found a number of miRNAs could inhibit the expression of proteins, such as ACE2, TMPRSS2, spike, and Nsp12, involved in the life cycle of SARS-CoV-2. Accordingly, miRNAs represent potential biomarkers and therapeutic targets for this devastating viral disease. Therefore, in this current review, we present the recent discoveries regarding the clinical relevance and biological roles of miRNAs in COVID-19.

SUMMARY Assigning behavioral roles to genetically defined neurons within the lateral hypothalamus (LH) is an ongoing challenge. We demonstrate that a subpopulation of LH GABAergic neurons expressing leptin receptors (LHLEPR) specifically drives appetitive behaviors in mice. Ablation of LH GABAergic neurons (LHVGAT) decreases weight gain and food intake, whereas LHLEPR ablation does not. Appetitive learning in a Pavlovian conditioning paradigm is delayed in LHVGAT-ablated mice but prevented entirely in LHLEPR-ablated mice. Both LHVGAT and LHLEPR neurons bidirectionally modulate reward-related behaviors, but only LHVGAT neurons affect feeding. In the Pavlovian paradigm, only LHLEPR activity discriminates between conditioned cues. Optogenetic activation or inhibition of either population in this task disrupts discrimination. However, manipulations of LHLEPR→VTA projections evoke divergent effects on responding. Unlike food-oriented learning, chemogenetic inhibition of LHLEPR neurons does not alter cocaine-conditioned place preference but attenuates cocaine sensitization. Thus, LHLEPR neurons may specifically regulate appetitive behaviors toward non-drug reinforcers., Graphical Abstract, In brief Siemian et al. investigate the functional contributions of leptin receptor-expressing lateral hypothalamic (LHLEPR) neurons to the broader population of GABAergic LH (LHVGAT) neurons in mice. They show that LHLEPR neurons specifically mediate appetitive behaviors, whereas LHVGAT neurons affect appetitive, consummatory, and displacement behaviors.

Polyamines (PAs) are polycationic biomolecules containing multiple amino groups. Patients with HIV-associated neurocognitive disorder (HAND) have high concentrations of the polyamine N-acetylated spermine in their brain and cerebral spinal fluid (CSF) and have increased PA release from astrocytes. These effects are due to the exposure to HIV-Tat. In healthy adult brain, PAs are accumulated but not synthesized in astrocytes, suggesting that PAs must enter astrocytes to be N-acetylated and released. Therefore, we tested if Cx43 hemichannels (Cx43-HCs) are pathways for PA flux in control and HIV-Tat-treated astrocytes. We used biotinylated spermine (b-SPM) to examine polyamine uptake. We found that control astrocytes and those treated with siRNA-Cx43 took up b-SPM, similarly suggesting that PA uptake is via a transporter/channel other than Cx43-HCs. Surprisingly, astrocytes pretreated with both HIV-Tat and siRNA-Cx43 showed increased accumulation of b-SPM. Using a novel polyamine transport inhibitor (PTI), trimer 44NMe, we blocked b-SPM uptake, showing that PA uptake is via a PTI-sensitive transport mechanism such as organic cation transporter. Our data suggest that Cx43 HCs are not a major pathway for b-SPM uptake in the condition of normal extracellular calcium concentration but may be involved in the release of PAs to the extracellular space during viral infection.

While energy balance is critical to survival, many factors influence food intake beyond caloric need or “hunger.” Despite this, some neurons that drive feeding in mice are routinely referred to as “hunger neurons,” whereas others are not. To understand how specific hypothalamic circuits control interoceptive hunger, we trained mice to discriminate fasted from sated periods. We then manipulated three hypothalamic neuronal populations with well-known effects on feeding while mice performed this task. While activation of ARC(AGRP) neurons in sated mice caused mice to report being food-restricted, LH(VGAT) neuron activation or LH(VGLUT2) neuron inhibition did not. In contrast, LH(VGAT) neuron inhibition or LH(VGLUT2) neuron activation in fasted mice attenuated natural hunger, whereas ARC(AGRP) neuron inhibition did not. Each neuronal population evoked distinct effects on food consumption and reward. After satiety- or sickness-induced devaluation, ARC(AGRP) neurons drove calorie-specific feeding, while LH(VGAT) neurons drove calorie-indiscriminate food intake. Our data support a role for ARC(AGRP) neurons in homeostatic feeding and implicate them in driving a hunger-like internal state that directs behavior toward caloric food sources. Moreover, manipulations of LH circuits did not evoke hunger-like effects in sated mice, suggesting that they may govern feeding more related to reward, compulsion, or generalized consumption than to energy balance but also that these LH circuits can be powerful negative appetite modulators in fasted mice. This study highlights the complexity of hypothalamic feeding regulation and can be used as a framework to characterize how other neuronal circuits affect hunger and identify potential therapeutic targets for eating disorders.

Vaccines against SARS-CoV-2 have been distributed at massive scale in developed countries, and have been effective at preventing COVID-19. Access to vaccines is limited, however, in low- and middle-income countries (LMICs) due to insufficient supply, high costs, and cold storage requirements. New vaccines that can be produced in existing manufacturing facilities in LMICs, can be manufactured at low cost, and use widely available, proven, safe adjuvants like alum, would improve global immunity against SARS-CoV-2. One such protein subunit vaccine is produced by the Serum Institute of India Pvt. Ltd. and is currently in clinical testing. Two protein components, the SARS-CoV-2 receptor binding domain (RBD) and hepatitis B surface antigen virus-like particles (VLPs), are each produced in yeast, which would enable a low-cost, high-volume manufacturing process. Here, we describe the design and preclinical testing of the RBD-VLP vaccine in cynomolgus macaques. We observed titers of neutralizing antibodies (>104) above the range of protection for other licensed vaccines in non-human primates. Interestingly, addition of a second adjuvant (CpG1018) appeared to improve the cellular response while reducing the humoral response. We challenged animals with SARS-CoV-2, and observed a ~3.4 and ~2.9 log10reduction in median viral loads in bronchoalveolar lavage and nasal mucosa, respectively, compared to sham controls. These results inform the design and formulation of current clinical COVID-19 vaccine candidates like the one described here, and future designs of RBD-based vaccines against variants of SARS-CoV-2 or other betacoronaviruses.