Microorganism response to stressed terrestrial environments: a raman spectroscopic perspective of extremophilic life strategies

Raman spectroscopy is a valuable analytical technique for the identification of biomolecules and minerals in natural samples, which involves little or minimal sample manipulation. In this paper, we evaluate the advantages and disadvantages of this technique applied to the study of extremophiles. Furthermore, we provide a review of the results published, up to the present point in time, of the bio- and geo-strategies adopted by different types of extremophile colonies of microorganisms. We also show the characteristic Raman signatures for the identification of pigments and minerals, which appear in those complex sample

Combined study of archaeomagnetism and Raman spectroscopy of experimentally burnt limestones from the middle-palaeolithic site of Pinilla del Valle (Madrid, Spain)

Se presentan los resultados arqueomagnéticos, de propiedades magnéticas y espectroscopia Raman obtenidos de analizar muestras de dos tipos de calizas calentadas experimentalmente bajo con-diciones controladas. Estas calizas han sido fre-cuentemente identificadas en los yacimientos de paleolítico medio de Cueva Descubierta y Abrigo de Navalmaíllo (Pinilla del Valle, Madrid, Es-paña). Mediante un sistema de termopares, se re-gistraron temperaturas máximas de calentamiento de 400-450 ºC durante una hora. El objetivo es caracterizar las variaciones en la mineralogía fe-rromagnética y los compuestos (in)orgánicos, así como estudiar el registro de la dirección del campo magnético terrestre en el momento del enfriamien-to. Esto permitirá obtener un patrón extrapolable a las calizas arqueológicas de cara a identificar estructuras de combustión en el yacimiento. Los experimentos de ambas técnicas fueron realizados en muestras representativas de ambas litologías tanto antes como después de la quema experimen-tal. Antes de la quema, ambos tipos de caliza es-taban dominados por dolomita y calcita y, como principales óxidos de hierro, goetita y magnetita. Tras la quema, la goetita original pervivió en las muestras calentadas <200 ºC mientras que se transformó a hematites y/o magnetita en las ca-lentadas en torno a 450 ºC. Las alteraciones mi-neralógicas se centraron mayoritariamente en el 1ercm de profundidad. La neoformación de magnetita como paso intermedio en el proceso de deshidra-tación de la goetita hacia hematites ha sido poco documentada y tiene implicaciones paleomagnéti-cas. Se discuten los resultados mineralógicos, sus implicaciones para la identificación de fuego en el yacimiento así como sus potenciales aplicaciones a casos de estudio similare.Here we report the archaeomagnetic, rock-magnetic and Raman spectroscopy results of sample analysis from two types of limestones that were experimentally heated under controlled conditions. These lithologies have been commonly identified at the middle-palaeolithic sites of Descubierta Cave and Navalmaillo rock-shelter (Pinilla del Valle, Madrid, Spain). By means of a thermocouple system, maximum heating temperatures of 400 − 450 ºC were recorded during one hour. The main goal was to characterize the variations in the magnetic mineralogy and (in)organic compounds, and to study the record of the Earth´s magnetic field direction at the time of cooling. This allowed us to obtain a pattern that is potentially applicable to archaeological limestones in order to identify combustion structures at the site. All experiments were carried out on representative samples both before and after experimental heating. Before heating, both types of limestones were dominated by dolomite and calcite and as main iron oxides, goethite and magnetite. After heating, the original goethite persisted in samples heated to low temperatures (<200 ºC) whereas it transformed to haematite and/or magnetite in those heated to 450 ºC. Mineralogical alterations mostly focused on the outermost 1st cm of depth. The neoformation of magnetite as intermediate step in the dehydration process of goethite to haematite has been barely documented and has palaeomagnetic implications. The mineralogical results obtained, their implications for the identification of fire at the site as well as potential applications to similar cases studies are discussed.Junta de Castilla y León (projects BU066U16 and BU235P18) and the European Fund for Economic and Regional Development (FEDER

Raman spectroscopic analysis of the effect of the lichenicolous fungus Xanthoriicola physciae on its lichen host

YesLichenicolous (lichen-dwelling) fungi have been extensively researched taxonomically over many years, and phylogenetically in recent years, but the biology of the relationship between the invading fungus and the lichen host has received limited attention, as has the effects on the chemistry of the host, being difficult to examine in situ. Raman spectroscopy is an established method for the characterization of chemicals in situ, and this technique is applied to a lichenicolous fungus here for the first time. Xanthoriicola physciae occurs in the apothecia of Xanthoria parietina, producing conidia at the hymenium surface. Raman spectroscopy of apothecial sections revealed that parietin and carotenoids were destroyed in infected apothecia. Those compounds protect healthy tissues of the lichen from extreme insolation and their removal may contribute to the deterioration of the apothecia. Scytonemin was also detected, but was most probably derived from associated cyanobacteria. This work shows that Raman spectroscopy has potential for investigating changes in the chemistry of a lichen by an invading lichenicolous fungus.This work was completed while D.L.H. was in receipt of an award from the Ministerio de Economica y Competitividad of Spain (Proyectos CGL 2014-55542-P)

Identification of morphological biosignatures in martian analogue field specimens using in situ planetary instrumentation

We have investigated how morphological biosignatures (i.e., features related to life) might be identified with an array of viable instruments within the framework of robotic planetary surface operations at Mars. This is the first time such an integrated lab-based study has been conducted that incorporates space-qualified instrumentation designed for combined in situ imaging, analysis, and geotechnics (sampling). Specimens were selected on the basis of feature morphology, scale, and analogy to Mars rocks. Two types of morphological criteria were considered: potential signatures of extinct life (fossilized microbial filaments) and of extant life (crypto-chasmoendolithic microorganisms). The materials originated from a variety of topical martian analogue localities on Earth, including impact craters, high-latitude deserts, and hydrothermal deposits. Our in situ payload included a stereo camera, microscope, M&ouml;ssbauer spectrometer, and sampling device (all space-qualified units from Beagle 2), and an array of commercial instruments, including a multi-spectral imager, an X-ray spectrometer (calibrated to the Beagle 2 instrument), a micro-Raman spectrometer, and a bespoke (custom-designed) X-ray diffractometer. All experiments were conducted within the engineering constraints of in situ operations to generate realistic data and address the practical challenges of measurement

Variations of algal communities cause darkening of a Greenland glacier

We have assessed the microbial ecology on the surface of Mittivakkat glacier in SE-Greenland during the exceptional high melting season in July 2012 when the so far most extreme melting rate for the Greenland Ice Sheet has been recorded. By employing a complementary and multi-disciplinary field sampling and analytical approach, we quantified the dramatic changes in the different microbial surface habitats (green snow, red snow, biofilms, grey ice, cryoconite holes). The observed clear change in dominant algal community and their rapidly changing cryo-organic adaptation inventory was linked to the high melting rate. The changes in carbon and nutrient fluxes between different microbial pools (from snow to ice, cryoconite holes and glacial forefronts) revealed that snow and ice algae dominate the net primary production at the onset of melting, and that they have the potential to support the cryoconite hole communities as carbon and nutrient sources. A large proportion of algal cells is retained on the glacial surface and temporal and spatial changes in pigmentation contribute to the darkening of the snow and ice surfaces. This implies that the fast, melt-induced algal growth has a high albedo reduction potential, and this may lead to a positive feedback speeding up melting processes

Optimization of universal allogeneic CAR-T cells combining CRISPR and transposon-based technologies for treatment of acute myeloid leukemia

Despite the potential of CAR-T therapies for hematological malignancies, their efficacy in patients with relapse and refractory Acute Myeloid Leukemia has been limited. The aim of our study has been to develop and manufacture a CAR-T cell product that addresses some of the current limitations. We initially compared the phenotype of T cells from AML patients and healthy young and elderly controls. This analysis showed that T cells from AML patients displayed a predominantly effector phenotype, with increased expression of activation (CD69 and HLA-DR) and exhaustion markers (PD1 and LAG3), in contrast to the enriched memory phenotype observed in healthy donors. This differentiated and more exhausted phenotype was also observed, and corroborated by transcriptomic analyses, in CAR-T cells from AML patients engineered with an optimized CAR construct targeting CD33, resulting in a decreased in vivo antitumoral efficacy evaluated in xenograft AML models. To overcome some of these limitations we have combined CRISPR-based genome editing technologies with virus-free gene-transfer strategies using Sleeping Beauty transposons, to generate CAR-T cells depleted of HLA-I and TCR complexes (HLA-IKO/TCRKO CAR-T cells) for allogeneic approaches. Our optimized protocol allows one-step generation of edited CAR-T cells that show a similar phenotypic profile to non-edited CAR-T cells, with equivalent in vitro and in vivo antitumoral efficacy. Moreover, genomic analysis of edited CAR-T cells revealed a safe integration profile of the vector, with no preferences for specific genomic regions, with highly specific editing of the HLA-I and TCR, without significant off-target sites. Finally, the production of edited CAR-T cells at a larger scale allowed the generation and selection of enough HLA-IKO/TCRKO CAR-T cells that would be compatible with clinical applications. In summary, our results demonstrate that CAR-T cells from AML patients, although functional, present phenotypic and functional features that could compromise their antitumoral efficacy, compared to CAR-T cells from healthy donors. The combination of CRISPR technologies with transposon-based delivery strategies allows the generation of HLA-IKO/TCRKO CAR-T cells, compatible with allogeneic approaches, that would represent a promising option for AML treatment

Hyperoxemia and excess oxygen use in early acute respiratory distress syndrome : Insights from the LUNG SAFE study

Publisher Copyright: © 2020 The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Background: Concerns exist regarding the prevalence and impact of unnecessary oxygen use in patients with acute respiratory distress syndrome (ARDS). We examined this issue in patients with ARDS enrolled in the Large observational study to UNderstand the Global impact of Severe Acute respiratory FailurE (LUNG SAFE) study. Methods: In this secondary analysis of the LUNG SAFE study, we wished to determine the prevalence and the outcomes associated with hyperoxemia on day 1, sustained hyperoxemia, and excessive oxygen use in patients with early ARDS. Patients who fulfilled criteria of ARDS on day 1 and day 2 of acute hypoxemic respiratory failure were categorized based on the presence of hyperoxemia (PaO2 > 100 mmHg) on day 1, sustained (i.e., present on day 1 and day 2) hyperoxemia, or excessive oxygen use (FIO2 ≥ 0.60 during hyperoxemia). Results: Of 2005 patients that met the inclusion criteria, 131 (6.5%) were hypoxemic (PaO2 < 55 mmHg), 607 (30%) had hyperoxemia on day 1, and 250 (12%) had sustained hyperoxemia. Excess FIO2 use occurred in 400 (66%) out of 607 patients with hyperoxemia. Excess FIO2 use decreased from day 1 to day 2 of ARDS, with most hyperoxemic patients on day 2 receiving relatively low FIO2. Multivariate analyses found no independent relationship between day 1 hyperoxemia, sustained hyperoxemia, or excess FIO2 use and adverse clinical outcomes. Mortality was 42% in patients with excess FIO2 use, compared to 39% in a propensity-matched sample of normoxemic (PaO2 55-100 mmHg) patients (P = 0.47). Conclusions: Hyperoxemia and excess oxygen use are both prevalent in early ARDS but are most often non-sustained. No relationship was found between hyperoxemia or excessive oxygen use and patient outcome in this cohort. Trial registration: LUNG-SAFE is registered with ClinicalTrials.gov, NCT02010073publishersversionPeer reviewe

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality