Contribution of microscopy for understanding the mechanism of action against trypanosomatids

Transmission electron microscopy (TEM) has proved to be a useful tool to study the ultrastructural alterations and the target organelles of new antitrypanosomatid drugs. Thus, it has been observed that sesquiterpene lactones induce diverse ultrastructural alterations in both T. cruzi and Leishmania spp., such as cytoplasmic vacuolization, appearance of multilamellar structures, condensation of nuclear DNA, and, in some cases, an important accumulation of lipid vacuoles. This accumulation could be related to apoptotic events. Some of the sesquiterpene lactones (e.g., psilostachyin) have also been demonstrated to cause an intense mitochondrial swelling accompanied by a visible kinetoplast deformation as well as the appearance of multivesicular bodies. This mitochondrial swelling could be related to the generation of oxidative stress and associated to alterations in the ergosterol metabolism. The appearance of multilamellar structures and multiple kinetoplasts and flagella induced by the sesquiterpene lactone psilostachyin C indicates that this compound would act at the parasite cell cycle level, in an intermediate stage between kinetoplast segregation and nuclear division. In turn, the diterpene lactone icetexane has proved to induce the external membrane budding on T. cruzi together with an apparent disorganization of the pericellar cytoskeleton. Thus, ultrastructural TEM studies allow elucidating the possible mechanisms and the subsequent identification of molecular targets for the action of natural compounds on trypanosomatids.Fil: Lozano, Esteban Sebastián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Spina Zapata, Renata María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Barrera, Patricia Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; ArgentinaFil: Tonn, Carlos Eugenio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Investigaciones en Tecnología Química. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Investigaciones en Tecnología Química; ArgentinaFil: Sosa Escudero, Miguel Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Instituto de Histología y Embriología de Mendoza Dr. Mario H. Burgos; Argentin

PIP5KIβ Selectively Modulates Apical Endocytosis in Polarized Renal Epithelial Cells

Localized synthesis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] at clathrin coated pits (CCPs) is crucial for the recruitment of adaptors and other components of the internalization machinery, as well as for regulating actin dynamics during endocytosis. PtdIns(4,5)P2 is synthesized from phosphatidylinositol 4-phosphate by any of three phosphatidylinositol 5-kinase type I (PIP5KI) isoforms (α, β or γ). PIP5KIβ localizes almost exclusively to the apical surface in polarized mouse cortical collecting duct cells, whereas the other isoforms have a less polarized membrane distribution. We therefore investigated the role of PIP5KI isoforms in endocytosis at the apical and basolateral domains. Endocytosis at the apical surface is known to occur more slowly than at the basolateral surface. Apical endocytosis was selectively stimulated by overexpression of PIP5KIβ whereas the other isoforms had no effect on either apical or basolateral internalization. We found no difference in the affinity for PtdIns(4,5)P2-containing liposomes of the PtdIns(4,5)P2 binding domains of epsin and Dab2, consistent with a generic effect of elevated PtdIns(4,5)P2 on apical endocytosis. Additionally, using apical total internal reflection fluorescence imaging and electron microscopy we found that cells overexpressing PIP5KIβ have fewer apical CCPs but more internalized coated structures than control cells, consistent with enhanced maturation of apical CCPs. Together, our results suggest that synthesis of PtdIns(4,5)P2 mediated by PIP5KIβ is rate limiting for apical but not basolateral endocytosis in polarized kidney cells. PtdIns(4,5)P2 may be required to overcome specific structural constraints that limit the efficiency of apical endocytosis. © 2013 Szalinski et al

Key Lessons Learned from Moffitt's Molecular Tumor Board: The Clinical Genomics Action Committee Experience

The increasing practicality of genomic sequencing technology has led to its incorporation into routine clinical practice. Successful identification and targeting of driver genomic alterations that provide proliferative and survival advantages to tumor cells have led to approval and ongoing development of several targeted cancer therapies. Within many major cancer centers, molecular tumor boards are constituted to shepherd precision medicine into clinical practice

2021 Tajogaite eruption records infiltration of crustal fluids within the upper mantle beneath La Palma, Canary Islands

The 2021 Tajogaite eruption at La Palma has represented a unique opportunity to investigate the characteristics of the mantle source feeding modern volcanism in the Canary Islands. With the aim of track the fingerprint of carbon in the local oceanic lithosphere-asthenosphere system, we report the isotopic composition of CO2 (δ13C values versus Vienna Pee Dee Belemnite) in olivine- and clinopyroxene-hosted fluid inclusions (FI) from the 2021 Tajogaite lavas and from lavas/ultramafic xenoliths (olivine-clinopyroxenites, clinopyroxenites, dunites and harzburgites) from the nearby 1677 San Antonio eruption cone/lavas, in an attempt to characterize the origin and evolution of carbon within the local mantle source. Our results indicate that the 2021 and 1677 lavas exhibit δ13C values ranging from −4.94‰ to −2.71‰ and CO2/3He ratios from 3.37 to 6.14 × 109. Ultramafic xenoliths fall in a comparable range of values despite showing higher CO2 concentrations. Our δ13C values fall within the range of carbon isotope results previously reported for the Dos Aguas cold spring located in the Taburiente Caldera (northern La Palma), suggesting an apparent carbon isotope homogeneity at the scale of the entire island. The (relatively narrow) δ13C vs. CO2/3He ratio range of La Palma samples is interpreted to reflect either i) variable extents of open-system degassing of a common mantle endmember having δ13C of ∼1.7‰, or ii) mixing between depleted mantle-like carbon (−6‰ < δ13C < −4‰) and crustal carbon (δ13C = 0‰) endmembers. Both models testify a crustal carbon component recycled in the local mantle. This component, also detected in mantle xenoliths from the neighboring island of El Hierro and the easternmost Lanzarote, indicates a regional characteristic of the mantle beneath the Canary Islands, interpreted as a result of infiltration of carbon-rich melts during past metasomatic events in the local mantle

The noble gas signature of the 2021 Tajogaite eruption (La Palma, Canary Islands)

Here, we characterize the temporal evolution of volatiles during the Tajogaite eruption by analyzing the elemental (He-Ar-CO2-N2) and isotopic (He-Ar-Ne) composition of fluid inclusions (FI) in phenocrysts (olivine+pyroxene) identified in erupted lavas. Our 2021 lava samples identify substantial temporal variations in volatile composition. We show that, during the 2021 Tajogaite eruption, the He-CO2-N2 concentrations in FI increased since October 15th; this increase was accompanied by increasing 40Ar/36Ar ratios (from ~300 to >500), and paralleled a major shift in bulk lava chemistry, with increasing Mg contents (Mg#, from 47 to 52 to 55–59), CaO/Al2O3 (from 0.65 to 0.74 to 0.75–0.90), Ni and Cr, and decreasing TiO2, P2O5 and incompatible elements. The olivine core composition also became more forsteritic (from Mg# = 80–81 to Mg# = 84–86). Mineral thermobarometry and FI barometry results indicate that the eruption was sustained by magmas previously stored in at least two magma accumulation zones, at respectively ~6–12 km and 15–30 km, corroborating previous seismic and FI evidence. We therefore propose that the compositional changes seen throughout the eruption can be explained by an increased contribution - since early/mid-October - of more primitive, lessdegassed magma from the deeper (mantle) reservoir. Conversely, Rc/Ra values (3He/4He ratios corrected for atmospheric contamination) remained constant throughout the whole eruption at MORB-like values (7.38 ± 0.22 Ra, 1σ), suggesting an isotopically homogeneous magma feeding source. The Tajogaite He isotope signature is within the range of values observed for the 1677 San Antonio lavas (7.37 ± 0.17Ra, 1σ), but is more radiogenic than the 3He/4He values (>9 Rc/Ra) observed in the Caldera de Taburiente to the north. The 3He/4He ratios (6.75 ± 0.20 Ra, 1σ) measured in mantle xenoliths from the San Antonio volcano indicate a relatively radiogenic nature of the mantle beneath the Cumbre Vieja ridge. Based on these results and mixing modeling calculations, we propose that the homogeneous He isotopic signatures observed in volatiles from the Tajogaite/San Antonio lavas reflect three component mixing between a MORB-like source, a radiogenic component and small additions (6–15%) of a high 3He/4He reservoir-derived (>9Ra) fluid components. The simultaneous occurrence of high 3 He/4 He (>9Ra)- and MORB-like He signatures in northern and southern La Palma is interpreted to reflect smallscale heterogeneities in the local mantle, arising from spatially variable proportions of MORB, radiogenic, and high 3He/4He component

Proposed diagnostic criteria for classical CMML, CMML variants and pre-CMML conditions

Chronic myelomonocytic leukemia (CMML) is a myeloid neoplasm characterized by dysplasia, abnormal production and accumulation of monocytic cells and an elevated risk to transform into acute leukemia. Over the past two decades, our knowledge about the pathogenesis and molecular mechanisms in CMML has increased substantially. In parallel, better diagnostic criteria and therapeutic strategies have been developed. However, many questions remain regarding prognostication and optimal therapy. In addition, there is a need to define potential pre-phases of CMML and special CMML variants, and to separate these entities from each other and from conditions mimicking CMML. To address these unmet needs, an international consensus group met in a Working Conference in August 2018 and discussed open questions and issues around CMML, its variants, and pre-CMML conditions. The outcomes of this meeting are summarized herein and include diagnostic criteria and a proposed classification of pre-CMML conditions as well as refined minimal diagnostic criteria for classical CMML and special CMML variants, including oligomonocytic CMML and CMML associated with systemic mastocytosis. Moreover, we propose diagnostic standards and tools to delineate between normal, pre-CMML and CMML entities. These criteria and standards should facilitate diagnostic and prognostic evaluations in daily practice and clinical studies in applied hematology

Data equivalence in cross-cultural international business research: Assessment and guidelines

Data equivalence refers to the extent to which the elements of a research design have the same meaning, and can be applied in the same way, in different cultural contexts. Failure to establish data equivalence in cross-cultural studies may bias empirical results and theoretical inferences. Although several authors have encouraged researchers to ensure high levels of data equivalence, no study has assessed the status of the field in relation to compliance with data equivalence standards. Accordingly, this study examines three aspects of data equivalence (construct equivalence, measurement equivalence, and data collection equivalence) within 167 studies that involve cross-cultural data published in the Journal of International Business Studies, Management International Review, Journal of World Business, Strategic Management Journal and the Academy of Management Journal from 1995 to 2005. The findings indicate that international business researchers report insufficient information in relation to data equivalence issues, thus limiting confidence in the findings of many cross-cultural studies. To enhance future research, a guideline for procedures for researchers to follow and report in establishing data equivalence is offered. © 2008 Academy of International Business