Compositional Evolution of the Variscan Intra-Orogenic Extensional Magmatism in the Valencia del Ventoso Plutonic Complex, Ossa-Morena Zone (SW Iberia): A View from Amphibole Compositional Relationships

The Ossa-Morena Zone (OMZ), SW Iberia, has numerous Lower Carboniferous compositionally zoned plutons that formed in a Variscan intra-orogenic extensional setting. This magmatism shows a wide compositional variation comprising alkaline, transitional, and calc-alkaline suites. The calc-alkaline suite was produced by hybridization of alkaline magmas with felsic melts generated by crustal anatexis related to the intrusion of mafic magmas in the middle crust. In this work, we present a textural and mineralogical study of the Variscan Valencia del Ventoso main pluton from the OMZ to track the compositional evolution of magmas during hybridization using constraints from amphibole compositions and to determine the P-T conditions of emplacement using amphibolebased thermobarometry. This pluton exhibits reverse zoning with an inner facies containing alkaline dolerites, gabbros, and quartz diorites, an intermediate facies with transitional diorites, and an outer facies with calc-alkaline quartz diorites to monzogranites. Magmas from the intermediate and border facies crystallized under oxidizing conditions at relatively low temperatures (range: 640–760 ◦C) and ca. 280–300 MPa, implying near H2O-saturated conditions. These rock facies show mineralogical evidence of hybridization between alkaline to mildly alkalic and calc-alkaline magmas. The former is inferred from the occurrence of antecrysts of labradorite-andesine, high-Ti pargasite-hastingsite, and biotite with deficiency in tetrahedral-site occupancy, a distinctive feature of biotite from the inner facies alkaline dolerites. This contrasts with later crystallization from the calc-alkaline magma of andesine-oligoclase, low-Ti magnesiohornblende-edenite, and biotite with full tetrahedral-site occupancy. Constraints from amphibole-melt compositional relationships in antecrystic high-Ti amphibole suggest that the alkaline magmatic component could have a high- to ultra-K affinity.Ministerio de Economía y Competitividad (Gobierno de España), CGL2017-84469-

Deep Phenotypic Characterisation of CTCs by Combination of Microfluidic Isolation (IsoFlux) and Imaging Flow Cytometry (ImageStream)

Ines Aznar-Peralta holds a "Garantia Juvenil" fellowship (contract number 8040), and M. Carmen Garrido-Navas has a postdoctoral fellowship funded by the Ministry of Economy, Competitiveness, Enterprises and Universities (DOC_01682).The isolation of circulating tumour cells (CTCs) in colorectal cancer (CRC) mostly relies on the expression of epithelial markers such as EpCAM, and phenotypic characterisation is usually performed under fluorescence microscopy with only one or two additional markers. This limits the ability to detect different CTC subpopulations based on multiple markers. The aim of this work was to develop a novel protocol combining two platforms (IsoFluxTM and ImageStream®X) to improve CTC evaluation. Cancer cell lines and peripheral blood from healthy donors were used to evaluate the efficiency of each platform independently and in combination. Peripheral blood was extracted from 16 early CRC patients (before loco-regional surgery) to demonstrate the suitability of the protocol for CTC assessment. Additionally, peripheral blood was extracted from nine patients one month after surgery to validate the utility of our protocol for identifying CTC subpopulation changes over time. Results: Our protocol had a mean recovery efficiency of 69.5% and a limit of detection of at least four cells per millilitre. We developed an analysis method to reduce noise from magnetic beads used for CTC isolation. CTCs were isolated from CRC patients with a median of 37 CTCs (IQ 13.0–85.5) at baseline. CTCs from CRC patients were significantly (p < 0.0001) larger than cytokeratin (CK)-negative cells, and patients were stratified into two groups based on BRAFV600E and PD-L1 expression on CK-positive cells. The changes observed over time included not only the number of CTCs but also their distribution into four different subpopulations defined according to BRAFV600E and PD-L1 positivity. We developed a novel protocol for semi-automatic CTC isolation and phenotypic characterisation by combining two platforms. Assessment of CTCs from early CRC patients using our protocol allowed the identification of two clusters of patients with changing phenotypes over time."Garantia Juvenil" fellowship 8040Ministry of Economy, Competitiveness, Enterprises and Universities DOC_0168

Lu-Hf ratios of crustal rocks and their bearing on zircon Hf isotope model ages: The effects of accessories

All other factors being equal, the calculation of zircon Hf two stage model ages (TDM Hf) depends on the par- ticular Lu/Hf value assumed for the magmatic source, the effect being more pronounced as the age difference between zircon and magmatic source increases. It is generally considered that the Lu/Hf measured in the zircon- hosting rock does not represent the composition of the source because of potential garnet or zircon fractionation. Accordingly, most authors either assume a single fixed value for Lu/Hfsource, often Lu/Hf ≈ 0.079 to 0.108, or use two alternative models, one for felsic sources, often Lu/Hf ≈ 0.09, and the other for mafic sources, often Lu/ Hf ≈ 0.165. In contrast with these opinions, however, here we show that partial melting of peraluminous sources causes little decoupling of Lu from Hf because of similar solubilities of zircon and monazite. Furthermore, the effects of residual garnet are largely compensated by the numerous zircon inclusions that garnet and other residual minerals almost always contain. Partial melting of metaluminous sources may sig- nificantly decouple Lu from Hf if allanite and/or titanite are not present in the source, but the effect decreases as the melt fraction increases. Similarly, fractional crystallization of metaluminous magmas may decouple Lu from Hf if amphibole or clinopyroxene begin to crystallize before zircon saturation. The Lu/Hf distribution in 4784 rocks from different regions and ages is lognomal rather than normal, and the calculated medians, i.e. the maximum of the probability density function for the logarithmically transformed Lu/Hf, are Lu/Hfmafic rocks ≈ 0.08, Lu/Hffelsic rocks ≈ 0.05, i.e. notably lower than the above-mentioned felsic and mafic magmatic source averages. Magmatic sources may be remarkably heterogeneous with respect to Lu/Hf. Our calculations show that fixed Lu/Hfsource values translate the Lu/Hf heterogeneity of the source to the TDM Hf thus producing an artificial distribution of model ages that may be erroneously interpreted as different episodes of crustal growth. Therefore, we propose that the best strategy to calculate two stage Hf model ages of zircon is to use the analytically determined whole-rock Lu/Hf ratio as a proxy of the source. In the case of detrital or inherited zircons, for which no whole-rock information is available, it is advisable first to determine whether they come from a mafic or felsic rock by interpreting cathodoluminescence images, Th/U ratios and other chemical parameters, and then venture an estimate of the Lu/Hfsource from the SiO2 average

Precise Measurement of Magnesium Isotopes in Fe-Mg Minerals Using a Multi-collector SHRIMP Ion Microprobe

The distribution of Mg isotopes in minerals is becoming increasingly relevant in Earth science. Usually, they are determined by dissolving mineral concentrates and, after purifying Mg with ion exchange resins, analysing the resulting solutions by TIMS or, most often, MC-ICP-MS. When applied to individual minerals, these methods are slow and prone to contamination from impurities in the concentrates, inconveniences that may be avoided using spot analysis techniques such as LA-MC-ICP-MS or SIMS, albeit at the price of a large instrumental mass fractionation (IMF) and isobaric interferences, most prominent in the former. Here, we studied the potential of the multi-collector SHRIMP II ion microprobe for measuring Mg isotopes in Fe-Mg silicates and oxides. We found that, when corrected for the divergence of the Mg ion paths within the sample chamber caused by the Earth’s magnetic field, the SHRIMP’s IMF overwhelmingly depends on the mineral species, and the effects of variable chemical composition are negligible. We propose that the IMF is caused by the force constant difference, ΔF, between "hard" and "soft" bonds linking the ions of the studied element to the mineral lattice. Given that ΔF is a constant for each mineral species, we calculated IMF-correction factors for the most common Mg-bearing minerals. The thus-calculated correction factors permit the analysis in the same session, and with reasonable accuracy (within ~0.3‰ of the δ26Mg determined by SN-MC-ICP-MS analyses of concentrates), of samples from different mineral species, facilitating the application of Mg isotopes to terrestrial studies.Andalucian-FEDER grant P18-FR-1696Spanish grant PID2020-114872GB-I0

Hepatocellular carcinoma risk-stratification based on ASGR1 in circulating epithelial cells for cancer interception

Purpose: Lack of diagnostic and prognostic biomarkers in hepatocellular carcinoma impedes stratifying patients based on their risk of developing cancer. The aim of this study was to evaluate phenotypic and genetic heterogeneity of circulating epithelial cells (CECs) based on asialoglycoprotein receptor 1 (ASGR1) and miR-122-5p expression as potential diagnostic and prognostic tools in patients with hepatocellular carcinoma (HCC) and liver cirrhosis (LC). Methods: Peripheral blood samples were extracted from LC and HCC patients at different disease stages. CECs were isolated using positive immunomagnetic selection. Genetic and phenotypic characterization was validated by double immunocytochemistry for cytokeratin (CK) and ASGR1 or by in situ hybridization with miR-122-5p and CECs were visualized by confocal microscopy. Results: The presence of CECs increased HCC risk by 2.58-fold, however, this was only significant for patients with previous LC (p = 0.028) and not for those without prior LC (p = 0.23). Furthermore, the number of CECs lacking ASGR1 expression correlated significantly with HCC incidence and absence of miR-122-5p expression (p = 0.014; r = 0.23). Finally, overall survival was significantly greater for patients at earlier cancer stages (p = 0.018), but this difference was only maintained in the group with the presence of CECs (p = 0.021) whereas progression-free survival was influenced by the absence of ASGR1 expression. Conclusion: Identification and characterization of CECs by ASGR1 and/or miR- 122-5p expression may be used as a risk-stratification tool in LC patients, as it was shown to be an independent prognostic and risk-stratification marker in LC and early disease stage HCC patients.Regional Ministry of Health of AndalusiaMinistry of Economy, Competitiveness, Enterprises and Universities PC-0522-2016 PC-0267-2017 PC-0033-2017Health Institute Carlos III (ISCIII) DOC_01682 CD18/0012

Starkeya nomas sp. nov., a prosthecate and budding bacterium isolated from an immunocompromized patient

Strain HF14-78462T is an environmental bacterium found in clinical samples from an immunocompromized patient in 2014 at Hospital Universitari i Politècnic La Fe (Valencia, Spain). Phenotypically, strain HF14-78462T cells were Gram-stain-negative, aerobic, non-spore forming and non-motile small rods which formed mucous and whitish-translucent colonies when incubated at 20-36 °C. Phylogenetic analyses based on the 16S rRNA genes and the whole genomes of closest sequenced relatives confirmed that strain HF14-78462T is affiliated with the genus Starkeya. The strain was oxidase, catalase and urease positive; but indole, lysine decarboxylase, ornithine decarboxylase and DNase negative, did not produce H2S and was able to utilize a wide variety of carbon sources including acetamide, adonitol, amygdalin, l-arabinose, citric acid, glucose, mannitol and melibiose. Unlike Starkeya novella and Starkeya koreensis, strain HF14-78462T failed to grow in thiosulphate-oxidizing media and had a narrower temperature growth range. Its genome was characterized by a size of 4.83 Mbp and a C+G content of 67.75 mol%. Major fatty acids were C18:1 ω7c, cyclo C19 : 0 and C16 : 0, its polar acids were diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol and an aminophospholipid; while the ubiquinones were Q9 (1.8 %) and Q10 (98.2 %). Digital DNA-DNA hybridization values were 41 and 41.4 against S. novella and S. koreensis, respectively, while average nucleotide identity values were around 84 %. Phenotypic, average nucleotide identity and phylogenomic comparative studies suggest that strain HF14-78462T is a new representative of the genus Starkeya and the name Starkeya nomas sp. nov. is proposed. The type strain is HF14-78462T (=CECT 30124T=LMG 31874T).Financial support was obtained by the IIS project 2013/0437.S

Allele and haplotype frequencies of HLA-A, -B, -C, -DRB1, -DQB1 and -DQA1 in Castile and Leon region from North West of Spain

HLA studies have been used to determine the admixture of different populations within the Iberian Peninsula including neighbouring regions with shared origins, such as Portugal and Castile and Leon. These studies certainly can be used to study human migration that could establish populations currently settled according to genetic distant analysis based on the HLA diversity and language variety.This work was supported by the “Gerencia Regional de Salud de Castilla y Leon” (GRS 2080/A/19, 2019) and (GRS COVID 70/A/20, 2020)

Using Interpretable Machine Learning to Identify Baseline Predictive Factors of Remission and Drug Durability in Crohn’s Disease Patients on Ustekinumab

Ustekinumab has shown efficacy in Crohn's Disease (CD) patients. To identify patient profiles of those who benefit the most from this treatment would help to position this drug in the therapeutic paradigm of CD and generate hypotheses for future trials. The objective of this analysis was to determine whether baseline patient characteristics are predictive of remission and the drug durability of ustekinumab, and whether its positioning with respect to prior use of biologics has a significant effect after correcting for disease severity and phenotype at baseline using interpretable machine learning. Patients' data from SUSTAIN, a retrospective multicenter single-arm cohort study, were used. Disease phenotype, baseline laboratory data, and prior treatment characteristics were documented. Clinical remission was defined as the Harvey Bradshaw Index <= 4 and was tracked longitudinally. Drug durability was defined as the time until a patient discontinued treatment. A total of 439 participants from 60 centers were included and a total of 20 baseline covariates considered. Less exposure to previous biologics had a positive effect on remission, even after controlling for baseline disease severity using a non-linear, additive, multivariable model. Additionally, age, body mass index, and fecal calprotectin at baseline were found to be statistically significant as independent negative risk factors for both remission and drug survival, with further risk factors identified for remission

Long-Term Real-World Effectiveness and Safety of Ustekinumab in Crohn’s Disease Patients: The SUSTAIN Study

Background Large real-world-evidence studies are required to confirm the durability of response, effectiveness, and safety of ustekinumab in Crohn’s disease (CD) patients in real-world clinical practice. Methods A retrospective, multicentre study was conducted in Spain in patients with active CD who had received ≥1 intravenous dose of ustekinumab for ≥6 months. Primary outcome was ustekinumab retention rate; secondary outcomes were to identify predictive factors for drug retention, short-term remission (week 16), loss of response and predictive factors for short-term efficacy and loss of response, and ustekinumab safety. Results A total of 463 patients were included. Mean baseline Harvey-Bradshaw Index was 8.4. A total of 447 (96.5%) patients had received prior biologic therapy, 141 (30.5%) of whom had received ≥3 agents. In addition, 35.2% received concomitant immunosuppressants, and 47.1% had ≥1 abdominal surgery. At week 16, 56% had remission, 70% had response, and 26.1% required dose escalation or intensification; of these, 24.8% did not subsequently reduce dose. After a median follow-up of 15 months, 356 (77%) patients continued treatment. The incidence rate of ustekinumab discontinuation was 18% per patient-year of follow-up. Previous intestinal surgery and concomitant steroid treatment were associated with higher risk of ustekinumab discontinuation, while a maintenance schedule every 12 weeks had a lower risk; neither concomitant immunosuppressants nor the number of previous biologics were associated with ustekinumab discontinuation risk. Fifty adverse events were reported in 39 (8.4%) patients; 4 of them were severe (2 infections, 1 malignancy, and 1 fever). Conclusions Ustekinumab is effective and safe as short- and long-term treatment in a refractory cohort of CD patients in real-world clinical practice