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Remdesivir (RDV) was the first antiviral drug approved by the FDA to treat severe coronavirus disease-2019 (COVID-19) patients. RDV inhibits SARS-CoV-2 replication by stalling the non structural protein 12 (nsp12) subunit of the RNA-dependent RNA polymerase (RdRp). No evidence of global widespread RDV-resistance mutations has been reported, however, defining genetic pathways to RDV resistance and determining emergent mutations prior and subsequent antiviral therapy in clinical settings is necessary. This study identified 57/149 (38.3%) patients who did not respond to one course (5-days) (n = 36/111, 32.4%) or prolonged (5 to 20 days) (n = 21/38, 55.3%) RDV therapy by subgenomic RNA detection. Genetic variants in the nsp12 gene were detected in 29/49 (59.2%) non responder patients by Illumina sequencing, including the de novo E83D mutation that emerged in an immunosuppressed patient after receiving 10 + 8 days of RDV, and the L838I detected at baseline and/or after prolonged RDV treatment in 9/49 (18.4%) non responder subjects. Although 3D protein modeling predicted no interference with RDV, the amino acid substitutions detected in the nsp12 involved changes on the electrostatic outer surface and in secondary structures that may alter antiviral response. It is important for health surveillance to study potential mutations associated with drug resistance as well as the benefit of RDV retreatment, especially in immunosuppressed patients and in those with persistent replication. IMPORTANCE This study provides clinical and microbiologic data of an extended population of hospitalized patients for COVID-19 pneumonia who experienced treatment failure, detected by the presence of subgenomic RNA (sgRNA). The genetic variants found in the nsp12 pharmacological target of RDV bring into focus the importance of monitoring emergent mutations, one of the objectives of the World Health Organization (WHO) for health surveillance. These mutations become even more crucial as RDV keeps being prescribed and new molecules are being repurposed for the treatment of COVID-19. The present article offers new perspectives for the clinical management of non responder patients treated and retreated with RDV and emphasizes the need of further research of the benefit of combinatorial therapies and RDV retreatment, especially in immunosuppressed patients with persistent replication after therapy. This work was financed by a Gilead Sciences grant (IN-ES-540-6089) and CIBER Enfermedades Infecciosas (CIBERINFEC), Instituto de Salud Carlos III, Madrid, España (CB21/13/00081). This work was financed by ad hoc patronage funds for research on COVID-19 from donations from citizens and organizations to the Hospital Clínic de Barcelona-Fundació Clínic per a la Recerca Biomèdica. Sí

We present high resolution spectra of comet C/2017 K2 (PanSTARRS) (hereafter 17K2), obtained using the upgraded high resolution spectrometer of the VLT, CRIRES+. We will show our findings in the (2.8 – 5.3) μm range, searching for primary volatiles (e.g., H2O, HCN, NH3, CO, C2H6, CH4, ...) and studying their evolution as the comet approach the Sun. 17K2 is a long period comet, very active already at record heliocentric distances of 16 au, and represents a unique opportunity to study the composition of a mostly unaltered comet. Comets formed from the material surrounding the proto-Sun about 4.6 billion years ago, and after their formation they were scattered into their current reservoirs [1,2], where the frozen nuclei have preserved most of the chemical and mineralogical properties linked to their formation site until today. Probing the chemical diversity in comets may thus unveil the processes that were in effect within the mid-plane of our proto-planetary disk, and test the hypothesis that comets may have contributed in delivering water and prebiotics to the early Earth [3]. Among other techniques, the composition of active comets can be studied from ground based telescopes using high resolution spectroscopy in the infrared (IR - 3 to 5 μm), where it is possible to observe emission lines produced by solar-pumped fluorescence of primary species, i.e., molecules released directly from the nucleus. High spectral and spatial resolutions are necessary to resolve different molecular species in the spectra, to study their distribution within the coma and to separate emission lines of the comet from their counterpart in the atmosphere. Comet 17K2 is in excellent observing conditions in 2022, allowing infrared high resolution studies. The comet shows already activity, probably driven by CO and other hyper-volatiles that can sublimate at distances from the Sun larger than 5 au [4,5]. Discovered in 2017 at about 16 au from the Sun [6], it is most likely entering the inner solar system for the first time, and its observation offers a unique opportunity to study its mostly unaltered material. We will present the results obtained from different spectra acquired using CRIRES+ at ESO-VLT at various epochs. We acquired comprehensive high-resolution spectra of the comet as it progressively moved towards the Sun, with the goal of monitoring the evolution of sublimating material with the heliocentric distance. In particular, we have granted time at the beginning of May, beginning of July, and end of August 2022, with the Sun-17K2 distance varying from about 3.5 to 2.3 au. In this heliocentric range, the comet is crossing the CO to H2O ice sublimation regime [7]. Data are reduced using custom semi-automated procedures (see [8] and references therein) that allow a fast analysis of the spectra. Spectral calibration and compensation for telluric absorption are achieved by comparing the data with highly accurate atmospheric radiance and transmittance models obtained with PUMAS/PSG [9]. Flux calibration is obtained using the spectra of a standard star observed closely in time with the comet, and reduced with the same algorithms. Production rates and relative abundances (i.e. mixing ratios with respect to water) of different primary species in the coma are obtained using state-of-the-art fluorescence models (see for example [10] and [11]). The molecular abundances found in this comet will be compared to reference median values retrieved for the comet population [12] and with the abundances found in other Oort Cloud Comets. References: 1. Gomes, R., et al., 2005, Nature, 435, 446 – 2. Morbidelli, A., et al., 2007, AJ, 134, 1790 – 3. Mumma, M. J., Charnley, S.B. 2011, Ann. Rev. Astron. Astroph., 49 – 4. Jewitt, D., et al., 2019, AJ, 157, 65 – 5. Yang, B., et al., 2021, ApJL, 914, L17 – 6. Wainscoat, R. J., et al., 2017, CBET, 4393, 1 – 7. Jewitt, D., et al., 2007, Protostars and Planets V. Univ. Arizona Press, Tucson, 863 – 8. Lippi, M., et al., 2020, AJ, 159, 157 – 9. Villanueva, G. L., et al., 2018, JQSRT, 217, 86 – 10. Villanueva, G. L., et al., 2012b, JQSRT, 113, 202 – 11. Villanueva, G. L., et al., 2011b, JGRE, 116, E08012 – 12. Lippi, M., et al., 2021, AJ, 162, 74.

NOMAD [1] (Nadir and Occultation for MArs Discovery) is a multi-channel spectrometer onboard the ExoMars 2016 Trace Gas Orbiter (TGO), which began its observations in April 2018. The Solar Occultation (SO) channel has a spectral range coverage from 2.3 to 4.3 μm (2320 to 4350 cm-1), where numerous ro-vibrational bands of H2O are located. An Acousto-Optical Tunable Filter (AOTF) is used to select different spectral windows (with a typical width from 20 to 35 cm-1) corresponding to the desired diffraction orders to be used during an atmospheric scan. The SO channel has a sample rate of ∼1 s, that allows a vertical sampling of typically 1 km. In order to obtain as much information as possible from the Martian atmosphere, for this study we have analyzed data from diffraction orders 134 (3011-3035 cm-1), 136 (3056-3081 cm-1), 168 (3775-3805 cm-1) and 169 (3798-3828 cm-1), taken during different solar occultation scans since the beginning of the science operations phase of the TGO/ExoMars mission. This combination of diffraction orders allowed us to obtain an optimal mapping of the distribution of the water vapor at altitudes from ∼10 km up to about ∼100 km and in different atmospheric conditions, and hence, permitting the study of its spatial and seasonal variability.The data presented here have been analyzed with pre-processing and cleaning tools developed entirely at the IAA (see companion contributions to this conference [2-4]), and then, inverted using a state-of-the-art retrieval scheme [5]. For the inversion, we have introduced a novel strategy of combining (when coincident in the same solar occultation) pairs of NOMAD orders (in contrast to a post-processing strategy), using orders 134 or 136 for the lower atmosphere, commonly below 60 km, and orders 168 or 169 for the upper atmosphere. This combination procedure allows us to exclude heavily saturated absorption lines present at low altitudes in orders 168-169, which contain strong absorption lines, permitting the exploitation of these orders for the analysis of the atmosphere up to 100 km. Here we present and discuss detailed maps showing the vertical distribution of the water vapor abundance of a subset of the solar occultations observed during the first two years of TGO/ExoMars. This research confirms some findings also obtained by other teams within the NOMAD consortium [6-8] and ACS instrument [9], showing that dust storms allow water vapor to reach higher altitudes in the atmosphere. This is important for the escape of atomic hydrogen [10] and therefore, for the long-term evolution of the Martian atmosphere. Detailed studies on the characterization of the Martian hygropause and saturation events are also presented. AcknowledgmentsThe IAA/CSIC team acknowledges financial support from the State Agency for Research of the Spanish MCIU through the 'Center of Excellence Severo Ochoa' award for the Instituto de Astrofísica de Andalucía (SEV- 2017-0709) and funding by grant PGC2018-101836-B-100 (MCIU/AEI/FEDER, EU). ExoMars is a space mission of the European Space Agency (ESA) and Roscosmos. The NOMAD experiment is led by the Royal Belgian Institute for Space Aeronomy (IASB-BIRA), assisted by Co-PI teams from Spain (IAA-CSIC), Italy (INAF-IAPS), and the United Kingdom (Open University). US investigators were supported by the National Aeronautics and Space Administration. References1. Vandaele, A. C. et al. NOMAD, an integrated suite of three spectrometers for the ExoMars Trace Gas mission: technical description, science objectives and expected performance. Space Science Reviews 214, 1-47 (2018).2. López-Valverde, M. A. et al. Temperature and density profiles from the two solar occultation instruments NOMAD and ACS during a full Martian Year. EPSC (2022).3. Modak, A. et al. Mapping of Martian CO from NOMAD solar occultation measurements for MY35 and 36. EPSC (2022).4. Stolzenbach, A. et al. Composition and size of Martian aerosols as seen in the IR by NOMAD-SO experiment onboard TGO. EPSC (2022).5. Jurado Navarro, A. A. et al. Retrieval of CO2 and collisional parameters from the MIPAS spectra in the earth atmosphere (2016).6. Aoki, S et al. Water vapor vertical profiles on Mars in dust storms observed by TGO/NOMAD. Journal of Geophysical Research: Planets (2019).7. Aoki, S. et al. Global vertical distribution of water vapor on Mars: Results from 3.5 years of ExoMarsTGO/NOMAD science operations. submitted (2022).8. Villanueva, G. L. et al. Water heavily fractionated as it ascends on Mars as revealed by ExoMars/NOMAD. Science Advances 7, eabc8843 (2021).9. Fedorova, A. A. et al. Stormy water on Mars: The distribution and saturation of atmospheric water during the dusty season. Science 367, 297-300 (2020).10. Chaffin, M., Deighan, J, Schneider, N. & Stewart, A. Elevated atmospheric escape of atomic hydrogen from Mars induced by high-altitude water. Nature geoscience 10, 174-178 (2017).

Protein citrullination (deimination) is the post-translational modification of arginine to the non-coded amino acid citrulline and is catalyzed by the peptidyl arginine deiminase (PADI) enzyme family. The most widely expressed of the PADIs family, PADI2, regulates cellular processes that impact several diseases. How PADI2 has evolved in mammals to gain its fundamental function is not fully understood. By performing a systematic evolutionary analysis, we now identify 20 positively selected residues in PADI2, 16 of which are structurally exposed mainly on the N-terminal and middle domains of PADI2. Our integrated evolutionary and structural analyses suggest that these residues have roles in maintaining the PADI2 interactions with cognate proteins. We experimentally demonstrate that one of the loops in the middle domain participates in the interactions of PADI2 with the positive-transcription elongation factor (P-TEFb), which is essential for active transcription and cellular proliferation. This interpretation is supported by our finding that L162 within this loop evolved under positive selection. Our work demonstrates the power of combining sequence-based phylogenetic methods with structural information for investigating the selective function of the middle domain of PADI2 in modulating transcription. This in-depth knowledge could be key to understanding the role of PADI2 overexpression in disease and points to potential targetable regions of the protein.Significance StatementA systematic evolutionary analysis identified positively selected residues in the non-catalytic domain of PADI2. We established a link between the positive evolution of key residues in the PADI2 and their role in transcription. Specifically, we identified the structurally exposed loop encompassing the positively selected L162 in the PADI2 middle domain and its role in transcription and cellular proliferation. This loop contributes to the PADI2 interaction with the P-TEFb complex and cellular proliferation. Our results showcase the use of combining evolutionary and experimental approaches to dissect the dynamic of evolutionary processes.

Spreading depolarization, usually termed cortical spreading depression has been proposed as the pathophysiological substrate of migraine aura and as an endogenous trigger of headache pain. The links between neurovascular coupling and cortical craniofacial nociceptive activities modulated by SD were assessed by combining in vivo local field potential (LFP) recordings in the primary somatosensory cortex (S1) with functional ultrasound imaging of S1 and caudal insular (INS) cortices of anesthetized male rats. A single SD wave triggered in the primary visual cortex elicited an ipsilateral, quadriphasic hemodynamic and electrophysiological response in S1 with an early phase consisting of concomitant increases of relative cerebral blood volume (rCBV) and LFPs. A transient hypoperfusion was then correlated with the beginning of the neuronal silence, followed by a strong increase of rCBV, whereas synaptic activities remained inhibited. LFPs and rCBV recovery period was followed by a progressive increase in S1 and INS baseline activities and facilitation of cortical responses evoked by periorbital cutaneous receptive field stimulation. Sensitization of cortical ophthalmic fields by SD was bilateral, occurred with precise spatiotemporal profiles, and was significantly reduced by pretreatment with an NMDA antagonist. Combined high-resolution assessing of neurovascular coupling and electrophysiological activities has revealed a useful preclinical tool for deciphering central sensitization mechanisms involved in migraine attacks. SIGNIFICANCE STATEMENT A crucial unsolved issue is whether visual aura and migraine headache are parallel or sequential processes. Here, we show that a single spreading depolarization wave triggered from the primary visual cortex is powerful enough to elicit progressive, sustained increases of hemodynamic and sensory responses to percutaneous periorbital noxious stimuli recorded in S1 and insular ophthalmic fields. Sensitization of cortical ophthalmic fields by SD was bilateral, occurred with precise spatiotemporal profiles, and was significantly reduced by pretreatment with an NMDA antagonist. Combined high-resolution assessing of neurovascular coupling and electrophysiological activities has revealed a useful preclinical tool for deciphering central sensitization mechanisms involved in migraine attacks.

We study the employment effects of the collapse of Mexican exports to the USA due to the Covid-19 crisis and its recovery. The employment outcomes are broken down by gender and skill, which enables the investigation to assess how these employment categories were affected. The article finds that 33.6% of all the jobs lost at the height of the Covid-19 pandemic in Mexico were exclusively due to the collapse of exports to the USA. Trade collapse was biased against female and unskilled workers, although all the jobs lost were re-gained during the recovery, being again biased against female workers. Gender-responsive policies are needed to ensure an inclusive recovery and resilience against future trade shocks.

Lethal congenital contracture syndrome 11 (LCCS11) is caused by homozygous or compound heterozygous variants in the GLDN gene on chromosome 15q21. GLDN encodes gliomedin, a protein required for the formation of the nodes of Ranvier and development of the human peripheral nervous system. We report a fetus with ultrasound alterations detected at 28 weeks of gestation. The fetus exhibited hydrops, short long bones, fixed limb joints, absent fetal movements, and polyhydramnios. The pregnancy was terminated and postmortem studies confirmed the prenatal findings: distal arthrogryposis, fetal growth restriction, pulmonary hypoplasia, and retrognathia. The fetus had a normal chromosomal microarray analysis. Exome sequencing revealed two novel compound heterozygous variants in the GLDN associated with LCCS11. This manuscript reports this case and performs a literature review of all published LCCS11 cases.

Neurological disorders (ND) are diseases that affect the brain and the central and autonomic nervous systems, such as neurodevelopmental disorders, cerebellar ataxias, Parkinson’s disease, or epilepsies. Nowadays, recommendations of the American College of Medical Genetics and Genomics strongly recommend applying next generation sequencing (NGS) as a first-line test in patients with these disorders. Whole exome sequencing (WES) is widely regarded as the current technology of choice for diagnosing monogenic ND. The introduction of NGS allows for rapid and inexpensive large-scale genomic analysis and has led to enormous progress in deciphering monogenic forms of various genetic diseases. The simultaneous analysis of several potentially mutated genes improves the diagnostic process, making it faster and more efficient. The main aim of this report is to discuss the impact and advantages of the implementation of WES into the clinical diagnosis and management of ND. Therefore, we have performed a retrospective evaluation of WES application in 209 cases referred to the Department of Biochemistry and Molecular Genetics of the Hospital Clinic of Barcelona for WES sequencing derived from neurologists or clinical geneticists. In addition, we have further discussed some important facts regarding classification criteria for pathogenicity of rare variants, variants of unknown significance, deleterious variants, different clinical phenotypes, or frequency of actionable secondary findings. Different studies have shown that WES implementation establish diagnostic rate around 32% in ND and the continuous molecular diagnosis is essential to solve the remaining cases.

To compare the short-term outcomes of patients undergoing open DP (ODP) and laparoscopic DP (LDP); and to analyze the association between the section site of the pancreas and pancreatic fistula.Clinical, perioperative, and histopathologic data of patients who underwent ODP and LDP between 2009 and 2019 were retrospectively analyzed.70 patients were included. 39 (56%) underwent ODP and 31 (44%) underwent LDP. The tumor size in ODP group was 70mm and in LDP group was 45mm (p = 0,032) Blood loss was lower in LDP group (229mL versus 498mL) (p = 0,001). Operative time, spleen preservation, B/C pancreatic fistula, major morbidity, reoperation, and length of hospital stay, were similar in both groups. There was no postoperative mortality. No differences were found in B/C pancreatic fistula rate regarding to pancreatic transection site.LDP is a safe procedure, with perioperative outcomes similar to ODP and with less blood loss. The pancreatic transection site did not influence post-operative pancreatic fistula rate.Comparar los resultados a corto plazo de pacientes intervenidos mediante pancreatectomía distal abierta (PDA) y laparoscópica (PDL); y analizar si el lugar de la sección del páncreas tiene relación con la formación de fístula pancreática.Serie retrospectiva y descriptiva de las PD realizadas, desde enero del 2009 a diciembre del 2019. Se compararon las características clínicas, perioperatorias e histopatológicas de pacientes con PDA y PDL.Se incluyeron 70 pacientes. Treinta y nueve casos (56%) con PDA y 31 casos (44%) con PDL. El tamaño tumoral promedio en la PDA fue de 70 mm y en la PDL 45 mm (p = 0.032). La pérdida sanguínea fue menor en la PDL (229 vs. 498 ml) (p = 0.001). No se encontró diferencia significativa en tiempo operatorio, porcentaje de preservación esplénica, fístula pancreática B/C, reoperación, morbilidad mayor y estancia hospitalaria. No hubo mortalidad postoperatoria. No hubo diferencias en la formación de fístula pancreática con respecto al lugar de sección del páncreas.La PDL es un procedimiento seguro, con resultados perioperatorios similares a la PDA y con menor pérdida sanguínea. El lugar de sección del páncreas no tuvo relación con la formación de fístula pancreática.

Background: The first interim analysis of the KEYNOTE-564 study showed improved disease-free survival with adjuvant pembrolizumab compared with placebo after surgery in patients with clear cell renal cell carcinoma at an increased risk of recurrence. The analysis reported here, with an additional 6 months of follow-up, was designed to assess longer-term efficacy and safety of pembrolizumab versus placebo, as well as additional secondary and exploratory endpoints. Methods: In the multicentre, randomised, double-blind, placebo-controlled, phase 3 KEYNOTE-564 trial, adults aged 18 years or older with clear cell renal cell carcinoma with an increased risk of recurrence were enrolled at 213 hospitals and cancer centres in North America, South America, Europe, Asia, and Australia. Eligible participants had an Eastern Cooperative Oncology Group performance status of 0 or 1, had undergone nephrectomy 12 weeks or less before randomisation, and had not received previous systemic therapy for advanced renal cell carcinoma. Participants were randomly assigned (1:1) via central permuted block randomisation (block size of four) to receive pembrolizumab 200 mg or placebo intravenously every 3 weeks for up to 17 cycles. Randomisation was stratified by metastatic disease status (M0 vs M1), and the M0 group was further stratified by ECOG performance status and geographical region. All participants and investigators involved in study treatment administration were masked to the treatment group assignment. The primary endpoint was disease-free survival by investigator assessment in the intention-to-treat population (all participants randomly assigned to a treatment). Safety was assessed in the safety population, comprising all participants who received at least one dose of pembrolizumab or placebo. As the primary endpoint was met at the first interim analysis, updated data are reported without p values. This study is ongoing, but no longer recruiting, and is registered with ClinicalTrials.gov, NCT03142334. Findings: Between June 30, 2017, and Sept 20, 2019, 994 participants were assigned to receive pembrolizumab (n=496) or placebo (n=498). Median follow-up, defined as the time from randomisation to data cutoff (June 14, 2021), was 30·1 months (IQR 25·7–36·7). Disease-free survival was better with pembrolizumab compared with placebo (HR 0·63 [95% CI 0·50–0·80]). Median disease-free survival was not reached in either group. The most common all-cause grade 3–4 adverse events were hypertension (in 14 [3%] of 496 participants) and increased alanine aminotransferase (in 11 [2%]) in the pembrolizumab group, and hypertension (in 13 [3%] of 498 participants) in the placebo group. Serious adverse events attributed to study treatment occurred in 59 (12%) participants in the pembrolizumab group and one (

Background: The Sequential Organ Failure Assessment (SOFA) is a scoring system used for the evaluation of disease severity and prognosis of critically ill patients. The impedance ratio (Imp-R) is a novel mortality predictor. Aims: This study aimed to evaluate the combination of the SOFA + Imp-R in the prediction of mortality in critically ill patients admitted to the Emergency Department (ED). Methods: A retrospective cohort study was performed in adult patients with acute illness admitted to the ED of a tertiary-care referral center. Baseline SOFA score and bioelectrical impedance analysis to obtain the Imp-R were performed within the first 24 h after admission to the ED. A Cox regression analysis was performed to evaluate the mortality risk of the initial SOFA score plus the Imp-R. Harrell’s C-statistic and decision curve analyses (DCA) were performed. Results: Out of 325 patients, 240 were included for analysis. Overall mortality was 31.3%. Only 21.3% of non-surviving patients died after hospital discharge, and 78.4% died during their hospital stay. Of the latter, 40.6% died in the ED. The SOFA and Imp-R values were higher in non-survivors and were significantly associated with mortality in all models. The combination of the SOFA + Imp-R significantly predicted 30-day mortality, in-hospital mortality, and ED mortality with an area under the curve (AUC) of 0.80 (95% CI: 74–0.86), 0.79 (95% CI: 0.74–0.86) and 0.75 (95% CI: 0.66–0.84), respectively. The DCA showed that combining the SOFA + Imp-R improved the prediction of mortality through the lower risk thresholds. Conclusions: The addition of the Imp-R to the baseline SOFA score on admission to the ED improves mortality prediction in severely acutely ill patients admitted to the ED.

SUMMARY OBJECTIVE: The aim of this study was to describe the medical nutritional therapy (MNT) of adult non-critically ill hospitalization patients. METHODS: In a retrospective study, adults hospitalized for more than 48 h in non-intensive care unit medical and surgical areas that were classified as being at nutritional risk were included. Malnutrition was defined according to Global Leadership Initiative on Malnutrition (GLIM) criteria. RESULTS: A total of 255 patients, aged 54.13±18.4 years, who were at risk of malnutrition were included in this study. Of these, 50% were males. Notably, 52.5% received oral nutrition supplementation (ONS), 23.5% enteral nutrition (EN), 15% parenteral nutrition (PN), and 9% received enteral and parenteral nutrition (EPN). Patients with EPN presented the highest frequency of malnutrition (52%), and therefore they received more than 100% of energy and protein requirements. The median length of stay was 25 days. Among patients with nutritional risk receiving EPN, no deaths occurred. Patients, identified at nutritional risk, but without malnutrition according to GLIM, and receiving ONS had significantly lower mortality than patients receiving other MNT. CONCLUSIONS: Oral nutrition supplementation was the more frequent MNT prescribed. The frequency of malnutrition and percentage of prescribed energy and protein were higher in patients receiving PN and EPN compared with those receiving ONS.

Cancer patient care requires a multi-disciplinary approach and multiple medical and ethical considerations. Clinical care during a pandemic health crisis requires prioritising the use of resources for patients with a greater chance of survival, especially in developing countries. The coronavirus disease 2019 crisis has generated new challenges given that cancer patients are normally not prioritised for admission in critical care units. Nevertheless, the development of new cancer drugs and novel adjuvant/neoadjuvant protocols has dramatically improved the prognosis of cancer patients, resulting in a more complex decision-making when prioritising intensive care in pandemic times. In this context, it is essential to establish an effective and transparent communication between the oncology team, critical care, and emergency units to make the best decisions, considering the principles of justice and charity. Concurrently, cancer treatment protocols must be adapted to prioritise according to oncologic response and prognosis. Communication technologies are powerful tools to optimise cancer care during pandemics, and we must adapt quickly to this new scenario of clinical care and teaching. In this new challenging pandemic scenario, multi-disciplinary work and effective communication between clinics, technology, science, and ethics is the key to optimising clinical care of cancer patients.

Background Fluid overload (FO) in critically ill patients is associated with increased adverse events. This study aims to compare different bioelectrical impedance analysis (BIA) parameters that demonstrate FO and their association with 30-day mortality in critical patients admitted to the emergency department (ED). Methods Five components of the BIA were obtained by multifrequency device-total body water (TBW), extracellular water (ECW), intracellular water (ICW), resistance (R), and reactance (Xc)-to calculate parameters (impedance vectors, impedance ratio, and the ratios of ECW to TBW, ECW to ICW, ECW to body surface area, TBW to height2 , ICW to height2 , Xc to height, and R to height) that have been used for the detection of FO. A concordance analysis (κ) was performed comparing every parameter with each other. Furthermore, different regression models (Cox regression) were created associating the FO for each parameter with 30-day mortality, adjusted for age, body mass index, sex, Sequential Organ Failure Assessment score, and serum albumin level. Results A total of 142 patients were included in the study. Only FO by impedance vector analysis (relative risk [RR] = 6.4; 95% CI, 1.5-27.9; P = .01), impedance ratio (RR = 2.7; 95% CI, 1.1-7.1; P = .04), and R (RR = 2.6; 95% CI, 1.2-5.5; P = .02) increased the probability of 30-day mortality. Conclusions Different parameters that determine FO by BIA were associated with the mortality of patients admitted to the ED, but the impedance vector analysis was superior to any other parameter of the BIA.

Estrogen (E2) and Progesterone (Pg), via their specific receptors (ERalpha and PR), are major determinants in the development and progression of endometrial carcinomas, However, their precise mechanism of action and the role of other transcription factors involved are not entirely clear. Using Ishikawa endometrial cancer cells, we report that E2 treatment exposes a set of progestin-dependent PR binding sites which include both E2 and progestin target genes. ChIP-seq results from hormone-treated cells revealed a non-random distribution of PAX2 binding in the vicinity of these estrogen-promoted PR sites. Altered expression of hormone regulated genes in PAX2 knockdown cells suggests a role for PAX2 in fine-tuning ERalpha and PR interplay in transcriptional regulation. Analysis of long-range interactions by Hi-C coupled with ATAC-seq data showed that these regions, that we call ‘progestin control regions’ (PgCRs), exhibited an open chromatin state even before hormone exposure and were non-randomly associated with regulated genes. Nearly 20% of genes potentially influenced by PgCRs were found to be altered during progression of endometrial cancer. Our findings suggest that endometrial response to progestins in differentiated endometrial tumor cells results in part from binding of PR together with PAX2 to accessible chromatin regions. What maintains these regions open remains to be studied., Consejo Nacional de Investigaciones Científicas y Técnicas (PIP 2015-682) Patricia Saragüeta. Fondo para la Investigación Científica y Tecnológica (PICT 2015-3426). Patricia Saragüeta H2020 European Research Council (FP7/2007-2013 grant agreement 609989)

BACKGROUND: Pembrolizumab is active in head and neck squamous cell carcinoma (HNSCC), with programmed cell death ligand 1 (PD-L1) expression associated with improved response.METHODS: KEYNOTE-048 was a randomised, phase 3 study of participants with untreated locally incurable recurrent or metastatic HNSCC done at 200 sites in 37 countries. Participants were stratified by PD-L1 expression, p16 status, and performance status and randomly allocated (1:1:1) to pembrolizumab alone, pembrolizumab plus a platinum and 5-fluorouracil (pembrolizumab with chemotherapy), or cetuximab plus a platinum and 5-fluorouracil (cetuximab with chemotherapy). Investigators and participants were aware of treatment assignment. Investigators, participants, and representatives of the sponsor were masked to the PD-L1 combined positive score (CPS) results; PD-L1 positivity was not required for study entry. The primary endpoints were overall survival (time from randomisation to death from any cause) and progression-free survival (time from randomisation to radiographically confirmed disease progression or death from any cause, whichever came first) in the intention-to-treat population (all participants randomly allocated to a treatment group). There were 14 primary hypotheses: superiority of pembrolizumab alone and of pembrolizumab with chemotherapy versus cetuximab with chemotherapy for overall survival and progression-free survival in the PD-L1 CPS of 20 or more, CPS of 1 or more, and total populations and non-inferiority (non-inferiority margin: 1·2) of pembrolizumab alone and pembrolizumab with chemotherapy versus cetuximab with chemotherapy for overall survival in the total population. The definitive findings for each hypothesis were obtained when statistical testing was completed for that hypothesis; this occurred at the second interim analysis for 11 hypotheses and at final analysis for three hypotheses. Safety was assessed in the as-treated population (all participants who received at least one dose of allocated treatment). This study is registered at ClinicalTrials.gov, number NCT02358031.FINDINGS: Between April 20, 2015, and Jan 17, 2017, 882 participants were allocated to receive pembrolizumab alone (n=301), pembrolizumab with chemotherapy (n=281), or cetuximab with chemotherapy (n=300); of these, 754 (85%) had CPS of 1 or more and 381 (43%) had CPS of 20 or more. At the second interim analysis, pembrolizumab alone improved overall survival versus cetuximab with chemotherapy in the CPS of 20 or more population (median 14·9 months vs 10·7 months, hazard ratio [HR] 0·61 [95% CI 0·45-0·83], p=0·0007) and CPS of 1 or more population (12·3 vs 10·3, 0·78 [0·64-0·96], p=0·0086) and was non-inferior in the total population (11·6 vs 10·7, 0·85 [0·71-1·03]). Pembrolizumab with chemotherapy improved overall survival versus cetuximab with chemotherapy in the total population (13·0 months vs 10·7 months, HR 0·77 [95% CI 0·63-0·93], p=0·0034) at the second interim analysis and in the CPS of 20 or more population (14·7 vs 11·0, 0·60 [0·45-0·82], p=0·0004) and CPS of 1 or more population (13·6 vs 10·4, 0·65 [0·53-0·80], pINTERPRETATION: Based on the observed efficacy and safety, pembrolizumab plus platinum and 5-fluorouracil is an appropriate first-line treatment for recurrent or metastatic HNSCC and pembrolizumab monotherapy is an appropriate first-line treatment for PD-L1-positive recurrent or metastatic HNSCC.FUNDING: Merck Sharp & Dohme.

In breast cancer cells, some topologically associating domains (TADs) behave as hormonal gene regulation units, within which gene transcription is coordinately regulated in response to steroid hormones. Here we further describe that responsive TADs contain 20- to 100-kb-long clusters of intermingled estrogen receptor (ESR1) and progesterone receptor (PGR) binding sites, hereafter called hormone-control regions (HCRs). In T47D cells, we identified more than 200 HCRs, which are frequently bound by unliganded ESR1 and PGR. These HCRs establish steady long-distance inter-TAD interactions between them and organize characteristic looping structures with promoters in their TADs even in the absence of hormones in ESR1-PGR cells. This organization is dependent on the expression of the receptors and is further dynamically modulated in response to steroid hormones. HCRs function as platforms that integrate different signals, resulting in some cases in opposite transcriptional responses to estrogens or progestins. Altogether, these results suggest that steroid hormone receptors act not only as hormone-regulated sequence-specific transcription factors but also as local and global genome organizers., Spanish Ministry of Economy and Competitiveness, ‘Centro de Excelencia Severo Ochoa 2013–2017’ and Plan Nacional (SAF2016-75006-P), as well as support of the CERCA Programme/ Generalitat de Catalunya

Background Gastric cancer is the fifth cause of cancer incidence worldwide. Multidisciplinary approaches that improve the survival are needed. Perioperative chemotherapies show improvement in pathological complete remission (pCR) and overall survival (OS), but less than 50% of the patients completed the chemotherapeutic regimen. The recent 5-fluorouracil, leucovorin, oxaliplatin, docetaxel-4 (FLOT4) study shows OS 50 months and pCR 16.6%, but only 46% of the patients completed pre- and postoperative treatment. This case series report evaluated pCR and safety in patients that received complete preoperative chemotherapeutic with FLOT. Methods Patients received eight cycles FLOT regimen before surgery. Each cycle comprised 50 mg/m2 docetaxel intravenous (iv) on day 1, 85 mg/m2 oxaliplatin iv on day 1, 200 mg/m2 leucovorin iv on day 1 and 2,600 mg/m2 5-fluorouracil iv in a 24-hour infusion on day 1, every 2 weeks. Results Fifty-nine patients were evaluated, 58 patients received preoperative cycles. Thirty-one patients received all eight cycles of preoperative therapy. 65.5% patients presented any major adverse event. Thirty-nine patients underwent surgery. Thirty-three biopsy reports were obtained. Six patients (18.2%) presented pCR, 13 patients (39.4%) had no lymph node involvement. OS was 21.32 months. Patients with histology of signet ring carcinoma cells had a shorter survival than other histologies. Conclusion Total neoadjuvant with FLOT chemotherapy presents an adequate safety profile, a similar pathologic regression rate, and a slightly higher rate of completing treatment to report in perioperative FLOT regimen studies. A prospective clinical study with suitable diagnostic, staging tools and an adequate follow-up may prove total neoadjuvant chemotherapy's efficacy.

The aim of this study was to generate and analyze the atlas of the loggerhead turtle blood transcriptome by RNA-seq, as well as identify and characterize thioredoxin (Tnxs) and peroxiredoxin (Prdxs) antioxidant enzymes of the greatest interest in the control of peroxide levels and other biological functions. The transcriptome of loggerhead turtle was sequenced using the Illumina Hiseq 2000 platform and de novo assembly was performed using the Trinity pipeline. The assembly comprised 515,597 contigs with an N50 of 2,631 bp. Contigs were analyzed with CD-Hit obtaining 374,545 unigenes, of which 165,676 had ORFs encoding putative proteins longer than 100 amino acids. A total of 52,147 (31.5%) of these transcripts had significant homology matches in at least one of the five databases used. From the enrichment of GO terms, 180 proteins with antioxidant activity were identified, among these 28 Prdxs and 50 putative Tnxs. The putative proteins of loggerhead turtles encoded by the genes Prdx1, Prdx3, Prdx5, Prdx6, Txn and Txnip were predicted and characterized in silico. When comparing Prdxs and Txns of loggerhead turtle with homologous human proteins, they showed 18 (9%), 52 (18%) 94 (43%), 36 (16%), 35 (33%) and 74 (19%) amino acid mutations respectively. However, they showed high conservation in active sites and structural motifs (98%), with few specific modifications. Of these, Prdx1, Prdx3, Prdx5, Prdx6, Txn and Txnip presented 0, 25, 18, three, six and two deleterious changes. This study provides a high quality blood transcriptome and functional annotation of loggerhead sea turtles.

Microbial fuel cells (MFCs) can convert chemical energy into electrical current by utilizing the biocatalytic microbes, capable to use a wide range of organic matters as fuel feedstocks. The MFCs can mitigate the overreliance on fossil fuels and expensive metal catalysts and reduce environmental pollution due to their near-zero carbon emission. That electrochemically active microorganism can generate protons and electrons in the anodic compartment makes them useful in energy production. In this chapter, the authors summarized the recent developments in the advanced synthesis of cathode and anode catalysts and polymeric membrane electrolytes for MFCs. The electrochemical performance and stability of the MFC devices were also evaluated and compared with published results. Our study indicated that the improvement of electrochemical performance of the MFC devices can be fulfilled using various nanoscaled electrocatalysts (particularly the cathodic catalyst) and homogeneity of the membranes. These different materials were produced using colloidal chemistry, radiofrequency deposition, and plasma-assisted chemical vapor deposition. Series of materials include biota-inspired metal oxides, single atomic metals, carbon nanotubes, graphene quantum dots, and metal–organic frameworks as cathode and anode catalysts, and different formulations of modified polymer membranes as electrolytes. Based on the structural characterization and performance of the devices, we further optimize the synthesis variable to enhance the stability and reactivities of these nanocatalysts.

De novo gene origination has been recently established as an important mechanism for the formation of new genes. In organisms with a large genome, intergenic and intronic regions provide plenty of raw material for new transcriptional events to occur, but little is know about how de novo transcripts originate in more densely-packed genomes. Here, we identify 213 de novo originated transcripts in Saccharomyces cerevisiae using deep transcriptomics and genomic synteny information from multiple yeast species grown in two different conditions. We find that about half of the de novo transcripts are expressed from regions which already harbor other genes in the opposite orientation; these transcripts show similar expression changes in response to stress as their overlapping counterparts, and some appear to translate small proteins. Thus, a large fraction of de novo genes in yeast are likely to co-evolve with already existing genes., Genome-wide studies of de novo genes have tended to focus on genomic open reading frames (ORFs). Here, Blevins et al. use deep transcriptomics and synteny information to identify de novo transcripts in the yeast Saccharomyces cerevisiae, many of which are expressed from the alternative DNA strand.

The current global pandemic due to the SARS-CoV-2 has pushed the limits of global health systems across all aspects of clinical care, including laboratory diagnostics. Supply chain disruptions and rapidly-shifting markets have resulted in flash-scarcity of commercial laboratory reagents; this has motivated health care providers to search for alternative workflows to cope with the international increase in demand for SARS-CoV-2 testing. The aim of this study is to present a reproducible workflow for real time RT-PCR SARS-CoV-2 testing using OT-2 open-source liquid-handling robots (Opentrons, NY). We have developed a framework that includes a code template which is helpful for building different stand-alone robotic stations, capable of performing specific protocols. Such stations can be combined together to create a complex multi-stage workflow, from sample setup to real time RT-PCR. Using our open-source code, it is easy to create new stations or workflows from scratch, adapt existing templates to update the experimental protocols, or to fine-tune the code to fit specific needs. Using this framework, we developed the code for two different workflows and evaluated them using external quality assessment (EQA) samples from the European Molecular Genetics Quality Network (EMQN). The affordability of this platform makes automated SARS-CoV-2 PCR testing accessible for most laboratories and hospitals with qualified bioinformatics personnel. This platform also allows for flexibility, as it is not dependent on any specific commercial kit, and thus it can be quickly adapted to protocol changes, reagent, consumable shortages, or any other temporary material constraints.

Current global pandemic due to the SARS-CoV-2 has struggled and pushed the limits of global health systems. Supply chain disruptions and scarce availability of commercial laboratory reagents have motivated worldwide actors to search for alternative workflows to cope with the demand.We have used the OT-2 open-source liquid-handling robots (Opentrons, NY), RNA extraction and RT-qPCR reagents to set-up a reproducible workflow for RT-qPCR SARS-CoV-2 testing. We developed a framework with a template and several functions and classes that allow the creation of customized RT-qPCR automated circuits. As a proof of concept, we provide data obtained by a fully-functional tested code using the MagMax™ Pathogen RNA/DNA kit and the MagMax™ Viral/Pathogen II kit (Thermo Fisher Scientific, MA) on the Kingfisher™ Flex instrument (Thermo Fisher Scientific, MA). With these codes available is easy to create new stations or circuits from scratch, adapt existing ones to changes in the experimental protocol, or perform fine adjustments to fulfil special needs.The affordability of this platform makes it accessible for most laboratories and hospitals with a bioinformatician, democratising automated SARS-CoV-2 PCR testing and increasing, temporarily or not, the capacity of carrying out tests. It also confers flexibility, as this platform is not dependant on any particular commercial kit and can be quickly adapted to protocol changes or other special needs.