Search

Abstract Coronavirus disease 2019 (COVID-19) is caused by Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Recent research has demonstrated how epigenetic mechanisms regulate the host–virus interactions in COVID-19. It has also shown that microRNAs (miRNAs) are one of the three fundamental mechanisms of the epigenetic regulation of gene expression and play an important role in viral infections. A pilot study published by our research group identified, through next-generation sequencing (NGS), that miR-4433b-5p, miR-320b, and miR-16–2-3p are differentially expressed between patients with COVID-19 and controls. Thus, the objectives of this study were to validate the expression of these miRNAs using quantitative real-time polymerase chain reaction (qRT-PCR) and to perform in silico analyses. Patients with COVID-19 (n = 90) and healthy volunteers (n = 40) were recruited. MiRNAs were extracted from plasma samples and validated using qRT-PCR. In addition, in silico analyses were performed using mirPath v.3 software. MiR-320b was the only miRNA upregulated in the case group com-pared to the control group. The in silico analyses indicated the role of miR-320b in the regulation of the KITLG gene and consequently in the inflammatory process. This study confirmed that miR-320b can distinguish patients with COVID-19 from control participants; however, further research is needed to determine whether this miRNA can be used as a target or a biomarker.

Abstract Efficient predictive biomarkers are needed for immune checkpoint inhibitor (ICI)-based immunotherapy in non-small cell lung cancer (NSCLC). Testing the predictive value of single nucleotide polymorphisms (SNPs) in programmed cell death 1 (PD-1) or its ligand 1 (PD-L1) has shown contrasting results. Here, we aim to validate the predictive value of PD-L1 SNPs in advanced NSCLC patients treated with ICIs as well as to define the molecular mechanisms underlying the role of the identified SNP candidate. rs822336 efficiently predicted response to anti-PD-1/PD-L1 immunotherapy in advanced non-oncogene addicted NSCLC patients as compared to rs2282055 and rs4143815. rs822336 mapped to the promoter/enhancer region of PD-L1, differentially affecting the induction of PD-L1 expression in human NSCLC cell lines as well as their susceptibility to HLA class I antigen matched PBMCs incubated with anti-PD-1 monoclonal antibody nivolumab. The induction of PD-L1 expression by rs822336 was mediated by a competitive allele-specificity binding of two identified transcription factors: C/EBPβ and NFIC. As a result, silencing of C/EBPβ and NFIC differentially regulated the induction of PD-L1 expression in human NSCLC cell lines carrying different rs822336 genotypes. Analysis by binding microarray further validated the competitive allele-specificity binding of C/EBPβ and NFIC to PD-L1 promoter/enhancer region based on rs822336 genotype in human NSCLC cell lines. These findings have high clinical relevance since identify rs822336 and induction of PD-L1 expression as novel biomarkers for predicting anti-PD-1/PD-L1-based immunotherapy in advanced NSCLC patients.

The purpose of the study was to evaluate dry matter phytomass production, percentage of soil cover, and dry matter phytomass decomposition curve of cover crops in single and mix cultivation, in a low-altitude subtropical environment. The study was conducted in Santa Maria - RS, in agricultural years 2019/2020 and 2021/2022. The experimental design used was randomized blocks, with ten treatments and four replications. The experiments were conducted with four winter cover crops: White oat (WO), black oat (BO), forage turnip (FT), and common vetch (CV), in single cultivation and as a mix of cover plants. For the total production of dry matter phytomass, the FT and BO+CV+FT treatments stand out which present the highest averages between the experimental years, with values ​​4365.32 and 4109.87 kg ha-1. FT in single cultivation has a high potential for soil coverage in periods of 45 DAS, and the common vetch crop showed greater soil coverage in the late period, with 84% coverage at 90 days after sowing. The BO and WO treatments showed greater maintenance of dry phytomass in soil cover at the end of the 150-day period, presenting values greater than 40% of the initial volume of dry phytomass present in the production system. Recommending the exclusive cultivation of the species that meets the rural producer's objective.

Chronic obstructive pulmonary disease (COPD) represents an independent risk factor for lung cancer development. Accelerated cell senescence, induced by oxidative stress and inflammation, is a common pathogenic determinant of both COPD and lung cancer. The post transcriptional regulation of genes involved in these processes is finely regulated by RNA-binding proteins (RBPs), which regulate mRNA turnover, subcellular localization, splicing and translation. Multiple pro-inflammatory mediators (including cytokines, chemokines, proteins, growth factors and others), responsible of lung microenvironment alteration, are regulated by RBPs. Several mouse models have shown the implication of RBPs in multiple mechanisms that sustain chronic inflammation and neoplastic transformation. However, further studies are required to clarify the role of RBPs in the pathogenic mechanisms shared by lung cancer and COPD, in order to identify novel biomarkers and therapeutic targets. This review will therefore focus on the studies collectively indicating the role of RBPs in oxidative stress and chronic inflammation as common pathogenic mechanisms shared by lung cancer and COPD.

Strawberry (Fragaria × ananassa Duch.) production can be evaluated as repeated measurements, since the same plant is harvested multiple times during the production season. The objectives were to evaluate the production of fresh mass and fruit number in successive harvests and compare three strawberry clones in two cultivation conditions. Two experiments were carried out in two environmental cultivations: the rural property and the experimental area of the Plant Science Department, Federal University of Santa Maria, Brazil. The parameters of the nonlinear logistic model and their critical points were estimated via bootstrap for each condition and clone for fresh mass and fruit number with accumulated values, depending on the thermal sum accumulated during the production season. For nonlinear regression analysis, the ordinary least squares method was used with the Gauss–Newton algorithm. Confidence intervals were obtained for each parameter and estimated critical points, and they did not cross; the treatments were considered different. There were significant differences between clones and cultivation conditions for fruit mass and number. The nonlinear logistic models, adjusted for mass and number of strawberry fruits, detailed the production season, highlighting the main differences between cultivation conditions and clones.

Abstract Macroautophagy, hereafter referred to as autophagy, represents a highly conserved catabolic process that maintains cellular homeostasis. At present, the role of autophagy in cutaneous melanoma (CM) is still controversial, since it appears to be tumor-suppressive at early stages of malignant transformation and cancer-promoting during disease progression. Interestingly, autophagy has been found to be often increased in CM harboring BRAF mutation and to impair the response to targeted therapy. In addition to autophagy, numerous studies have recently conducted in cancer to elucidate the molecular mechanisms of mitophagy, a selective form of mitochondria autophagy, and secretory autophagy, a process that facilitates unconventional cellular secretion. Although several aspects of mitophagy and secretory autophagy have been investigated in depth, their involvement in BRAF-mutant CM biology has only recently emerged. In this review, we aim to overview autophagy dysregulation in BRAF-mutant CM, along with the therapeutic advantages that may arise from combining autophagy inhibitors with targeted therapy. In addition, the recent advances on mitophagy and secretory autophagy involvement in BRAF-mutant CM will be also discussed. Finally, since a number of autophagy-related non-coding RNAs (ncRNAs) have been identified so far, we will briefly discussed recent advances linking ncRNAs to autophagy regulation in BRAF-mutant CM.

Chronic obstructive pulmonary disease (COPD) is an independent risk factor for the development of lung cancer. Several pathogenic pathways such as oxidative stress and chronic inflammation of the lower airways are shared by these two diseases. The RNA-binding proteins (RBPs) can modulate the post-transcriptional expression of genes involved in several biological processes, such as cell cycle, cell proliferation and injury/stress responses. The bonding between RBPs and targeted transcripts regulates mRNA turnover, subcellular localization, splicing and translation, through the formation of ribonucleoproteins (RNPs) complexes. Several pro-inflammatory mediators, that induce and maintain chronic lung inflammation and carcinogenesis, are modulate by RBPs. RBPs can have a key role in lung cancer onset in COPD patients, sharing several common pathogenic processes, including oxidative stress, inflammation of lower airways, DNA damage and impaired DNA-repair mechanisms. Further studies are mandatory to clarify the application of RBPs as novel biomarkers and potential therapeutic targets. This review will investigate recent evidences showing the role of RBPs in the pathogenic mechanisms sharing by lung cancer and COPD

IntroductionThe RNA-binding protein AU-rich-element factor-1 (AUF-1) participates to posttranscriptional regulation of genes involved in inflammation and cellular senescence, two pathogenic mechanisms of chronic obstructive pulmonary disease (COPD). Decreased AUF-1 expression was described in bronchiolar epithelium of COPD patients versus controls and in vitro cytokine- and cigarette smoke-challenged human airway epithelial cells, prompting the identification of epithelial AUF-1-targeted transcripts and function, and investigation on the mechanism of its loss.ResultsRNA immunoprecipitation-sequencing (RIP-Seq) identified, in the human airway epithelial cell line BEAS-2B, 494 AUF-1-bound mRNAs enriched in their 3’-untranslated regions for a Guanine-Cytosine (GC)-rich binding motif. AUF-1 association with selected transcripts and with a synthetic GC-rich motif were validated by biotin pulldown. AUF-1-targets’ steady-state levels were equally affected by partial or near-total AUF-1 loss induced by cytomix (TNFα/IL1β/IFNγ/10 nM each) and siRNA, respectively, with differential transcript decay rates. Cytomix-mediated decrease in AUF-1 levels in BEAS-2B and primary human small-airways epithelium (HSAEC) was replicated by treatment with the senescence- inducer compound etoposide and associated with readouts of cell-cycle arrest, increase in lysosomal damage and senescence-associated secretory phenotype (SASP) factors, and with AUF-1 transfer in extracellular vesicles, detected by transmission electron microscopy and immunoblotting. Extensive in-silico and genome ontology analysis found, consistent with AUF-1 functions, enriched RIP-Seq-derived AUF-1-targets in COPD-related pathways involved in inflammation, senescence, gene regulation and also in the public SASP proteome atlas; AUF-1 target signature was also significantly represented in multiple transcriptomic COPD databases generated from primary HSAEC, from lung tissue and from single-cell RNA-sequencing, displaying a predominant downregulation of expression.DiscussionLoss of intracellular AUF-1 may alter posttranscriptional regulation of targets particularly relevant for protection of genomic integrity and gene regulation, thus concurring to airway epithelial inflammatory responses related to oxidative stress and accelerated aging. Exosomal-associated AUF-1 may in turn preserve bound RNA targets and sustain their function, participating to spreading of inflammation and senescence to neighbouring cells.

Abstract Phospholipid scramblase 1 (PLSCR1) is the most studied protein of the scramblase family. Originally, it was identified as a membrane protein involved in maintaining plasma membrane asymmetry. However, studies conducted over the past few years have shown the involvement of PLSCR1 in several other cellular pathways. Indeed, PLSCR1 is not only embedded in the plasma membrane but is also expressed in several intracellular compartments where it interacts with a diverse repertoire of effectors, mediators, and regulators contributing to distinct cellular processes. Although most PLSCR1 interactors are thought to be cell-type specific, PLSCR1 often exerts its regulatory functions through shared mechanisms, including the trafficking of different molecules within intracellular vesicles such as endosomes, liposomes, and phagosomes. Intriguingly, besides endogenous proteins, PLSCR1 was also reported to interact with exogenous viral proteins, thereby regulating viral uptake and spread. This review aims to summarize the current knowledge about the multiple roles of PLSCR1 in distinct cellular pathways. Video Abstract

Abstract Introduction Paracoccidioidomycosis is a systemic mycosis endemic to Latin America, with frequent reports of poor treatment adherence resulting in worse clinical outcomes. The present study evaluated factors related to treatment adherence in patients with paracoccidioidomycosis. Methods This cross‐sectional study included data from medical records of patients with paracoccidioidomycosis treated between 2005 and 2018 in a reference hospital in southeastern Brazil. Clinical variables, demographic characteristics, and smoking and alcohol consumption were collected and analyzed. Results Of the 198 cases included, most were adults (median age = 51 years), males (93.4%), presented chronic paracoccidioidomycosis (83.8%), reported smoking (90.4%) and alcohol consumption (84.8%), and 56.6% adhered to treatment. Treatment adherence was associated with the discontinuation of alcohol consumption (odds ratios (OR) = 5.227; 95% confidence interval (CI) = 2.697–10.129) and smoking (OR = 3.365; 95% CI = 1.781–6.358) during therapy. Conclusions Patients' continued consumption of alcohol and tobacco was associated with non‐adherence to treatment, suggesting that a joint therapeutic approach that addresses alcohol and tobacco use alongside paracoccidioidomycosis treatment may hold promise for improving adherence and patients' quality of life.

Purpose: Current World Health Organization/International Society of Urological Pathology (2004 WHO/ISUP) grading of bladder urothelial carcinoma relies on the highest pathologic grade of the specimen and does not reflect the inherent qualitative and quantitative heterogeneity of disease. Materials and Methods: We retrospectively studied consecutive urothelial high-grade cT1 (cT1HG) carcinomas submitted to adjuvant bacille Calmette–Guérin between 2008 and 2015 to evaluate the prognostic potential of grade 3 (presence or predominance) according to the 1973 WHO system concerning disease progression and cancer-specific death. Results: Among 253 patients, grading distribution was 34.4% 1+2, 7.5% 2+1, 20.2% 2+2, 19.0% 2+3, 5.1% 3+2, and 13.8% 3+3. Recurrence was diagnosed in 115 (45.5%), progression in 83 (32.8%), and cancer-specific death in 50 patients (19.8%). Mean time to recurrence, progression, and death from disease were 35.9±31.7, 47.6±44.5, and 51.2±50.4 months, respectively. Grade 3 presence (2+3, 3+2, or 3+3) occurred in 96 (37.9%) and independently predicted time to progression (p

Vascular aging is linked to reduce NO bioavailability, endothelial dysfunction, oxidative stress, and inflammation. We previously showed that a 4-week treatment of middle-aged Wistar rats (MAWRs, 46 weeks old) with Moringa oleifera seed powder (MOI, 750 mg/kg/day) improved vascular function. Here, we investigated the involvement of SIRT1 in MOI-induced vascular improvement. MAWRs were treated with a standard or MOI-containing diet. Young rats (YWR, 16 weeks old) were the controls and received a standard diet. The hearts and aortas were harvested to evaluate SIRT1 and FOXO1 expression via Western blot and/or immunostaining, SIRT1 activity via a fluorometric assay, and oxidative stress using the DHE fluorescent probe. In the hearts and aortas, SIRT1 expression, reduced in MAWRs compared to YWRs, was enhanced in MOI MAWRs. In the hearts, SIRT1 activity did not differ between YWRs and MAWRs, whereas it was increased in MOI MAWRs compared with them. In the aortas, SIRT1 activity decreased in MAWRs, and it was similar in the MOI MAWRs and YWRs. FOXO1 expression increased in the nuclei of MAWR aortas compared to YWR and was reversed in MOI MAWRs. Interestingly, MOI treatment normalized oxidative stress enhanced in MAWRs, in both the heart and aorta. These results demonstrate the protective role of MOI against cardiovascular dysfunction due to aging via enhanced SIRT1 function and subsequently reduced oxidative stress.

The article presents a brief historiography of choral dances in the light of Rudolf Laban's ideological premises in comparison with the National Socialism ideology in Germany in the 1920s and 1930s. Laban choral dances were a form of collective dance that the choreographer proposed as a means of recovering a sense of community, and which occupied an essential place in his project of propagation of dance. Considering the background of choral dances and, more specifically, the episode of the choreography that Laban created for the 1936 Olympic games activities, the article deals with the approximations and distances between Laban's cultural projects and National Socialism. The article shows how the collective power contained in choral dances, which were part of the cultural policy instituted in the early years of the Nazi regime in Germany, ended up being a key point for both Laban's rise and fall within the German government. Finally, the text seeks to reflect on the responsibilities and consequences of concessions in the relations between art, politics and ethics that arose in this specific relationship between Laban and Nazism. Keywords Rudolf Laban. Choral Dance. Dance History. Nazism.

Introdução: Atualmente, a alface (Lactuca sativa L.) destaca-se por ser a folhosa mais consumida no Brasil. Em saúde pública, grande parte dos agentes etiológicos de enfermidades entéricas é veiculada por meio de hortaliças, legumes e frutas contaminadas. Objetivo: Verificar a qualidade de alfaces, através da análise microscópica, comparando os resultados dos diferentes tipos de cultivo e da alface minimamente processada. Método: Foram analisadas 84 amostras de alfaces produzidas por três métodos de cultivo (tradicional, orgânico, hidropônico) e alfaces minimamente processadas, comercializadas em municípios das regiões nordeste e metropolitana do estado de São Paulo. Realizou-se primeiramente a pesquisa de matérias estranhas macroscópicas e, para verificar a presença de estruturas parasitárias e outras matérias estranhas microscópicas, foi utilizada a técnica de sedimentação espontânea. Resultados: Os resultados revelaram que as amostras de alfaces dos três tipos de cultivo apresentaram alta ocorrência de matérias estranhas (total de 87,0% de amostras positivas), bem como presença de parasitas (total de 20,0% de amostras positivas). Em relação às alfaces minimamente processadas, a porcentagem de positividade para presença de matérias estranhas e parasitas foi de 58,0%. Conclusões: Os resultados do presente estudo alertam para a importância dos procedimentos de higienização dos diferentes tipos de cultivo de alface previamente ao seu consumo. Em relação às alfaces minimamente processadas, os resultados demonstram que devem ser tomadas ações corretivas no processo de sanitização por parte dos produtores e/ou comerciantes. Quanto às análises laboratoriais, ainda se faz necessário o aprimoramento constante de métodos analíticos que possibilitem melhor detecção de parasitas em alimentos.

The surgical treatment of intracranial aneurysms is a routine operation in the neurosurgeon practice. Complex aneurysms are those with morphological irregularities, usually large or giant; thrombosed, partially thrombosed or calcified; with aberrant fundus/neck ratio and near eloquent neurological structures. These cases demand special skills by the surgical team. The present article is a report of two cases of complex aneurysms successfully treated, with a discussion on the role of neurophysiological monitoring. In these two cases of supra- and infratentorial complex giant aneurysms, intraoperative monitoring was extremely relevant. Thus, we believe that treating complex and giant aneurysms carries several pitfalls, and the use of multimodal intraoperative monitoring is mandatory to mitigate risks and deliver the best result to the patient.

Immunogenic cell death (ICD) in cancer represents a functionally unique therapeutic response that can induce tumor-targeting immune responses. ICD is characterized by the exposure and release of numerous damage-associated molecular patterns (DAMPs), which confer adjuvanticity to dying cancer cells. The spatiotemporally defined emission of DAMPs during ICD has been well described, whereas the epigenetic mechanisms that regulate ICD hallmarks have not yet been deeply elucidated. Here, we aimed to examine the involvement of miRNAs and their putative targets using well-established in vitro models of ICD. To this end, B cell lymphoma (Mino) and breast cancer (MDA-MB-231) cell lines were exposed to two different ICD inducers, the combination of retinoic acid (RA) and interferon-alpha (IFN-α) and doxorubicin, and to non ICD inducers such as gamma irradiation. Then, miRNA and mRNA profiles were studied by next generation sequencing. Co-expression analysis identified 16 miRNAs differentially modulated in cells undergoing ICD. Integrated miRNA-mRNA functional analysis revealed candidate miRNAs, mRNAs, and modulated pathways associated with Immune System Process (GO Term). Specifically, ICD induced a distinctive transcriptional signature hallmarked by regulation of antigen presentation, a crucial step for proper activation of immune system antitumor response. Interestingly, the major histocompatibility complex class I (MHC-I) pathway was upregulated whereas class II (MHC-II) was downregulated. Analysis of MHC-II associated transcripts and HLA-DR surface expression confirmed inhibition of this pathway by ICD on lymphoma cells. miR-4284 and miR-212-3p were the strongest miRNAs upregulated by ICD associated with this event and miR-212-3p overexpression was able to downregulate surface expression of HLA-DR. It is well known that MHC-II expression on tumor cells facilitates the recruitment of CD4+ T cells. However, the interaction between tumor MHC-II and inhibitory coreceptors on tumor-associated lymphocytes could provide an immunosuppressive signal that directly represses effector cytotoxic activity. In this context, MHC-II downregulation by ICD could enhance antitumor immunity. Overall, we found that the miRNA profile was significantly altered during ICD. Several miRNAs are predicted to be involved in the regulation of MHC-I and II pathways, whose implication in ICD is demonstrated herein for the first time, which could eventually modulate tumor recognition and attack by the immune system.

Anti-CK7 and anti-CK20 immunohistochemistry is sometimes used to establish a diagnosis of primary lung cancer. We performed a retrospective study on the value of anti-CK7 and anti-CK20 immunohistochemistry in 359 biopsies of patients with suspected lung carcinoma in order to assess the usefulness of these antibodies in the evaluation of lung tumors in biopsies. Our results showed TTF-1 positivity in 73.3% of patients. EGFR mutations and ALK rearrangements were significantly different between TTF-1 positive and TTF-1 negative tumors (p < 0.001 and p = 0.023, respectively). Our results show a significant difference (p < 0.001) between TTF-1 positive and TTF-1 negative carcinomas with a median survival of 21.97 months (CI95% = 17.48–30.9 months) and 6.52 months (CI95% = 3.34–10.3 months), respectively. In the group of TTF-1 negative patients, anti-CK7 and CK20 immunohistochemistry was performed in 70 patients and showed CK7+/CK20- staining in 61 patients (87.1%), CK7-/CK20- in 4 patients (5.7%), CK7+/CK20+ in 3 patients (4.3%), and CK7-/CK20- in 2 patients (2.8%). No specific or molecular pattern was found in these groups of CK7/CK20 combinations. In total, this work brings arguments concerning the uselessness of anti-CK7/CK20 immunohistochemistry in the case of suspicion of primary lung cancer in biopsies.

Background: Posterior fossa extradural hematomas are an uncommon condition with more than half of cases appearing in the pediatric population. Although the majority of cases are caused by substantial trauma, it may also be related to systemic diseases and bleeding disorders without the history of head trauma. Case report: We present the case of a 6-year-old female affected by Sensenbrenner syndrome with a non-traumatic subacute posterior fossa epidural hematoma. The patient was surgically treated and medically discharged without further neurological alteration. This is the first report of spontaneous cerebral hematoma associated with this syndrome in the literature. A literature review of the main aspects of both topics was performed. Conclusions: Although spontaneous posterior fossa epidural hematomas are rare, clinicians should be aware and highly suspicious of this differential diagnosis in certain situations such as coagulopathies and other specific systemic diseases, as fast recognition and assertive treatment are important for better outcomes.

Introduction Eye movement deficits in psychiatric patients have often been investigated with linear models, which fail to fully capture the complex dynamics characterizing eye movements. Objectives The present work aims to investigate the deficits in fixational eye movements in psychiatric patients according as non-linear chaotic dynamic. Methods We recruited 191 patients (91 males, average age 45 years) diagnosed with schizophrenia, bipolar disorder, depression and personality disorder. The control sample consisted of 22 healthy subjects (12 males, mean age 41 years). Fixational eye movements were recorded with the Eytribe infrared system and off-line analyzed using Matlab. The dynamics of fixation eye movements were investigated using a phase space graph, which refers to chaotic system analysis. This analysis allows to evaluate how the changes in space during fixation as a function of their speed. Results A major difference emerged: psychiatric patients showed larger and faster eye movements gravitating around a single point of density, while control subjects exhibited slower and smaller eye movements with multiple drifts and microtremors. Conclusions In conclusion, the dynamics of fixational eye movements in psychiatric patients seemed to be characterized by poorer efficiency in space exploration. These differences could be attributed to a worse coordination between the perceptual and the oculomotor system. Disclosure No significant relationships.

Introduction Although eye-movement disorders are one of the most replicated deficits in the psychiatric literature, the strong heterogeneity of results is still an unexplained issue that could be effectively addressed with a quantitative review of evidence. Objectives For this reason, a large-scale metanalytic study comprising more than 200 studies was conducted to analyse the presence of eye-movement deficits in schizophrenia patients, as compared to healthy controls. Methods To this aim, saccadic eye movements were grouped based on the type of task required (e.g., standard, predictive) and the quantification method used (e.g., number, duration, amplitude). For each sub-group separate meta-analysis were computed. Cohen’s d was used as measure of effect size. Risk of bias within and between studies and heterogeneity were also analysed. Results indicated low Cohen’s d with the exception of the number of correct antisaccades – where schizophrenia patients reportedless correct anti-saccadesthan healthy controls - and antisaccades error rate – where schizophrenia patients reported a higher number of errors than healthy controls. Conclusions Antisaccades emerged as better suited to differentiate between patients and healthy controls, thus making them the most promising candidate as a possible biomarker for schizophrenia.

Background and Objectives: To present a series of brain metastases from gynecologic primaries and provide a summary of the relevant literature. Materials and Methods: We retrospectively review 18 patients with histologically confirmed brain metastases from gynecologic primaries and summarize the largest series of relative reports. Results: Six brain metastases were of endometrial primary and 12 of ovarian primary. In 3 cases (16.7%), diagnosis of brain metastases was made at presentation of the gynecologic primary; in the others, median time to development of brain metastasis was 34 (range, 6–115) months. Median survival after brain metastasis diagnosis was 5 (range, 1–89) months. Favorable prognostic factors were better performance status (p = 0.04) and, marginally, smaller metastasis size (p = 0.06). No differences in brain metastases between endometrial and ovarian primaries were found, except for the time interval from primary to brain metastases diagnosis, which was shorter for endometrial tumors (p = 0.05). A comprehensive summary of previous studies is provided. Conclusions: Performance status and smaller brain metastases size are good prognostic factors. Endometrial cancer brain metastases develop earlier than ovarian cancer brain metastases.

Background: Posttranscriptional gene regulation (PTGR) contributes to inflammation through alterations in messenger RNA (mRNA) turnover and translation rates. RNA-binding proteins (RBPs) coordinate these processes but their role in lung inflammatory diseases is ill-defined. We evaluated the expression of a curated list of mRNA-binding RBPs (mRBPs) in selected Gene Expression Omnibus (GEO) transcriptomic databases of airway epithelium isolated from chronic obstructive pulmonary disease (COPD), severe asthma (SA) and matched control subjects, hypothesizing that global changes in mRBPs expression could be used to infer their pathogenetic roles and identify novel disease-related regulatory networks.Methods: A published list of 692 mRBPs [Nat Rev Genet 2014] was searched in GEO datasets originated from bronchial brushings of stable COPD patients (C), smokers (S), non-smokers (NS) controls with normal lung function (n = 6/12/12) (GEO ID: GSE5058) and of (SA) and healthy control (HC) (n = 6/12) (GSE63142). Fluorescence intensity data were extracted and normalized on the medians for fold change (FC) comparisons. FCs were set at ≥ |1.5| with a false discovery rate (FDR) of ≤ 0.05. Pearson correlation maps and heatmaps were generated using tMEV tools v4_9_0.45. DNA sequence motifs were searched using PScan-ChIP. Gene Ontology (GO) was performed with Ingenuity Pathway Analysis (IPA) tool.Results: Significant mRBP expression changes were detected for S/NS, COPD/NS and COPD/S (n = 41, 391, 382, respectively). Of those, 32% of genes changed by FC ≥ |1.5| in S/NS but more than 60% in COPD/NS and COPD/S (n = 13, 267, 257, respectively). Genes were predominantly downregulated in COPD/NS (n = 194, 73%) and COPD/S (n = 202, 79%), less so in S/NS (n = 4, 31%). Unsupervised cluster analysis identified in 4 out of 12 S the same mRBP pattern seen in C, postulating subclinical COPD. Significant DNA motifs enrichment for transcriptional regulation was found for downregulated RBPs. Correlation analysis identified five clusters of co-expressed mRBPs. GO analysis revealed significant enrichments in canonical pathways both specific and shared among comparisons. Unexpectedly, no significant mRBPs modulation was found in SA compared to controls.Conclusions: Airway epithelial mRBPs profiling reveals a COPD-specific global downregulation of RBPs shared by a subset of control smokers, the potential of functional cooperation by coexpressed RBPs and significant impact on relevant pathogenetic pathways in COPD. Elucidation of PTGR in COPD could identify disease biomarkers or pathways for therapeutic targeting.

Thin melanomas are tumors less than 1 mm thick according to Breslow classification. Their prognosis is in most cases excellent. However, a small subset of these tumors relapses. These clinical findings emphasize the need of novel prognostic biomarkers to identify this subset of tumors. Characterization of tumor immune microenvironment (TIME) is currently investigated as a prognostic and predictive biomarker for cancer immunotherapy in several solid tumors including melanoma. Here, taking into account the limited availability of tumor tissues, by characterizing some of the characteristics of TIME such as number of infiltrating lymphocytes, HLA class I antigen and PD-L1 expression, we show that number of infiltrating CD8+ and FOXP3+ T cells as well as CD8+/FOXP3+ T cell ratio can represent a useful prognostic biomarker in thin melanoma. Although further investigations in a larger patient cohort are needed, these findings have potential clinical significance since they can be used to define subgroups of thin melanoma patients who have a worse prognosis and might need different treatment modalities.

Mutual interactions between cancer cells and the tumor microenvironment importantly contribute to the development of tyrosine kinase inhibitor (TKI) resistance in patients affected by EGFR-mutant NSCLC. In particular, immune recognition-associated proteins with impact on tumor cell clearance through phagocytosis, such as CD47 and calreticulin, could contribute to adaptive resistance and immune escape. Preclinical studies targeting the anti-phagocytic CD47 molecule showed promising results in different cancer types including lung cancer, but no data are available on its role in patients acquiring resistance to EGFR TKI treatment. We analyzed the functional contribution of CD47 and calreticulin to immune surveillance and evasion in a panel of NSCLC cell lines carrying sensitizing or resistant mutations in the EGFR gene, following treatment with the TKI gefitinib and after in vitro development of gefitinib resistance. While CD47 and calreticulin protein levels were markedly variable in both EGFR-mutant and wild-type cell lines, analysis of NSCLC transcriptomic dataset revealed selective overexpression of CD47 in patients carrying EGFR mutations. EGFR inhibition significantly reduced CD47 expression on the surface of pre-apoptotic cells, favoring more efficient engulfment of cancer cells by monocyte-derived dendritic cells. This was not necessarily associated with augmented surface exposure of calreticulin or other molecular markers of immunogenic cell death. Moreover, CD47 expression became up-regulated following in vitro drug resistance development, and blocking of this protein by a specific monoclonal antibody increased the clearance of EGFR-TKI resistant cells by phagocytes. Our study supports CD47 neutralization by specific monoclonal antibody as a promising immunotherapeutic option for naïve and resistant EGFR-mutant NSCLCs.

Luca Ricciardi,1,* Jessica Dal Col,1,* Paolo Casolari,2 Domenico Memoli,1 Valeria Conti,1 Alessandro Vatrella,1 Becky M Vonakis,3 Alberto Papi,2 Gaetano Caramori,4 Cristiana Stellato1,3 1Department of Medicine, Surgery and Dentistry “Scuola Medica Salernitana”, University of Salerno, Salerno, Italy; 2Interdipartimental Study Center for Inflammatory and Smoke-related Airway Diseases (CEMICEF), Cardiorespiratory and Internal Medicine Section, University of Ferrara, Ferrara, Italy; 3Division of Allergy and Clinical Immunology, Johns Hopkins University School of Medicine, Baltimore, MD, USA; 4Department of Biomedical Sciences, Dentistry and Morphological and Functional Imaging (BIOMORF), University of Messina, Messina, Italy *These authors contributed equally to this work Purpose: Inflammatory gene expression is modulated by posttranscriptional regulation via RNA-binding proteins (RBPs), which regulate mRNA turnover and translation by binding to conserved mRNA sequences. Their role in COPD is only partially defined. This study evaluated RBPs tristetraprolin (TTP), human antigen R (HuR), and AU-rich element-binding factor 1 (AUF-1) expression using lung tissue from COPD patients and control subjects and probed their function in epithelial responses in vitro. Patients and methods: RBPs were detected by immunohistochemistry in bronchial and peripheral lung samples from mild-to-moderate stable COPD patients and age/smoking history-matched controls; RBPs and RBP-regulated genes were evaluated by Western blot, ELISA, protein array, and real-time PCR in human airway epithelial BEAS-2B cell line stimulated with hydrogen peroxide, cytokine combination (cytomix), cigarette smoke extract (CSE), and following siRNA-mediated silencing. Results were verified in a microarray database from bronchial brushings of COPD patients and controls. RBP transcripts were measured in peripheral blood mononuclear cell samples from additional stable COPD patients and controls. Results: Specific, primarily nuclear immunostaining for the RBPs was detected in structural and inflammatory cells in bronchial and lung tissues. Immunostaining for AUF-1, but not TTP or HuR, was significantly decreased in bronchial epithelium of COPD samples vs controls. In BEAS-2B cells, cytomix and CSE stimulation reproduced the RBP pattern while increasing expression of AUF-1-regulated genes, interleukin-6, CCL2, CXCL1, and CXCL8. Silencing expression of AUF-1 reproduced, but not enhanced, target upregulation induced by cytomix compared to controls. Analysis of bronchial brushing-derived transcriptomic confirmed the selective decrease of AUF-1 in COPD vs controls and revealed significant changes in AUF-1-regulated genes by genome ontology. Conclusion: Downregulated AUF-1 may be pathogenic in stable COPD by altering posttranscriptional control of epithelial gene expression. Keywords: airway epithelium, AUF-1, COPD, inflammation, posttranscriptional gene regulation

We undertook a structured review of stereotactic radiosurgery (SRS) using linear particle accelerator (linac) equipment, focusing on volumetric modulated arc therapy (VMAT) technology, and frameless image-guided radiotherapy (IGRT), for the treatment of brain metastases. We analyzed the role of linac SRS and its clinical applications, exploring stereotactic localization. Historically, there was a shift from fixed frames to frameless approaches, moving toward less invasive treatments. Thus, we reviewed the concepts of VMAT for multiple-target applications, comparing its dosimetric and technical features to those of other available techniques. We evaluated relevant technical issues and discussed the planning parameters that have gained worldwide acceptance to date. Thus, we reviewed the current literature on the clinical aspects of SRS, especially its main indications and how the advantages of VMAT may achieve clinical benefits in such scenarios. Finally, we reported our institutional results on IGRT-VMAT for SRS treatments for patients with multiple brain metastases.

ABSTRACT Paraclinoid aneurysms are lesions located adjacent to the clinoid and ophthalmic segments of the internal carotid artery. In recent years, flow diverter stents have been introduced as a better endovascular technique for treatment of these aneurysms. Method From 2009 to 2014, a total of 43 paraclinoid aneurysms in 43 patients were surgically clipped. We retrospectively reviewed the records of these patients to analyze clinical outcomes. Results Twenty-six aneurysms (60.5%) were ophthalmic artery aneurysms, while 17 were superior hypophyseal artery aneurysms (39.5%). The extradural approach to the clinoid process was used to clip these aneurysms. One hundred percent of aneurysms were clipped (complete exclusion in 100% on follow-up angiography). The length of follow-up ranged from 1 to 60 months (mean, 29.82 months). Conclusion Surgical clipping continues to be a good option for the treatment of paraclinoid aneurysms.

Recent advances in cancer immunotherapy have clearly shown that checkpoint-based immunotherapy is effective in a small subgroup of cancer patients. However, no effective predictive biomarker has been identified so far. The major histocompatibility complex, better known in humans as human leukocyte antigen (HLA), is a very polymorphic gene complex consisting of more than 200 genes. It has a crucial role in activating an appropriate host immune response against pathogens and tumor cells by discriminating self and non-self peptides. Several lines of evidence have shown that down-regulation of expression of HLA class I antigen derived peptide complexes by cancer cells is a mechanism of tumor immune escape and is often associated to poor prognosis in cancer patients. In addition, it has also been shown that HLA class I and II antigen expression, as well as defects in the antigen processing machinery complex, may predict tumor responses in cancer immunotherapy. Nevertheless, the role of HLA in predicting tumor responses to checkpoint-based immunotherapy is still debated. In this review, firstly, we will describe the structure and function of the HLA system. Secondly, we will summarize the HLA defects and their clinical significance in cancer patients. Thirdly, we will review the potential role of the HLA as a predictive biomarker for checkpoint-based immunotherapy in cancer patients. Lastly, we will discuss the potential strategies that may restore HLA function to implement novel therapeutic strategies in cancer patients.

The safety and feasibility of dendritic cell (DC)-based immunotherapies in cancer management have been well documented after more than twenty-five years of experimentation, and, by now, undeniably accepted. On the other hand, it is equally evident that DC-based vaccination as monotherapy did not achieve the clinical benefits that were predicted in a number of promising preclinical studies. The current availability of several immune modulatory and targeting approaches opens the way to many potential therapeutic combinations. In particular, the evidence that the immune-related effects that are elicited by immunogenic cell death (ICD)-inducing therapies are strictly associated with DC engagement and activation strongly support the combination of ICD-inducing and DC-based immunotherapies. In this review, we examine the data in recent studies employing tumor cells, killed through ICD induction, in the formulation of anticancer DC-based vaccines. In addition, we discuss the opportunity to combine pharmacologic or physical therapeutic approaches that can promote ICD in vivo with in situ DC vaccination.

Although promising, the clinical benefit provided by dendritic cell (DC)-based vaccines is still limited and the choice of the optimal antigen formulation is still an unresolved issue. We have developed a new DC-based vaccination protocol for aggressive and/or refractory lymphomas which combines the unique features of interferon-conditioned DC (IFN-DC) with highly immunogenic tumor cell lysates (TCL) obtained from lymphoma cells undergoing immunogenic cell death. We show that treatment of mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL) cell lines with 9-cis-retinoic acid and IFNα (RA/IFNα) induces early membrane exposure of Calreticulin, HSP70 and 90 together with CD47 down-regulation and enhanced HMGB1 secretion. Consistently, RA/IFNα-treated apoptotic cells and -TCLs were more efficiently phagocytosed by DCs compared to controls. Notably, cytotoxic T cells (CTLs) generated with autologous DCs pulsed with RA/IFNα-TCLs more efficiently recognized and specifically lysed MCL or DLBCL cells or targets loaded with several HLA-A*0201 cyclin D1 or HLA-B*0801 survivin epitopes. These cultures also showed an expansion of Th1 and Th17 cells and an increased Th17/Treg ratio. Moreover, DCs loaded with RA/IFNα-TCLs showed enhanced functional maturation and activation. NOD/SCID mice reconstituted with human peripheral blood lymphocytes and vaccinated with autologous RA/IFNα-TCL loaded-IFN-DCs showed lymphoma-specific T-cell responses and a significant decrease in tumor growth with respect to mice treated with IFN-DC unpulsed or loaded with untreated TCLs. This study demonstrates the feasibility and efficacy of the use of RA/IFNα to generate a highly immunogenic TCL as a suitable tumor antigen formulation for the development of effective anticancer DC-based vaccines.

O objetivo deste estudo foi determinar o tamanho ótimo de amostra para avaliar a massa, o comprimento e o diâmetro de frutos de abacaxieiro, usando parcelas grandes submetidas a diferentes adubações. No experimento com abacaxizeiro (cultivar Pérola), foram avaliadas duas fontes de nitrogênio (ureia e sulfato de amônio) e cinco doses de cloreto de potássio (zero;350; 700; 1.050 e 1.400 kg ha-1). Cada parcela continha cinco fileiras duplas de quatro metros de comprimento, sendo que as três fileiras centrais foram consideradas como área útil. O espaçamento de plantio foi 1,2 x 0,4 x 0,4 m, correspondente a 60 plantas por parcela útil. O delineamento experimental foi o de blocos completos casualizados, com quatro repetições. Foi calculado o tamanho de amostra ( η ) para as semiamplitudes do intervalo de confiança (SA) iguais a 2; 4; 6;8 e 10% da estimativa da média, com grau de confiança (1-α) de 95%, usando a distribuição t de Student. Posteriormente, fixou-se η como o total de frutos colhidos por parcela para o cálculo da SA em percentagem da estimativa da média para cada um dos caracteres. Com um erro de estimação de 4% da média, devem ser amostrados, respectivamente, 83; 35 e 10 frutos em cada uma das parcelas experimentais, para a avaliação da massa, do comprimento e do diâmetro de frutos de abacaxizeiro, cultivar Pérola, em experimentos com adubação.The aim of this study was to determine the sample size needed to evaluated the mass, length and diameter of pineapple fruits in large experimental units under different fertilizations. In the experiment with pineapple (cultivar Perola) it was evaluated two sources of nitrogen (urea and ammonium sulfate) and five doses of potassium (zero, 350, 700, 1,050 and 1,400 kg ha-1). Each plot had five double strips of four meters length, being the three central strips taken as useful area. The plant spacing was 1,2 x 0,4 x 0,4 m, making 60 plants per useful area of each plot. It was used a randomized blocks design with four replications. The sample size ( η ) was calculated for the semiamplitude of the confidence interval (SA) equal to: 2, 4, 6, 8 and 10% of the average estimate, with degree of confidence (1-α) of 95%, using the Student's t distribution. Afterwards, η was set as the total amount of fruits per plot for calculating SA as percentage of the average estimate for each character. With an average estimate error of 4%, should be sample 83, 35 and 10 fruits in each experimental plot for evaluation of mass, length and diameter, respectively, for pineapple fruits, cultivar Perola, in experiments with fertilization.

A incorporação da palha após a colheita do arroz dificulta a redução do banco de sementes; por outro lado, favorece a decomposição da palha e, consequentemente, possibilita o preparo antecipado da área, viabilizando a semeadura do arroz no período recomendado. O objetivo deste trabalho foi avaliar a interferência de diferentes manejos após a colheita em áreas cultivadas com arroz irrigado na redução da viabilidade de sementes de arroz-vermelho. O delineamento experimental utilizado foi de blocos ao acaso, em parcelas subdivididas no tempo, com quatro repetições. Os tratamentos foram dispostos em esquema fatorial (7x6), sendo o fator A composto por: [1] lâmina de água permanente, [2] incorporação da palha com preparo do solo seco logo após a colheita do arroz, [3] incorporação da palha com preparo do solo alagado após a colheita, [4] incorporação da palha com o solo seco somente em julho, [5] incorporação da palha com solo alagado logo após a colheita e gradagem da área em julho com solo seco, [6] incorporação da palha com solo seco logo após a colheita e gradagem da área em julho com solo seco e [7] sem incorporação. O fator B foi composto por seis épocas de amostragem do banco de sementes quanto à presença de arroz-vermelho; a amostragem iniciou-se a partir da colheita, ocorrendo a cada 30 dias e finalizando no mês de outubro. O maior efeito sobre a redução do banco de sementes ocorreu quando se utilizaram implementos que incorporaram superficialmente a palha, como o rolo-faca, o qual atua sobre a quebra da dormência, a inviabilidade e o estímulo à germinação das sementes de arroz-vermelho.The incorporation of straw after rice harvest makes rice seed reduction difficult. On the other hand, it favors the decomposition of straw, enabling the preparation of the area in advance, thus allowing the preparation of the recommended planting period. The objective of this work was to assess the interference of different management techniques after harvest in rice-cultivated areas in reducing the viability of red rice seeds. The experiment was arranged in a randomized block design, in split-plots in time, with four replications. The treatments were arranged in a factorial design (7x6), consisting of factor A: [a] permanent water blade, [2] straw incorporation with dry soil tillage after rice harvest, [3] incorporation of straw with flooded soil after harvest, [4] incorporation of dry soil only in July, [5] incorporation of straw with flooded soil after harvest and area disking in July with dry soil, [6] incorporation of straw in dry soil immediately after harvest and area disking only in July with dry soil and [7] without incorporation. Factor B was composed of different timings of seed bank sampling for the presence of red rice. Sampling was initiated at harvest, occurring every thirty days until October. The greatest effect on seed bank reduction was verified when using implements that superficially incorporated the straw, such as the rolling knife, which acts on dormancy breaking, non-viability and stimulation of red rice germination.

Neste trabalho são analisadas imagens microtomográficas do Arenito da Formação Rio do Rasto, obtidas pela metodologia de Microtomografia Computadorizada por raios X. As medidas foram realizadas com o microtomógrafo SkyScan, modelo 1172, e propriedades físicas de interesse foram obtidas, tais como: porosidade (fechada e conectada), distribuição de tamanho de poros, VER (Volume Elementar Representativo) e reconstrução 3D. Esta é uma técnica de análise não destrutiva e, por fornecer informações internas das rochas sem destruí-las, é foco de atenção de empresas petrolíferas. Para a aquisição de imagens foi utilizada uma resolução espacial de 5,87 micrometro com uma tensão de 70 kV no tubo de raios X. As projeções foram reconstruídas - gerando as seções 2D – onde, então, foram minimizados alguns dos artefatos nas imagens tomográficas. Com os softwares Imago e CTAn foram analisados os parâmetros físicos sobre uma região de interesse (ROI). Aplicado o limiar médio de 67(2) em uma escala de cores de 8 bits, se obteve – pelo software Imago – uma porosidade absoluta de 13,54(66) %, onde 12,58(74) % são poros conectados. Baseando-se na porcentagem de poros conectados e na distribuição de tamanho de poros, infere-se que esta amostra poderia conter uma razoável permeabilidade. O VER foi alcançado com uma aresta de 2,900 mm. Comparando com trabalhos anteriores, notou-se que esta amostra possui um algo grau de variabilidade.

A cravina é uma planta excelente para compor jardins por possuir florescimento precoce, floração abundante e ótimo desempenho na primavera e outono. O objetivo do trabalho foi estimar o tamanho de amostra para plantas de cravina de jardim, cultivadas em diferentes substratos, e verificar a variabilidade do tamanho de amostra entre variáveis de crescimento e produção e substratos. Utilizaram-se sete tratamentos (substratos): S1 = 50% solo + 50% cinzas de casca de arroz; S2 = 80% solo + 20% húmus de minhoca; S3 = 80% cinzas de casca de arroz + 20% húmus de minhoca; S4 = 40% solo + 40% cinzas de casca de arroz + 20% húmus de minhoca; S5 = 100% turfa preta; S6 = 100% substrato comercial Mecplant®; S7 = 50% turfa preta + 50% cinzas de casca de arroz, com 56 repetições cada um, totalizando 392 plantas de cravina de jardim, nas quais se avaliaram 17 variáveis de crescimento e produção. Utilizou-se a metodologia de reamostragem bootstrap, com reposição, para cada variável dentro de cada substrato, com erro preestabelecido de: 5, 10, 20 e 40% da média (D%). Para um intervalo de confiança de 95%, com D%=20, o substrato com 50% de solo e 50% de cinzas de casca de arroz apresentou o maior tamanho de amostra em 11 variáveis; quando se comparou as variáveis, o número de botões florais apresentou o maior tamanho de amostra, em média 113 plantas. A utilização de amostras de 44 plantas de cravina de jardim para o substrato comercial Mecplant® atenderá a precisões inferiores ou iguais a 20%, para todas as variáveis avaliadas. Há variação no tamanho de amostra em relação ao substrato utilizado e a variável avaliada em plantas de cravina de jardim.

Este trabalho teve por objetivo propor padrões para a padronização dos índices das análises de sementes de espécies florestais. Para tanto, foram utilizados resultados de análises de sementes dessas espécies a partir do ano de 1997 até 2007, pertencentes aos arquivos do Laboratório de Análise de Sementes da Fundação Estadual de Pesquisa Agropecuária - FEPAGRO, localizado em Santa Maria, RS. Os limites de confiabilidade foram construídos a partir de uma análise de normalidade dos dados e da utilização do critério do pseudossigma. Os limites de confiança estabeleceram os valores mínimos aceitáveis para que um lote de sementes seja comercializado, propondo que valores de germinação dentro dos limites de confiabilidade muito baixos ou baixos devem ser descartados, pois não apresentam índices germinativos aceitáveis.