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Leishmania are kinetoplastid pathogens that cause leishmaniasis, a debilitating and potentially life-threatening infection if untreated. Unusually, Leishmania regulate their gene expression largely post-transcriptionally due to the arrangement of their coding genes into polycistronic transcription units that may contain 100s of functionally unrelated genes. Yet, Leishmania are capable of rapid and responsive changes in gene expression to challenging environments, often instead correlating with dynamic changes in their genome composition, ranging from chromosome and gene copy number variations to the generation of extrachromosomal DNA and the accumulation of point mutations. Typically, such events indicate genome instability in other eukaryotes, coinciding with genetic abnormalities, but for Leishmania, exploiting these products of genome instability can provide selectable substrates to catalyse necessary gene expression changes by modifying gene copy number. Unorthodox DNA replication, DNA repair, replication stress factors and DNA repeats are recognised in Leishmania as contributors to this intrinsic instability, but how Leishmania regulate genome plasticity to enhance fitness whilst limiting toxic under- or over-expression of co-amplified and co-transcribed genes is unclear. Herein, we focus on fresh, and detailed insights that improve our understanding of genome plasticity in Leishmania. Furthermore, we discuss emerging models and factors that potentially circumvent regulatory issues arising from polycistronic transcription. Lastly, we highlight key gaps in our understanding of Leishmania genome plasticity and discuss future studies to define, in higher resolution, these complex regulatory interactions.

The intake of microplastics by freshwater fishes has been scarcely studied, and it is not yet clear whether the amount of particles these vertebrates ingest is associated with their feeding strategies. Hence, this study is focused on the suckermouth armored catfish (Pterygoplichthys spp.) under the hypothesis that, due to its detritivorous habits and demersal distribution, it may show evidence of the presence of microplastics in the sediments of a lake located in the Metropolitan Area of Villahermosa. A total of 21 organisms contained plastic microfibers throughout their digestive systems. These were treated with three solutions (hydrogen peroxide, potassium hydroxide, and sodium hypochlorite) to separate the microplastics. A total of 147 plastic particles of two categories were collected: microfibers (92%) and fragments (8%). Considering these results, as well as evidence in the literature, these species may constitute a global indicator of the incorporation into the trophic chain of microfibers deposited in the sediments of freshwater ecosystems.

The Neotropical region hosts 4225 freshwater fish species, ranking first among the world's most diverse regions for freshwater fishes. Our NEOTROPICAL FRESHWATER FISHES data set is the first to produce a large-scale Neotropical freshwater fish inventory, covering the entire Neotropical region from Mexico and the Caribbean in the north to the southern limits in Argentina, Paraguay, Chile, and Uruguay. We compiled 185,787 distribution records, with unique georeferenced coordinates, for the 4225 species, represented by occurrence and abundance data. The number of species for the most numerous orders are as follows: Characiformes (1289), Siluriformes (1384), Cichliformes (354), Cyprinodontiformes (245), and Gymnotiformes (135). The most recorded species was the characid Astyanax fasciatus (4696 records). We registered 116,802 distribution records for native species, compared to 1802 distribution records for nonnative species. The main aim of the NEOTROPICAL FRESHWATER FISHES data set was to make these occurrence and abundance data accessible for international researchers to develop ecological and macroecological studies, from local to regional scales, with focal fish species, families, or orders. We anticipate that the NEOTROPICAL FRESHWATER FISHES data set will be valuable for studies on a wide range of ecological processes, such as trophic cascades, fishery pressure, the effects of habitat loss and fragmentation, and the impacts of species invasion and climate change. There are no copyright restrictions on the data, and please cite this data paper when using the data in publications.

Along with structural and non-structural proteins, SARS-CoV-2, SARS-CoV, and MERS-CoV can also express accessory proteins. During the past few years, there have been only a few studies focusing on this set of proteins. Despite available data on these proteins, there are still a lot of questions on the functions of these proteins during infection that must be answered. With these three betacoronaviruses causing outbreaks in humans during the past few years, the need for a thorough understanding of the roles of these proteins is becoming more important and relevant. This review provides a survey of the existing knowledge on the roles of these proteins during infection. In addition to current evidence, a more comprehensive view of the functions of these proteins is presented together with their potential as therapeutic targets, which were determined by using different bioinformatics platforms. This information may help test effective therapeutic regimens against these viruses and in preparing for future pandemics.

International audience; Assembly bowing in a PWR core can be a serious problem for managing the power during normal operation or for periodical maintenance. One of the causes of this phenomenon is the transverse flow in the core, generated by the non-uniformity of the axial flow. Modelling the mechanical behaviour of a PWR core is made difficult by the complex geometry and the numerous friction points that lead to non-linearities. A simple but efficient way to deal with these issues is the porous medium model proposed by Ricciardi et al. (2009). In this model, the equations used at the fluid–structure interface require empirical parameters such as the added mass, or axial and normal drag coefficients. Using a new experimental setup at CEA, Eudore, which hosts 3 half-scale fuel assemblies in a line, the forces acting on the assembly by a non-uniform flow profile were measured. An analytical model to retrieve the normal drag coefficient was then proposed. This coefficient is then used in FSCORE, a numerical software based on the porous medium approach. The experimental and numerical results are presented and show good agreement.

Background The data about the influence of chemotherapy on right ventricular (RV) function and mechanics are scarce and conflicting. Due to the special geometric structure and unique wall motion of the RV, routine techniques for echocardiographic assessment of the RV has challenges. However, 3-dimensional echocardiography (3DE) tracks changes in RV volumes in real time, including the inflow and outflow tracts and the apex throughout the cardiac cycle. 3DE evaluation of RV function and morphology has been compared to cardiac magnetic resonance and is accurate, feasible, and reproducible Methods This study conducted was a retrospective cohort study which included 75 breast cancer patients who underwent chemotherapy at a tertiary hospital from January 2019 to May 2021 to determine clinical outcomes of patients among those who developed an abnormal right ventricular longitudinal free wall strain and global longitudinal strain in terms of all-cause mortality, development of overt right ventricular systolic dysfunction, and adverse cardiovascular events. Results There was a striking statistically significant difference in terms of right ventricular ejection fraction among those who developed an abnormal right ventricular global longitudinal strain at 6 and 12 months post-chemotherapy with a p-value of 0.001 and 0.000 respectively. The difference between the 2 groups of patients in terms of right ventricular global longitudinal strain 6 and 12 months after chemotherapy was also statistically significant (p=0.000). As for the left ventricle, the LV ejection fraction and global longitudinal strain were also statistically significantly different at 12 months post-chemotherapy. Among those who developed an abnormal RV global longitudinal strain, the LV global longitudinal strain was also subsequently lower compared to the normal subgroup at 12 months post-treatment. Conclusion Although the sample size was small, the development of abnormal RV GLS post-chemotherapy was generally associated with subsequent development of overt heart failure, increased all-cause mortality, and CV death. The use of echocardiographic RV GLS surveillance to guide chemotherapy is crucial to enhance clinical outcomes and survival. Funding Acknowledgement Type of funding sources: None.

Opuntia species grow in arid and semiarid lands, where water for irrigation is scarce. However, supplemental irrigation can be a feasible strategy for commercial cactus pear orchards. From 2018 to 2020, a commercial cactus pear orchard was managed to validate the effect of supplemental irrigation on fruit yield, crop water use efficiency, fruit quality, and storability of ‘Roja Lisa’ cactus pear grown in the semiarid region of Mexico. The irrigation treatments were no irrigation and supplemental irrigation, with four replications. Crop water use was less and, therefore, water productivity greater in non-irrigated plants than in plants with supplemental irrigation. Mean fruit yield, mean fruit mass, and proportion of commercial fruit increased with supplemental irrigation. These differences were more pronounced in growing seasons with less rainfall. Fruit quality at harvest or after room temperature or cold storage was examined. Fruit mass loss rate was reduced in fruit receiving supplemental irrigation in both storage conditions. In addition, supplemental irrigation was consistent with water savings and food security programs in marginal areas: this irrigation strategy improved both pre- and postharvest some quality components of cactus pear fruit. Therefore, this irrigation strategy is suggested for cactus pear growers, depending on the availability of water for irrigation.

Until 2015, loss-of-function studies to elucidate protein function in Leishmania relied on gene disruption through homologous recombination. Then, the CRISPR/Cas9 revolution reached these protozoan parasites allowing efficient genome editing with one round of transfection. In addition, the development of LeishGEdit, a PCR-based toolkit for generating knockouts and tagged lines using CRISPR/Cas9, allowed a more straightforward and effective genome editing. In this system, the plasmid pTB007 is delivered to Leishmania for episomal expression or integration in the β-tubulin locus and for the stable expression of T7 RNA polymerase and Cas9. In South America, and especially in Brazil, Leishmania (Viannia) braziliensis is the most frequent etiological agent of tegumentary leishmaniasis. The L. braziliensis β-tubulin locus presents significant sequence divergence in comparison with Leishmania major, which precludes the efficient integration of pTB007 and the stable expression of Cas9. To overcome this limitation, the L. major β-tubulin sequences, present in the pTB007, were replaced by a Leishmania (Viannia) β-tubulin conserved sequence generating the pTB007_Viannia plasmid. This modification allowed the successful integration of the pTB007_Viannia cassette in the L. braziliensis M2903 genome, and in silico predictions suggest that this can also be achieved in other Viannia species. The activity of Cas9 was evaluated by knocking out the flagellar protein PF16, which caused a phenotype of immobility in these transfectants. Endogenous PF16 was also successfully tagged with mNeonGreen, and an in-locus complementation strategy was employed to return a C-terminally tagged copy of the PF16 gene to the original locus, which resulted in the recovery of swimming capacity. The modified plasmid pTB007_Viannia allowed the integration and stable expression of both T7 RNA polymerase and Cas9 in L. braziliensis and provided an important tool for the study of the biology of this parasite.

Antecedentes: La interconexion lateral en el sistema rio-llanura de inundacion Maluco fue reducida parcialmente por una carretera construida sobre una barrera natural. Objetivo: Determinar si las concentraciones de clorofila-a (Cla-a) y nutrientes contrastan en diferentes niveles de agua (NA) durante un ano hidrologico en el humedal fluvial. Metodos: El NA, Cla-a y nutrientes (NO2-, NO3-, NH4+ y PT) se midieron simultaneamente seis veces en dos estaciones durante dos anos. Las diferencias esperadas de las concentraciones de Cla-a y nutrientes, relacionadas a la fluctuacion intra-anual del NA, fueron analizadas por medio de tecnicas multivariadas. La hipotesis sostiene que no se detectaran diferencias intra anuales de Cla-a y nutrientes debido a la reduccion de la interconexion lateral por el efecto fisico del terraplen. Resultados: Ninguna variable fue descartada con el ACP. La distribucion intra-anual de las muestras resulto similar segun la rutina de perfiles de similitud. Coincidentemente: 1) mas del 54% de los valores de Cla-a y PT indicaron condiciones hipereutroficas durante todo el ano y 2) ninguno de los valores de WL sobrepaso la altura del terraplen (4.5 msnm). Ademas, las concentraciones maximas de Cla-a se registraron en los WL minimos. Conclusiones: La similitud intra-anual de la Cla-a y los nutrientes inorganicos se puede vincular a que la llanura de inundacion estuvo desconectada del rio Grijalva, ya que la inundacion no sobrepaso la altura del terraplen durante todo el tiempo de muestreo. La tendencia intra-anual independiente al tiempo de la persistencia de las altas concentraciones de Cla-a y PT en el sistema rio llanura de inundacion abre la oportunidad de estimar el caudal de entrada minimo requerido para restaurar los procesos biogeoquimicos en un estudio a largo plazo.

In Brazil, cutaneous leishmaniasis is caused predominantly by L. (V.) braziliensis. The few therapeutic drugs available exhibit several limitations, mainly related to drug toxicity and reduced efficacy in some regions. Miltefosine (MF), the only oral drug available for leishmaniasis treatment, is not widely available and has not yet been approved for human use in Brazil. Our group previously reported the existence of differential susceptibility among L. (V.) braziliensis clinical isolates. In this work, we further characterized three of these isolates of L. (V.) braziliensis chosen because they exhibited the lowest and the highest MF half maximal inhibitory concentrations and were therefore considered less tolerant or more tolerant, respectively. Uptake of MF, and also of phosphocholine, were found to be significantly different in more tolerant parasites compared to the less sensitive isolate, which raised the hypothesis of differences in the MF transport complex Miltefosine Transporter (MT)-Ros3. Although some polymorphisms in those genes were found, they did not correlate with the drug susceptibility phenotype. Drug efflux and compartmentalization were similar in the isolates tested, and amphotericin B susceptibility was retained in MF tolerant parasites, suggesting that increased fitness was also not the basis of observed differences. Transcriptomic analysis revealed that Ros3 mRNA levels were upregulated in the sensitive strain compared to the tolerant ones. Increased mRNA abundance in more tolerant isolates was validated by quantitative PCR. Our results suggest that differential gene expression of the MT transporter complex is the basis of the differential susceptibility in these unselected, naturally occurring parasites., Graphical abstract Image 1, Highlights • Brazilian L. (V.) braziliensis isolates vary in mitefosine susceptibility. • Diminished drug internalization was observed in more tolerant isolates. • Drug susceptibility did not correlate with SNPs in MT-Ros3 genes. • Drug efflux and compartmentalization were similar in the isolates tested. • Increased drug sensitivity is accompanied by Ros3 mRNA upregulation.

In trypanosomatids, regulation of gene expression occurs mainly at the posttranscriptional level, and RNA-binding proteins (RBPs) are key players in determining the fates of transcripts. RBPs are major targets of protein arginine methyltransferases (PRMTs), which posttranslationally regulate the RNA-binding capacity and other macromolecular interactions of RBPs by transferring methyl groups to protein arginine residues. Herein, we present the results of a study that functionally characterized the five predicted PRMTs inLeishmania braziliensisby gene knockout and endogenous protein HA tagging using CRISPR/Cas9 gene editing. We report that arginine methylation profiles vary amongLeishmaniaspecies and that target protein methylation changes across differentL. braziliensislife cycle stages, with higher PRMT expression in the promastigote stages than in the axenic amastigote stage. Knockout of some of theL. braziliensisPRMTs led to significant changes in global arginine methylation patterns without affecting promastigote axenic growth. Deletion of either PRMT1 or PRMT3 disrupted most type I PRMT activity, resulting in a global increase in monomethyl arginine (MMA) levels, which is mainly catalyzed by PRMT7. Putative targets and/or PRMT-interacting proteins were identified by coimmunoprecipitation using HA-tagged PRMTs, revealing a network of target RBPs and suggesting functional interactions between them and a relevant participation in epigenetic control of gene expression. Finally, we demonstrate thatL. braziliensisPRMT1 and PRMT5 are required for efficient macrophage infectionin vitro, and that in the absence of PRMT1 and PRMT5, axenic amastigote proliferation is impaired. The results indicate that arginine methylation is modulated across life cycle stages inL. braziliensisand show possible functional overlap and cooperation among the different PRMTs in targeting proteins. Overall, our data suggest important regulatory roles of these proteins throughout theL. braziliensislife cycle, showing that arginine methylation is important for parasite-host cell interactions.

Se presenta una lista sistemática de peces recolectados durante 2014 y 2015 en la Reserva de la Biosfera Pantanos de Centla con un incremento del 25% de su diversidad principalmente por la presencia de peces diádromos. El inventario comprende 75 especies, que representa el 37% de la ictiofauna en la cuenca Grijalva-Usumacinta, de las cuales cinco especies fueron no nativas invasoras donde el bagre armado (Pterygoplichthys spp.) fue el que presentó la mayor frecuencia de captura. Se identificaron 48 especies migratorias (38 diádromas y 10 potamodromas). La dominancia de peces diádromos y su presencia en sistemas de lagunas y fluviales nos indica el mantenimiento de la interconexión hidráulica entre ríos y lagunas. En los muestreos, se observó la ausencia de siete especies dulceacuícolas que fueron comunes en publicaciones previas, lo anterior puede ser debido a la disminución de hábitats como consecuencia de las actividades humanas y la introducción de especies invasoras. En este estudio se resalta la necesidad de mayores estudios enfocados a la biología de varias especies, con el objetivo de aclarar su condición de potamodromia y comprender la migración lateral que algunas especies presentan durante su ontogenia para satisfacer sus necesidades eco-fisiológicas.

Leishmania major is the main causative agent of cutaneous leishmaniasis in the Old World. In Leishmania parasites, the lack of transcriptional control is mostly compensated by post-transcriptional mechanisms. Methylation of arginine is a conserved post-translational modification executed by Protein Arginine Methyltransferase (PRMTs). The genome from L. major encodes five PRMT homologs, including the cytosolic protein associated with several RNA-binding proteins, LmjPRMT7. It has been previously reported that LmjPRMT7 could impact parasite infectivity. In addition, a more recent work has clearly shown the importance of LmjPRMT7 in RNA-binding capacity and protein stability of methylation targets, demonstrating the role of this enzyme as an important epigenetic regulator of mRNA metabolism. In this study, we unveil the impact of PRMT7-mediated methylation on parasite development and virulence. Our data reveals that higher levels of LmjPRMT7 can impair parasite pathogenicity, and that deletion of this enzyme rescues the pathogenic phenotype of an attenuated strain of L. major. Interestingly, lesion formation caused by LmjPRMT7 knockout parasites is associated with an exacerbated inflammatory reaction in the tissue correlated with an excessive neutrophil recruitment. Moreover, the absence of LmjPRMT7 also impairs parasite development within the sand fly vector Phlebotomus duboscqi. Finally, a transcriptome analysis shed light onto possible genes affected by depletion of this enzyme. Taken together, this study highlights how post-transcriptional regulation can affect different aspects of the parasite biology., Author summary Understanding the genetics of Leishmania, a protozoan parasite causing leishmaniasis, is relevant for understanding fundamental questions on the pathogen’s biology and its interaction with hosts. We explore mechanisms used by Leishmania to promptly adapt to different hosts investigating the control of gene expression occurring at the post-transcriptional level in the parasite. Methylation of arginine performed by Protein Arginine Methyltransferase (PRMTs), among other post-translational modifications, may alter the function and interactions of target proteins, some of them are RNA binding proteins, known regulators of gene expression. In this study, we unveil the impact of PRMT7 on parasite development and pathogenicity. In addition to a negative correlation between the levels of LmjPRMT7 and parasite pathogenicity, we observed an impairment of the parasite development in the sand fly vector. Remarkably, despite a severe lesion development in mice, we observed no differences in parasite burden between infections with the pathogenic LmjPRMT7 knockout parasite or the attenuated parental line. Instead, the severe pathology observed is associated with an exacerbated inflammatory response correlated with excessive neutrophil recruitment.

The availability of Trypanosomatid genomic data in public databases has opened myriad experimental possibilities that have contributed to a more comprehensive understanding of the biology of these parasites and their interactions with hosts. In this review, after brief remarks on the history of the Trypanosoma cruzi and Leishmania genome initiatives, we present an overview of the relevant contributions of genomics, transcriptomics and functional genomics, discussing the primary obstacles, challenges, relevant achievements and future perspectives of these technologies.

A large number of eukaryotic proteins are processed by single or combinatorial post-translational covalent modifications that may alter their activity, interactions and fate. The set of modifications of each protein may be considered a “regulatory code”. Among the PTMs, arginine methylation, catalyzed by protein arginine methyltransferases (PRMTs), can affect how a protein interacts with other macromolecules such as nucleic acids or other proteins. In fact, many RNA-binding (RBPs) proteins are targets of PRMTs. The methylation status of RBPs may affect the expression of their bound RNAs and impact a diverse range of physiological and pathological cellular processes. Unlike most eukaryotes, Kinetoplastids have overwhelmingly intronless genes that are arranged within polycistronic units from which mature mRNAs are generated by trans-splicing. Gene expression in these organisms is thus highly dependent on post-transcriptional control, and therefore on the action of RBPs. These genetic features make trypanosomatids excellent models for the study of post-transcriptional regulation of gene expression. The roles of PRMTs in controlling the activity of RBPs in pathogenic kinetoplastids have now been studied for close to 2 decades with important advances achieved in recent years. These include the finding that about 10% of the Trypanosoma brucei proteome carries arginine methylation and that arginine methylation controls Leishmania:host interaction. Herein, we review how trypanosomatid PRMTs regulate the activity of RBPs, including by modulating interactions with RNA and/or protein complex formation, and discuss how this impacts cellular and biological processes. We further highlight unique structural features of trypanosomatid PRMTs and how it contributes to their singular functionality.

RNA binding proteins (RBPs) are the primary gene regulators in kinetoplastids as transcriptional control is nearly absent, making Leishmania an exceptional model for investigating methylation of non-histone substrates. Arginine methylation is an evolutionarily conserved protein modification catalyzed by Protein aRginine Methyl Transferases (PRMTs). The chromatin modifier PRMT7 is the only Type III PRMT found in higher eukaryotes and a restricted number of unicellular eukaryotes. In Leishmania major, PRMT7 is a cytoplasmic protein implicit in pathogenesis with unknown substrates. Using comparative methyl-SILAC proteomics for the first time in protozoa, we identified 40 putative targets, including 17 RBPs hypomethylated upon PRMT7 knockout. PRMT7 can modify Alba3 and RBP16 trans-regulators (mammalian RPP25 and YBX2 homologs, respectively) as direct substrates in vitro. The absence of PRMT7 levels in vivo selectively reduces Alba3 mRNA-binding capacity to specific target transcripts and can impact the relative stability of RBP16 in the cytoplasm. RNA immunoprecipitation analyses demonstrate PRMT7-dependent methylation promotes Alba3 association with select target transcripts and thus indirectly stabilizes mRNA of a known virulence factor, δ-amastin surface antigen. These results highlight a novel role for PRMT7-mediated arginine methylation of RBP substrates, suggesting a regulatory pathway controlling gene expression and virulence in Leishmania. This work introduces Leishmania PRMTs as epigenetic regulators of mRNA metabolism with mechanistic insight into the functional manipulation of RBPs by methylation.

The free-flowing Usumacinta River maintains an average annual water-level fluctuation of 6.7 m. This study evaluated relationships between 14 physicochemical and biological variables and key factors in four water-level conditions in a river–floodplain system. The analysis incorporated intra-annual variation in all variables, with each selected in accordance with multiple statistical and physicochemical criteria. Possible correlations were examined as a function of various physicochemical and biological factors at each water level. Within the study area, defined by a temporal gradient in principal component 1, the yearly river overflow above the bankfull stage is characterized by water with a low level of total suspended solids. Data on riverine wetlands suggest that seasonal changes are key to determining intra- and inter-annual chlorophyll-a levels and water clarity. However, opposite trends are observed for high and low water-level conditions. Nutrient enrichment cannot be taken as the key physicochemical factor of water level, under either water-level conditions, due to the lack of a temporal gradient in principal component 2 and the high biochemical variability of nitrate and orthophosphate levels. In conclusion, the hypothesis was accepted for the production of phytoplankton biomass and light attenuation at low and high water levels, as both were dependent on intra-annual changes. The increases in chlorophyll-a related to the minimal variability at the lowest water level open the opportunity to gauge this relationship as a possible environmental predictor for river–floodplain systems.

We present new records for Fleischmann’s Glassfrog, Hyalinobatrachium fleischmanni (Boettger, 1893), which extendthe distribution of this species into central and southwestern Ecuador and thus represent the southernmost knownlocalities. These new reports are based on specimens collected at Cerro de Hayas, province of Guayas, and Macul,province of Los Ríos. These new localities extend the known geographic range of H. fleischmanni by approximately250 km south of previous known occurrences.

Non-coding RNAs (ncRNAs) are regulatory elements present in a wide range of organisms, including trypanosomatids. ncRNAs transcribed from the untranslated regions (UTRs) of coding genes have been described in the transcriptomes of several eukaryotes, including Trypanosoma brucei. To uncover novel putative ncRNAs in two Leishmania species, we examined a L. major cDNA library and a L. donovani non-polysomal RNA library. Using a combination of computational analysis and experimental approaches, we classified 26 putative ncRNA in L. major, of these, 5 arising from intergenic regions and 21 from untranslated regions. In L. donovani, we classified 37 putative ncRNAs, of these, 7 arising from intergenic regions, and 30 from UTRs. Our results suggest, for the first time, that UTR-transcripts may be a common feature in the eukaryote Leishmania similarly to those previously shown in T. brucei and other eukaryotes.

Migratory birds are demonstrating changes in phenology linked to climate change. Understanding these changes requires connecting events that occur over the multiple regions occupied during their annual cycle. The Burrowing Owl (Athene cunicularia) is a species of concern in North America, with pronounced declines in regions of the Great Plains. Using a dataset that spanned 10 breeding sites from South Dakota to northern Mexico in various years during 1989–2017, we observed both advances and delays in nesting along with increasing variation in nest initiation dates. We examined the effects of a large-scale climate system (El Niño Southern Oscillation), drought, and local weather patterns throughout the annual cycle as potential predictors of early and late nesting. Moisture conditions during the winter and spring migratory period had the greatest influence on nest phenology. Years with more intense drought on winter and migratory grounds increased the probability of nests initiating late relative to early. Correspondingly, wet conditions were associated with an increased probability of early nest initiation. Drought likely has cascading ecological effects that negatively influence food abundance for Burrowing Owls, resulting in delays in the ability of individuals to meet energetic demands required for migration. How climate change will impact Burrowing Owl phenology is important considering a projected increase in the magnitude and frequency of drought and declining owl population trends.

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Background Prostaglandins (PG) are lipid mediators derived from arachidonic acid metabolism. They are involved in cellular processes such as inflammation and tissue homeostasis. PG production is not restricted to multicellular organisms. Trypanosomatids also synthesize several metabolites of arachidonic acid. Nevertheless, their biological role in these early-branching parasites and their role in host-parasite interaction are not well elucidated. Prostaglandin F2α synthase (PGF2S) has been observed in the Leishmania braziliensis secreted proteome and in L. donovani extracellular vesicles. Furthermore, we previously reported a positive correlation between L. braziliensis PGF2S (LbrPGF2S) expression and pathogenicity in mice. Methods LbrPGF2S gene expression and PGF2α synthesis in promastigotes were detected and quantified by western blotting and EIA assay kit, respectively. To investigate LbrPGF2S localization in amastigotes during bone marrow-derived macrophage infection, parasites expressing mCherry-LbrPGF2S were generated and followed by time-lapse imaging for 48 h post-infection. PGF2S homolog sequences from Leishmania and humans were analyzed in silico using ClustalW on Geneious v6 and EMBOSS Needle. Results Leishmania braziliensis promastigotes synthesize prostaglandin F2α in the presence of arachidonic acid, with peak production in the stationary growth phase under heat stress. LbrPGF2S is a cytoplasmic protein enriched in the secretory site of the parasite cell body, the flagellar pocket. It is an enzyme constitutively expressed throughout promastigote development, but overexpression of LbrPGF2S leads to an increase of infectivity in vitro. The data suggest that LbrPGF2S may be released from intracellular amastigotes into the cytoplasm of bone marrow-derived macrophages over a 48-hour infection period, using time-lapse microscopy and mCherry-PGF2S (mChPGF2S)-expressing parasites. Conclusions LbrPGF2S, a parasite-derived protein, is targeted to the host cell cytoplasm. The putative transfer of this enzyme, involved in pro-inflammatory lipid mediator synthesis, to the host cell suggests a potential role in host-parasite interaction and may partially explain the increased pathogenicity associated with overexpression of LbrPGF2S in L. braziliensis. Our data provide valuable insights to help understand the importance of parasite-derived lipid mediators in pathogenesis.

Leishmania RNA virus (LRV) is an important virulence factor associated with the development of mucocutaneous Leishmaniasis, a severe form of the disease. LRV-mediated disease exacerbation relies on TLR3 activation, but downstream mechanisms remain largely unexplored. Here, we combine human and mouse data to demonstrate that LRV triggers TLR3 and TRIF to induce type I IFN production, which induces autophagy. This process results in ATG5-mediated degradation of NLRP3 and ASC, thereby limiting NLRP3 inflammasome activation in macrophages. Consistent with the known restricting role of NLRP3 for Leishmania replication, the signaling pathway triggered by LRV results in increased parasite survival and disease progression. In support of this data, we find that lesions in patients infected with LRV+ Leishmania are associated with reduced inflammasome activation and the development of mucocutaneous disease. Our findings reveal the mechanisms triggered by LRV that contribute to the development of the debilitating mucocutaneous form of Leishmaniasis., NLRP3 activation by Leishmania parasites is critical to the outcome of the disease. Here the authors show that LRV, a virus infecting Leishmania strains associated with more severe human disease, enables the parasite to suppress the inflammasome by activating type 1 interferon through TLR3, which leads to autophagy-mediated NLRP3 degradation.

Adhesion of T. cruzi trypomastigotes to components of the extracellular matrix (ECM) is an important step in mammalian host cell invasion. We have recently described a significant increase in the tyrosine nitration levels of histones H2A and H4 when trypomastigotes are incubated with components of the ECM. In this work, we used chromatin immunoprecipitation (ChIP) with an anti-nitrotyrosine antibody followed by mass spectrometry to identify nitrated DNA binding proteins in T. cruzi and to detect alterations in nitration levels induced upon parasite incubation with the ECM. Histone H1, H2B, H2A and H3 were detected among the 9 most abundant nitrated DNA binding proteins using this proteomic approach. One nitrated tyrosine residue (Y29) was identified in Histone H2B in the MS/MS spectrum. In addition, we observed a significant increase in the nitration levels of histones H1, H2B, H2A and H4 upon parasite incubation with ECM. Finally, we used ChIP-Seq to map global changes in the DNA binding profile of nitrated proteins. We observed a significant change in the binding pattern of nitrated proteins to DNA after parasite incubation with ECM. This work provides the first global profile of nitrated DNA binding proteins in T. cruzi and additional evidence for modification in the nitration profile of histones upon parasite incubation with ECM. Our data also indicate that the parasite interaction with the ECM induces alterations in chromatin structure, possibly affecting nuclear functions., Author summary T. cruzi, the etiological agent of Chagas’ disease, interacts with ECM in order to invade cells and establish the infection in the mammalian host. Studying this interaction, our group has recently described changes in the nitration profile of histones upon incubation of the parasite with ECM. Herein we used a proteomic approach to identify nitrated DNA binding proteins in T. cruzi. We also used ChIP-Seq to investigate whether the contact of the parasite with host ECM is capable of promoting changes in the DNA binding pattern of nitrated proteins. We observed significant alterations in the nitration levels and DNA binding profile of T. cruzi histones upon parasite incubation with ECM.

Frameworks for limiting ecosystem exposure to excess nutrients and acidity require accurate and complete deposition budgets of reactive nitrogen (Nr). While much progress has been made in developing total Nr deposition budgets for the U.S., current budgets remain limited by key data and knowledge gaps. Analysis of National Atmospheric Deposition Program Total Deposition (NADP/TDep) data illustrates several aspects of current Nr deposition that motivate additional research. Averaged across the continental U.S., dry deposition contributes slightly more (55%) to total deposition than wet deposition and is the dominant process (> 90%) over broad areas of the Southwest and other arid regions of the West. Lack of dry deposition measurements imposes a reliance on models, resulting in a much higher degree of uncertainty relative to wet deposition which is routinely measured. As nitrogen oxide (NO(x)) emissions continue to decline, reduced forms of inorganic nitrogen (NH(x) = NH(3) + NH(4)(+)) now contribute > 50% of total Nr deposition over large areas of the U.S. Expanded monitoring and additional process-level research are needed to better understand NH(x) deposition, its contribution to total Nr deposition budgets, and the processes by which reduced N deposits to ecosystems. Urban and suburban areas are hotspots where routine monitoring of oxidized and reduced Nr deposition is needed. Finally, deposition budgets have incomplete information about the speciation of atmospheric nitrogen; monitoring networks do not capture important forms of Nr such as organic nitrogen. Building on these themes, we detail the state of the science of Nr deposition budgets in the U.S. and highlight research priorities to improve deposition budgets in terms of monitoring and flux measurements, leaf- to regional-scale modeling, source apportionment, and characterization of deposition trends and patterns.

In the last 30 years, significant advances in genetic manipulation tools along with complete genome and transcriptome sequencing have advanced our understanding of the biology of Leishmania parasites and their interplay with the sand fly and mammalian hosts. High-throughput sequencing in association with CRISPR/Cas9 have prepared the ground for significant advances. Given the richness of the progress made over the last decade, in this article, we focused on the most recent contributions of genome-wide and transcriptome analyses of Leishmania spp., which permit the comparison of life cycle stages, the evaluation of different strains and species in their natural niches and in the field and the simultaneously comparison of the gene expression profiles of parasites and hosts.

This work was developed aiming at evaluating the effect of different corn grains milling grades under the characterisation and quantification of laying hens’ waste. For this, 128 Hisex Brown strain 30 weeks old hens (laying hens) were used, in a period of time of 112 days (4 cycles of 28 days). The hens were distributed in a completely randomized experimental design, with four treatments, four replicates and eight hens per each experimental unit. The treatments consisted of four diets with different milling diameters (mm): 2.0 (average geometrical diameter AGD= 565 μm); 4.0 (AGD= 620 μm), 6.0 (AGD= 781 μm), and 8.0 (AGD= 1085 μm). The variables analyzed were: waste production (kg·hen.day-1), dry matter (kg and %), percentages of phosphorus and nitrogen, and residue coefficient (kg). The results were submitted to analysis of polynomial regression (p

Highlights • Mutations in the Wnt, Notch, and Ras-ERK signaling pathways in C. elegans lead to infertility, sterility, and multivulva formation. • Phenotypic assays using C. elegans mutant strains can be used as in vivo models for drug repurposing. • Itraconazole, disulfiram, etodolac, and ouabain have anticancer potential that can specifically target the Wnt, Notch, and RAS-ERK signaling pathways., Drug repurposing is used as a strategy for finding new drugs for cancer. The process is a faster and a more cost-effective way of providing new indications for drugs that can address emerging drug resistance and numerous side effects of chemotherapeutic drugs. In this study, the in vivo anticancer potential of itraconazole, disulfiram, etodolac, and ouabain were assessed using five different C. elegans mutant strains. Each strain contains mutations in genes involved in different signaling pathways such as Wnt (JK3476), Notch (JK1107 and BS3164), and Ras-ERK (SD939 and MT2124) that result to phenotypes of sterility, infertility, and multivulva formation. These same signaling pathways have been shown to be defective in several human cancer types. The four candidate drugs were tested on the C. elegans mutant strains to determine if they rescue the mutant phenotypes. Both ouabain and etodolac significantly reduced the sterile and infertile phenotypes of JK3476, JK1107, BS3164, and SD939 strains (p=0.0010). Finally, itraconazole and etodolac significantly reduced multivulva formation (p=0.0021). The degrees of significant phenotypic rescues of each mutant were significantly higher than vehicle only (1% DMSO). Therefore, this study demonstrated that the four candidate drugs have anticancer potential in vivo, and etodolac had the highest anticancer potential.

Leishmaniasis is a worldwide public health problem caused by protozoan parasites of the genus Leishmania. Leishmania braziliensis is the most important species responsible for tegumentary leishmaniases in Brazil. An understanding of the molecular mechanisms underlying the success of this parasite is urgently needed. An in-depth study on the modulation of gene expression across the life cycle stages of L. braziliensis covering coding and noncoding RNAs (ncRNAs) was missing and is presented herein. Analyses of differentially expressed (DE) genes revealed that most prominent differences were observed between the transcriptomes of insect and mammalian proliferative forms (6,576 genes). Gene ontology (GO) analysis indicated stage-specific enriched biological processes. A computational pipeline and 5 ncRNA predictors allowed the identification of 11,372 putative ncRNAs. Most of the DE ncRNAs were found between the transcriptomes of insect and mammalian proliferative stages (38%). Of the DE ncRNAs, 295 were DE in all three stages and displayed a wide range of lengths, chromosomal distributions and locations; many of them had a distinct expression profile compared to that of their protein-coding neighbors. Thirty-five putative ncRNAs were submitted to northern blotting analysis, and one or more hybridization-positive signals were observed in 22 of these ncRNAs. This work presents an overview of the L. braziliensis transcriptome and its adjustments throughout development. In addition to determining the general features of the transcriptome at each life stage and the profile of protein-coding transcripts, we identified and characterized a variety of noncoding transcripts. The novel putative ncRNAs uncovered in L. braziliensis might be regulatory elements to be further investigated.

espanolEl estudio determino el capacidad de sustancias naturales de Moringa, Cactus, Neem y Maiz en el tratamiento del agua del rio Magdalena que consumen las comunidades del corregimiento de Palermo, en Colombia. Por medio de la prueba de jarras, se establecieron las dosis optimas de coagulante, tiempo y velocidad de floculacion. El sulfato de aluminio (0.02 g L-1) se utilizo como control. Los resultados mostraron que Moringa (2g L-1) redujo la turbidez en 96.8% y el color en 97.8% mientras que el control logro 95.7% y 96.5%, respectivamente. Por otro lado, el tratamiento con Neem (0.8g L-1) disminuyo 99.4% de los coliformes totales y 99.2% de los coliformes fecales mientras que el Maiz (2.5g L-1), redujo 94.4% de los coliformes totales. El uso de sustancias naturales es una alternativa asequible y sostenible de potabilizacion del agua del rio Magdalena. EnglishThe study determined the capability of natural substances such as Moringa, Cactus, Neem and Corn on the treatment of water from Magdalena rive, water used by communities in the municipality of Palermo in Colombia. Through jars tests, optimal doses of coagulant, time and flocculation rate were established, aluminum sulfate (0.02 g L-1) was used as control. The results showed that Moringa (2g L-1) attenuated in 96.8% the turbidity and 97.8% of color while the conventional one obtained 95.7% and 96.5%, respectively; In turn, Neem (0.8g L-1) decreased 99.4% total coliforms and 99.2% of fecal coliforms. Also, Corn (2.5g L-1), reduced 94.4% of total coliforms. The use of natural substances is a feasible and sustainable alternative to water purification of the Magdalena river.

Leishmania contains two phosphoglycerate kinase (PGK) genes, PGKB and PGKC, which code for the cytosolic and glycosomal isoforms of the enzyme, respectively. Although differences in PGKB and PGKC transcript and protein levels and isoform activities have been well documented, the mechanisms of control of both transcript and protein abundance have not been described to date. To better understand the regulation of Leishmania PGK expression, we investigated the stabilities of both PGK transcripts using reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) in combination with transcription and trans-splicing inhibitors. Cells were treated with sinefungin and actinomycin D, and RNA decay kinetics were assessed. In addition, immunoblotting and protein synthesis inhibition by cycloheximide were employed to evaluate protein steady states and degradation. We observed increased stabilities of both PGKB mRNA and protein compared with the glycosomal isoform (PGKC). Our results indicate that both post-transcriptional and post-translational events contribute to the distinct expression levels of the PGKB and PGKC isoforms in Leishmania major.

Orbital biopsy of nonspecific orbital inflammation, commonly referred to as “orbital pseudotumor,” typically shows a combination of polyclonal lymphocytes, plasmocytes, leukocytes, macrophages, and variable degrees of collagen deposition. Herein, we report a patient with a positive history of mucocutaneous leishmaniasis who presented with an orbital mass with a histological profile of idiopathic orbital inflammation. Immunohistochemical and molecular analysis of the orbital specimens demonstrated that the orbital inflammation was associated with the presence of antigens of Leishmania braziliensis and DNA from the parasite.

Objective. To determine the frequency and distribution of ABO and Rh (D) antigens and, additionally, investigate gene diversity and the structure of Mexican populations.Materials and Methods. Blood groups were tested in 271,164 subjects from 2014 to 2016. The ABO blood group was determined by agglutination using the antibodies anti-A, Anti-B, and Anti-D for the Rh factor, respectively.Results. The overall distribution of ABO and Rh (D) groups in the population studied was as follows: O: 61.82%; A: 27.44%; B: 8.93%; and AB: 1.81%. For the Rh group, 95.58% of people were Rh (D), and 4.42% were Rh (d). Different distributions of blood groups across regions were found; additionally, genetic analysis revealed that theIOandIDallele showed an increasing trend from the north to the center, while theIAandIdallele tended to increase from the center to the north. Also, we found more gene diversity in bothlociin the north compared with the center, suggesting population structure in Mexico.Conclusion. This work could help health institutions to identify where they can obtain blood products necessary for medical interventions. Moreover, this piece of information contributes to the knowledge of the genetic structure of the Mexican populations which could have significant implications in different fields of biomedicine.

Common beans have been associated with anti-diabetic effects, due to its high content of bioactive compounds. Nevertheless, its consumption has decreased worldwide. Therefore, there is an increasing interest in the development of novel functional foods elaborated with common beans. The aim of this study was to evaluate the anti-diabetic effect of oat-bean flour cookies, and to analyze its bioactive composition, using commercial oat-wheat cookies for comparative purposes.Oat-bean cookies (1.2 g kgThese results suggest that common beans can be used as functional ingredients for the elaboration of cookies with anti-diabetic effects. © 2017 Society of Chemical Industry.

Through whole-genome sequencing analysis, we identified non-Leishmania parasites isolated from a man with a fatal visceral leishmaniasis–like illness in Brazil. The parasites infected mice and reproduced the patient’s clinical manifestations. Molecular epidemiologic studies are needed to ascertain whether a new infectious disease is emerging that can be confused with leishmaniasis.

This study focused on the design and construction of wave energy generation system. Wave energy is produced once the electric generators is placed parallel on the ocean surface and rides in different height of waves along the length of the device. It follows the concept of Faraday’s law of induction in the linear generator which an alternating current is induced in the loops of copper wire whenever the magnet slides back and forth through it. Using Axially magnetic type linear generator that will utilized the horizontal forces brought by the waves and placed on a waterproof floating boat with a data logger using microcontroller. The wave converter is deployed in a controlled and uncontrolled environment to observe the behaviour of the prototype and compared the measured parameters such as voltage, current, power, and energy at different time and wave height.

Trichoderma species are fungi widely employed as plant-growth-promoting agents and for biological control. Several commercial and laboratory-made solid formulations for mass production of Trichoderma have been reported. In this study, we evaluated a solid kaolin-based formulation to promote the absortion/retention of Trichoderma asperellum in the substrate for growing tomato plants. The unique implementation of this solid formulation resulted in an increased growth of the tomato plants, both in roots and shoots after 40 days of its application. Plants were challenged with two fungal pathogens, Fusarium oxysporum and Botrytis cinerea, and pretreatment with T. asperellum resulted in less severe wilting and stunting symptoms than non-treated plants. Treatment with T. asperellum formulation inhibited Reactive Oxygen Species (ROS) production in response to the pathogens in comparison to plants that were only challenged with both pathogens. These results suggest that decrease in ROS levels contribute to the protective effects exerted by T. asperellum in tomato.

Aims: To test and compare relative efficiencies of Guava Leaf Ethanolic Extract (GLEEx) and antibiotic standards against two pathogenic gram-negative bacteria in planktonic and biofilm lifestyles. Study Design: In vitro antibacterial study. Place and Duration of Study: Department of Biochemistry & Molecular Biology in the University of the Philippines Manila, between January 2014 and April 2014. Methodology: The susceptibility of planktonic and biofilm forms of Escherichia coli and Original Research Article Annual Research & Review in Biology, 4(24): 4370-4380, 2014 4371 Pseudomonas aeruginosa to GLEEx and to appropriate antibiotic standards were evaluated using the Kirby-Bauer disc diffusion assay (KB) and the Bio Timer Assay (BTA), respectively. Results: In KB, GLEEx exhibited comparable antibacterial strengths against both bacteria (P > .05), but was significantly less than the antibiotic standards (P .05). Conclusion: The relative antibacterial potencies of GLEEx and standard antibiotics vary depending on bacterial lifestyle, thus suggesting the need to use BTA alongside KB when assessing the antibacterial properties of natural products.

There is growing and compelling evidence demonstrating the extra-skeletal role of vitamin D and the importance of maintaining adequate levels of this nutrient. Currently, there is very limited information available on the vitamin D status in children and adults in underserved groups, including Puerto Ricans. We assessed the vitamin D status of 4,090 Puerto Ricans living in six geographical regions in the island. Only 31.5% of the studied population had sufficient vitamin D levels (>30 ng/ml). The 18–39 year age group and the females showed inadequate (30 ng/ml). Future studies are certainly warranted to understand the prevalence of vitamin D deficiency and influencing factors (including obesity) in Puerto Ricans.

Aluminum pieces are often used in various industrial processes like automotive and aerospace manufacturing, as well as in ornamental applications, so it is necessary to develop processes to protect these materials, processes that can be industrialized to protect the aluminum as well or better than chromate treatments. The purpose of this research is to evaluate boric acid as an accelerator by optimizing its concentration in cerium conversion coatings (CeCC) with 10-minute immersion time with a concentration of 0.1 g L −1 over aluminum to protect it. The evaluation will be carried out by measuring anticorrosion properties with electrochemical techniques (polarization resistance, , polarization curves, PC, and electrochemical impedance spectroscopy, EIS) in NaCl 3.5% wt. aqueous solution and surface characterization with scanning electron microscopy (SEM).