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Technological and mathematical advances have provided opportunities to investigate new approaches for the holistic quantification of complex biological systems. One objective of these approaches, including the multi-inverse deterministic approach proposed in this paper, is to deepen the understanding of biological systems through the structural development of a useful, best-fitted inverse mechanistic model. The objective of the present work was to evaluate the capacity of a deterministic approach, that is, the multi-inverse approach (MIA), to yield meaningful quantitative nutritional information. To this end, a case study addressing the effect of diet composition on sheep weight was performed using data from a previous experiment on saccharina (a sugarcane byproduct), and an inverse deterministic model (named Paracoa) was developed. The MIA successfully revealed an increase in the final weight of sheep with an increase in the percentage of corn in the diet. Although the soluble fraction also increased with increasing corn percentage, the effective nonsoluble degradation increased fourfold, indicating that the increased weight gain resulted from the nonsoluble substrate. A profile likelihood analysis showed that the potential best-fitted model had identifiable parameters, and that the parameter relationships were affected by the type of data, number of parameters and model structure. It is necessary to apply the MIA to larger and/or more complex datasets to obtain a clearer understanding of its potential.

Paciente de 78 años, varón, que acude al servicio de Seguimiento Farmacoterapéutico de nuestra farmacia, refiriendo una sensación de ahogo desde hace un tiempo, que le imposibilita el desarrollo de las actividades cotidianas.

La línea de investigación con que el Departamento de Historia Moderna de la Universidad de Salamanca ha venido abordando las diferentes perspectivas de su propia institución académica está comenzando ya a producir frutos. Refirámonos someramente al «corpus» de los libros de claustros entre 1503 y 1555 que ha dirigido el Dr. Manuel Fernández Álvarez, a los estudios sobre Colegios Mayores de la Dr? Ana Carabias Torres, o a los míos propios sobre la Universidad Salmantina del Barroco.

Banana, the main export fruit for Colombia, is threatened by Fusarium wilt (FWB), caused by Fusarium oxysporum f. sp. cubense (Foc), tropical race 4 (TR4). Pathogen containment through disinfecting tools, machinery, shoes, and any means that may carry contaminated soil particles with proper disinfectants is at the forefront of disease management. In this study, the biocide efficacy of 10 commercial quaternary ammonium compounds (QACs) products and one based on glutaraldehyde (GA) were evaluated on both reproductive structures (microconidia and macroconidia) and survival spores (chlamydospores) of Foc TR4 (strain 140038) isolated from La Guajira, Colombia. QACs were evaluated at 1200 ppm and two exposure times: Foc TR4 chlamydospores, respectively, after Foc TR4 propagules after 15 min for all concentrations tested.

El objetivo de este trabajo fue realizar un diagnóstico de las empresas agropecuarias que ofertan sus productos por Internet. El área de estudio consistió en empresas agropecuarias mexicanas que se anuncian por Internet. Una búsqueda en línea se llevó a cabo para generar una base de datos de 481 empresas agropecuarias anunciadas en sitios mexicanos que tuvieran como requisito principal un correo electrónico. Posteriormente, se realizó una encuesta con preguntas orientadas a conocer la situación actual del comercio electrónico. Para incrementar su confiabilidad, se envió varias veces la encuesta utilizando los conceptos del método Delphi y el muestreo simple aleatorio. La información se analizó y se obtuvo el tamaño de muestra para estimar proporciones o porcentajes de acuerdo a la fórmula empleada por el diseño de muestra y muestreo aleatorio simple. El tamaño de muestra calculado por este procedimiento fue de 40 empresas, mismas que representaron a la población con un 95% de confianza, y una varianza estimada de alrededor de 0.56%. El 95.0% de las empresas que se anuncian por Internet se originaron fuera de él y no han conseguido sus objetivos que se plantearon al momento de moverse hacia el Internet.

El estudio fue realizado en el trópico húmedo del sureste mexicano con el objetivo de estimar la partici-pación de las Asociaciones en el acopio y comercialización de cacao en Tabasco, México, durante el ciclo de cosecha 1999 - 2000. Para analizar el acopio y comercialización del cacao se aplicó un censo y entrevistas participativas a los Presidentes de las 27 Asociaciones ubicadas en el estado de Tabasco. Las variables consideradas fueron: acopio de cacao, precio de compra y venta de cacao, crédito, infraestructura vehicular, años de estudio y de trabajo de los Presidentes; estas variables se analizaron mediante correlación lineal simple. Con las variables de acopio y venta, se obtuvieron algunos estadísticos descriptivos, tales como la media aritmética y porcentajes. Los resultados indicaron que del volumen estatal (22 289 t) de grano seco cosechado durante el ciclo de producción 1999 - 2000; la Unión Nacional de Productores de Cacao (UNPC) acopió 17.7%. Del total del cacao beneficiado (8 142 t) por las Asociaciones, éstas vendieron a la UNPC 37.3 %, y 62.7% fue comercializado a empresas privadas.

Se aislaron, identificaron y caracterizaron 20 cepas de bacterias ácido lácticas (BAL) de un “queso crema tropical” tradicionalmente fabricado con leche bronca. Las 20 cepas de BAL utilizadas crecieron en leche descremada y cada una de ellas fue evaluada mediante su poder acidificante (pH) en cuatro tiempos de incubación (6, 24, 48 y 72 h) y tres temperaturas de fermentación (30, 37 y 42oC). En base al pH más bajo, se seleccionaron seis cepas para fabricar un “queso tipo crema tropical” con leche pasteurizada. Las cepas fueron caracterizadas mediante pruebas bioquímicas (fermentación de carbohidratos y producción de amonio a partir de arginina) y de secuenciación del 16S rDNA. Se identificaron cinco especies de Lactobacillus fermentum y una especie de Lactobacillus pentosus. Entre las temperaturas de fermentación y el tiempo de incubación se estimaron diferencias significativas (p < 0.05) en cada una de las cepas. En las temperaturas de 37oC y 42oC se produjeron los valores de pH más bajos a las 72 horas de incubación. Además, el ácido láctico, ácido acético, ácido propiónico y ácido butírico en leche fermentada se evaluaron con cada cepa y no hubo diferencias significativas en los niveles de producción de dichos metabolitos (p > 0.05). Asimismo, se realizaron pruebas que incluyeron cuentas totales de flora no láctica, cuenta de BAL, coliformes totales, Escherichia coli, hongos y levaduras en el queso crema elaborado con leche pasteurizada y como referencia se uso al queso crema artesanal.

El objetivo del presente estudio fue evaluar la variabilidad disponible en 21 genotipos de Brachiaria humi-dícola en cuanto a la concentración de proteína y producción de materia seca, durante la época seca del año. El estudio se realizó durante los meses de marzo, abril y mayo de los años 2004 y 2005, sin aplicar riego ni fertilizante al suelo. Se realizaron tres cosechas por genotipo en cada año, con un tiempo de rebrote de cada cosecha que varió de 33 a 35 d. El experimento fue establecido bajo un diseño de bloques completos al azar en arreglo de parcelas subdivididas. Los resultados mostraron una importante variabilidad entre los genotipos (p ≤ 0.0001) para la concentración de proteína dentro del año, la cual varió de 101.1 a 119.5 g kg-1 de MS, valores que corresponden a los genotipos 26 160 y 26 413. Los genotipos 26 413 y 16 891 fueron los que presentaron las concentraciones de proteína más elevadas pero, sin ser significativamente superiores (p > 0.05) al genotipo 679 (Humidicola comercial). Además, la concentración de proteína mostró una estabilidad importante al ambiente al no interactuar ni con el año ni con la fecha de corte (p > 0.05).

Se evaluó el potencial de un consorcio microbiano constituido por bacterias (Pseudomonas sp y Serratia marcescens y hongos (Aspergillus sp y Trichoderma sp) y de fertilizante inorgánico incorporados al suelo en la biorremediación, y el potencial del pasto egipto (Brachiaria mutica) y de la leguminosa guaje (Leucaena leucocephala) en la fitorremediación de un suelo contaminado con petróleos nuevo (PN) recién incorporado al suelo y el intemperizado (PI) procedente de derrames crónicos de petróleo desde hace 25 años. El experimento fue factorial 4x3x2x2, estos fueron petróleo [150 mg kg-1 de hidrocarburos totales del petróleo (HTP) de origen biogénico, 50 000 y 78 000 mg kg-1 HTP de PN y 79 457 mg kg-1 HTP de PI], planta (sin planta, con planta de pasto egipto o de guaje),consorcio microbiano (sin inóculo y con inóculo, 5 mL de inóculo bacteriano con 250 x 10 6 UFC mL-1 + 40x105 UFC de hongos mL-1) y fertilizante (sin fertilizante y con fertilizante, dosis 160-50-00). Se evaluaron 48 tratamientos. Se cuantificaron al inicio (día 1) y al final (día 158) la degradación de los HTP y las poblaciones de bacterias y hongos. La medición de bacterias y hongos se realizó con el método de recuento en cajas Petri. La extracción de HTP se efectuó con diclorometano, procedimiento EPA 418.1 y la cuantificación por gravimetría según la NMX-AA-134-SCFI-2006. La interacción de rizósfera, consorcio microbiano y el fertilizante originaron efectos positivos significativos (Tukey p ≤ 0.05), promovieron la degradación del PN y el PI.

The application of a 3D domain decomposition finite-element and spherical mode expansion for the design of planar ESPAR (electronically steerable passive array radiator) made with probe-fed circular microstrip patches is presented in this work. A global generalized scattering matrix (GSM) in terms of spherical modes is obtained analytically from the GSM of the isolated patches by using rotation and translation properties of spherical waves. The whole behaviour of the array is characterized including all the mutual coupling effects between its elements. This procedure has been firstly validated by analyzing an array of monopoles on a ground plane, and then it has been applied to synthesize a prescribed radiation pattern optimizing the reactive loads connected to the feeding ports of the array of circular patches by means of a genetic algorithm.

The article presents a study describing a method for evaluating the control of the welding process. This investigation is aimed at presenting a qualitative and quantitative means of analyzing the welding process control. Further, this study also addresses the effects of different technology in various applications and the elimination of the typical defects.

In this paper we explore the effects of self-obscuration in protostellar disks with a radially decreasing temperature gradient and a colder midplane. We are motivated by recent reports of resolved dark lanes (“hamburgers”) and (sub)millimeter spectral indices systematically below the ISM value for optically thin dust, αISM = 3.7. We explore several model grids, scaling disk mass and varying inclination angle i and observing frequency ν from the VLA Ka band (∼37 GHz) to ALMA Band 8 (∼405 GHz). We also consider the effects of decreasing the index of the (sub-)millimeter dust opacity power-law β from 1.7 to 1. We find that a distribution of disk masses in the range Mdisk = 0.01–2 M⊙ is needed to reproduce the observed distribution of spectral indices, and that assuming a fixed β = 1.7 gives better results than β = 1. A wide distribution of disk masses is also needed to produce some cases with α < 2, as reported for some sources in the literature. Such extremely low spectral indices arise naturally when the selected observing frequencies sample the appropriate change in the temperature structure of the optically thick model disk. Our results show that protostellar disk masses could often be underestimated by > ×10, and are consistent with recent hydrodynamical simulations. Although we do not rule out the possibility of some grain growth occurring within the short protostellar timescales, we conclude that self-obscuration needs to be taken into account. [ABSTRACT FROM AUTHOR]

Massive star-forming regions exhibit a rich chemistry with complex gas distributions, especially on small scales. While surveys have yielded constraints on typical gas conditions, they often have coarse spatial resolution and limited bandwidths. Thus, to establish an interpretative framework for these efforts, detailed observations that simultaneously provide high sensitivity, spatial resolution, and large bandwidths for a subset of diverse sources are needed. Here, we present wide-band (≈32 GHz) Submillimeter Array (SMA) observations of four high-mass star-forming regions (G28.20-0.05, G20.08-0.14 N, G35.58-0.03, and W33 Main) at subarcsecond resolution, where we detect and spatially resolve hundreds of lines from over 60 molecules, including many complex organic molecules (COMs). The chemical richness of our sample is consistent with an evolutionary sequence from the line-rich hot cores and hypercompact H ii regions of G28.20-0.05 and G20.08-0.14 N to the more chemically modest ultracompact H ii regions in G35.58-0.03, followed by the molecule-poor H ii region W33 Main. We detect lines across a range of excitation conditions ( E _u ≈ 20 to ≳800 K) and from numerous isotopologues, which enables robust estimates of gas properties. We derive nearly constant COM column density ratios that agree with literature values in other low- and high-mass protostellar cores, supporting the idea that COM abundances are set during the pre-stellar phase. In all regions, we identify spatial offsets among different molecular families, due to a combination of source physical structure and chemistry. In particular, we find potential evidence of carbon grain sublimation in G28.20-0.05 and identify an elemental oxygen gradient and rich sulfur chemistry in G35.58-0.03. Overall, these results demonstrate that the SMA's wide bandwidth is a powerful tool to untangle the complex molecular gas structures associated with massive star formation.

Earth construction is a revisited topic due to the low-cost self-production employed in communities and locations where conventional materials (concrete and steel) are not available. One of the properties that are popularly recognized for earth-based buildings is the pleasant indoor thermal sensation. This article presents the development of a model to characterize the temperature of the indoor air applied to adobe houses located in San Carlos, Tamaulipas, Mexico. The model is developed from the heat transfer equations in an ideal case where the temperature is modified by conduction through the wall, and the interior air is heated by convection. In the ideal case, it is considered that there are no doors or windows open because it is what commonly occurs in the earth-based houses during the warmest period of the year. It was found that for a thickness greater than 10 cm there is no significant increase in temperature assuming an outside temperature above 40 °C and an initial indoor temperature of 25 °C. The model characterizes and predict a strong relation between the physical properties of the air and the wall, the thickness of the wall, the dimensions and shape of the room, and the roughness of the outer wall. This consideration makes it possible to quantitatively estimate comfortability based on indoor temperature and wall thickness, which is an expected behavior according to literature. This is important because not only has an impact on the construction of new buildings but also in the critical maintenance of existing houses.

ABSTRACT Objective Ultrasonography (US) is the best diagnostic tool for initial assessment of thyroid nodule. Recently, data reporting systems for thyroid lesions, such as the Thyroid Imaging Reporting and Data System (TI-RADS) and American Thyroid Association (ATA), which stratifies the risk for malignancy, have demonstrated good performance in differentiating malignant thyroid nodules. The purpose of this study is to determine the reliability of both data reporting systems in predicting thyroid malignancy in a tertiary care hospital. Materials and methods We evaluated 195 thyroid nodules using modified TI-RADS and ATA risk stratification. The results were compared to the cyto-pathology analysis. Histopathological results were available for 45 cases after surgery, which is considered the golden standard for diagnosis of thyroid cancer. Results When compared with cytological results, sensitivity, specificity, negative predictive value (NPV), and accuracy were 100, 61.1, 100, and 63%, respectively, for TI-RADS; and 100, 75, 100, and 76%, respectively, for ATA. When compared with histopathological results, sensitivity, specificity, NPV, and accuracy were 90, 51.4, 94.7, and 60% respectively, for TI-RADS; and 100, 60, 100, and 68%, respectively, for ATA. All patients with malignant nodules were classified in the categories 4 or 5 of TI-RADS and in the intermediate or high suspicion risk according to the ATA system. Conclusion Both TI-RADS and the ATA guidelines have high sensitivity and NPV for the diagnosis of thyroid carcinoma. These systems are feasible for clinical application, allowing to better select patients to undergo fine-needle aspiration biopsies.

Se presenta una aplicación del método de análisis de criticidad en efectos y modos de fallos, paraevaluar cualitativa y cuantitativamente el control del proceso de soldadura y la incidencia de la tecnología,en diferentes momentos y en la eliminación de los defectos típicos. An application of the method of criticality analysis is presented in effects and ways of shortcomings,to evaluate qualitative and quantitatively the control of the welding process and the incidence ofthe technology, in different moments and in the elimination of the typical defects

Two microsporidian species infect honeybees worldwide, Nosema apis and Nosema ceranae. Two different clinical patterns are considered: nosemosis type A (N. apis) and nosemosis type C (N. ceranae). Nosemosis type A is characterized in acute forms and nosemosis type C shows no clear outward clinical signs. The development of a rapid and simple tool for Nosema detection could allow beekeepers or veterinarians to carry out diagnostic tests in situ. Currently, PCR and microscopy are expensive techniques that require qualified staff and may not be available in every laboratory. The present study describes the production and characterization of four monoclonal antibodies (mAbs) against N. ceranae and N. apis, and the development of an IFAT. An IFAT using the mAbs was compared with microscopy and PCR for 180 beehive samples. The diagnostic test revealed similar sensitivity and specificity percentages to IFAT (97.79% and 93.18%, respectively) and microscopy (97.79% and 95.45%), considering 100% for the PCR as the 'gold standard'. A mAb (7D2) was patented for its high specificity for N. ceranae. The IFAT using the mAbs is a good alternative to microscopy and PCR in laboratories where PCR is not available for the detection and identification of both Nosema spp., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Using density-functional theory calculations, we determined the pressure domain of stability of beryllium metasilicate, BeSiO 3 , an elusive compound for which no stable polymorph is known until now. We found that BeSiO 3 is stable at pressures above 9 GPa, a condition that makes it accessible with a large-volume press. After considering the cubic, orthorhombic, and hexagonal perovskite structures and the Ilmenite structure, known from related compounds, we propose that the most stable structure among them is the orthorhombic perovskite structure described by space group Pnma . The unit-cell parameters of this structure are a = 4.966 Å, b = 7.160 Å, and c = 4.374 Å. We also determined the frequencies of Raman and infrared phonons, the elastic constants and modulus, and the electronic band structure for the orthorhombic perovskite structure. Finally, the pressure dependence of unit-cell parameters was calculated. Compression was found to be slightly anisotropic, with the axial compressibilities decreasing following the sequence κ b > κ c > κ a . In addition, we found that BeSiO 3 is quite incompressible with a bulk modulus of 242 GPa, which makes it one of the less compressible silicates. To understand, such a large bulk modulus, both the quantum theory of atoms in molecules and the electron localization function decomposition were utilized to analyze the bonding and to relate it to the mechanical properties.

HMGA1 is a structural epigenetic chromatin factor that has been associated with tumor progression and drug resistance. Here, we reported the prognostic/predictive value of HMGA1 for trabectedin in advanced soft-tissue sarcoma (STS) and the effect of inhibiting HMGA1 or the mTOR downstream pathway in trabectedin activity. The prognostic/predictive value of HMGA1 expression was assessed in a cohort of 301 STS patients at mRNA (n = 133) and protein level (n = 272), by HTG EdgeSeq transcriptomics and immunohistochemistry, respectively. The effect of HMGA1 silencing on trabectedin activity and gene expression profiling was measured in leiomyosarcoma cells. The effect of combining mTOR inhibitors with trabectedin was assessed on cell viability in vitro studies, whereas in vivo studies tested the activity of this combination. HMGA1 mRNA and protein expression were significantly associated with worse progression-free survival of trabectedin and worse overall survival in STS. HMGA1 silencing sensitized leiomyosarcoma cells for trabectedin treatment, reducing the spheroid area and increasing cell death. The downregulation of HGMA1 significantly decreased the enrichment of some specific gene sets, including the PI3K/AKT/mTOR pathway. The inhibition of mTOR, sensitized leiomyosarcoma cultures for trabectedin treatment, increasing cell death. In in vivo studies, the combination of rapamycin with trabectedin downregulated HMGA1 expression and stabilized tumor growth of 3-methylcholantrene-induced sarcoma-like models. HMGA1 is an adverse prognostic factor for trabectedin treatment in advanced STS. HMGA1 silencing increases trabectedin efficacy, in part by modulating the mTOR signaling pathway. Trabectedin plus mTOR inhibitors are active in preclinical models of sarcoma, downregulating HMGA1 expression levels and stabilizing tumor growth., (© 2024. The Author(s).)

Subvalent compounds raise the question of where those valence electrons not belonging to chemical bonds are. In the limiting case of Ag 7 Pt 2 O 7 , there is just one-electron excess in the chemical formula requiring the presence of Ag atoms with oxidation states below +1, assuming conventional Pt 4+ and O 2- ions. Such a situation challenges the understanding of the semiconducting and diamagnetic behavior observed in this oxide. Previous explanations that localize pairwise the electron excess in tetrahedral Ag 4 interstices do not suffice in this case, since there are six silver tetrahedral voids and only an excess of nine electrons in the unit cell. Here, we provide an alternative explanation for the subvalent nature of this compound by combining interatomic distances, electron density-based descriptors, and orbital energetic analysis criteria. As a result, Ag atoms that do not participate in their valence electron are revealed. We identify excess electrons located in isolated subvalent silver clusters with electron-deficient multicenter bonds resembling pieces of metallic bonding in fcc-Ag and Ag 7 Pt 2 alloy. Our analysis of the electronic band structure also supports the multicenter bonding picture. This combined approach from the real and reciprocal spaces reconciles existing discrepancies and is key to understanding the new chemistry of silver subvalent compounds.

In the search for new active pharmaceutical ingredients, the precise control of the chemistry of cocrystals becomes essential. One crucial step within this chemistry is proton migration between cocrystal coformers to form a salt, usually anticipated by the empirical ΔpK a rule. Due to the effective role it plays in modifying intermolecular distances and interactions, pressure adds a new dimension to the ΔpK a rule. Still, this variable has been scarcely applied to induce proton-transfer reactions within these systems. In our study, high-pressure X-ray diffraction and Raman spectroscopy experiments, supported by DFT calculations, reveal modifications to the protonation states of the 4,4'-bipyridine (BIPY) and malonic acid (MA) cocrystal (BIPYMA) that allow the conversion of the cocrystal phase into ionic salt polymorphs. On compression, neutral BIPYMA and monoprotonated (BIPYH + MA - ) species coexist up to 3.1 GPa, where a phase transition to a structure of P2 1 /c symmetry occurs, induced by a double proton-transfer reaction forming BIPYH 2 2+ MA 2- . The low-pressure C2/c phase is recovered at 2.4 GPa on decompression, leading to a 0.7 GPa hysteresis pressure range. This is one of a few studies on proton transfer in multicomponent crystals that shows how susceptible the interconversion between differently charged species is to even slight pressure changes, and how the proton transfer can be a triggering factor leading to changes in the crystal symmetry. These new data, coupled with information from previous reports on proton-transfer reactions between coformers, extend the applicability of the ΔpK a rule incorporating the pressure required to induce salt formation., (open access.)

Microsporidia are widely spread obligate intracellular fungal pathogens from vertebrate and invertebrate organisms, mainly transmitted by contaminated food and water. This study aims to detect the presence of major human-pathogenic microsporidia, i.e., Enterocytozoon bieneusi , Encephalitozoon intestinalis , Encephalitozoon hellem , and Encephalitozoon cuniculi , in the gastrointestinal tract of commercially harvested marine fish from Mediterranean coast of the Comunidad Valenciana, Eastern Spain. A total of 251 fish, 138 farmed fish and 113 wild fish from commercial fishing were tested by SYBR Green real-time PCR, enabling the simultaneous detection of the four targeted species. E. intestinalis/hellem was found in 1.45% of farmed fish and 7.96% of wild fish, while Enterocytozoonidae was detected in 2.90% and 18.58% of farmed and wild fish, respectively. E. cuniculi was not detected in any of the analyzed specimens. To the authors' knowledge, this is the first report of E. intestinalis/hellem in fish, particularly in marine fish. Although the role of fish in these species' epidemiology remains unknown, this finding points out a potential public health risk linked to fish consumption. Further studies are necessary to characterize these microsporidia in fish hosts better and to elucidate their epidemiological role.

Covalent crystals such as diamonds are a class of fascinating materials that are challenging to fabricate in the form of thin films. This is because spatial kinetic control of bond formation is required to create covalently bonded crystal films. Directional crystal growth is commonly achieved by chemical vapor deposition, an approach that is hampered by technical complexity and associated high cost. Here we report on a liquid-liquid interfacial approach based on physical-organic considerations to synthesize an ultrathin covalent crystal film. By distributing reactants into separate phases using hydrophobicity, the chemical reaction is confined to an interface that orients the crystal growth. A molecular-smooth interface combined with in-plane isotropic conditions enables the synthesis of films on a centimeter size scale with a uniform thickness of 13 nm. The film exhibits considerable mechanical robustness enabling a free-standing length of 37 µm, as well as a clearly anisotropic chemical structure and crystal lattice alignment., (© 2023. The Author(s).)

Microsporidia are opportunistic intracellular parasites, generating serious pathology in individuals with a compromised immune system. Infection by microsporidia inhibits p53 and Caspase 3, proteins involved in apoptosis and the cell cycle, which are vital in the malignant process of epithelial cells. The presence of microsporidia in the intestinal tissues of 87 colon cancer (CC) patients and 25 healthy controls was analyzed by real-time PCR and an immunofluorescence antibody test. Anti-Encephalitozoon antibodies were analyzed in serum samples by ELISA (enzyme linked immunosorbent assay). In 36 (41.3%) CC cases, microsporidia infections were identified in their tissues vs. no cases among control subjects (p < 0.0001). An increase in IgG and IgE anti-Encephalitozoon antibodies was found in patients with CC, which would demonstrate continuous and previous contact with the parasite. The high prevalence of microsporidia in tissues and the seroprevalence in patients with CC suggest a relationship between microsporidia and the etiopathogenesis of CC.

Lynx pardinus is one of the world's most endangered felines inhabiting the Iberian Peninsula. The present study was performed to identify the presence of microsporidia due to the mortality increase in lynxes. Samples of urine ( n = 124), feces ( n = 52), and tissues [spleen ( n = 13), brain ( n = 9), liver ( n = 11), and kidney ( n = 10)] from 140 lynxes were studied. The determination of microsporidia was evaluated using Weber's chromotrope stain and Real Time-PCR. Of the lynxes analyzed, stains showed 10.48% and 50% positivity in urine and feces samples, respectively. PCR confirmed that 7.69% and 65.38% belonged to microsporidia species. The imprints of the tissues showed positive results in the spleen (38.46%), brain (22.22%), and liver (27.27%), but negative results in the kidneys. PCR confirmed positive microsporidia results in 61.53%, 55.55%, 45.45%, and 50%, respectively. Seroprevalence against Encephalitozoon cuniculi was also studied in 138 serum samples with a positivity of 55.8%. For the first time, the results presented different species of microsporidia in the urine, feces, and tissue samples of Lynx pardinus . The high titers of anti- E. cuniculi antibodies in lynx sera confirmed the presence of microsporidia in the lynx environment. New studies are needed to establish the impact of microsporidia infection on the survival of the Iberian lynx.

The direct chemical reactivity between phosphorus and nitrogen was induced under high-pressure and high-temperature conditions (9.1 GPa and 2000-2500 K), generated by a laser-heated diamond anvil cell and studied by synchrotron X-ray diffraction, Raman spectroscopy, and DFT calculations. α-P 3 N 5 and γ-P 3 N 5 were identified as reaction products. The structural parameters and vibrational frequencies of γ-P 3 N 5 were characterized as a function of pressure during room-temperature compression and decompression to ambient conditions, determining the equation of state of the material up to 32.6 GPa and providing insight about the lattice dynamics of the unit cell during compression, which essentially proceeds through the rotation of the PN 5 square pyramids and the distortion of the PN 4 tetrahedra. Although the identification of α-P 3 N 5 demonstrates for the first time the direct synthesis of this compound from the elements, its detection in the outer regions of the laser-heated area suggests α-P 3 N 5 as an intermediate step in the progressive nitridation of phosphorus toward the formation of γ-P 3 N 5 with increasing coordination number of P by N from 4 to 5. No evidence of a higher-pressure phase transition was observed, excluding the existence of predicted structures containing octahedrally hexacoordinated P atoms in the investigated pressure range.

Using Lifshitz theory, we assess the role of van der Waals forces at interfaces of ice and water. The results are combined with measured structural forces from computer simulations to develop a quantitative model of the surface free energy of premelting films. This input is employed within the framework of wetting theory and allows us to predict qualitatively the behavior of quasi-liquid layer thickness as a function of ambient conditions. Our results emphasize the significance of vapor pressure. The ice-vapor interface is shown to exhibit only incomplete premelting, but the situation can shift to a state of complete surface melting above water saturation. The results obtained serve also to assess the role of subsurface freezing at the water-vapor interface, and we show that intermolecular forces favor subsurface ice nucleation only in conditions of water undersaturation. We show that ice regelation at ambient pressure may be explained as a process of capillary freezing, without the need to invoke the action of bulk pressure melting. Our results for van der Waals forces are exploited in order to gauge dispersion interactions in empirical point charge models of water.

Predictive biomarkers of trabectedin represent an unmet need in advanced soft-tissue sarcomas (STS). DNA damage repair (DDR) genes, involved in homologous recombination or nucleotide excision repair, had been previously described as biomarkers of trabectedin resistance or sensitivity, respectively. The majority of these studies only focused on specific factors (ERCC1, ERCC5, and BRCA1) and did not evaluate several other DDR-related genes that could have a relevant role for trabectedin efficacy. In this retrospective translational study, 118 genes involved in DDR were evaluated to determine, by transcriptomics, a predictive gene signature of trabectedin efficacy. A six-gene predictive signature of trabectedin efficacy was built in a series of 139 tumor samples from patients with advanced STS. Patients in the high-risk gene signature group showed a significantly worse progression-free survival compared with patients in the low-risk group (2.1 vs 6.0 months, respectively). Differential gene expression analysis defined new potential predictive biomarkers of trabectedin sensitivity (PARP3 and CCNH) or resistance (DNAJB11 and PARP1). Our study identified a new gene signature that significantly predicts patients with higher probability to respond to treatment with trabectedin. Targeting some genes of this signature emerges as a potential strategy to enhance trabectedin efficacy., (© 2021 The Authors. Molecular Oncology published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Here an all-carbon linked 3D covalent organic framework (COF) is introduced by employing a templated surface reaction in a continuous flow (TSRCF). The presented method of synthesis provides spatial control over the reaction chemistry and allows for the creation of ultrasmooth COF films of desired thickness and significant crystallinity. The films show high electrical conductivity (≈3.4 S m -1 ) after being doped with tetracyanoquinodimethane (TCNQ), setting a new record for 3D COF materials. The concurrence of 3D nanosized channels and high conductivity opens up for a number of hitherto unexplored applications for this class of materials, such as high surface area electrodes, electrochemical transistors, and for electronic sensing., (© 2021 The Authors. Small published by Wiley-VCH GmbH.)

L. feeleii is one of the most frequent Legionella species isolated from natural pools of the central region of Spain. This study aimed to evaluate its ecology and to identify this Legionella species as a respiratory pathogen. A PCR assay for detecting the L. feeleii mip gene was developed to identify it in clinical and environmental samples. Culture and PCR were performed in environmental samples from four drinking water treatment plants (DWTPs). Free L. feeleii was only detected in raw water samples (3.4%), while L. feeleii as an Acanthamoeba endosymbiont was found in 30.7% of raw water, 11.5% of decanter biofilm, and 32% of finished water samples. Therefore, Acanthamoeba spp. plays an essential role in the multiplication, persistence, and spread of Legionella species in the environment. The first case of Legionnaires' disease caused by L. feeleii in Spain is described in this study. The case was diagnosed in an older woman through PCR and sequencing from urine and sputum samples. A respiratory infection could be linked with health care procedures, and the patient presented several risk factors (age, insulin-dependent diabetes, and heart disease). The detection of non- L . pneumophila , such as L. feeleii , is a factor that must be considered when establishing or reviewing measures for the control and prevention of legionellosis., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Vaccaro, Gomes, Izquierdo, Magnet, Llorens Berzosa, Ollero, Salso, Alhambra, Gómez, López Cano, Pelaz, Bellido Samaniego, del Aguila, Fenoy and Hurtado-Marcos.)

The present study analyzes at the national level, the presence of circulating Legionella in the artificial aquatic systems of different facilities of all of them state-owned centers throughout Spain for 12 months. 1754 water samples from various state-owned centers were collected from January to December 2014. Samples were collected from the cooling towers and evaporative condensers (CTC), and water distribution networks such as domestic hot water (DHW), cold water for human consumption (CW), sprinkler irrigation systems (SIS), fire sprinkler systems (FSS), and water from decorative fountains (DF). All these facilities are considered, according to current regulations, as potential amplifying systems for bacteria and possible sources of infection by the generation of droplets and aerosols. The isolation and counting of Legionella in water samples was carried out using microbiological culture following the international normative UNE-EN-ISO 11,731:2007 (ISO 11,731:1998) and UNE-EN ISO 8199:2008 (ISO 8199:2005).The quantification of Legionella colonization, the annual distribution, and the geographical distribution of the Legionella isolates recovered in the water were analyzed. Besides, molecular techniques were used for the characterization of the Legionella non-pneumophila isolates. Legionella was recovered from 15.79% of the analyzed water samples. High colonization was more frequently detected in water samples from CTC, DHW, CW, and DF. Regarding the geographic distribution, positive samples of Legionella were obtained in 14 of the 18 Spanish locations analyzed. Legionella non-pneumophila was the most prevalent and was isolated from water samples from 13 different geographical locations (72%). Legionella anisa and Legionella jordanis were the most frequently non-pneumophila species isolated. Legionella donaldsonii was isolated for the first time in the water distribution networks in Spain. Legionella pneumophila sg 2-14 was detected in 13 locations and Legionella pneumophila sg 1 in 11 locations. Therefore, our study concludes that the presence of Legionella pneumophila and Legionella non-pneumophila species in these systems can be a potential threat to public health and should be examined thoroughly with complementary techniques, such as molecular techniques as a screen for routine diagnosis., Competing Interests: Declaration of Competing Interest All authors declare no actual or potential conflict of interest including any financial, personal or other relationships with founding organizations., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Objectives: This work aimed to assess the diagnostic validity of two approaches for the screening of gestational diabetes mellitus (GDM) with less discomfort for pregnant women., Methods: A prospective diagnostic validation study was conducted with 2007 pregnant women. According to risk factors for GDM, women were classified into high-risk and low-risk groups. The current diagnostic procedure, based on oral glucose overload, was followed; simultaneously HbA 1c was tested and an algorithm combining both biomarkers was applied., Results: In the low-risk group, the Glucose challenge test (GCT) showed a higher area under the curve (AUC 0.953; 95% CI 0.915-0.992) than the HbA 1c test (0.688; 95% CI 0.541-0.834). The best GCT cut-off, 153.5 mg/dL (8.52 mmol/L), showed higher diagnostic validity than that for HbA 1c , 28 mmol/mol (4.75%), and that the algorithm using both tests. In the high-risk group, the GCT showed better diagnostic performance than the HbA 1c and the algorithm; the optimal GCT cut-offs were higher than those recommended in current protocols. 13th week: GCT AUC 0.882 (95% CI 0.843-0.921), HbA 1c AUC 0.624 (95% CI 0.562-0.686), GCT cut-off 140.5 mg/dL (7.8 mmol/L), HbA 1c cut-off 33 mmol/mol (5.15%). 24th week: GCT AUC 0.944 (95% CI 0.925-0.962), HbA 1c AUC 0.642 (95% CI 0.575-0.709), GCT cut-off, 145.5 mg/dL (8.08 mmol/L), HbA 1c cut-off 29 mmol/mol (4.85%)., Conclusions: The GDM diagnostic approach using as the first step the GCT with higher cut-offs showed the best diagnostic validity. Applying these thresholds, 55.6 and 13.7% of 100 g. Oral glucose overloads would have been avoided in low-risk and high-risk pregnant women., Competing Interests: Competing interests: Authors state no conflict of interest., (© 2021 Miguel Calero Rojas et al., published by De Gruyter, Berlin/Boston.)

Microsporidia are obligate intracellular protist-like fungal pathogens that infect a broad range of animal species, including humans. This study aimed to assess the presence of zoonotic microsporidia ( Enterocytozoon bieneusi , Encephalitozoon intestinalis , Encephalitozoon hellem , and Encephalitozoon cuniculi ) in organ meats of European wild rabbit ( Oryctolagus cuniculus ) and Iberian hare ( Lepus granatensis ) consumed by humans in Spain. Between July 2015 and December 2018, kidney samples from 383 wild rabbits and kidney and brain tissues from 79 Iberian hares in southern Spain were tested by species-specific PCR for the detection of microsporidia DNA. Enterocytozoon bieneusi infection was confirmed in three wild rabbits (0.8%; 95% CI: 0.0-1.7%) but not in hares (0.0%; 95% CI: 0.0-4.6%), whereas E. intestinalis DNA was found in one wild rabbit (0.3%; 95% CI: 0.0-0.8%) and three Iberian hares (3.8%; 95% CI: 0.0-8.0%). Neither E. hellem nor E. cuniculi infection were detected in the 462 (0.0%; 95% CI: 0.0-0.8%) lagomorphs analyzed. The absence of E. hellem and E. cuniculi infection suggests a low risk of zoonotic foodborne transmission from these wild lagomorph species in southern Spain. To the authors' knowledge, this is the first report of E. intestinalis infection in wild rabbits and Iberian hares. The presence of E. bieneusi and E. intestinalis in organ meats from wild lagomorphs can be of public health concern. Additional studies are required to determine the real prevalence of these parasites in European wild rabbit and Iberian hare.

Objective: To estimate the prevalence of burnout and psychological distress among trainee General Practitioners of two training placements and its relationship with the year of residence, lifestyle and working conditions., Design: Cross-sectional study., Location: Primary Care., Participants: On-site surveys for all trainees of two training placements (n = 278), including all trainees of the sample. The response percentage is 76.9%., Main Measurements: Prevalence of burnout using the Maslach Burnout Inventory (MBI) scale and prevalence of psychological distress using the General Health Questionnaire-12 scale (GHQ-12). Also variables on lifestyles, labor organization and drug use., Outcomes: 72% of participants are women. The prevalence of burnout is 33.8% (CI 95% 27.0-40.6), being 25.6% (CI 95% 19.5-31.7) emotional exhaustion, 37.9% (CI 95% 31.0-44.7)depersonalization and 52.9% (CI 95% 45.9-59.9) the low personal accomplishment. Prevalence increases significantly in all dimensions after the second year of training, except in personal accomplishment. The prevalence of emotional distress is 40.4% (CI 95% 33.5-47.3), without significant differences according to year of training. In the group of trainees with psychological distress, 50.0% also have burnout, while this percentage is 22.1% among those who do not present psychological distress., Conclusions: The prevalence of emotional distress and burnout (especially in low personal accomplishment) is high in trainee General Practitioners. The prevalence in higher than the described in previous studies for other groups of health professionals and should be taken into account to develop preventive measures., (Copyright © 2020 The Authors. Publicado por Elsevier España, S.L.U. All rights reserved.)

Over the past decade, there has been a considerable increase in the utilization of cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) in the management of patients with peritoneal metastases. This is due to improved safety and favorable oncologic outcomes, including curative potential. CRS/HIPEC has a steep learning curve and requires familiarity with peritonectomy procedures. This review will outline the technical aspects and learning curve of CRS/HIPEC., (© 2020 Wiley Periodicals LLC.)

Free-living amoebae (FLA) are ubiquitous and many isolates have been shown to be infected with amoeba-resisting bacteria, as the example of Acanthamoeba and Legionella interaction. Due to the high environmental prevalence of Acanthamoeba. in the Castilian Plateau (Spain), the aims of this work were to investigate the occurrence of Acanthamoeba and other FLA in water from several sampling points from four Drinking Water Treatment Plants (DWTP) and to investigate the presence of Legionella spp. and other amoeba-resisting bacteria in biofilms in raw and finished water, taking into account that no legislation exists for this protozoa control. Acanthamoeba was detected at different sampling points, and sand filters seemed to contribute to amoebic enrichment. After ozonation, a temporary decrease in viable amoebae was observed. The genotypes detected were T3, T4, and T5, revealing the first report of genotype T5 in waters from this region. Moreover, Balamuthia mandrillaris, Vermamoeba vermiformis and Paravahlkampfia sp. were detected. Regarding Legionella, PCR detection in raw and finished water was higher than by agar culture, but even higher after Acanthamoeba co-culture. Also, Legionella's presence was higher in raw water than in finished water. The decrease of free Legionella observed from raw (27.5%, by PCR) to finished water (3.4% by PCR) contrasted with the increase of Legionella-infected FLA from raw (30.7%) to finished water (52%). At biofilm, free Legionella was not detected, and the percentage of infected FLA was low (3.8%). Legionella species identified in these samples were L. drozanskii, L. donaldsonii and L. feeleii. Additionally, Acanthamoeba co-culture led to the isolation of Pseudomonas aeruginosa, P. stutzeri, P. fluorecens, Achromobacter xylosoxidans and Stenotrophomonas maltophilia. The highly disseminated presence of Acanthamoeba and the detection of amoeba-resisting bacteria inside amoebae highlight the importance of developing methods for controlling FLA in order to limit human pathogenic amoeba-resisting bacteria survival to the water purification processes., Competing Interests: Declaration of competing interest All authors declare no actual or potential conflict of interest including any financial, personal or other relationships with founding organizations., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Chemical nomenclature is perceived to be a closed topic. However, this work shows that the identification of polyanionic groups is still ambiguous and so is the nomenclature for some ternary compounds. Two examples, boron phosphate (BPO 4 ) and boron arsenate (BAsO 4 ), which were assigned to the large phosphate and arsenate families, respectively, nearly a century ago, are explored. The analyses show that these two compounds should be renamed phosphorus borate (PBO 4 ) and arsenic borate (AsBO 4 ). Beyond epistemology, this has pleasing consequences at several levels for the predictive character of chemistry. It paves the way for future work on the possible syntheses of SbBO 4 and BiBO 4 , and it also renders previous structure field maps completely predictive, allowing us to foresee the structure and phase transitions of NbBO 4 and TaBO 4 . Overall, this work demonstrates that quantum mechanics calculations can contribute to the improvement of current chemical nomenclature. Such revisitation is necessary to classify compounds and understand their properties, leading to the main final aim of a chemist: predicting new compounds, their structures and their transformations.

Background: The malignant mechanisms that control the development of cutaneous T-cell lymphoma (CTCL) are beginning to be identified. Recent evidence suggests that disturbances in specific intracellular signalling pathways, such as RAS-mitogen-activated protein kinase, T-cell receptor (TCR)-phospholipase C gamma 1 (PLCG1)-nuclear factor of activated T cells (NFAT) and Janus kinase (JAK)-signal transducer and activator of transcription (STAT), may play an essential role in the pathogenesis of CTCL., Objectives: To investigate the mechanisms controlling disease development and progression in mycosis fungoides (MF), the most common form of CTCL., Methods: We collected 100 samples that were submitted for diagnosis of, or a second opinion regarding, MF between 2001 and 2018, 80% of which were in the early clinical stages of the disease. Formalin-fixed paraffin-embedded tissues were used for histological review and to measure the expression by immunohistochemistry of surrogate markers of activation of the TCR-PLCG1-NFAT, JAK-STAT and NF-κB pathways. Folliculotropism and large-cell transformation were also examined., Results: NFAT and nuclear factor kappa B (NF-κB) markers showed a comparable activation status in early and advanced stages, while STAT3 activation was more frequent in advanced stages and was associated with large-cell transformation. Consistently with this observation, STAT3 activation occurred in parallel with MF progression in two initially MF-negative cases. A significant association of NFAT with NF-κB markers was also found, reflecting a common mechanism of activation in the two pathways. Genomic studies identified nine mutations in seven genes known to play a potential role in tumorigenesis in T-cell leukaemia/lymphoma, including PLCG1, JAK3 and STAT3, which underlies the activation of these key cell-survival pathways. A higher mutational allele frequency was detected in advanced stages., Conclusions: Our results show that STAT3 is activated in advanced cases and is associated with large-cell transformation, while the activation of NFAT and NF-κB is maintained throughout the disease. These findings could have important diagnostic and therapeutic implications. What's already known about this topic? Mycosis fungoides is characterized by a clonal expansion of T cells in the skin. The mechanisms controlling disease development and progression are not fully understood. What does this study add? An association of the nuclear factor of activated T cells and nuclear factor kappa B pathways was found, which could reflect a common mechanism of activation. These pathways were activated in early and advanced stages at the same level. Signal transducer and activator of transcription 3 activation was associated with large-cell transformation and was more frequent in advanced stages. A genomic analysis of cutaneous T-cell lymphoma-associated genes was performed. Nine mutations were detected. What is the translational message? These results could have important implications for the treatment of MF in the near future., (© 2019 British Association of Dermatologists.)

The genus Legionella comprises more than 60 species, and about half are associated with infection. Legionella pneumophila is the most commonly associated with these infections and by far the most studied, but L. non- pneumophila species, such as L. feeleii , L. anisa , etc., may also present clinical importance. Free-living amoebae are their preferred environmental host, where these bacteria not only survive but also succeed in multiplying, and this relationship can lead to an increase in bacterial virulence. The goal of this study was to evaluate the alterations of Legionella pathogenicity due to its interaction with Acanthamoeba . For this, the expression of protein effectors SdhA, LegK2, and SidK were evaluated in L. pneumophila and L. feeleii , before and after infecting Acanthamoeba . Additionally, the host response was evaluated by measuring the production of IL-6, IL-8, and IFN-γ in infected macrophages. Regarding the virulence factors, an increase in SdhA expression was observed after these bacteria infected Acanthamoeba , with a higher increase in the macrophage cultures infected with L. feeleii . Also, an increase in the expression of LegK2 was observed after infecting Acanthamoeba , but it was more intense in the cultures infected with L. pneumophila . With regard to SidK, it was increased in L. feeleii after infecting Acanthamoeba , however the same effect was not observed for L. pneumophila . In cytokine production, the effect on IL-6 and IL-8 was similar for both cytokines, increasing their concentration, but higher production was observed in the cultures infected with L. feeleii , even though it demonstrated slightly lower production with the inoculum obtained from Acanthamoeba . Concerning IFN-γ, induction was observed in both species but higher in the infection by L. pneumophila . Nevertheless, it is not known if this induction is enough to promote an efficient immune response against either L. pneumophila or L. feeleii . Altogether, these alterations seem to increase L. feeleii virulence after infecting Acanthamoeba . However, this increase does not seem to turn L. feeleii as virulent as L. pneumophila . More studies are necessary to understand the aspects influenced in these bacteria by their interaction with Acanthamoeba and, thus, identify targets to be used in future therapeutic approaches.

Background: The use of contact lenses has increased in recent years as has the incidence of Dry Eye Syndrome, partly due to their use. Artificial tears are the most common treatment option. Since these changes can facilitate Acanthamoeba infection, the present study has been designed to evaluate the effect of three artificial tears treatments in the viability of Acanthamoeba genotype T4 trophozoites. Optava Fusion™, Oculotect®, and Artelac® Splash were selected due to their formulation., Methods: Viability was assessed using two staining methods, Trypan Blue stain and CTC stain at different time intervals (2, 4, 6, 8 and 24 h). Trypan Blue viability was obtained by manual count with light microscopy while the CTC stain was determined using flow cytometry., Results: Trypan Blue staining results demonstrated a decrease in viability for Optava Fusion™ and Artelac® Splash during the first 4 h of incubation. After, this effect seems to lose strength. In the case of Oculotect®, complete cell death was observed after 2 h. Using flow cytometry analysis, Optava Fusion™ and Oculotect® exhibited the same effect observed with Trypan Blue staining. However, Artelac® Splash revealed decreasing cell respiratory activity after four hours, with no damage to the cell membrane., Conclusions: The present study uses, for the first time, CTC stain analyzed by flow cytometry to establish Acanthamoeba viability demonstrating its usefulness and complementarity with the traditional stain, Trypan Blue. Artelac® Splash, with no preservatives, and Optava Fusion TM, with Purite®, have not shown any useful amoebicidal activity. On the contrary, promising results presented by Ocultect®, with BAK, open up a new possibility for Acanthamoeba keratitis prophylaxis and treatment although in vivo studies should be carried out.

We evaluated the performance of the MinION DNA sequencer in-flight on the International Space Station (ISS), and benchmarked its performance off-Earth against the MinION, Illumina MiSeq, and PacBio RS II sequencing platforms in terrestrial laboratories. Samples contained equimolar mixtures of genomic DNA from lambda bacteriophage, Escherichia coli (strain K12, MG1655) and Mus musculus (female BALB/c mouse). Nine sequencing runs were performed aboard the ISS over a 6-month period, yielding a total of 276,882 reads with no apparent decrease in performance over time. From sequence data collected aboard the ISS, we constructed directed assemblies of the ~4.6 Mb E. coli genome, ~48.5 kb lambda genome, and a representative M. musculus sequence (the ~16.3 kb mitochondrial genome), at 100%, 100%, and 96.7% consensus pairwise identity, respectively; de novo assembly of the E. coli genome from raw reads yielded a single contig comprising 99.9% of the genome at 98.6% consensus pairwise identity. Simulated real-time analyses of in-flight sequence data using an automated bioinformatic pipeline and laptop-based genomic assembly demonstrated the feasibility of sequencing analysis and microbial identification aboard the ISS. These findings illustrate the potential for sequencing applications including disease diagnosis, environmental monitoring, and elucidating the molecular basis for how organisms respond to spaceflight.

Background: Microsporidia are intracellular obligate parasites traditionally associated with immunosuppressed patients; their detection in immunocompetent patients has increased, highlighting their possible importance as emerging pathogens. Detection of spores in stools, urine, body fluids and tissues is difficult and immunological techniques such as immunofluorescence have proved to be a useful and reliable tool in the diagnosis of human microsporidiosis. For this reason, we have produced and characterized monoclonal antibodies (MAbs) specific for Encephalitozoon intestinalis (the second most frequent microsporidian infecting humans), and other Encephalitozoon species, that can be used in different diagnostic techniques., Results: Seven MAbs were selected in accordance with their optical density (OD). Four (4C4, 2C2, 2E5 and 2H2) were isotype IgG2a; two (3A5 and 3C9) isotype IgG3, and one Mab, 1D7, IgM isotype. The selected monoclonal antibody-secreting hybridomas were characterized by indirect immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA), Western blot, immunoelectron microscopy (Immunogold) and in vitro cultures. The study by IFAT showed different behavior depending on the MAbs studied. The MAbs 4C4, 2C2, 2E5 and 2H2 showed reactivity against epitopes in the wall of the spore (exospore and endospore) epitopes located in Encephalitozoon sp. spores, whereas the MAbs 3A5, 1D7 and 3C9 showed reactivity against internal epitopes (cytoplasmic contents or sporoplasm) of E. intestinalis spores. All MAbs recognized the developing parasites in the in vitro cultures of E. intestinalis. Additionally, 59 formalin-fixed stool samples that had been previously analyzed were screened, with 26 (44%) presenting microsporidian spores (18 samples with E. intestinalis and 8 samples with Enterocytozoon bieneusi). Detection of microsporidian spores by microscopy was performed using Calcofluor stain, Modified Trichrome, Quick-Hot Gram Chromotrope, as well as IFAT using MAbs 4C4, 2C2, 2E5 and 2H2. The 4 MAbs tested clearly recognized the larger spores corresponding to E. intestinalis, but showed no reactivity with Enterocytozoon bieneusi spores. The mass spectrometry and proteomic study revealed that the Mabs 4C4, 2C2, 2E5 and 2H2 recognized the Spore Wall Protein 1 (SWP1) as the antigenic target., Conclusions: The IFAT-positive MAbs exhibited excellent reactivity against spores and developmental stages, permitting their use in human and animal diagnosis. The epitopes recognized (exospore, endospore and cytoplasmic contents) by the different MAbs developed need further study, and may reveal potential targets for vaccine development, immunotherapy and chemotherapy.

This paper analyzes the structural, energetic and mechanical properties of carbon dioxide hydrate clathrates calculated using finite cluster and periodic ab initio density-functional theory methodologies. Intermolecular interactions are described by the exchange-hole dipole moment method. The stability, gas saturation energetics, guest-host interactions, cage deformations, vibrational frequencies, and equation of state parameters for the low-pressure sI cubic phase of the CO₂@H₂O clathrate hydrate are presented. Our results reveal that: (i) the gas saturation process energetically favors complete filling; (ii) carbon dioxide molecules prefer to occupy the larger of the two cages in the sI structure; (iii) blue shifts occur in both the symmetric and antisymmetric stretching frequencies of CO₂ upon encapsulation; and (iv) free rotation of guest molecules is restricted to a plane parallel to the hexagonal faces of the large cages. In addition, we calculate the librational frequency of the hindered rotation of the guest molecule in the plane perpendicular to the hexagonal faces. Our calculated spectroscopic data can be used as signatures for the detection of clathrate hydrates in planetary environments.

[This corrects the article DOI: 10.1371/journal.pone.0159726.].

Legionnaires' disease is a severe form of pneumonia, with worldwide relevance, caused by Legionella spp. Approximately 90% of all cases of legionellosis are caused by Legionella pneumophila, but other species can also be responsible for this infection. These bacteria are transmitted by inhalation of aerosols or aspiration of contaminated water. In Spain, environmental studies have demonstrated the presence of Legionella non-pneumophila species in drinking water treatment plants and water distribution networks. Aware that this evidence indicates a risk factor and the lack of routine assays designed to detect simultaneously diverse Legionella species, we analyzed 210 urine samples from patients presenting clinical manifestations of pneumonia using a semi-nested PCR for partial amplification of the 16S rDNA gene of Legionella and a diagnostic method used in hospitals for Legionella antigen detection. In this study, we detected a total of 15 cases of legionellosis (7.1%) and the first case of Legionnaires' disease caused by L. anisa in Spain. While the conventional method used in hospitals could only detect four cases (1.9%) produced by L. pneumophila serogroup 1, using PCR, the following species were identified: Legionella spp. (10/15), L. pneumophila (4/15) and L. anisa (1/15). These results suggest the need to change hospital diagnostic strategies regarding the identification of Legionella species associated with this disease. Therefore, the detection of Legionella DNA by PCR in urine samples seems to be a suitable alternative method for a sensitive, accurate and rapid diagnosis of Legionella pneumonia, caused by L. pneumophila and also for L. non-pneumophila species.

A thorough investigation of pressure effects on the structural properties of crystalline cesium uranyl chloride was performed by means of first-principles calculations within the density functional theory framework. Total energies, equilibrium geometries and vibrational frequencies were computed at selected pressures up to 50 GPa. Zero pressure results present good agreement with available experimental and theoretical data. Our calculated equation of state parameters reveal that Cs2UO2Cl4 is a high compressible material, similar to other ionic compounds with cesium cations, and displays a structural anisotropic behavior guided by the uranyl moiety. An unexpected variation of the U-O bond length, dUO, is detected as pressure is applied. It leads to a dUO-stretching frequency relationship that cannot be described by the traditional Badger's rule. Interestingly enough, it can be explained in terms of a change in the main factor controlling dUO. At low pressure, the charge transferred to the uranyl cation induces an increase of the bond length and a red shift of the stretching frequencies, whereas it is the mechanical effect of the applied pressure above 10 GPa that is the dominant factor that leads to a shortening of dUO and a blue shift of the stretching frequencies.

Molecular characterization of Cryptosporidium spp. and Enterocytozoon bieneusi has improved our understanding of the transmission of both organisms in humans. In this study, to infer possible infection sources, Cryptosporidium spp. and E. bieneusi in fecal specimens from 90 HIV-infected patients attending antiretroviral clinics in Lagos, Nigeria were detected and genotyped by PCR and DNA sequencing. Cryptosporidium spp. and E. bieneusi were identified in four and five patients, respectively, including the occurrence of subtype IeA11T3G3 of Cryptosporidium hominis in two patients, subtype IIcA5G3k of Cryptosporidium parvum in one patient, and Type IV of E. bieneusi in four patients. Among the remaining positive patients, one had mixed infection of Cryptosporidium meleagridis and C. hominis and one had mixed E. bieneusi genotypes. These data highlight a possible difference in major transmission routes (anthroponotic vs. zoonotic) between Cryptosporidium spp. and E. bieneusi in HIV+ patients in the study area., (© 2015 The Author(s) Journal of Eukaryotic Microbiology © 2015 International Society of Protistologists.)