Key role of the 3' untranslated region in the cell cycle regulated expression of the Leishmania infantum histone H2A genes: minor synergistic effect of the 5' untranslated region

<p>Abstract</p> <p>Background</p> <p>Histone synthesis in <it>Leishmania </it>is tightly coupled to DNA replication by a post-transcriptional mechanism operating at the level of translation.</p> <p>Results</p> <p>In this work we have analyzed the implication of the 5' and 3' untranslated regions (UTR) in the cell cycle regulated expression of the histone <it>H2A </it>in <it>Leishmania infantum</it>. For that purpose, <it>L. infantum </it>promastigotes were stably transfected with different plasmid constructs in which the <it>CAT </it>coding region used as a reporter was flanked by the 5' and 3' UTR regions of the different <it>H2A </it>genes. We report that in spite of their sequence differences, histone <it>H2A </it>5' and 3' UTRs conferred a cell cycle dependent pattern of expression on the CAT reporter since <it>de novo </it>synthesis of CAT increased when parasites enter the S phase. Using one established <it>L. infantum </it>cell line we showed that CAT expression is controlled by the same regulatory events that control the endogenous histone gene expression. Thus, although we did not detect changes in the level of <it>CAT </it>mRNAs during cell cycle progression, a drastic change in the polysome profiles of <it>CAT </it>mRNAs was observed during the progression from G1 to S phase. In the S phase <it>CAT </it>mRNAs were on polyribosomal fractions, but in the G1 phase the association of <it>CAT </it>transcripts with ribosomes was impaired. Furthermore, it was determined that the addition of just the <it>H2A </it>3' UTR to the <it>CAT </it>reporter gene is sufficient to achieve a similar pattern of post-transcriptional regulation indicating that this region contains the major regulatory sequences involved in the cell cycle dependent expression of the <it>H2A </it>genes. On the other hand, although CAT transcripts bearing the <it>H2A </it>5' alone were translated both in the G1 and S phase, higher percentages of transcripts were detected on polyribosomes in the S phase correlating with an increase in the <it>de novo </it>synthesis of CAT. Thus, it can be concluded that this region also contributes, although to a minor extent than the 3' UTR, in the enhancement of translation in the S phase relative to the G1 phase.</p> <p>Conclusion</p> <p>Our findings indicate that both, the 5' and the 3' UTRs contain sequence elements that contribute to the cell cycle expression of <it>L. infantum </it>H2A. The 3' UTR region is essential for cell cycle dependent translation of the <it>L. infantum </it>H2A transcripts whereas the 5' UTR has a minor contribution in their S phase dependent translation.</p

Coadministration of the Three Antigenic Leishmania infantum Poly (A) Binding Proteins as a DNA Vaccine Induces Protection against Leishmania major Infection in BALB/c Mice

Highly conserved intracellular proteins from Leishmania have been described as antigens in natural and experimental infected mammals. The present study aimed to evaluate the antigenicity and prophylactic properties of the Leishmania infantum Poly (A) binding proteins (LiPABPs). Three different members of the LiPABP family have been described. Recombinant tools based on these proteins were constructed: recombinant proteins and DNA vaccines. The three recombinant proteins were employed for coating ELISA plates. Sera from human and canine patients of visceral leishmaniasis and human patients of mucosal leishmaniasis recognized the three LiPABPs. In addition, the protective efficacy of a DNA vaccine based on the combination of the three Leishmania PABPs has been tested in a model of progressive murine leishmaniasis: BALB/c mice infected with Leishmania major. The induction of a Th1-like response against the LiPABP family by genetic vaccination was able to down-regulate the IL-10 predominant responses elicited by parasite LiPABPs after infection in this murine model. This modulation resulted in a partial protection against L. major infection. LiPABP vaccinated mice showed a reduction on the pathology that was accompanied by a decrease in parasite burdens, in antibody titers against Leishmania antigens and in the IL-4 and IL-10 parasite-specific mediated responses in comparison to control mice groups immunized with saline or with the non-recombinant plasmid. The results presented here demonstrate for the first time the prophylactic properties of a new family of Leishmania antigenic intracellular proteins, the LiPABPs. The redirection of the immune response elicited against the LiPABP family (from IL-10 towards IFN-γ mediated responses) by genetic vaccination was able to induce a partial protection against the development of the disease in a highly susceptible murine model of leishmaniasisThe study was supported in Spain by grants from Ministerio de Ciencia e Innovación FIS PI11/00095 and FISPI14/00366 from the Instituto de Salud Carlos III within the Network of TropicalDiseases Research (VI P I+D+I 2008-2011, ISCIII -Subdirección General de Redes y Centros de Investigación Cooperativa (RD12/0018/0009)). This work was also supported in Brazil by a grant from CNPq (Ciencia sem Fronteiras-PVE 300174/2014-4). A CBMSO institutional grant from Fundación Ramón Areces is also acknowledged. EAFC is a grant recipient of CNPq. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscrip

Effectiveness of habitat management in the recovery of low-density populations of wild rabbit.

Understanding the relationship between spatial patterns of landscape attributes and population presence and abundance is essential for understanding population processes as well as supporting management and conservation strategies. This study evaluates the influence of three factors: environment, habitat management, and season on the presence and abundance of the wild rabbit (Oryctolagus cuniculus), an important prey species for Mediterranean endangered predator species. To address this issue, we estimated wild rabbit presence and abundance by latrine counting in transects located in 45 plots within a 250×250 m grid from June 2007 until June 2009 in a 1,200 ha hunting area in southern Portugal.We then analyzed how wild rabbit presence and abundance correlatewith the aforementioned factors. Our results showed that the main variable influencing wild rabbit presence and abundance was the distance to the artificial warrens. North and northeast slope directions were negatively related to wild rabbit presence. Conversely, rabbit presence was positively correlated with short distances to ecotone, artificial warrens, and spring. Regarding rabbit abundance, in addition to artificial warrens, soft soils, bushes, and season also had a positive effect. We found that environmental variables, management practices, and season each affect wild rabbit presence and abundance differently at a home range scale in low-density population. Thus, our major recommendations are reducing the distance to artificial warrens and ecotone, ideally to less than 100 m, and promoting habitat quality improvement on slopes with plenty of sun exposure

Disruption of the CCL1-CCR8 axis inhibits vascular Treg recruitment and function and promotes atherosclerosis in mice

The CC chemokine 1 (CCL1, also called I-309 or TCA3) is a potent chemoattractant for leukocytes that plays an important role in inflammatory processes and diseases through binding to its receptor CCR8. Here, we investigated the role of the CCL1-CCR8 axis in atherosclerosis. We found increased expression of CCL1 in the aortas of atherosclerosis-prone fat-fed apolipoprotein E (Apoe)-null mice; moreover, in vitro flow chamber assays and in vivo intravital microscopy demonstrated an essential role for CCL1 in leukocyte recruitment. Mice doubly deficient for CCL1 and Apoe exhibited enhanced atherosclerosis in aorta, which was associated with reduced plasma levels of the anti-inflammatory interleukin 10, an increased splenocyte Th1/Th2 ratio, and a reduced regulatory T cell (Treg) content in aorta and spleen. Reduced Treg recruitment and aggravated atherosclerosis were also detected in the aortas of fat-fed low-density lipoprotein receptor-null mice treated with CCR8 blocking antibodies. These findings demonstrate that disruption of the CCL1-CCR8 axis promotes atherosclerosis by inhibiting interleukin 10 production and Treg recruitment and function.This study was supported by the Spanish Ministerio de Ciencia, Innovación y Universidades (MCIU, grants SAF2016-79490-R and SAF2014-57845-R) and the Instituto de Salud Carlos III (ISCIII, grants PI14/00526, PI17/01395, CP11/00145, and CPII16/00022) with co-funding from the European Regional Development Fund (ERDF, “Una manera de hacer Europa”), the Fundación Ramón Areces, European Union (EuroCellNet COST Action CA15214) and the INSERM. VZG is supported by the ISCIII, JMG-G by the ISCIII Miguel Servet Program and the Instituto de Investigación Sanitaria Hospital 12 de Octubre (imas12), AdMM by the MCIU (predoctoral contract BES-2014-06779), and ZM by a British Heart Foundation Professorship. The CNIC is supported by the MCIU and the Pro CNIC Foundation and is a Severo Ochoa Center of Excellence (SEV-2015-0505).S

Spatiotemporal Characteristics of the Largest HIV-1 CRF02_AG Outbreak in Spain: Evidence for Onward Transmissions

Background and Aim: The circulating recombinant form 02_AG (CRF02_AG) is the predominant clade among the human immunodeficiency virus type-1 (HIV-1) non-Bs with a prevalence of 5.97% (95% Confidence Interval-CI: 5.41–6.57%) across Spain. Our aim was to estimate the levels of regional clustering for CRF02_AG and the spatiotemporal characteristics of the largest CRF02_AG subepidemic in Spain.Methods: We studied 396 CRF02_AG sequences obtained from HIV-1 diagnosed patients during 2000–2014 from 10 autonomous communities of Spain. Phylogenetic analysis was performed on the 391 CRF02_AG sequences along with all globally sampled CRF02_AG sequences (N = 3,302) as references. Phylodynamic and phylogeographic analysis was performed to the largest CRF02_AG monophyletic cluster by a Bayesian method in BEAST v1.8.0 and by reconstructing ancestral states using the criterion of parsimony in Mesquite v3.4, respectively.Results: The HIV-1 CRF02_AG prevalence differed across Spanish autonomous communities we sampled from (p &lt; 0.001). Phylogenetic analysis revealed that 52.7% of the CRF02_AG sequences formed 56 monophyletic clusters, with a range of 2–79 sequences. The CRF02_AG regional dispersal differed across Spain (p = 0.003), as suggested by monophyletic clustering. For the largest monophyletic cluster (subepidemic) (N = 79), 49.4% of the clustered sequences originated from Madrid, while most sequences (51.9%) had been obtained from men having sex with men (MSM). Molecular clock analysis suggested that the origin (tMRCA) of the CRF02_AG subepidemic was in 2002 (median estimate; 95% Highest Posterior Density-HPD interval: 1999–2004). Additionally, we found significant clustering within the CRF02_AG subepidemic according to the ethnic origin.Conclusion: CRF02_AG has been introduced as a result of multiple introductions in Spain, following regional dispersal in several cases. We showed that CRF02_AG transmissions were mostly due to regional dispersal in Spain. The hot-spot for the largest CRF02_AG regional subepidemic in Spain was in Madrid associated with MSM transmission risk group. The existence of subepidemics suggest that several spillovers occurred from Madrid to other areas. CRF02_AG sequences from Hispanics were clustered in a separate subclade suggesting no linkage between the local and Hispanic subepidemics

Breeding dispersal movements of Dupont’s Lark Chersophilus duponti in fragmented landscape

Understanding patterns of dispersal behaviour of threatened species is important in conservation biology and population ecology, especially in fragmented landscapes. Dupont’s Lark Chersophilus duponti is a threatened passerine whose European population has declined by around 40% in the last decade. To study natal and breeding dispersal of the species, we used a long-term mark-recapture dataset (2011–2019) and analysed the records of 32 males (39 dispersal movements) and five females (five dispersal movements) captured during the breeding season at Rincón de Ademuz (Valencia, eastern Spain). Adult birds had a median breeding dispersal of 154 m (Q25-Q75 = 70.0–300.3). Among these captures, two adult males dispersed to a new patch of habitat separated by more than 5,800 m. Only one out of 26 nestlings ringed was trapped as an adult bird, which occurred at a site different from the natal territory (4,500 m). Our results show a low breeding dispersal for the species and are, in essence, in agreement with previous studies carried out in the Ebro Valley metapopulation, one of the core areas for the species in Europe. Nonetheless, unlike in the Ebro Valley, we detected movements of adult birds between habitat patches. The low recovery rate of young birds suggests that they left their natal sites and moved outside the study area or that their survival rate was very low. Future studies focused on the post-fledging survival rate and natal dispersal movements are essential to determine effective conservation measures for the species. Habitat management actions in occupied and potential sites should be carried out close to the areas inhabited by the species to increase the success rate of the interventions and the effective dispersal and therefore population connectivity.This study was supported by Levantina y Asociados de Minerales, S.A., as part of the project “Estudios de investigación aplicados a la conservación de las poblaciones de alondra ricotí (Chersophilus duponti) en el entorno del municipio de Vallanca”

Supervivencia y probabilidad de recaptura del avión zapador en una colonia de Aspe: análisis del periodo 2004-2011

En este trabajo se analizan los anillamientos y recapturas de adultos en una colonia de avión zapador próxima a Aspe (Alicante) entre 2004 y 2011. El objetivo es estimar la variación de las tasas de supervivencia entre sexos y a lo largo del tiempo. La probabilidad de recaptura (p) muestra una tendencia a aumentar a lo largo del tiempo, pero de forma más marcada en los machos. La tasa de supervivencia difiere entre sexos, siendo ligeramente mayor en los machos. El mejor modelo incluye la variabilidad temporal de la supervivencia de machos, pero considera la de las hembras constante a lo largo del tiempo. La mayor variabilidad de la supervivencia de los machos es un patrón no detectado en estudios previos

Microhabitat use and seasonality of mayflies (Ephemeroptera) in two streams in eastern Cuba

We identified groups of mayfly species that share microhabitats in eastern Cuban streams by sampling during rainy and dry seasons in four microhabitats in pools (cobbles, sand, leaf litter and bank vegetation) and one microhabitat (cobbles) in riffles. A total of 20 species and three morphospecies were found. Species similarity in habitat use was analysed using k-means clustering. Overall, the highest number of individuals was collected in cobbles in riffles, followed by cobbles in pools. Five species groups, labelled from I to V, were identified based on their primary microhabitat use. Two groups (I and V) included species with a generalist pattern of habitat use, but differing in seasonality. Species in remaining groups tend to concentrate in specific microhabitats. Species of groups II and IV use mostly cobbles in riffles and in pools, respectively, while species in group III concentrate in sand during the rainy season. Our results allowed to identify the species most vulnerable to alterations of the hydrologic regimen.The University of Alicante funded the research stays of PLC in the Ecology department through the ‘Proyecto Habana’

Chemotherapy induces cell plasticity; controlling plasticity increases therapeutic response

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