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Immune checkpoint inhibitors (ICIs) and targeted therapies have revolutionised the diagnostic and treatment paradigm of advanced non-small cell lung cancer (NSCLC). However, treatment advances in squamous cell subtype have been much slower than those occurring in adenocarcinoma, mainly due to the lack of targetable oncogenic aberrations in squamous tumours. The unprecedented durable response and favourable toxicity profile observed with ICIs in advanced squamous NSCLC, represents a true therapeutic milestone in a population previously limited to cytotoxic chemotherapy (Cht). The inhibition of programmed cell death ligand 1 (PD-1/PD-L1) pathway has been extensively investigated in NSCLC and currently dominates the treatment landscape of advanced NSCLC. The first approval of ICIs came from several randomized trials conducted in platinum-treated advanced NSCLC patients (any histology) (1-4) where overall survival (OS) and toxicity profile favoured PD-1/PD-L1 inhibitors over docetaxel. Subsequently, KEYNOTE-024 (5) showed that, in advanced NSCLC patients (all histologies) with a PD-L1 tumour proportion score (TPS) ≥50%, first-line pembrolizumab achieved higher objective response rates (ORR) and longer progression-free survival (PFS) and OS when compared to standard platinum-based Cht. However, high PD-L1 expression only occurs in a third of NSCLC patients and, until recently, platinum-based Cht remained the only approved option for fit patients with a PD-L1 TPS

The advent of PD-(L)1 and CTLA-4 immune check point inhibitors (CPIs) has dramatically changed the treatment landscape of advanced non-small-cell lung cancer (NSCLC). For up to a quarter of patients with advanced NSCLC, CPIs have the potential to induce durable responses with long-term survival outcomes. Since the approval of first-line pembrolizumab for patients whose tumors express a PD-L1 ≥ 50%, several pivotal first-line CPI-based phase 3 studies have been conducted investigating combination treatments combining CPIs with chemotherapy (ChT) or combining different CPIs with or without ChT. As a result, there has been an increase in front-line treatment options for advanced NSCLC, and treatment algorithms are changing very quickly. In fit patients with advanced NSCLC, combination treatments including CPI and ChT are considered the new standard of care with improved clinical outcomes. CPI combination treatments are well tolerated and quality of life also seems to be better when CPIs are implemented in the first-line setting. The aim of this review is to provide a summary of the recently published first-line phase 3 studies investigating CPIs as monotherapy or in combination with other CPIs or ChT in advanced NSCLC, and to suggest possible treatment algorithms.

Background Incorporating in the Intensive Care Unit (ICU) a clinical pharmacist who performs interventions on antimicrobials may be cost-effective. Objectives To evaluate the clinical and economic impact of clinical pharmacist interventions on antimicrobials in an ICU. To identify drug related problems and medication errors detected by the pharmacist. Methods A retrospective observational study was performed to analyze drug related problems, medication errors and clinical pharmacist interventions related to antimicrobials in adults admitted to an ICU in a 5-month period. The economic impact of pharmacist interventions was estimated considering difference in cost derived from antimicrobial treatment, adverse drug events and clinical pharmacist time. Results A total of 212 drug related problems were detected in 114 patients, 18 being medication errors. Clinical pharmacist developed one intervention for each problem identified. 204 interventions (96.2%) were considered important with improved patient care and 7 (3.3%) very important. No negative impact of any intervention was identified. Physicians accepted 97.6% of the interventions. A potential saving of 10,905 € was estimated as a result of pharmacist interventions and 4.8 € were avoided per euro invested in a clinical pharmacist. Conclusions A clinical pharmacist performing interventions on antimicrobials in the ICU has a positive impact on patient care and decreases costs.

Objectives To analyze the prevalence of May-Thurner syndrome (MTS) among patients with deep vein thrombosis (DVT) of left lower limb (LLL), and outcomes after long-term follow-up. Method Retrospective observational study that included patients older than 18 diagnosed with acute DVT and MTS. Results Among 760 patients diagnosed with DVT in the LLL, 8 patients were diagnosed with MTS (1.05%), with a mean age of 39 years. All patients received long-term anticoagulation, with a mean of 31.9 (±25.2) months. Invasive treatment with pharmaco-mechanical thrombectomy was performed in 5 patients (62.5%). During follow-up (mean of 32.4 months), 25% of patients suffered DVT recurrence. Only 1 case presented major bleeding, and no deaths were registered. Conclusions May-Thurner syndrome constitutes a permanent and underdiagnosed risk factor for the development of DVT of LLL. Our findings suggest that long-term anticoagulation therapy might be considered in selected cases.

Background: Epidermolysis bullosa (EB) corresponds to a series of conditions characterized by extreme fragility of the skin and/or mucous membranes. Of the four main types of EB, junctional EB (JEB) is the most associated with alterations in the teeth. The purposes of this study were to determine the clinical, histopathological and ultrastructural characteristics of teeth with amelogenesis imperfecta (AI) in a patient with JEB, to compare them with control teeth and to correlate the findings with the mutations present in the patient. Case report: The study was conducted on a 10-year-old patient with JEB carring two recessive mutations in the LAMB3 gene and the absence of the laminin-332 protein (LM-332), determined by immunofluorescence on a skin biopsy. The patient presents hypoplastic amelogenesis imperfecta with very thin and yellow-brown colored enamel. required eExtractionodontia of two permanent molars were performed due to pain and soft tissue over the crown, compatible resembling with pulp polyp or hyperplastic gingiva. The molars were studied by Light and scanning electron microscopy (SEM) revealed very thing enamel varying from complete absence to 60 microns, absence of normal prismatic structure and presence of a cross-banding with a laminated appearance. The studied teeth had very thin and yellow-brown colored enamel. The histopathological study revealed soft tissue on the crown corresponded to granulation tissue causing generating external crown tooth resorption. With both light and electron microscopy, bands semi-parallel to the dentino-enamel junction were observed in the enamel and absence of prismatic structure. Conclusion: Although coronary resorption has been reported in patients with syndromic and non-syndromic amelogenesis imperfecta, this is the first clinicopathological report of coronary resorption in partially erupted teeth in patients with JEB with mutations in the LAMB3 gene and hypoplastic amelogenesis imperfecta. In patients with this condition, the presence of partially erupted teeth with soft tissue covering part of the crown, without a periodontal pocket and with a radiographic image of abrupt loss of partial coronal radiolucency should lead to suspicion of external coronary resorption.

Transcriptomic signature of XPO1 was highly expressed and inversely related to left ventricular function in ischemic cardiomyopathy patients. We hypothesized that treatment with AAV9-shXPO1 attenuates left ventricular dysfunction and remodeling in a myocardial infarction rat model. We induced myocardial infarction by coronary ligation in Sprague-Dawley rats (n = 10), which received AAV9-shXPO1 (n = 5) or placebo AAV9-scramble (n = 5) treatment. Serial echocardiographic assessment was performed throughout the study. After myocardial infarction, AAV9-shXPO1-treated rats showed partial recovery of left ventricular fractional shortening (16.8 ± 2.8 vs 24.6 ± 4.1%, P P P XPO1.

Background:The main reason for using a two-stage exchange in Prosthetic Joint Infection (PJI) is that bacteria are completely eradicated in reimplantation surgery. However, reports of a positive culture in the second surgery are growing. The number of positive intraoperative cultures and their influence on final results is not well-established.Objectives:To compare epidemiological characteristics, infection recurrence and clinical evolution of patients with only onevs.at least two positive cultures based on our series of cases with positive cultures in reimplantation surgery.Material and Methods:Retrospective study of 55 patients was conducted prospectively. They were diagnosed with chronic PJI, treated with a two-stage protocol and at least three intraoperative cultures were obtained in the second stage. These cultures were negative in 28 patients. Fourteen patients showed two or more cultures with the same microorganism and they were denominated patients with positive cultures. Thirteen patients showed only one positive culture, and they were considered contaminated. Both groups of patients (positive cultures and contaminated ones) received the second cycle of oral antibiotics for 6 months. Functional results were evaluated with the Harris Hip Score (hips) or Knee Society Clinical Rating Score (KSCRS) (knees).Results:There were no significant differences between patients with positive or contaminated cultures for age (p=0.420) and sex (p=0.385). The knee was involved in 13/14 positive and in only 6/13 contaminated patients (p=0.013).Staphylococcus epidermidiswas the predominant isolate, but there were differences between positive (methicillin-resistant in 7/14 patients) and contaminated cultures (methicillin-sensitive in 6/13). There were no differences in the prevalence of polymicrobial cultures (p=0.785) or coincidence with cultures from the first stage (p=0.257). Three infection recurrences have appeared in patients with positive cultures (3/13, 21%) and none in patients with contaminated cultures. There are no differences in HSS or KSCRS when comparing final functional results between groups (p=0.411).Conclusion:The prevalence of positive cultures in reimplantation surgery is higher than expected (25%), and more frequent in women and in knee arthroplasties. The most frequently involved microorganism isStaphylococcus epidermidis, but antibiotic sensitivity varies between patients with positive cultures (methicillin-resistant) and those with contaminated cultures (methicillin-sensitive). There were no infection recurrences in patients with contaminated cultures, but those with positive cultures present a risk of over 20%.

Prognosis of early stage non-small cell lung cancer (eNSCLC) is poor even when treated radically with surgery and (neo)adjuvant chemotherapy (Cht). The discovery of tyrosine kinase inhibitors (TKIs) for oncogene addicted NSCLC and immune checkpoint inhibitors (ICIs) have revolutionised the therapeutic paradigm and improved survival of advanced NSCLC. The unprecedented impact of these drugs has shifted the focus of investigation to early stage disease aiming at improving cure. In this context, several single arm phase II studies evaluating neoadjuvant ICI alone or in combination with platinum-based Cht have shown encouraging rates of pathological response which have spurred several ongoing randomized trials with (neo)adjuvant ICI. More recently, ADAURA study evaluating adjuvant osimertinib demonstrated a profound reduction of the risk of recurrence in patients with stage I (>4 cm)-IIIA eNSCLC harbouring EGFR sensitizing mutations. ICIs and TKIs represent a true revolution in the treatment of eNSCLC call to challenge the current standard of care. However, questions regarding drug resistance, recurrence patterns, biomarker identification, optimal treatment duration and sequencing need be answered to effectively integrate new drugs in the rapidly evolving therapeutic landscape of NSCLC. In this review we critically review new developments and future perspectives of TKIs and ICI as (neo)adjuvant strategies for eNSCLC.

Background Small-cell lung cancer (SCLC) is an aggressive disease with high metastatic potential and poor prognosis. Due to its low prevalence, epidemiological and clinical information of SCLC patients retrieved from lung cancer registries is scarce. Patients and methods This was an observational multicenter study that enrolled patients with lung cancer and thoracic tumors, recruited from August 2016 to January 2020 at 50 Spanish hospitals. Demographic and clinical data, treatment patterns and survival of SCLC patients included in the Thoracic Tumor Registry (TTR) were analyzed. Results With a total of 956 cases, the age of 64.7 ± 9.1 years, 78.6% were men, 60.6% smokers, and ECOG PS 0, 1 or ≥ 2 in 23.1%, 53.0% and 23.8% of cases, respectively. Twenty percent of patients had brain metastases at the diagnosis. First-line chemotherapy (CT), mainly carboplatin or cisplatin plus etoposide was administered to >90% of patients. In total, 36.0% and 13.8% of patients received a second and third line of CT, respectively. Median overall survival was 9.5 months (95% CI 8.8–10.2 months), with an estimated rate of 70.3% (95% CI 67.2–73.4%), 38.9% (95% CI 35.4–42.4%), and 14.8% (95% CI 11.8–17.8%) at 6, 12 and 24 months respectively. Median progression-free survival was 6.3 months. Higher mortality and progression rates were significantly associated with male sex, older age, smoking habit, and ECOG PS 1–2. Long-term survival (> 2 years) was confirmed in 6.6% of patients, showing a positive correlation with better ECOG PS, poor smoking and absence of certain metastases at diagnosis. Conclusion This study provides an updated overview of the clinical situation and treatment landscape of ES-SCLC in Spain. Our results might assist oncologists to improve current clinical practice towards a better prognosis for these patients.

Sirtuins have become important players in renal damage in hypertension and diabetes, but their value as biomarkers is poorly assessed. The aims of the study were to evaluate the levels of sirtuin1 (SIRT1), and two miRNAs that regulate SIRT1 expression in hypertensive patients with incipient renal damage with and without diabetes. We quantified urinary SIRT1 and claudin 1 (CLDN1) mRNA and miR34-a and miR-200a levels by quantitative real-time polymerase chain reaction (RT-qPCR) from patients and in cultured podocytes treated with high glucose and angiotensin II. Western blot and fluorescence analyses were also performed. We found decreased SIRT1 levels in patients with increased urinary albumin excretion (UAE), the lowest with diabetes presence, and a strong association with UAE, discriminating incipient renal damage. In vitro experiments also showed SIRT1 overall decreases in podocyte cultures under treatment conditions. In urine samples, miR-34a was reduced and miR-200a increased, both related to UAE levels. However, both miRNAs were generally increased in podocyte cultures under high glucose and angiotensin-II treatment. These results show a significant urinary SIRT1 decrease in albuminuric hypertensive patients, strongly associated with albuminuria, suggesting that SIRT1 could be a potential and non-invasive method to assess incipient renal damage in hypertensive patients.

Small-cell lung cancer (SCLC) is an aggressive and rapidly growing disease with poor prognosis. Despite intense efforts to improve clinical outcomes, platinum/etoposide chemotherapy has remained the most effective regimen for first-line extensive disease SCLC for decades. The addition of immune checkpoint inhibitors, and specifically programmed death-ligand 1 inhibitors, to standard platinum/etoposide, significantly improves survival and represents a promising advance in this field. However, identification of a predictive biomarker to refine patient selection is an area of unmet need. Further understanding of tumour immunity and mechanism of resistance is required to design novel strategies that improve survival. In this review, we describe recent developments and future directions on first-line immune checkpoint blockade for extensive disease-SCLC., Highlights • The addition of PD-L1 inhibitors but not CTLA-4 inhibitors to platinum/etoposide improves survival in first line SCLC. • A small proportion of patients derive long-term survival benefit which highlights the relevance of tumour heterogeneity. • At present, maintenance therapy with immune checkpoint inhibitors following platinum/etoposide can't be recommended. • Predictive biomarkers to aid patient selection for immune checkpoint inhibitors in SCLC remain an area of unmet need. • Upcoming trials will evaluate PD-L1 inhibitors plus chemotherapy in combination with drugs that promote tumour immunity.

Kidney injury in hypertension and diabetes entails, among in other structures, damage in a key cell of the glomerular filtration barrier, the podocyte. Podocytes are polarized and highly differentiated cells in which vesicular transport, partly driven by Rab GTPases, is a relevant process. The aim of the present study was to analyze Rab GTPases of the Rab-Rabphilin system in human immortalized podocytes and the impact of high glucose and angiotensin II. Furthermore, alterations of the system in urine cell pellets from patients with hypertension and diabetes were studied. Apoptosis was analyzed in podocytes, and mRNA level quantification, Western blot analysis, and immunofluorescence were developed to quantify podocyte-specific molecules and Rab-Rabphilin components (Rab3A, Rab27A, and Rabphilin3A). Quantitative RT-PCR was performed on urinary cell pellet from patients. The results showed that differentiated cells had reduced protein levels of the Rab-rabphillin system compared with undifferentiated cells. After glucose overload and angiotensin II treatment, apoptosis was increased and podocyte-specific proteins were reduced. Rab3A and Rab27A protein levels were increased under glucose overload, and Rabphilin3A decreased. Furthermore, this system exhibited higher levels under stress conditions in a manner of angiotensin II dose and time treatment. Immunofluorescence imaging indicated different expression patterns of podocyte markers and Rab27A under treatments. Finally, Rab3A and Rab27A were increased in patient urine pellets and showed a direct relationship with albuminuria. Collectively, these results suggest that the Rab-Rabphilin system could be involved in the alterations observed in injured podocytes and that a mechanism may be activated to reduce damage through the vesicular transport enhancement directed by this system.

Urinary exosomes, especially microRNAs (miRNAs) packaged within, are ideal sources of renal damage markers. We investigated the association between exosomal miR-146a, (anti-inflammatory regulator) and disease activity, proteinuria and systemic lupus erythematosus (SLE) flares over a 36-month follow-up period. We isolated urinary exosomes from 41 SLE patients, 27 with lupus nephritis (LN) and 20 healthy controls, and exosomal miR-146a, quantified by the real-time quantitative polymerase chain reaction (RT-qPCR), was correlated with histological features in 13 renal biopsies. We also analysed the association between the exosomal miR-146a and TRAF6 axis. Exosomal miR-146a showed an inverse association with circulating C3 and C4 complement components, proteinuria, and with histological features such as chronicity index. This marker was able to identify LN with an AUC of 0.82 (p = 0.001). Basal exosomal miR-146a was associated with disease activity and proteinuria changes and was an independent marker of 36-month follow-up flares (OR 7.08, p = 0.02). Pathway analysis identified IRAK1 and TRAF6 as miR-146a target genes. Finally, in vitro experiments suggested that miR-146a exerts a protective effect through negative regulation of inflammation by suppressing IRAK1 and TRAF6. Urinary exosomal miR-146a levels are correlated with lupus activity, proteinuria and histological features, discriminating patients with LN and being a good baseline marker of SLE flares. We have identified a relevant biological miR-146a-TRAF6 axis association in LN renal fibrosis progression.

BACKGROUND The proper dosage of antibiotics is a key element in the effective treatment of infection, especially in critically ill patients. This study aimed to evaluate the efficacy of optimized meropenem regimens based on pharmacokinetic/pharmacodynamic criteria in patients admitted to the intensive care unit. METHODS This observational, naturalistic, retrospective, unicentric cohort study was performed between May 2011 and December 2017. The clinical and bacteriologic responses of 77 control intensive care unit patients receiving meropenem were compared with those of 77 propensity score-balanced patients who received meropenem dose adjusted by therapeutic drug monitoring. The primary end point of clinical response was a reduction at the end of treatment of at least 80% of the maximum procalcitonin (PCT) value recorded during the meropenem treatment. RESULTS The primary end point was met by 55 patients (71.4%) in the adjusted group compared with 41 (53.3%) patients in the control group (mean difference 18.1%, P = 0.02). Fifty-one patients (66.2%) in the adjusted group required a meropenem dose adjustment, being necessary in 46 of them (90.2%) to decrease the dose. The reduction of PCT was the greatest in the adjusted group compared with the unadjusted group (93% versus 85%, P = 0.004); a greater percentage of patients reached a PCT level < 0.5 ng/mL (63.6% versus 41.6%, P = 0.006), and there was a trend toward an improved bacteriologic response (relative risk = 1.27; 95% confidence interval: 0.92-1.56). There were no differences in early mortality or safety between groups. CONCLUSIONS Adjustment of meropenem therapy by monitoring is a useful strategy for improving meropenem effectiveness in the treatment of infection in critically ill patients, with no impact on safety.

On March 11, 2020, the Director-General of the World Health Organization (WHO) declared the disease caused by SARS-CoV-2 (COVID-19) as a pandemic. The spread and evolution of the pandemic is overwhelming the healthcare systems of dozens of countries and has led to a myriad of opinion papers, contingency plans, case series and emerging trials. Covering all this literature is complex. Briefly and synthetically, in line with the previous recommendations of the Working Groups, the Spanish Society of Intensive, Critical Medicine and Coronary Units (SEMICYUC) has prepared this series of basic recommendations for patient care in the context of the pandemic.

Background and importance Meropenem dose adjustment following pharmacokinetic/pharmacodynamic monitoring (TDM) in critical patients (CP) presents a clinical benefit. An economic analysis of this activity could facilitate its use in clinical practice. Aim and objectives To conduct a cost effectiveness analysis of meropenem TDM in CP versus standard dose (SD) according to the package insert recommendations. Material and methods We conducted a naturalistic, retrospective, observational cohort study of CP receiving meropenem between May 2011 and December 2017 in a university hospital. Two cohorts were analysed: patients with meropenem TDM (cohort A) and patients with SD meropenem (cohort B). The main effectiveness variable was the percentage of patients with a reduction of at least 80% in the procalcitonin value at the end of meropenem treatment compared with the maximum value during meropenem treatment. Costs included in the analysis were: meropenem, material for drug preparation, TDM, time for preparation, administration and infusion surveillance, meropenem adverse drug reactions (ADR), critical care hospitalisation days and re-entries. Propensity score (PS) matching was applied for patient selection. The χ2 was used to compare effectiveness and bootstrap to calculate the difference in costs between cohorts. A cost effectiveness analysis with deterministic and probabilistic sensitivity analyses was performed. Results A total of 154 patients were included (77 per cohort) after PS matching. Meropenem dose was changed in 51 (66.2%) patients with TDM, in most (90.2%) because they were overdosed. In cohort A, 71.4% of patients had reduced procalcitonin by at least 80% compared with 53.2% in cohort B (difference 18.2% (95% CI 3.1; 33.2; p=0.020)). No significant differences were found in ADR between the two cohorts. An average decrease in cost per patient of −1454€ (95% CI −4627;1720€) with TDM was observed, with lower cost per patient for meropenem −62€ (95% CI −116; −4), disposable material −12€ (95% CI −29; 4) and nursing time −38€ (95% CI −71; −4) in cohort A, that offset the TDM cost (47€). Mean hospitalisation cost in patients with TDM was 8912€ versus 10 325€ in cohort B. There was a 75% probability that TDM was more effective and cheaper (dominant) than SD according to the sensitivity analysis. Conclusion and relevance Meropenem dose adjustment following pharmacokinetic/pharmacodynamic criteria was more effective, with similar safety and lower costs, than dosing according to the package insert recommendations. References and/or acknowledgements 1. Schuetz P, et al. Procalcitonin-guided antibiotic stewardship. Clin Chem Lab Med 2019. No conflict of interest.

BACKGROUND Levetiracetam (LEV) is a second-generation antiepileptic drug extensively used in therapeutics. The aim of this study was to evaluate the influence that sex, age, and weight exert on LEV pharmacokinetics in clinical practice. METHODS We conducted a 6-year retrospective observational study. Patients were classified in subgroups according to sex, weight (normal range, overweight, and obese), and age (young adult: 16-30 years old, middle-aged adult: 31-50 years old, advanced adult: 51-64 years old, and elderly adult: ≥65 years old). We compared LEV apparent oral clearance (LEV CL/F) between the subgroups. RESULTS A total of 238 LEV basal serum concentrations (LEV C0) corresponding to 156 patients were identified. Significant differences were observed in LEV CL/F between males and females when LEV CL/F was expressed as L/h [mean (SD): 4.79 (1.84) L/h in males versus 4.13 (1.64) L/h in females; P < 0.001]. These differences were not significant when LEV CL/F was normalized by weight [mean (SD): 60.64 (24.90) mL/h/kg in males versus 64.10 (28.87) mL/h/kg in females; n.s.]. Weight in females was 17% lower compared with males. A progressive reduction in LEV CL/F was observed with increasing age, in a proportion that was similar to the decline in renal function. The elderly patients presented 30% lower LEV CL/F (mL/h/kg) and 43% lower creatinine clearance (CCr) in comparison with adults. No statistically significant differences were observed in LEV CL/F calculated in L/h between weight subgroups. However, when LEV CL/F was expressed in mL/h/kg, a progressive reduction was observed [normal weight: 72.21 (28.97); overweight: 57.84 (25.38); obese: 49.45 (14.50); P < 0.001]. A significant and positive correlation between CCr and LEV CL/F was observed, confirming the important role of the renal function in LEV CL/F. The CCr increased in each sex group when weight increased; however, LEV CL/F (L/h) remained constant. CONCLUSIONS Sex, age, and weight affect LEV pharmacokinetics, having an impact on the individual dosage regimen needed to achieve the therapeutic objective. Sex is a conditioning factor of LEV CL/F, although its influence is principally due to the weight. LEV CL/F decreases with advancing age, proportionally to the decline in renal function. It is confirmed that LEV dosage per body weight is not required, and prescribing higher doses of LEV in obese patients is not justified. These data suggest that routine LEV therapeutic drug monitoring in the elderly patients, patients with renal dysfunction, and obese patients is indicated.

Aims Ischaemic cardiomyopathy (ICM) leads to impaired contraction and ventricular dysfunction, causing high rates of morbidity and mortality. Epigenomics allows the identification of epigenetic signatures in human diseases. We analyse the differential epigenetic patterns of the ASB gene family in ICM patients and relate these alterations to their haemodynamic and functional status. Methods and results Epigenomic analysis was carried out using 16 left ventricular (LV) tissue samples, eight from ICM patients undergoing heart transplantation and eight from control (CNT) subjects without cardiac disease. We increased the sample size up to 13 ICM and 10 CNT for RNA sequencing and to 14 ICM for pyrosequencing analyses. We found a hypermethylated profile (cg11189868) in the ASB1 gene that showed a differential methylation of 0.26Δβ (P = 0.016). This result was validated by a pyrosequencing technique (0.23Δβ, P = 0.048). Notably, the methylation pattern was strongly related to LV ejection fraction (r = -0.849, P = 0.008), stroke volume (r = -0.929, P = 0.001), and end-systolic and diastolic LV diameters (r = -0.743, P = 0.035 for both). ASB1 showed a down-regulation in messenger RNA levels (-1.2-fold, P = 0.039). Conclusions Our findings link a specific ASB1 methylation pattern to LV structure and performance in end-stage ICM, opening new therapeutic opportunities and providing new insights regarding which is the functionally relevant genome in the ischaemic failing myocardium.

Orofacial granulomatosis is a nonspecific term that contains a wide variety of granulomatous entities, which share a clinical and histopathological presentation. It manifests as persistent or recurrent orofacial swelling, amongst other findings. Idiopathic orofacial granulomatosis, characterized by an absence of systemic granulomatous disease, is a diagnosis of exclusion. The main differential diagnosis is Crohn's disease. Its pathogenesis is unknown, however, it seems to be immune-mediated. Patch-test sensitivity to multiple allergens is well documented. Currently, therapeutic options consider restrictive diets, topical, intralesional, and systemic agents. First-line therapy is currently a matter of debate. We present a review of the value of diet therapy in this syndrome, along with two illustrative cases.

Background/purpose There are few studies comparing the expression of enamel proteins, such as amelogenin, and cytokeratins in cyst and odontogenic tumors like in ameloblastoma and odontogenic keratocyst, indicating that amelogenin could be a potential biomarker for the aggressiveness in the odontogenic tumors. The aim of this study was to evaluate if the expression of amelogenin, cytokeratin AE1/AE3 (CKAE1/AE3) and cytokeratin 14 (CK14) in cysts and odontogenic tumors with calcified matrices such as calcifying odontogenic cyst (COC), compound (CdO) and complex (CxO) odontomas, adenomatoid odontogenic tumor (AOT) and calcifying epithelial odontogenic tumor (CEOT) as an aggressiveness indicator. Materials and methods Three COC, eight CxO, three CdO, twelve AOT, two CEOT and three dental germs were submitted to an immunohistochemistry panel of antibodies composed of amelogenin, CKAE1/AE3 and CK14. Results CKAE1/AE3 and CK14 was present in all odontogenic epithelia. The amelogenin protein was detected in prismatic and amorphous calcified matrices of epithelial origin belonging to CxO, CdO, AOT, COC and the tooth germs used as controls. On the other hand, the CEOT was the only tumor or cyst studied that did not present immunostaining for amelogenin in calcified matrices. Conclusion Amelogenin was detected in pathologies with a low or absent recurrence rate and excellent prognosis. CEOT was the lesion of greater clinical aggressiveness which did not express amelogenin. The presence of amelogenin in calcified matrices of odontogenic arise could be an indicator of low aggressiveness.

Background: the standard treatment for locally advanced rectal cancer is neoadjuvant chemo-radiotherapy, surgery and adjuvant chemotherapy. Only 50% of patients receive the adjuvant treatment due to the surgical complications and toxicity of radiotherapy. Recently, neoadjuvant chemotherapy has been investigated in the locally advanced rectal cancer setting, with the aim of guaranteeing an uninterrupted systemic treatment. The objective of the present study was to assess the safety and efficacy of neoadjuvant chemotherapy in locally advanced rectal cancer. Methods and patients: patients treated with neoadjuvant chemotherapy and surgery were identified from a prospective database of patients with rectal cancer (cII-III). The primary outcomes were the assessment of the number of R0 resections, the degree of pathologic response, patterns of recurrence and overall and disease-free survival. Treatment schedule: patients received 6-8 cycles of oxaliplatin and fluoropyrimides based chemotherapy. Results: twenty-seven patients who received neoadjuvant chemotherapy were identified. Twenty-six anterior resections and one Hartmann intervention were performed. An R0 resection was performed in 27 (100%) patients and no involvement of the circumferential margin was observed. Complete pathologic response (ypT0N0) was confirmed in four (14.8%) patients. The median follow-up was 35 months (range: 10-81) and four distant recurrences were recorded. Overall and disease-free survival at five years was 85% and 84.7%, respectively. Twenty-seven (100%) patients received all the cycles of chemotherapy, with a mean of six cycles (range 5-8) per patient. Conclusions: neoadjuvant chemotherapy is a promising alternative in the locally advanced rectal cancer setting and further phase III clinical trials are clearly warranted.

The sensitivity of in vivo low-dose high-resolution micro-computed tomography imaging enables monitoring the lung damage caused by tuberculosis. Here, we propose a radiological score integrated in the experimental workflow that enables longitudinal monitoring for prospective efficacy studies in drug development programs. The score is based on an automatic measurement of total unaffected lung volume in vivo normalized for inter-subject comparison. It was validated on well-characterized progression of chronic tuberculosis in Erdman and H37Rv strains in C3HeB/FeJ-based models. We demonstrated that a decrease in the score value indicates increasing adverse effects and vice versa. The colony-forming units count confirmed the variability in the host response suggested by the score values. The correlation between changes in the mice's weight and the score is consistent with disease progression. The classification of disease extent by k-means clustering of the score values provided the definition of the lung damage severity according to the bacillus strain. The proposed score will reduce sources of bias and improve the statistical robustness of studies by the attrition of non-infected subjects or subjects with a weak immune response. Readily available quantifications allow for a fast assessment of the therapeutic potential in drug-resistant tuberculosis strains.

The purpose of the study was to analyze the effectiveness of adding nebulized antibiotics to systemic antimicrobials in critically ill patients with respiratory tract infections (pneumonia or tracheobronchitis) and the effect on renal function. A retrospective observational cohort study including critically ill patients with respiratory tract infections during a 2-year period was conducted. Intervention group included patients that received nebulized and systemic antimicrobials. Patients in the control group received only systemic antimicrobials. Clinical resolution was the primary endpoint. Secondary outcomes included change in fever, inflammatory parameters, and creatinine clearance; length of hospital stay, systemic therapy, and mechanical ventilation; hospital readmission; and mortality. Regression models were performed to estimate the effect of nebulized antibiotics on outcome variables adjusted by potential confounders. A total of 136 patients were included (93 in control group and 43 in intervention group). The intervention group had higher odds of clinical resolution (adjusted odds ratio (OR): 7.1; 95% confidence interval (95% CI): 1.2, 43.3). Nebulized antibiotic therapy was independently associated with reduction in procalcitonin (adjusted OR: 12.4; 95% CI: 1.4, 109.7). There were no significant differences in the rest of the secondary outcomes or in creatinine clearance reduction. Adding nebulized antibiotics for the management of respiratory tract infections has a positive impact on clinical resolution without increasing the risk of renal toxicity. Electronic supplementary material The online version of this article (10.1007/s10096-019-03733-6) contains supplementary material, which is available to authorized users.

Aims The kinase ion channel transient receptor potential melastatin 7 (TRPM7) is considered a modulator of cardiac fibrosis progression; nevertheless, we lack of studies analysing its role in human ischaemic cardiomyopathy (ICM). Our objective was to analyse the expression of genes encoding cardiac ion channels in human ICM, focusing on the alterations in mRNA levels of TRPM7 and its relationship with changes in the ventricular function. Methods and results RNA-sequencing was carried out in 13 left ventricular (LV) samples of patients with ICM compared with a control group (n = 10). The analysis revealed a total of 25 ion channel genes differentially expressed. We performed an RTqPCR analysis of the TRPM7 mRNA in LV and left atrial samples and found that it was down-regulated in both cavities (−1.43-fold and −1.52-fold, respectively). Atrial TRPM7 mRNA levels showed an excellent and inverse relationships with the depressed ejection fraction (r = −0.724, P = 0.042) and with the mitral A wave (r = −0.938, P = 0.006). Conclusions We report the down-regulation of TRPM7 in tissue samples from both left atria and left ventricle in patients with ICM. We found an inverse relationship between both cardiac chambers mRNA levels with LV dysfunction, suggesting an important role of TRPM7 in the left atrial and LV functional depression found in this cardiomyopathy.

Amelogenesis imperfecta (AI) is a group of enamel development disorders that alter the structure and chemical composition of the tissue. There is great variability in the clinical presentation; according to Witkop, AI can be categorized into 14 subtypes, which makes its diagnosis extremely complex. Objective: This study aimed to describe and determine the frequency of clinical and radiographic features and inheritance patterns found in 41 Chilean families diagnosed with diverse types of AI. Material and Methods: We analyzed the clinical records, photographs, pedigrees and radiographs of 121 individuals recruited between 2003 and 2016. All of the information was included in a database that was analyzed using the application Stata 14. Results: The 72 affected individuals had average age of 16 years, and no sex association with the presence of AI was found. The most frequent clinical subtypes were as follows: 43% hypomature, 25% hypoplastic, 21% hypomature/hypoplastic, 7% hypocalcified and 4% hypocalcified/hypoplastic. The number of severely affected teeth was 22, which occurred in the patients with hypocalcified and hypocalcified/hypoplasic AI who presented the highest number of damaged teeth. Caries and periodontal disease were found in 47 and 32% of the patients, respectively. Malocclusions were observed in 43% of the individuals with AI, with open bite being the most frequent. Radiographically, the thickness of the enamel decreased in 51% of the patients, and 80% showed decreased radiopacity of the enamel compared to that of dentin. Autosomal dominant inheritance pattern was found in 37% of the families with hypoplastic AI, and autosomal recessive pattern was present in 56% of the other clinical subtypes, but more frequently in those affected with hypomature and hypocalcified AI. Conclusion: Of the five clinical subtypes, autosomal recessive hypomature, autosomal dominant hypoplastic and autosomal recessive hypomature/hypoplastic AI were the most prevalent subtypes in this group.

Objective To compare cell proliferation and the presence of melanocytes in non-syndromic and syndromic odontogenic keratocyst (OK). Study Design We studied 20 cases of sporadic OK and 10 cases of OK associated with NBCC. To evaluate cell proliferation and the presence of melanocytes, immunohistochemistry was performed with antibodies against Ki-67 and Melan-A, respectively. Results The OK associated with NBCC presented 23.4% (range 8.6%-36.8%) and the sporadic OK 21.8% (range 7.6%-40.4%) of proliferating cells. This difference was not significant. Cells positive for Melan-A occurred exclusively in 4 of 9 cases of syndromic keratocyst. Conclusions No differences were found in the proliferation between OK associated with NBCC and sporadic. Only in the OK associated with the NBCC, the presence of melanocytes was detected.

Adenomatoid odontogenic tumor (AOT) is a benign (hamartomatous), noninvasive lesion with slow but progressive growth. The 3 variants—follicular, extrafollicular, and peripheral—present identical histologic findings. Objective: To present 27 cases of AOT, highlighting their clinical and histologic characteristics. Study Design: Twenty-seven AOT cases were retrieved from the archives of the Oral Pathology Biopsy Service from University of Chile, between 1976-2013. Clinical and histologic findings are described. Results: Twenty-seven AOT cases were reviewed, of which 25 were intraosseous and 2 were peripheral (gingiva). The cases came from 16 females and 11 males with an age range of 5 to 57 years. Of the 25 intraosseous cases, 17 were follicular (associated with impacted teeth), while 8 were extrafollicular (present between teeth). Thirteen of the 27 cases were in the maxilla, and the other 14 were in the mandible. The 2 peripheral cases were clinically diagnosed as a peripheral giant cell granuloma. Histologically, all specimens were similar in morphology, demonstrating a varied degree of duct-like structures and spindle-shaped epithelial cells. Conclusions: The follicular type was the most frequent variant and initially diagnosed as a dentigerous cyst, emphasizing the importance of the histopathologic study of pericoronary radiolucent lesions.

BACKGROUND A variety of local and systemic processes caused exogenous and endogenous pigmentation of the oral mucosa. Solitary melanotic pigmentation is rare, hence the scarce number of studies in children and adolescents. METHODS Clinical and histopathologic features of 10 Latin American children with solitary pigmented lesions of the oral mucosa were reviewed. RESULTS The area most affected was the gingiva, followed by the palate. All lesions were flat and

Introduction Clear cell carcinomas in the jaws are very infrequent neoplasms. Differential diagnosis includes metastatic carcinomas, mucoepidermoid carcinoma, clear cell odontogenic carcinoma, and others. We present two cases of jaw tumors and focus on the diagnostic challenge of each. Case Reports The first case is a 65 year-old man without diagnosis of a systemic disease, with a gingival red tumor and a radiolucent image with irregular borders in the incisor area. Histopathology showed proliferation of clear cells with round hyperchromatic nuclei, some with atypia. These cells formed solid nests separated by thin connective tissue septa with marked vascular proliferation. The clear cells presented diastase–periodic acid–Schiff, anti-Vimentin, anti-CD-10 and anti-PAX-8, anti-human Ki-67 positivity (30% of the cells) and it was negative for S-100 and CK-7. The diagnosis was clear cell carcinoma suggestive of clear cell metastatic carcinoma (MRCC). The second case is a 36 year-old woman with an asymptomatic radiolucent lesion in the periapical area of maxillary premolars. Histopathology showed a cellular proliferation formed by nests of clear oval and polygonal cells, with mild atypia separated by fibrous connective tissue septa. The immunohistochemical staining showed positivity for cytokeratin AE1/AE3 and negative for both S-100 and-smooth muscle actin. Mucicarmin and Congo-red stains were negative. This case was diagnosed as suggestive of clear cell odontogenic carcinoma (CCOC); it was indicated to rule out metastasis. The imaging evaluation confirmed a renal neoplasm in the first case and rule out the presence of lesions in the rest of the body in the second case. Conclusions CCOC and MRCC are histologically similar and immunohistochemistry studies play an important role in diagnosing clear cell tumors. So it is vital for the pathologist to know histomorphology and histo and immuno-histochemistry staining should be considered.

Herein, we report a case of post-transfusion purpura after the reversal of anticoagulation for surgical purposes in a 66-year old ethnic Asian man who was undergoing long-term warfarin therapy for antiphospholipid syndrome. The patient experienced a sudden decrease in platelet count, from 308,000 per μL from the day of admission to 38,000 per μL the following day. Follow-up testing revealed unremarkable red blood cell (RBC) morphology, no evidence of platelet clumping, and negative heparin-induced antibody test results. Platelet antibody testing revealed anti-HPA15a antibodies.

Background The association between dilated cardiomyopathy (DCM) and low thyroid hormone (TH) levels has been previously described. In these patients abnormal thyroid function is significantly related to impaired left ventricular (LV) function and increased risk of death. Although TH was originally thought to be produced exclusively by the thyroid gland, we recently reported TH biosynthesis in the human ischemic heart. Objectives Based on these findings, we evaluated whether the genes required for TH production are also altered in patients with DCM. Methods Twenty-three LV tissue samples were obtained from patients with DCM (n = 13) undergoing heart transplantation and control donors (n = 10), and used for RNA sequencing analysis. The number of LV DCM samples was increased to 23 to determine total T4 and T3 tissue levels by ELISA. Results We found that all components of TH biosynthesis are expressed in human dilated heart tissue. Expression of genes encoding thyroperoxidase (-2.57-fold, P < 0.05) and dual oxidase 2 (2.64-fold, P < 0.01), the main enzymatic system of TH production, was significantlyaltered in patients with DCM and significantly associated with LV remodeling parameters. Thyroxine (T4) cardiac tissue levels were significantly increased (P < 0.01), whilst triiodothyronine (T3) levels were significantly diminished (P < 0.05) in the patients. Conclusions Expression of TH biosynthesis machinery in the heart and total tissue levels of T4 and T3, are altered in patients with DCM. Given the relevance of TH in cardiac pathology, our results provide a basis for new gene-based therapeutic strategies for treating DCM.

Purpose To determine the frequency of limitations on life support techniques (LLSTs) on admission to intensive care units (ICU), factors associated, and 30-day survival in patients with LLST on ICU admission. Methods This prospective observational study included all patients admitted to 39 ICUs in a 45-day period in 2011. We recorded hospitals’ characteristics (availability of intermediate care units, usual availability of ICU beds, and financial model) and patients’ characteristics (demographics, reason for admission, functional status, risk of death, and LLST on ICU admission (withholding/withdrawing; specific techniques affected)). The primary outcome was 30-day survival for patients with LLST on ICU admission. Statistical analysis included multilevel logistic regression models. Results We recruited 3042 patients (age 62.5 ± 16.1 years). Most ICUs (94.8%) admitted patients with LLST, but only 238 (7.8% [95% CI 7.0–8.8]) patients had LLST on ICU admission; this group had higher ICU mortality (44.5 vs. 9.4% in patients without LLST; p

Cutting-edge translational research on preclinical models of lung infectious diseases, such as Tuberculosis disease uses computed tomography (CT) images for assessing infection burden and drug efficacy over treatment. Biomarkers which characterize the distribution and extent of the disease-associated tissue are commonly based on the analysis of the intensity histogram as the involved tissues present abnormal densities in the organ being diagnosed. Often the cellular composition of the tissue represented by those grey-levels is ignored. Our hypothesis is that an accurate CT segmentation of the disease-associate tissue components could be based on the histopathological analysis of the sample. Drug development studies would then benefit of the efficacy assessment by lesion compartment response. We present here a protocol that allows to segment the healthy parenchyma, foamy macrophages and neutrophil foci in excised lung samples of healthy and tuberculous animal models.

The purpose of this paper was to review the literature regarding the clinical and economic impact of pharmacist interventions (PIs) related to antimicrobials in the hospital setting. A PubMed literature search from January 2003 to March 2016 was conducted using the terms pharmacist* or clinical pharmacist* combined with antimicrobial* or antibiotic* or anti-infective*. Comparative studies that assessed the clinical and/or economic impact of PIs on antimicrobials in the hospital setting were reviewed. Outcomes were classified as: treatment-related outcomes (TROs), clinical outcomes (COs), cost and microbiological outcomes (MOs). Acceptance of pharmacist recommendations by physicians was collected. PIs were grouped into patient-specific recommendations (PSRs), policy, and education. Studies' risk of bias was analyzed using Cochrane's tool. Twenty-three studies were evaluated. All of them had high risk of bias. The design in most cases was uncontrolled before and after. PSRs were included in every study; five also included policy and four education. Significant impact of PI was found in 14 of the 18 studies (77.8%) that evaluated costs, 15 of the 20 studies (75.0%) that assessed TROs, 12 of the 22 studies (54.5%) that analyzed COs, and one of the two studies (50.0%) that evaluated MOs. None of the studies found significant negative impact of PIs. It could not be concluded that adding other strategies to PSRs would improve results. Acceptance of recommendations varied from 70 to 97.5%. Pharmacists improve TROs and COs, and decrease costs. Additional research with a lower risk of bias is unlikely to change this conclusion. Future research should focus on identifying the most efficient interventions.

Background A clinical pharmacist (CP), as part of the healthcare team (HT), can contribute to adequate anti-infective use. Few studies have evaluated the economic impact of CP´s interventions (CPI) in the intensive care unit (ICU), and most consider only drug costs. Purpose To analyse the economic impact of CPI regarding antimicrobial therapy (AT) in the ICU. Material and methods We conducted a retrospective analysis of CPI regarding AT in the ICU over a 5 month period. The CP spends 5 hours/day, 5 days/week in the ICU. 33% of CPI are anti-infective related. Information regarding CPI is recorded daily in the hospital´s information system and includes the drug involved, type of intervention, acceptance by physicians and estimated costs (incremental and avoided) as a consequence of the CPI. These costs include changes in drugs, time and products for drug preparations and administration, and the pharmacist’s time. To estimate costs (incremental or avoided) we assumed that the change to the recommended and accepted therapy would have happened 2 days later the without CPI (CPI contribute to earlier changes). For sensitivity analysis, we considered that the change would have happened in 1–4 days. The ratio ‘avoided cost to invested money’ was calculated. Results 212 interventions were recorded, corresponding to 114 patients. Most frequent types of CPI were: modification of drug dose and/or interval (MD) (50.9%), drug discontinuation (DD) (22.6%), change to a more cost effective administration route (CR) (14.6%), initiate a drug (7.5%) and change to a more cost effective drug (CD) (2.4%). Physicians´ acceptance rate was 97.6%. Over the 5-month period, we estimated a total decrease in costs as a consequence of CPI of €7013 (34.8% decrease), corresponding to €33.1/intervention and €61.5/patient. According to sensitivity analysis, total savings varied from €2779 to €19 011. This estimation included the cost of the CP’s time during the studied period (€2859). Therefore, €3.5 were avoided per €1 invested in the CP. Types of CPI associated with greater total savings were (in decreasing order): DD, MD and CR(and per intervention: CD, CR and DD). Conclusion Having a CP as a member of the HT in the ICU performing interventions related to antimicrobials is economically beneficial. References and/or acknowledgements Thanks to the pharmacy service. No conflict of interest

Background In hospital settings (HS), pharmacists´ interventions (PI) can contribute to rational use of antimicrobials. Due to scarce resources, the pharmacist’s time has to be dedicated to interventions with impact on patient outcomes and costs. Purpose A systematic review was conducted to summarise published evidence regarding clinical and/or economic outcomes of PI related to antimicrobials in HS to estimate the impact of PI and to identify strategies with higher impact. Material and methods A PubMed search of papers published from 2003 to March 2016 was conducted using the terms (’pharmacist*’ OR ‘clinical pharmacist*’) AND (’antimicrobial*’ OR ‘antibiotic*’ OR ‘anti infective*’). Additional references were identified from citations. Inclusion criteria were: comparative studies that assessed clinical or economic impact of PI regarding antimicrobials in HS or those that evaluated intermediate outcomes, microbiological impact or appropriate antimicrobial prescription (AAP). Exclusion criteria were: studies in paediatric, primary care or community settings, and evaluations of multidisciplinary teams. We collected: study design, type of PI, impact of PI and acceptance by physicians. Risk of bias of studies was analysed using Cochrane´s tool.1 Results 23 studies were included; all had a high risk of bias. Most frequent design was before and after without a control group (57%) and 83% were single centre studies. Identified PI were grouped into three types: specific recommendations (SR) (in 22 studies), policy (in 4) and education (in 3). Six studies combined various strategies. A significant positive impact of PI was found in 14 of 17 (82%) studies that evaluated costs, in 11 of 15 (73%) that studied AAP, in 9 of 18 (50%) that analysed clinical outcomes (CO), in 1 of 2 (50%) that assessed microbiological outcomes (MO) and in 3 of 7 (43%) that evaluated adverse drug events (ADE). A combination of SR, education and policy had the highest impact on AAP, CO and costs; SR and education on MO; and dose adjustment on ADE. 70–92% of recommendations were accepted. Conclusion Pharmacists´ interventions regarding antimicrobials had a positive impact on appropriate prescribing and patient outcomes, and decreased costs. Combinations of strategies seems to be superior to single strategies. Quality of published studies is poor and better studies are necessary to confirm these results. References and/or acknowledgements 1. Available at http://handbook.cochrane.org/chapter_8/8_5_the_cochrane_collaborations_tool_for_assessing_risk_of_bias.htm No conflict of interest

Changes in cardiomyocyte cytoskeletal components, a crucial scaffold of cellular structure, have been found in heart failure (HF); however, the altered cytoskeletal network remains to be elucidated. This study investigated a new map of cytoskeleton-linked alterations that further explain the cardiomyocyte morphology and contraction disruption in HF. RNA-Sequencing (RNA-Seq) analysis was performed in 29 human LV tissue samples from ischemic cardiomyopathy (ICM; n=13) and dilated cardiomyopathy (DCM, n=10) patients undergoing cardiac transplantation and six healthy donors (control, CNT) and up to 16 ICM, 13 DCM and 7 CNT tissue samples for qRT-PCR. Gene Ontology analysis of RNA-Seq data demonstrated that cytoskeletal processes are altered in HF. We identified 60 differentially expressed cytoskeleton-related genes in ICM and 58 genes in DCM comparing with CNT, hierarchical clustering determined that shared cytoskeletal genes have a similar behavior in both pathologies. We further investigated MYLK4, RHOU, and ANKRD1 cytoskeletal components. qRT-PCR analysis revealed that MYLK4 was downregulated (-2.2-fold; P

Ventricular remodeling, a process involving morphological changes in cardiomyocytes and the extracellular matrix (ECM), has been linked to worsening of clinical status and heart failure (HF) development. In HF, remodeling initially occurs as a compensatory response to preserve the structural

Background: Abnormal thyroid hormone (TH) metabolism is significantly associated with impaired left ventricular (LV) function and death. Although THwas traditionally thought to be produced exclusively by the thyroid gland, an increasing number of studies report TH production in other tissues. Based on these findings, we evaluated whether the genes required for TH biosynthesis are expressed in the human heart, and whether their expression is altered in patients with ischemic cardiomyopathy (ICM) and is related to epigenetic variations. Methods: Twenty-three LV tissue sampleswere obtained fromICMpatients (n=13) undergoing heart transplantation and control donors (n=10) for RNA sequencing analysis. We increased the LV samples to 27 for the ELISA determination of total T4 and T3 tissue levels. For epigenomic studies, 850 K InfiniumMethylationEPIC BeadChip platform was performed. Results: Using RNA-sequencing, we displayed the expression levels of all components required for TH biosynthesis in human heart tissue. We observed significantly altered expression of genes encoding thyroperoxidase (TPO; -2.48-fold, P < 0.05) and dual oxidase 2 (2.83-fold, P < 0.05), themain enzymatic systemof TH production, and significant relationships between their altered expression and LV remodeling parameters. In addition, epigenetic analysis revealed a differential methylation pattern in TPO, and triiodothyronine tissue levels were significantly decreased (P < 0.01). Conclusions: These results showed that the human heart expresses the TH biosynthesismachinery, being altered its main enzymatic system in patients with ICM. Given the relevance of TH in cardiac pathology, our results provide a foundation for new therapeutic approaches based on TPO for treating ICM. (C) 2017 Published by Elsevier Ireland Ltd.

BACKGROUND: The detection of heart transplant rejection by non-invasive methods remains a major challenge. Despite the well-known importance of the study of sarcoplasmic reticulum Ca2+-ATPase (SERCA2a) in the heart, its role as a rejection marker has never been analyzed. Our objective in this study was to determine whether circulating SERCA2a could be a good marker of cardiac rejection. METHODS: We collected 127 consecutive endomyocardial biopsies (EMBs) and serum samples from adult heart transplant recipients (49 without allograft rejection and 78 with the diagnosis of biopsy allograft rejection, including 48 Grade 1R, 21 Grade 2R and 9 Grade 3R). Serum concentrations of SERCA2a were determined using a specific sandwich enzyme-linked immunosorbent assay. We also analyzed SERCA2a expression changes on EMBs using immunofluorescence. RESULTS: SERCA2a cardiac tissue and serum levels were decreased in patients with cardiac rejection (p < 0.0001). A receiver-operating characteristic analysis showed that SERCA2a strongly discriminated between patients with and without allograft rejection: normal grafts vs all rejecting grafts (AUC = 0.804); normal grafts vs Grade 1R (AUC = 0.751); normal grafts vs Grade 2R (AUC = 0.875); normal grafts vs Grade 3R (AUC = 0.922); normal grafts vs Grade 2R and 3R (AUC = 0.889), with p < 0.0001 for all comparisons. CONCLUSIONS: We demonstrated that changes in SERCA2a cardiac tissue and serum levels occur in cardiac allograft rejection. Our findings suggest that SERCA2a concentration assessment may be a relatively simple, non-invasive test for heart transplant rejection, showing a strong capability for detection that improves progressively as rejection grades increase. (C) 2017 The Authors. Published by Elsevier Inc.

The role of nitric oxide synthase 1 (NOS1) as a major modulator of cardiac function has been extensively studied in experimental models; however, its role in human ischemic cardiomyopathy (ICM) has never been analysed. Thus, the objectives of this work are to study NOS1 and NOS-related counterparts involved in regulating physiological function of myocyte, to analyze NOS1 localisation, activity, dimerisation, and its relationship with systolic function in ICM. The study has been carried out on left ventricular tissue obtained from explanted human hearts. Here we demonstrate that the upregulation of cardiac NOS1 is not accompanied by an increase in NOS activity, due in part to the alterations found in molecules involved in the regulation of its activity. We observed partial translocation of NOS1 to the sarcolemma in ischemic hearts, and a direct relationship between its protein levels and systolic ventricular function. Our findings indicate that NOS1 may be significant in the pathophysiology of human ischemic heart disease with a preservative role in maintaining myocardial homeostasis.

13 páginas, 7 figuras, 7 figuras suplementarias, 7 tablas suplementarias.-- et al., Aging in worms and flies is regulated by the PI3K/Akt/Foxo pathway. Here we extend this paradigm to mammals. Ptentg mice carrying additional genomic copies of Pten are protected from cancer and present a significant extension of life span that is independent of their lower cancer incidence. Interestingly, Ptentg mice have an increased energy expenditure and protection from metabolic pathologies. The brown adipose tissue (BAT) of Ptentg mice is hyperactive and presents high levels of the uncoupling protein Ucp1, which we show is a target of Foxo1. Importantly, a synthetic PI3K inhibitor also increases energy expenditure and hyperactivates the BAT in mice. These effects can be recapitulated in isolated brown adipocytes and, moreover, implants of Ptentg fibroblasts programmed with Prdm16 and Cebpβ form subcutaneous brown adipose pads more efficiently than wild-type fibroblasts. These observations uncover a role of Pten in promoting energy expenditure, thus decreasing nutrient storage and its associated damage, Work in the laboratory of A.M.V. was funded by grant SAF2009-08114 and by the CIBERDEM (ISCIII), and work in the laboratory of E. Rial was funded by grants SAF2010-20256 and Consolider-Ingenio CSD2007-00020. A.O.-M. was recipient of a predoctoral contract of the Regional Government of Madrid. E.L.-G. was recipient of a predoctoral contract from the Spanish Ministry of Education. M.M.G.-B. was supported by the “Ramon y Cajal” program of the Spanish Ministry of Science and Innovation.