Cyto-Histological Profile of MicroRNAs as Diagnostic Biomarkers in Differentiated Thyroid Carcinomas.

INTRODUCTION: The repertoire of microRNAs (miRNAs) in thyroid carcinomas starts to be elucidated. Among differentiated thyroid carcinomas (DTCs), papillary thyroid carcinoma (PTC) is the most frequent. The assessment of miRNAs expression may contribute to refine the pre-surgical diagnosis in order to obtain a personalized and more effective treatment for patients. AIMS: This study aims to evaluate (1) the miRNAs in a series of DTCs, and their association with the presence of selected genetic mutations in order to improve diagnosis and predict the biologic behavior of DTC/PTC. (2) The reliability of molecular tests in Ultrasound-guided Fine Needle Aspiration Cytology (US-FNAC) for a more precise preoperative diagnosis. MATERIAL AND METHODS: This series includes 176 samples (98 cytology and 78 histology samples) obtained from 106 patients submitted to surgery, including 13 benign lesions (controls) and 93 DTCs (cases). The microRNA expression was assessed for miR-146b, miR-221, miR-222, and miR-15a through quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). The results were analyzed by the 2-ΔΔCT method, using miR16 as an endogenous control. Regarding PTC diagnosis, the discriminative ability of miRNAs expression was assessed by the area under the Receiver Operating Characteristic Curve (AUC). In PTCs, the association of miRNAs expression, clinicopathological features, and genetic mutations (BRAF, RAS, and TERTp) was evaluated. RESULTS/DISCUSSION: All the analyzed miRNAs presented a tendency to be overexpressed in DTCs/PTCs when compared with benign lesions, both in cytology and histology samples. In cytology, miRNAs expression levels were higher in malignant tumors than in benign tumors. In histology, the discriminative abilities regarding PTC diagnosis were as follows: miR-146b (AUC 0.94, 95% CI 0.87-1), miR-221 (AUC 0.79, 95% CI 0.68-0.9), miR-222 (AUC 0.76, 95% CI 0.63-0.89), and miR-15a (AUC 0.85, 95% CI 0.74-0.97). miR-146b showed 89% sensitivity (se) and 87% specificity (sp); miR-221 se = 68.4, sp = 90; miR-222 se = 73, sp = 70; and mi-R15a se = 72, sp = 80. MicroRNAs were associated with worst-prognosis clinicopathological characteristics in PTCs (p < 0.05), particularly for miR-222. Our data reveal a significant association between higher expression levels of miR-146b, miR-221, and miR-222 in the presence of the BRAF mutation (p < 0.001) and miR-146b (p = 0.016) and miR-221 (p = 0.010) with the RAS mutation, suggesting an interplay of these mutations with miRNAs expression. Despite this study having a relatively small sample size, overexpression of miRNAs in cytology may contribute to a more precise preoperative diagnosis. The miRNAs presented a good discriminative ability in PTC diagnosis. The association between the miRNAs expression profile and genetic alterations can be advantageous for an accurate diagnosis of DTCs/PTCs in FNAC.

Comparative Cyto-Histological Genetic Profile in a Series of Differentiated Thyroid Carcinomas.

INTRODUCTION: Molecular tests can contribute to improve the preoperative diagnosis of thyroid nodules. Tests available are expensive and not adapted to different populations. AIM: This study aimed to compare the cyto-histological genetic profile and to evaluate the reliability of molecular tests using ultrasound-guided fine needle aspiration cytology (US-FNAC) in accurately diagnosing differentiated thyroid carcinomas (DTCs) and predicting biologic behavior of papillary thyroid carcinomas (PTCs). MATERIALS AND METHODS: The series included 259 patients with paired cyto-histological samples totaling 518 samples. The genetic alterations were analyzed via PCR/Sanger sequencing. The association with clinicopathologic features was evaluated in PTCs. RESULTS/DISCUSSION: From the 259 patients included, histologies were 50 (19.3%) benign controls and 209 (80.7%) DTC cases, from which 182 were PTCs; cytologies were 5.8% non-diagnostic, 18.2% benign, 39% indeterminate, and 37.1% malignant. In histology, indeterminate nodules (n = 101) were 22.8% benign and 77.2% malignant. Mutation frequencies in cytology and histology specimens were, respectively, TERTp: 3.7% vs. 7.9%; BRAF: 19.5% vs. 25.1%; and RAS: 11% vs. 17.5%. The overall cyto-histological agreement of the genetic mutations was 94.9%, with Cohen's k = 0.67, and in indeterminate nodules agreement was 95.7%, k = 0.64. The identified mutations exhibited a discriminative ability in diagnosing DTC with a specificity of 100% for TERTp and BRAF, and of 94% for RAS, albeit with low sensitivity. TERTp and BRAF mutations were associated with aggressive clinicopathological features and tumor progression in PTCs (p < 0.001). The obtained good cyto-histological agreement suggests that molecular analysis via US-FNAC may anticipate the genetic profile and the behavior of thyroid tumors, confirming malignancy and contributing to referring patients to surgery.

Relationship between swimming speed, intra-cycle variation of horizontal speed, and Froude efficiency during consecutive stroke cycles in adolescent swimmers.

The purpose of this study was to investigate the relationship between swimming speed, intra-cycle variation of horizontal speed of displacement (dv), and Froude efficiency (ηF) in front-crawl during three consecutive stroke cycles. The sample consisted of 15 boys aged 16.07 ± 0.77 years and 15 girls aged 15.05 ± 1.07 years. Swimming speed, dv and ηF were measured during a 25 m front-crawl trial. Three consecutive stroke cycles were measured. Swimming speed showed a non-significant stroke-by-stroke effect (F = 2.55, p = 0.087, η2 = 0.08), but a significant sex effect (F = 90.46, p < 0.001, η2 = 0.76). The dv and ηF had the same trend as the swimming speed for the stroke-by-stroke effect, but a non-significant sex effect (p > 0.05). The Spearman correlation matrix between swimming speed and dv, and swimming speed and ηF showed non-significant correlations for the three stroke cycles in both sexes. However, the tendency of the former was not always inverse, and the latter was not always direct. Coaches and swimmers need to be aware that lower dvs are not always associated with faster swimming speeds and vice-versa, and that ηF is a predictor of swimming speed, not dv.

Unveiling the efficacy of pre-emergent application of young Eucalyptus globulus leaves as a weed control strategy: Bridging macroscopic effects and cellular responses.

Allelopathy, the inhibition of neighbouring plant growth by certain plants, can be particularly useful if applied in a targeted way for weed management. So, this study aimed to assess and characterize the herbicidal activity of fresh and dried leaves from young Eucalyptus globulus Labill. trees applied as a soil amendment. For this, fresh and dried leaves (FL and DL, respectively) were incorporated into the soil at different concentrations (0, 1, 5, and 10% w/w), where Portulaca oleracea L. seeds were sown. After 5 weeks of exposure, results revealed that the soil incorporation of DL at 10% (w/w) presented the strongest herbicidal properties, inhibiting seed germination by 63% and inducing the loss of cell viability. To unravel the possible mode of action and the main targets at both cellular and subcellular levels, an in vitro experiment was performed. Purslane seeds were sown in a nutritive medium containing different dilutions of an aqueous extract prepared with dried eucalyptus leaves. After 5 days of exposure, germinated seedlings were processed for transmission electron microscopy and histological analyses as well as for reactive oxygen species (ROS) in vivo detection by confocal laser scanning microscopy. Results revealed that the allelochemical release from DL induced ROS overproduction, resulting in the loss of cell integrity and organization, which was characterized by damage to several cellular sub-structures, along with enhanced accumulation of lipid droplets. Overall, the incorporation of DL into the soil can represent a sustainable alternative to reduce synthetic herbicide application and subsequent environmental contamination.

Eucalyptus globulus Leaf Aqueous Extract Differentially Inhibits the Growth of Three Bacterial Tomato Pathogens.

As available tools for crop disease management are scarce, new, effective, and eco-friendly solutions are needed. So, this study aimed at assessing the antibacterial activity of a dried leaf Eucalyptus globulus Labill. aqueous extract (DLE) against Pseudomonas syringae pv. tomato (Pst), Xanthomonas euvesicatoria (Xeu), and Clavibacter michiganensis michiganensis (Cmm). For this, the inhibitory activity of different concentrations of DLE (0, 15, 30, 45, 60, 75, 90, 105, 120, 135, and 250 g L-1) was monitored against the type strains of Pst, Xeu, and Cmm through the obtention of their growth curves. After 48 h, results showed that the pathogen growth was strongly inhibited by DLE, with Xeu the most susceptible species (15 g L-1 MIC and IC50), followed by Pst (30 g L-1 MIC and IC50), and Cmm (45 and 35 g L-1 MIC and IC50, respectively). Additionally, using the resazurin assay, it was possible to verify that DLE considerably impaired cell viability by more than 86%, 85%, and 69% after Pst, Xeu, and Cmm were incubated with DLE concentrations equal to or higher than their MIC, respectively. However, only the treatment with DLE at 120 g L-1 did not induce any hypersensitive response in all pathogens when treated bacterial suspensions were infiltrated onto tobacco leaves. Overall, DLE can represent a great strategy for the prophylactic treatment of tomato-associated bacterial diseases or reduce the application of environmentally toxic approaches.

Accumulation of Proline in Plants under Contaminated Soils-Are We on the Same Page?

Agricultural soil degradation is occurring at unprecedented rates, not only as an indirect effect of climate change (CC) but also due to intensified agricultural practices which affect soil properties and biodiversity. Therefore, understanding the impacts of CC and soil degradation on plant physiology is crucial for the sustainable development of mitigation strategies to prevent crop productivity losses. The amino acid proline has long been recognized for playing distinct roles in plant cells undergoing osmotic stress. Due to its osmoprotectant and redox-buffering ability, a positive correlation between proline accumulation and plants' tolerance to abiotic stress has been pointed out in numerous reviews. Indeed, proline quantification is used systematically by plant physiologists as an indicator of the degree of tolerance and a measurement of the antioxidant potential in plants under stressful conditions. Moreover, the exogenous application of proline has been shown to increase resilience to several stress factors, including those related to soil degradation such as salinity and exposure to metals and xenobiotics. However, recent data from several studies often refer to proline accumulation as a signal of stress sensitivity with no clear correlation with improved antioxidant activity or higher stress tolerance, including when proline is used exogenously as a stress reliever. Nevertheless, endogenous proline levels are strongly modified by these stresses, proving its involvement in plant responses. Hence, one main question arises-is proline augmentation always a sign of improved stress resilience? From this perspective, the present review aims to provide a more comprehensive understanding of the implications of proline accumulation in plants under abiotic stress induced by soil degradation factors, reinforcing the idea that proline quantification should not be employed as a sole indicator of stress sensitivity or resilience but rather complemented with further biochemical and physiological endpoints.

Untargeted metabolomic profiling of fresh and dried leaf extracts of young and mature Eucalyptus globulus trees indicates differences in the presence of specialized metabolites.

Aqueous extracts from Eucalyptus globulus leaves contain a wide variety of specialized metabolites, mainly polyphenols and appreciable amounts of volatile compounds, which are responsible for their diverse biological activities, such as antioxidant, antimicrobial, and allelopathic features. For this reason, several studies have been conducted to explore the composition of E. globulus leaf extracts for multiple therapeutic and commercial applications. However, so far, the available bibliographic reports only refer to the chemical composition of extracts prepared with leaves from mature trees, leaving much to clarify about the composition of juvenile eucalyptus leaf extracts. Furthermore, there is no consensus regarding the type of leaves, fresh or dried ones, to be used in the extraction procedure, considering the highest recovery of biologically active compounds. In this sense, this study aimed to characterize the chemical composition of aqueous extracts prepared with fresh and dried leaves from young and mature E. globulus trees. For this, leaf biomass from young and mature E. globulus trees was collected in three distinct places from a forest area, and after oven-drying a portion of the leaves, an extraction in hot water was carried out, followed by GC-MS and HPLC-MS/MS analyses. The results revealed that the maturity of eucalyptus trees and biomass drying significantly influenced the volatile and non-volatile composition of the aqueous extracts. Accordingly, while fresh leaf extracts of young trees had great levels of hydrolysable tannins, extracts prepared with fresh leaves from mature trees presented a wide range of terpenes. When dried leaf material was used, extracts had notorious contents of amino acids derivatives, C13 norisoprenoids, fatty and other organic acids. Overall, this study showed, for the first time, that plant maturity (young vs mature) and pre-processing (fresh vs dried) of foliar biomass of E. globulus trees need to be considered in the preparation of leaf aqueous extracts depending on the desired purposes, since major changes in what regards biologically active compounds were found.

Numerical and experimental methods used to evaluate active drag in swimming: A systematic narrative review.

Introduction: In swimming, it is necessary to understand and identify the main factors that are important to reduce active drag and, consequently, improve the performance of swimmers. However, there is no up-to-date review in the literature clarifying this topic. Thus, a systematic narrative review was performed to update the body of knowledge on active drag in swimming through numerical and experimental methods. Methods: To determine and identify the most relevant studies for this review, the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) approach was used. Results: 75 studies related to active drag in swimming and the methodologies applied to study them were analyzed and kept for synthesis. The included studies showed a high-quality score by the Delphi scale (mean score was 5.85 ± 0.38). Active drag was included in seven studies through numerical methods and 68 through experimental methods. In both methods used by the authors to determine the drag, it can be concluded that the frontal surface area plays a fundamental role. Additionally, the technique seems to be a determining factor in reducing the drag force and increasing the propulsive force. Drag tends to increase with speed and frontal surface area, being greater in adults than in children due to body density factors and high levels of speed. However, the coefficient of drag decreases as the technical efficiency of swimming increases (i.e., the best swimmers (the fastest or most efficient) are those with the best drag and swimming hydrodynamics efficiency). Conclusion: Active drag was studied through numerical and experimental methods. There are significantly fewer numerical studies than experimental ones. This is because active drag, as a dynamical phenomenon, is too complex to be studied numerically. Drag is greater in adults than in children and greater in men than in women across all age groups. The study of drag is increasingly essential to collaborate with coaches in the process of understanding the fundamental patterns of movement biomechanics to achieve the best performance in swimming. Although most agree with these findings, there is disagreement in some studies, especially when it is difficult to define competitive level and age. The disagreement concerns three main aspects: 1) period of the studies and improvement of methodologies; 2) discrimination of methodologies between factors observed in numerical vs. experimental methods; 3) evidence that drag tends to be non-linear and depends on personal, technical, and stylistic factors. Based on the complexity of active drag, the study of this phenomenon must continue to improve swimming performance.

Subpial Hemorrhage : A Distinctive Neonatal Stroke Pattern.

BACKGROUND AND PURPOSE: Subpial hemorrhage is a rare form of neonatal stroke, still poorly understood. The aim of this study was to characterize a cohort of term and preterm neonates with subpial hemorrhages and contribute to a better knowledge of this condition. MATERIAL AND METHODS: Clinical records and magnetic resonance (MR) imaging data of all neonates with subpial hemorrhage followed at a pediatric hospital between 2010 and 2020 were retrospectively reviewed. RESULTS: A total of 10 patients were included in the analysis, 40% of whom were term neonates. Operative vaginal delivery was registered in 30%. Temporal was the most common location of subpial hemorrhage (70%), and all patients displayed underlying brain infarction. A characteristic yin-yang pattern was present in 90% of the study cohort, and ingurgitation of medullary veins on susceptibility weighted imaging in 80%. Cerebellar microbleeds were observed in 60% of neonates, both term and preterm. When available, MR angiography and venography were unremarkable. Patients' clinical outcome was variable, with early prematurity not associated to worse outcomes. CONCLUSION: Subpial hemorrhage has a distinctive MR pattern, with underlying parenchymal venous infarction, and can occur in term and preterm neonates. This study results suggest an association between subpial hemorrhage and cerebellar microbleeds but further studies are required to confirm it and better understand the pathophysiology of subpial hemorrhage.

Educational Programs and Sexual Counselling for Postmenopausal Sexual Dysfunction: A Systematic Review and Meta-Analysis.

BACKGROUND: Female sexual dysfunction has a high prevalence in women in the postmenopausal period. Not only factors like comorbidities and chronic illnesses are responsible for this high prevalence, but also psychological and interpersonal factors play a significant role. Sexual counselling educational programs have shown positive sexual behavioral changes and they should be considered the first line approach to female sexual dysfunction in this group of women. AIM: To review the evidence of the efficacy of sexual counselling educational programs for sexual dysfunction in postmenopausal women. METHODS: A systematic search was performed in February 2021 from electronic databases (MEDLINE, CENTRAL, and Scopus), unpublished studies, ongoing clinical trials, conference abstracts and journal archives, dissertations and theses, gray literature and free search on the Google Scholar search engine. Studies that evaluate the effectiveness of sexual educational programs in postmenopausal women were selected. Study selection and data extraction were performed by 2 independent researchers. Extracted data included author identification, publication date, geographic location, study population and sample size, type of intervention, sexual function evaluation tool and outcome. Studies that evaluate sexual function by using the Female Sexual Function Index (FSFI) were included in the meta-analysis, calculating the mean difference. OUTCOMES: Improvement of sexual function in postmenopausal women. RESULTS: We included 8 studies in the systematic review, 6 randomized and 2 nonrandomized controlled trials, with a total sample size of 619 women, aged between 39 and 75 years old, all in menopause for less than 5 years. The studies described sexual education programs, with 4-10 sessions, 45-60 minutes each, including themes like sexual anatomy, physiological sexual response, menopause, methods of stimulation, and common sexual myths. Five studies also included cognitive-behavior therapy and 3 studies assessed mindfulness techniques. Six studies evaluated the effectiveness of sexual educational programs using FSFI. The results showed that sexual counselling educational programs had statistically significant effects on enhancing the total FSFI score (mean difference = +7.14, 95% confidence interval = 3.70-10.6, P < .0001) in comparison to routine care. Results were also significant in all evaluated sex domains: pain, arousal, lubrication, desire, orgasm and satisfaction (P < .05). CONCLUSION: Our meta-analysis shows that sexual counselling educational programs are effective in improving sexual dysfunction in postmenopausal women when compared to routine care. These are simple approaches, easily administered with minimal resources that help prevent the psychological and social consequences of sexual dysfunction at this age. IMS Silva, MP Pinto, D Gonçalves. Educational Programs and Sexual Counselling for Postmenopausal Sexual Dysfunction: A Systematic Review and Meta-Analysis. J Sex Med 2022;19:54-63.

Salivary cortisol and eye temperature changes during endurance competitions.

BACKGROUND: The purpose of this study was to investigate the usefulness of salivary cortisol (SC) and eye temperature measured by infrared thermography (IRTET) as biomarkers to manage competitions more effectively and monitor horse welfare in endurance competitions. Based on previous studies, it was hypothesised that pre-exercise baseline SC and IRTET would be higher in younger or less experienced horses, and that post-exercise variation from baseline would be higher in the top finishers. RESULTS: Salivary cortisol measured in 61 competing at qualifier 40 km and 80 km rides showed an abrupt variation (93-256% rise) of the baseline SC levels [median ± interquartile range (IQR) = 0.27 ng/dl ± 0.36] obtained at the Pre-Inspection (PI) into Vet Gate (VG)1 independently of the covered distance, but modest or even lower in the subsequent Vet Gates, e.g. VG2 or VG3. The IRTET measured concomitantly in 16 horses showed significant (p < 0.05) higher levels at the PI in less experienced horses participating in the 40 km ride (median ± IQR = 35.7 °C ± 1.4) than their counterparts in the 80 km ride (median ± IQR = 35.0 °C ± 1.5), but not SC. Baseline SC levels at the PI of horses classifying in the Top5 in the 40 km ride category were significantly (p < 0.05) higher median ± IQR = 0.90 ng/ml ±0.61) when compared to horses positioned from 10th position on (median ± IQR = 0.16 ng/ml ±0.40). A lower IRTET in the PI was correlated with better placement (p < 0.05) and those in the Top5 (median ± IQR = 33.9 °C ± 0.0) had a significantly (p < 0.5) higher variation (+ 10.65%) into the last VG. CONCLUSION: Pre-exercise baseline IRTET levels, but not SC, were higher in less experienced horses in the 40 compared to their counterparts in the 80 km ride competitions. SC and IRTET showed different indications according to the competition. In the40 km ride competition, higher baseline pre-exercise SC levels seemed to be linked to a better classification outcome. In contrast, in the 80 km ride horses, the higher IRTET variation from pre-exercise into final Vet Gate was the parameter associated with a better performance. A more controlled environment and a larger sample are needed to confirm these results and monitor horse welfare in competitions.

Herbicidal Effects and Cellular Targets of Aqueous Extracts from Young Eucalyptus globulus Labill. Leaves.

Eucalyptus globulus Labill. is a widespread exotic species that contributes to the formation of fire-prone environments, a great concern under climate change conditions. Therefore, sustainable practices to help locals managing eucalyptus stands are needed. In this perspective, harnessing eucalyptus' specialized metabolism as a source of allelochemicals can be a promising approach for weed control. Thus, the main goals of this work were to evaluate the herbicidal potential of post-fire regenerated E. globulus leaves against Portulaca oleracea L. and to unravel the physiological mechanisms behind this phytotoxic action. For this, aqueous extracts of fresh (FLE; 617 g FW L-1) or oven-dried leaves (DLE; 250 g DW L-1) were foliar-sprayed at different dilutions in purslane seedlings. After five weeks, results revealed that DLE at the highest dose detained the greatest herbicidal activity, affecting purslane growth and cellular viability. Moreover, biochemical data pointed towards an overproduction of reactive oxygen species, causing harsh oxidative damage in roots, where the upregulation of important cellular players, like sugars, amino acids, and proline, was not able to reestablish redox homeostasis. Overall, this study proved that dried leaves from young E. globulus had potent herbicidal properties against P. oleracea and can represent a feasible strategy for weed management.

Epigenomics in Hurthle Cell Neoplasms: Filling in the Gaps Towards Clinical Application.

It has been widely described that cancer genomes have frequent alterations to the epigenome, including epigenetic silencing of various tumor suppressor genes with functions in almost all cancer-relevant signalling pathways, such as apoptosis, cell proliferation, cell migration and DNA repair. Epigenetic alterations comprise DNA methylation, histone modification, and microRNAs dysregulated expression and they play a significant role in the differentiation and proliferation properties of TC. In this review, our group assessed the published evidence on the tumorigenic role of epigenomics in Hurthle cell neoplasms (HCN), highlighting the yet limited, heteregeneous and non-validated data preventing its current use in clinical practice, despite the well developed assessment techniques available. The identified evidence gaps call for a joint endeavour by the medical community towards a deeper and more systematic study of HCN, aiming at defining epigenetic markers in early diagnose, allowing for accurate stratification of maligancy and disease risk and for effective systemic treatment.

TERTp mutations and p53 expression in head and neck cutaneous basal cell carcinomas with different aggressive features.

Cutaneous basal cell carcinoma (cBCC) is an economic burden to health services, due to its great morbidity and increasing incidence in old people. Infiltrative cBCCs and cBCCs with micronodular pattern are considered as more aggressive. The role of p53 expression and TERTp mutation on cBCC behavior remains to be clarified. We aimed to assess TERTp mutations and p53 expression in relation to the cBCC histological subtype in a cohort of patients referred to an ENT Department of a tertiary Hospital of Northern Portugal. We performed a retrospective clinicopathological and histological review of the head and neck cBCCs followed-up at the otorhinolaryngology department of Trás-os-Montes e Alto Douro hospital (January 2007-June 2018). We assessed TERTp mutations in 142 cBCCs and p53 protein expression, through immunohistochemistry, in 157 cBCCs. We detected TERTp mutations in 43.7% of cBCCs and p53 overexpression in 60.5% of cBCCs. We spotted association of p53 overexpression and TERTp mutation with necrosis. In the infitrative-growth pattern cBCCs, there was no significant association with the clinical and histological features evaluated, except for necrosis. In the indolent-growth cBCCs, we identified a significant association of TERTp mutation status with female sex, necrosis, multiple cBCCs, and p53 positive expression. Our results suggest that TERTp mutation may be useful to identify more aggressive features in the indolent-growth pattern cBCCs (nodular and superficial subtypes). Further studies with larger cohorts are warranted to clarify the relevance of TERTp mutation in cBCCs.

Genetic Determinants for Prediction of Outcome of Patients with Papillary Thyroid Carcinoma.

Papillary thyroid carcinoma (PTC) usually presents an excellent prognosis, but some patients present with aggressive metastatic disease. BRAF, RAS, and TERT promoter (TERTp) genes are altered in PTC, and their impact on patient outcomes remains controversial. We aimed to determine the role of genetic alterations in PTC patient outcomes (recurrent/persistent disease, structural disease, and disease-specific mortality (DSM)). The series included 241 PTC patients submitted to surgery, between 2002-2015, in a single hospital. DNA was extracted from tissue samples of 287 lesions (primary tumors and metastases). Molecular alterations were detected by Sanger sequencing. Primary tumors presented 143 BRAF, 16 TERTp, and 13 RAS mutations. Isolated TERTpmut showed increased risk of structural disease (HR = 7.0, p < 0.001) and DSM (HR = 10.1, p = 0.001). Combined genotypes, BRAFwt/TERTpmut (HR = 6.8, p = 0.003), BRAFmut/TERTpmut (HR = 3.2, p = 0.056) and BRAFmut/TERTpwt (HR = 2.2, p = 0.023) showed increased risk of recurrent/persistent disease. Patients with tumors BRAFwt/TERTpmut (HR = 24.2, p < 0.001) and BRAFmut/TERTpmut (HR = 11.5, p = 0.002) showed increased risk of structural disease. DSM was significantly increased in patients with TERTpmut regardless of BRAF status (BRAFmut/TERTpmut, log-rank p < 0.001; BRAFwt/TERTpmut, log-rank p < 0.001). Our results indicate that molecular markers may have a role in predicting PTC patients' outcome. BRAFmut/TERTpwt tumors were prone to associate with local aggressiveness (recurrent/persistent disease), whereas TERTpmut tumors were predisposed to recurrent structural disease and DSM.

Translational Potential of Epigenetic-Based Markers on Fine-Needle Aspiration Thyroid Specimens.

The awareness of epigenetic alterations leading to neoplasia attracted the attention of researchers toward its potential use in the management of cancer, from diagnosis to prognosis and prediction of response to therapies. Our group has focused its attention on the epigenomics of thyroid neoplasms. Although most of the epigenetic studies have been applied on histological samples, the fact is that cytology, through fine-needle aspiration, is a primary diagnostic method for many pathologies, of which thyroid nodules are one of the most paradigmatic examples. This has led to an increasing literature report of epigenetic studies using these biological samples over the past decade. In this review, our group aimed to document recent research of epigenetic alterations and its associated assessment techniques, based on cytology material. Our review covers the main epigenetic categories-DNA methylation, histone modification, and RNA-silencing-whose evidence in thyroid cytology samples may represent solid soil for future prospectively designed studies aiming at validating patterns of epigenetic alterations and their potential use in the clinical management of thyroid neoplasms.

Prediction of disease progression and outcomes in multiple sclerosis with machine learning.

Multiple Sclerosis is a chronic inflammatory disease, affecting the Central Nervous System and leading to irreversible neurological damage, such as long term functional impairment and disability. It has no cure and the symptoms vary widely, depending on the affected regions, amount of damage, and the ability to activate compensatory mechanisms, which constitutes a challenge to evaluate and predict its course. Additionally, relapsing-remitting patients can evolve its course into a secondary progressive, characterized by a slow progression of disability independent of relapses. With clinical information from Multiple Sclerosis patients, we developed a machine learning exploration framework concerning this disease evolution, more specifically to obtain three predictions: one on conversion to secondary progressive course and two on disease severity with rapid accumulation of disability, concerning the 6th and 10th years of progression. For the first case, the best results were obtained within two years: AUC=[Formula: see text], sensitivity=[Formula: see text] and specificity=[Formula: see text]; and for the second, the best results were obtained for the 6th year of progression, also within two years: AUC=[Formula: see text], sensitivity=[Formula: see text], and specificity=[Formula: see text]. The Expanded Disability Status Scale value, the majority of functional systems, affected functions during relapses, and age at onset were described as the most predictive features. These results demonstrate the possibility of predicting Multiple Sclerosis progression by using machine learning, which may help to understand this disease's dynamics and thus, advise physicians on medication intake.

Mechanical thrombectomy in stroke in nonagenarians: useful or futile?

BACKGROUND: Mechanical thrombectomy is the standard treatment in acute ischemic stroke due to large vessel occlusion, but there is limited evidence about its efficacy in very old patients. We sought to analyse safety and effectiveness of mechanical thrombectomy in nonagenarian versus octagenarian patients. METHODS: We included consecutive patients with acute ischemic stroke due to large vessel occlusion subjected to mechanical thrombectomy, during 29 months in a tertiary center. Patients were divided into two sub-groups, according to age: 80-89 and >90 years old. Recanalization, complications, functional outcome and mortality at discharge and at 3 months were compared. Multivariable analysis was performed to identify independent predictors of functional outcome at 3 months of follow-up, assessed by the modified Rankin Scale. RESULTS: A total of 128 octogenarians (88.9%) and 16 nonagenarians (11.1%) met the inclusion criteria. Successful revascularization was achieved in 87.5% of octagenarians and in 81.3% of nonagenarians (p = 0.486). Symptomatic hemorrhage occurred in 3.1% and 6.3% of younger and older patients, respectively (p = 0.520). Cerebral edema occured in 35.2% of octagenarians versus 25.0% of nonagenarians (p = 0.419). Functional independence (mRS ≤ 2) at 3 months was achieved in 28 (22.6%) and 5 (31.3%) of octagenarians and nonagenarians, respectively (p = 0.445). Mortality at 3 months was not significantly higher in nonagenarians (37.5%) versus octagenarians (33.9%, p = 0.773). CONCLUSIONS: No significant diferences were found in functional outcome, mortality, recanalization and complication rates between octagenarians and nonagenarians submitted to mechanical thrombectomy, underlining that patients should not be excluded from mechanical thrombectomy based on age alone.

Development of a Gold Nanoparticle-Based Lateral-Flow Immunoassay for Pneumocystis Pneumonia Serological Diagnosis at Point-of-Care.

Pneumocystis jirovecii pneumonia (PcP) is a major human immunodeficiency virus (HIV)-related illness, rising among immunocompromised non-HIV patients and in developing countries. Presently, the diagnosis requires respiratory specimens obtained through invasive and costly techniques that are difficult to perform in all patients or implement in all economic settings. Therefore, the development of a faster, cost-effective, non-invasive and field-friendly test to diagnose PcP would be a significant advance. In this study, recombinant synthetic antigens (RSA) of P. jirovecii's major surface glycoprotein (Msg) and kexin-like serine protease (Kex1) were produced and purified. These RSA were applied as antigenic tools in immunoenzymatic assays for detection of specific anti-P. jirovecii antibodies (IgG and IgM) in sera of patients with (n = 48) and without (n = 28) PcP. Results showed that only IgM anti-P. jirovecii levels were significantly increased in patients with PcP compared with patients without P. jirovecii infection (p ≤ 0.001 with both RSA). Thus, two strip lateral flow immunoassays (LFIA), based on the detection of specific IgM anti-P. jirovecii antibodies in human sera samples, were developed using the innovative association of P. jirovecii's RSA with spherical gold nanoparticles (AuNPs). For that, alkanethiol-functionalized spherical AuNPs with ca. ~40 nm in diameter were synthetized and conjugated with the two RSA (Msg or Kex1) produced. These AuNP-RSA conjugates were characterized by agarose gel electrophoresis (AGE) and optimized to improve their ability to interact specifically with serum IgM anti-P. jirovecii antibodies. Finally, two LFIA prototypes were developed and tested with pools of sera from patients with (positive sample) and without (negative sample) PcP. Both LFIA had the expected performance, namely, the presence of a test and control red colored lines with the positive sample, and only a control red colored line with the negative sample. These results provide valuable insights into the possibility of PcP serodiagnosis at point-of-care. The optimization, validation and implementation of this strip-based approach may help to reduce the high cost of medical diagnosis and subsequent treatment of PcP both in industrialized and low-income regions, helping to manage the disease all around the world.

Phytotoxic effects of bulk and nano-sized Ni on Lycium barbarum L. grown in vitro - Oxidative damage and antioxidant response.

This study aimed to evaluate the effects of nickel oxide nanomaterial (nano-NiO) on goji berry (Lycium barbarum L.) shoots grown under in vitro conditions and to determine if the nanomaterial was more harmful than its bulk counterpart, nickel (II) sulphate (NiSO4). For this purpose, in vitro shoots of L. barbarum were cultured on MS medium supplemented with 15 mg L-1 of NiSO4 or nano-NiO. Nano-NiO was more harmful for shoots growth and photosynthetic pigments than NiSO4, with reductions up to 82% in comparison to the control. Shoots treated with nano-NiO presented an overproduction of hydrogen peroxide (H2O2; 130% increase) and superoxide anion (O2-; 110% increase), which led to higher levels of lipid peroxidation (LP; 57% increase) and the occurrence of oxidative stress. In opposition, bulk Ni seemed not to induce oxidative stress, once LP and reactive oxygen species content decreased in comparison with the control. The evaluation of the non-enzymatic antioxidant (AOX) system revealed that, under nano-NiO excess, proline, ascorbate, glutathione and phenols levels increased up to 4-fold, but did not change in response to the bulk treatment. With respect to the enzymatic AOX system, nano-NiO enhanced the activities of superoxide dismutase (203%) and ascorbate peroxidase (62%), though catalase activity was negatively affected, while bulk Ni did not majorly affect these enzymes' behavior. Overall, the data showed that Ni phytotoxicity in L. barbarum shoots depends on the metal source and that, in this case, nano-NiO seemed to be more deleterious to goji shoots grown under in vitro conditions than NiSO4.