Breast Tumor Localization by Prone to Supine Landmark Driven Registration for Surgical Planning

Breast cancer is the most common cancer in women worldwide. Screening programs and imaging improvements have increased the detection of clinically occult non-palpable lesions requiring preoperative localization. Wire guided localization (WGL) is the current standard of care for the excision of non-palpable carcinomas during breast conserving surgery. Due to the current limitations of intraoperative tumor localization approaches, the integration of multimodal imaging information may be especially relevant in surgical planning. This research proposes a novel method for performing preoperative image-to-surgical surface data alignment to determine the position of the tumor at the time of surgery and aid preoperative planning. First, the volume of the breast in the surgical position is reconstructed and a set of surface correspondences is defined. Then, the preoperative (prone) and intraoperative (supine) volumes are co-registered using landmark driven non-rigid registration methods. We compared the performances of diffeomorphic and Bspline based registration methods. Finally, our method was validated using clinical data from 67 patients considering as target registration error (TRE) the distance between the estimated tumor position and the reference surgical position. The proposed method achieved a TRE of 16.21 ± 8.18 mm and it could potentially assist the surgery planning and guidance of breast cancer treatment in the clinical practice.This work was supported in part by the Spanish Ministry of Science and Innovation under Project RTI2018-098682-B-I00 (MCIU/AEI/FEDER,UE), Project PI18/01625 (Instituto de Salud Carlos III) and Grant BGP18/00178 under Beatriz Galindo Programme; in part by the European Union's European Regional Development Fund (ERDF); and in part by the Madrid Government (Comunidad de Madrid-Spain) under the Multiannual Agreement with Universidad Politécnica de Madrid in the line Support for Research and Development Projects for Beatriz Galindo researchers, in the context of the V Plan Regional de Investigación Científíca e Innovación Tecnológica (PRICIT)

Distribution and genotype-phenotype correlation of GDAP1 mutations in Spain

Mutations in the GDAP1 gene can cause Charcot-Marie-Tooth disease. These mutations are quite rare in most Western countries but not so in certain regions of Spain or other Mediterranean countries. This cross-sectional retrospective multicenter study analyzed the clinical and genetic characteristics of patients with GDAP1 mutations across Spain. 99 patients were identified, which were distributed across most of Spain, but especially in the Northwest and Mediterranean regions. The most common genotypes were p.R120W (in 81% of patients with autosomal dominant inheritance) and p.Q163X (in 73% of autosomal recessive patients). Patients with recessively inherited mutations had a more severe phenotype, and certain clinical features, like dysphonia or respiratory dysfunction, were exclusively detected in this group. Dominantly inherited mutations had prominent clinical variability regarding severity, including 29% of patients who were asymptomatic. There were minor clinical differences between patients harboring specific mutations but not when grouped according to localization or type of mutation. This is the largest clinical series to date of patients with GDAP1 mutations, and it contributes to define the genetic distribution and genotype-phenotype correlation in this rare form of CMT

Non-replicative antibiotic resistance-free DNA vaccine encoding S and N proteins induces full protection in mice against SARS-CoV-2

17 p.-8 fig.SARS-CoV-2 vaccines currently in use have contributed to controlling the COVID-19 pandemic. Notwithstanding, the high mutation rate, fundamentally in the spike glycoprotein (S), is causing the emergence of new variants. Solely utilizing this antigen is a drawback that may reduce the efficacy of these vaccines. Herein we present a DNA vaccine candidate that contains the genes encoding the S and the nucleocapsid (N) proteins implemented into the non-replicative mammalian expression plasmid vector, pPAL. This plasmid lacks antibiotic resistance genes and contains an alternative selectable marker for production. The S gene sequence was modified to avoid furin cleavage (Sfs). Potent humoral and cellular immune responses were observed in C57BL/6J mice vaccinated with pPAL-Sfs + pPAL-N following a prime/boost regimen by the intramuscular route applying in vivo electroporation. The immunogen fully protected K18-hACE2 mice against a lethal dose (105 PFU) of SARS-CoV-2. Viral replication was completely controlled in the lungs, brain, and heart of vaccinated mice. Therefore, pPAL-Sfs + pPAL-N is a promising DNA vaccine candidate for protection from COVID-19.This work was funded by PTI-Salud Global (CSIC), Center for Technological and Industrial Development (CDTI), REACT-ANTICIPA-UCM (Comunidad de Madrid), and European Research Council (Advanced Grant VERDI, ERC2015AdG grant number 694160).Peer reviewe

Healthcare workers hospitalized due to COVID-19 have no higher risk of death than general population. Data from the Spanish SEMI-COVID-19 Registry

Aim To determine whether healthcare workers (HCW) hospitalized in Spain due to COVID-19 have a worse prognosis than non-healthcare workers (NHCW). Methods Observational cohort study based on the SEMI-COVID-19 Registry, a nationwide registry that collects sociodemographic, clinical, laboratory, and treatment data on patients hospitalised with COVID-19 in Spain. Patients aged 20-65 years were selected. A multivariate logistic regression model was performed to identify factors associated with mortality. Results As of 22 May 2020, 4393 patients were included, of whom 419 (9.5%) were HCW. Median (interquartile range) age of HCW was 52 (15) years and 62.4% were women. Prevalence of comorbidities and severe radiological findings upon admission were less frequent in HCW. There were no difference in need of respiratory support and admission to intensive care unit, but occurrence of sepsis and in-hospital mortality was lower in HCW (1.7% vs. 3.9%; p = 0.024 and 0.7% vs. 4.8%; p<0.001 respectively). Age, male sex and comorbidity, were independently associated with higher in-hospital mortality and healthcare working with lower mortality (OR 0.211, 95%CI 0.067-0.667, p = 0.008). 30-days survival was higher in HCW (0.968 vs. 0.851 p<0.001). Conclusions Hospitalized COVID-19 HCW had fewer comorbidities and a better prognosis than NHCW. Our results suggest that professional exposure to COVID-19 in HCW does not carry more clinical severity nor mortality

Treatment with tocilizumab or corticosteroids for COVID-19 patients with hyperinflammatory state: a multicentre cohort study (SAM-COVID-19)

Objectives: The objective of this study was to estimate the association between tocilizumab or corticosteroids and the risk of intubation or death in patients with coronavirus disease 19 (COVID-19) with a hyperinflammatory state according to clinical and laboratory parameters. Methods: A cohort study was performed in 60 Spanish hospitals including 778 patients with COVID-19 and clinical and laboratory data indicative of a hyperinflammatory state. Treatment was mainly with tocilizumab, an intermediate-high dose of corticosteroids (IHDC), a pulse dose of corticosteroids (PDC), combination therapy, or no treatment. Primary outcome was intubation or death; follow-up was 21 days. Propensity score-adjusted estimations using Cox regression (logistic regression if needed) were calculated. Propensity scores were used as confounders, matching variables and for the inverse probability of treatment weights (IPTWs). Results: In all, 88, 117, 78 and 151 patients treated with tocilizumab, IHDC, PDC, and combination therapy, respectively, were compared with 344 untreated patients. The primary endpoint occurred in 10 (11.4%), 27 (23.1%), 12 (15.4%), 40 (25.6%) and 69 (21.1%), respectively. The IPTW-based hazard ratios (odds ratio for combination therapy) for the primary endpoint were 0.32 (95%CI 0.22-0.47; p < 0.001) for tocilizumab, 0.82 (0.71-1.30; p 0.82) for IHDC, 0.61 (0.43-0.86; p 0.006) for PDC, and 1.17 (0.86-1.58; p 0.30) for combination therapy. Other applications of the propensity score provided similar results, but were not significant for PDC. Tocilizumab was also associated with lower hazard of death alone in IPTW analysis (0.07; 0.02-0.17; p < 0.001). Conclusions: Tocilizumab might be useful in COVID-19 patients with a hyperinflammatory state and should be prioritized for randomized trials in this situatio

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

Effects of Gαi2 and Gαz protein knockdown on alpha2A-adrenergic and cannabinoid CB1 receptor regulation of MEK-ERK and FADD pathways in mouse cerebral cortex

Background: Alpha-adrenergic (α-AR) and cannabinoid CB (CB-R) receptors exert their functions modulating multiple signaling pathways, including MEK-ERK (extracellular signal-regulated kinases) and FADD (Fas-associated protein with death domain) cascades. These molecules are relevant in finding biased agonists with fewer side effects, but the mechanisms involving their modulations by α-AR- and CB-R in vivo are unclear. This study investigated the roles of Gαi and Gαz proteins in mediating α-AR- and CB-R-induced alterations of MEK-ERK and FADD phosphorylation (p-) in mouse brain cortex. Methods: Gαi or Gαz protein knockdown was induced in mice with selective antisense oligodeoxinucleotides (ODNs; 3 nmol/day, 5 days) prior to UK-14,304 (UK or brimonidine; 1 mg/kg) or WIN55212-2 (WIN; 8 mg/kg) acute treatments. Inactivated (p-T) MEK1, activated (p-S) MEK1/2, activated (p-T/Y) ERK1/2, p-S FADD, and the corresponding total forms of these proteins were quantified by immunoblotting. Results: Increased (+ 88%) p-T MEK1 cortical density, with a concomitant reduction (–43%) of activated ERK was observed in UK-treated mice. Both effects were attenuated by Gαi or Gαz antisense ODNs. Contrastingly, WIN induced Gαi- and Gαz-independent upregulations of p-T MEK1 (+ 63%), p-S MEK1/2 (+ 86%), and activated ERK (+ 111%) in brain. Pro-apoptotic FADD was downregulated (− 34 to 39%) following UK and WIN administration, whereas the neuroprotective p-S FADD was increased (+ 74%) in WIN-treated mice only. None of these latter effects required from Gαi or Gαz protein integrity. Conclusion: The results indicate that α-AR (UK), but not CB-R (WIN), agonists use Gαi and Gαz proteins to modulate MEK-ERK, but not FADD, pathway in mouse brain cortex.The study was supported by grants RTI2018-094414-A-I00 (AR-M), SAF2011-29918 (JAG-S), and RT 2018-093677-B-100 (PSB) from the Spanish Ministry of Science, Innovation and Universities (MCIU) and the European Regional Development Fund (ERDF). AR-M is a ‘Ramón y Cajal’ Researcher (grant RYC-2016-19282) supported by MCIU. JAG-S is a member of the Institut d’Estudis Catalans (Barcelona, Catalonia, Spain). None of the above funding agencies took part in the elaboration of the study, formulation of the hypotheses, or interpretation of the results

Effects of Gαi2 and Gαz protein knockdown on alpha2A-adrenergic and cannabinoid CB1 receptor regulation of MEK-ERK and FADD pathways in mouse cerebral cortex

Background: Alpha-adrenergic (α-AR) and cannabinoid CB (CB-R) receptors exert their functions modulating multiple signaling pathways, including MEK-ERK (extracellular signal-regulated kinases) and FADD (Fas-associated protein with death domain) cascades. These molecules are relevant in finding biased agonists with fewer side effects, but the mechanisms involving their modulations by α-AR- and CB-R in vivo are unclear. This study investigated the roles of Gαi and Gαz proteins in mediating α-AR- and CB-R-induced alterations of MEK-ERK and FADD phosphorylation (p-) in mouse brain cortex. Methods: Gαi or Gαz protein knockdown was induced in mice with selective antisense oligodeoxinucleotides (ODNs; 3 nmol/day, 5 days) prior to UK-14,304 (UK or brimonidine; 1 mg/kg) or WIN55212-2 (WIN; 8 mg/kg) acute treatments. Inactivated (p-T) MEK1, activated (p-S) MEK1/2, activated (p-T/Y) ERK1/2, p-S FADD, and the corresponding total forms of these proteins were quantified by immunoblotting. Results: Increased (+ 88%) p-T MEK1 cortical density, with a concomitant reduction (–43%) of activated ERK was observed in UK-treated mice. Both effects were attenuated by Gαi or Gαz antisense ODNs. Contrastingly, WIN induced Gαi- and Gαz-independent upregulations of p-T MEK1 (+ 63%), p-S MEK1/2 (+ 86%), and activated ERK (+ 111%) in brain. Pro-apoptotic FADD was downregulated (− 34 to 39%) following UK and WIN administration, whereas the neuroprotective p-S FADD was increased (+ 74%) in WIN-treated mice only. None of these latter effects required from Gαi or Gαz protein integrity. Conclusion: The results indicate that α-AR (UK), but not CB-R (WIN), agonists use Gαi and Gαz proteins to modulate MEK-ERK, but not FADD, pathway in mouse brain cortex.The study was supported by grants RTI2018-094414-A-I00 (AR-M), SAF2011-29918 (JAG-S), and RT 2018-093677-B-100 (PSB) from the Spanish Ministry of Science, Innovation and Universities (MCIU) and the European Regional Development Fund (ERDF). AR-M is a ‘Ramón y Cajal’ Researcher (grant RYC-2016-19282) supported by MCIU. JAG-S is a member of the Institut d’Estudis Catalans (Barcelona, Catalonia, Spain). None of the above funding agencies took part in the elaboration of the study, formulation of the hypotheses, or interpretation of the results

Ethanol desensitizes cannabinoid CB1 receptors modulating monoamines synthesis in the rat brain in vivo

[eng] The endocannabinoid system and the cannabinoid CB1 receptors are involved in the development of ethanol tolerance and dependence. This study aimed to investigate the in vivo sensitivity of a CB1 receptor agonist (WIN 55,212-2) modulating the synthesis of 3,4-dihydroxyphenylalanine/dopamine/noradrenaline (DOPA/DA/NA) and that of 5-hydroxy-tryptophan/serotonin (5-HTP/5-HT) in rat brain after ethanol treatment and withdrawal. In control rats, WIN 55,212-2 (4 mg/kg, i.p., for 1 h), through a mechanism sensible to the CB1 antagonist SR 141716A, increased the synthesis of DOPA/NA in a slice of brainstem containing the locus ceruleus (250%) and in the hippocampus (64%), and it reduced DOPA/DA synthesis in the striatum (47%). WIN 55,212-2 also decreased the synthesis of 5-HTP/5-HT in the locus ceruleus (43%), hippocampus (35%) and striatum (35%). In the locus ceruleus of ethanol-treated rats, the stimulatory effect of WIN 55,212-2 on DOPA/NA synthesis was abolished (acute treatment) or markedly attenuated (53-55%, chronic treatment and withdrawal), whereas in the hippocampus this effect was reduced only in chronic ethanol-withdrawn rats (33%). In the striatum of ethanol-treated rats (acute, chronic and withdrawal), the inhibitory effect of WIN 55,212-2 on DOPA/DA synthesis was completely blunted or markedly reduced. Similarly, the inhibitory effect of WIN 55,212-2 on 5-HTP/5-HT synthesis was reduced or abolished in the three brain regions after chronic ethanol and during withdrawal. These results indicate that treatment with ethanol in rats induces a functional desensitization of CB1 receptors modulating the synthesis of brain monoamines

Compression Behavior of Sheets Metals of Pure Titanium 2 and Ti6Al4V Alloy under High Temperature: Evaluation of the Tension–Compression Asymmetry

Determining the intrinsic indices of sheet metals under compression states at high temperatures is vital to accurately predict the behavior of the material in warm/hot forming processes. Nevertheless, the literature contains little previous experimental data in this regard due to the difficulty of carrying out specific test methodologies in sheet metals. The authors of the present manuscript previously developed an approach to evaluate the in-plane compression behavior under a wide range of test conditions, which was applied here to characterize pure titanium and Ti6Al4V alloy until 750 °C. This procedure allowed us to quantify the asymmetric and anisotropic tension–compression (T-C) response of the materials involved and their evolution with temperature and strain rate. The asymmetry detected at room temperature showed a higher compression response in all cases, mostly reaching differences of around 10%. For the lowest strain rate studied, the typical assumed symmetric T-C behavior was observed from 300 and 450 °C onwards, for the rolling and transverse direction, respectively. In addition, stepped compression tests led us to deduce the anisotropy indices, which were different from those found under tension, in contrast to the r-values applied by most authors. Using the experimental results, a factor related to the asymmetry found was proposed to formulate an extended constitutive model. The asymmetry and anisotropy data supplied for compression under warm/hot conditions are the main novelty of this research