Unveiling the role of osteosarcoma-derived secretome in premetastatic lung remodelling.

BACKGROUND: Lung metastasis is the most adverse clinical factor and remains the leading cause of osteosarcoma-related death. Deciphering the mechanisms driving metastatic spread is crucial for finding open therapeutic windows for successful organ-specific interventions that may halt or prevent lung metastasis. METHODS: We employed a mouse premetastatic lung-based multi-omics integrative approach combined with clinical features to uncover the specific changes that precede lung metastasis formation and identify novel molecular targets and biomarker of clinical utility that enable the design of novel therapeutic strategies. RESULTS: We found that osteosarcoma-bearing mice or those preconditioned with the osteosarcoma cell secretome harbour profound lung structural alterations with airway damage, inflammation, neutrophil infiltration, and extracellular matrix remodelling with increased deposition of fibronectin and collagens by resident stromal activated fibroblasts, favouring the adhesion of disseminated tumour cells. Systemic-induced microenvironmental changes, supported by transcriptomic and histological data, promoted and accelerated lung metastasis formation. Comparative proteome profiling of the cell secretome and mouse plasma identified a large number of proteins involved in extracellular-matrix organization, cell-matrix adhesion, neutrophil degranulation, and cytokine-mediated signalling, consistent with the observed lung microenvironmental changes. Moreover, we identified EFEMP1, an extracellular matrix glycoprotein exclusively secreted by metastatic cells, in the plasma of mice bearing a primary tumour and in biopsy specimens from osteosarcoma patients with poorer overall survival. Depletion of EFEMP1 from the secretome prevents the formation of lung metastasis. CONCLUSIONS: Integration of our data uncovers neutrophil infiltration and the functional contribution of stromal-activated fibroblasts in ECM remodelling for tumour cell attachment as early pro-metastatic events, which may hold therapeutic potential in preventing or slowing the metastatic spread. Moreover, we identified EFEMP1, a secreted glycoprotein, as a metastatic driver and a potential candidate prognostic biomarker for lung metastasis in osteosarcoma patients. Osteosarcoma-derived secreted factors systemically reprogrammed the lung microenvironment and fostered a growth-permissive niche for incoming disseminated cells to survive and outgrow into overt metastasis. Daily administration of osteosarcoma cell secretome mimics the systemic release of tumour-secreted factors of a growing tumour in mice during PMN formation; Transcriptomic and histological analysis of premetastatic lungs revealed inflammatory-induced stromal fibroblast activation, neutrophil infiltration, and ECM remodelling as early onset pro-metastatic events; Proteome profiling identified EFEMP1, an extracellular secreted glycoprotein, as a potential predictive biomarker for lung metastasis and poor prognosis in osteosarcoma patients. Osteosarcoma patients with EFEMP1 expressing biopsies have a poorer overall survival.

Cx43-mediated hyphal folding counteracts phagosome integrity loss during fungal infection.

Phagolysosomes are crucial organelles during the elimination of pathogens by host cells. The maintenance of their membrane integrity is vital during stressful conditions, such as during Candida albicans infection. As the fungal hyphae grow, the phagolysosome membrane expands to ensure that the growing fungus remains entrapped. Additionally, actin structures surrounding the hyphae-containing phagosome were recently described to damage and constrain these pathogens inside the host vacuoles by inducing their folding. However, the molecular mechanism involved in the phagosome membrane adaptation during this extreme expansion process is still unclear. The main goal of this study was to unveil the interplay between phagosomal membrane integrity and folding capacity of C. albicans-infected macrophages. We show that components of the repair machinery are gradually recruited to the expanding phagolysosomal membrane and that their inhibition diminishes macrophage folding capacity. Through an analysis of an RNAseq data set of C. albicans-infected macrophages, we identified Cx43, a gap junction protein, as a putative player involved in the interplay between lysosomal homeostasis and actin-related processes. Our findings further reveal that Cx43 is recruited to expand phagosomes and potentiates the hyphal folding capacity of macrophages, promoting their survival. Additionally, we reveal that Cx43 can act as an anchor for complexes involved in Arp2-mediated actin nucleation during the assembly of actin rings around hyphae-containing phagosomes. Overall, this work brings new insights on the mechanisms by which macrophages cope with C. albicans infection ascribing to Cx43 a new noncanonical regulatory role in phagosome dynamics during pathogen phagocytosis. IMPORTANCE Invasive candidiasis is a life-threatening fungal infection that can become increasingly resistant to treatment. Thus, strategies to improve immune system efficiency, such as the macrophage response during the clearance of the fungal infection, are crucial to ameliorate the current therapies. Engulfed Candida albicans, one of the most common Candida species, is able to quickly transit from yeast-to-hypha form, which can elicit a phagosomal membrane injury and ultimately lead to macrophage death. Here, we extend the understanding of phagosome membrane homeostasis during the hypha expansion and folding process. We found that loss of phagosomal membrane integrity decreases the capacity of macrophages to fold the hyphae. Furthermore, through a bioinformatic analysis, we reveal a new window of opportunities to disclose the mechanisms underlying the hyphal constraining process. We identified Cx43 as a new weapon in the armamentarium to tackle infection by potentiating hyphal folding and promoting macrophage survival.

Telehealth Actions to Address COVID-19 in Latin American Countries.

Introduction: This study set out to examine the use of telehealth resources to tackle the coronavirus disease 2019 (COVID-19) pandemic in Latin America within the scope of national telehealth projects (NTPs). Methods: A qualitative study developed using ethnomethodology for appropriate understanding of how telehealth actions were carried out in practice during the COVID-19 pandemic within the scope of NTPs, in the following countries: Argentina, Colombia, Costa Rica, Ecuador, El Salvador, Guatemala, Honduras, Mexico, Peru, and Uruguay. The study was carried out from October to 2020 to March 2021. The number of participations in the discussion groups, formed by coordinating teams of NTPs, totaled 90. Results were described in the worksheet completed according to the script. Each country reviewed its respective data, three times on average, in an effort to clarify actions developed. Results: Three groups of countries were identified: (1) Countries with a telehealth background that used these resources to tackle COVID-19 and thereby refined telehealth activities. Countries with greater experience in NTP design, such as Mexico, Colombia, Peru, and Argentina, were able to use a wide range of telehealth activities to tackle the pandemic, with offers of teleconsultation, teleguidance, telemonitoring to patients, and training of health professionals; (2) Countries with some telehealth activities to address COVID-19. Uruguay, Ecuador, El Salvador, and Costa Rica; and (3) Countries with no evidence of telehealth resource use during the pandemic. Honduras and Guatemala. Discussion: Most NTPs in Latin America have improved their telehealth activities, contributing to address the COVID-19 pandemic in Latin America.

Condutas da enfermeira em centro cirúrgico no cenário da pandemia por COVID-19 / Conducta de enfermeros en el centro quirúrgico en el escenario de pandemia COVID-19 / Nurses conduct in the surgical center in the COVID-19 pandemic scenario

Objetivo: descrever o que tem sido escrito cientificamente sobre a adequação da assistência da enfermeira no centro cirúrgico no cenário da pandemia por Covid-19. Método: Trata-se de uma revisão integrativa de literatura. Como critérios de inclusão, elegeu-se: artigos completos disponíveis em português e inglês, publicados a partir de 2020, ano que deu início a pandemia até janeiro de 2022. Para organização e análise dos dados, recorreu-se ao Método de Análise de Conteúdo. Resultados: Foram selecionados 8 artigos. Como categorias de análise, emergiram os seguintes temas: o estabelecimento de protocolos operacionais específicos para a realização de cirurgias durante a pandemia da Covid-19 e a necessidade de readequação dos profissionais de saúde e a importância da enfermeira neste contexto. Conclusão: A enfermeira teve papel fundamental em todo o processo de estruturação e direcionamento do cuidado ao paciente, destacando seu potencial como protagonista no processo de cuidar em saúde(AU)

Objective: to describe what has been scientifically written about the adequacy of nurse assistance in the surgical center in the context of the Covid-19 pandemic. Methodology: This is an integrative literature review. As inclusion criteria, the following were chosen: full articles available in Portuguese and English, published from 2020, the year the pandemic started until January 2022. For data organization and analysis, the Content Analysis Method was used . Results: Eight articles were selected. As categories of analysis, the following themes emerged: the establishment of specific operational protocols for performing surgeries during the Covid-19 pandemic and the need to readjust health professionals and the importance of the nurse in this context. conclusion: The nurse played a fundamental role in the entire process of structuring and directing patient care, highlighting her potential as a protagonist in the health care process.(AU)

Objetivo: describir lo que científicamente se ha escrito sobre la adecuación de la asistencia de enfermería en el centro quirúrgico en el contexto de la pandemia de la Covid-19. Metodología: Esta es una revisión integrativa de la literatura. Como criterios de inclusión, se eligieron: artículos completos disponibles en portugués e inglés, publicados a partir de 2020, año de inicio de la pandemia, hasta enero de 2022. Para la organización y análisis de los datos, se utilizó el Método de Análisis de Contenido . Resultados: Se seleccionaron ocho artículos. Como categorías de análisis surgieron los siguientes temas: el establecimiento de protocolos operativos específicos para la realización de cirugías durante la pandemia de Covid-19 y la necesidad de readaptación de los profesionales de la salud y la importancia del enfermero en este contexto. Conclusión La enfermera jugó un papel fundamental en todo el proceso de estructuración y dirección del cuidado del paciente, destacando su potencial como protagonista en el proceso de atención a la salud.(AU)

Discovery of sustainable drugs for Alzheimer's disease: cardanol-derived cholinesterase inhibitors with antioxidant and anti-amyloid properties.

As part of our efforts to develop sustainable drugs for Alzheimer's disease (AD), we have been focusing on the inexpensive and largely available cashew nut shell liquid (CNSL) as a starting material for the identification of new acetylcholinesterase (AChE) inhibitors. Herein, we decided to investigate whether cardanol, a phenolic CNSL component, could serve as a scaffold for improved compounds with concomitant anti-amyloid and antioxidant activities. Ten new derivatives, carrying the intact phenolic function and an aminomethyl functionality, were synthesized and first tested for their inhibitory potencies towards AChE and butyrylcholinesterase (BChE). 5 and 11 were found to inhibit human BChE at a single-digit micromolar concentration. Transmission electron microscopy revealed the potential of five derivatives to modulate Aß aggregation, including 5 and 11. In HORAC assays, 5 and 11 performed similarly to standard antioxidant ferulic acid as hydroxyl scavenging agents. Furthermore, in in vitro studies in neuronal cell cultures, 5 and 11 were found to effectively inhibit reactive oxygen species production at a 10 µM concentration. They also showed a favorable initial ADME/Tox profile. Overall, these results suggest that CNSL is a promising raw material for the development of potential disease-modifying treatments for AD.

Micro-RNA Analysis in Pulmonary Arterial Hypertension: Current Knowledge and Challenges.

Pulmonary arterial hypertension (PAH) is a rare, chronic disease of the pulmonary vasculature that is associated with poor outcomes. Its pathogenesis is multifactorial and includes micro-RNA (miRNA) deregulation. The understanding of the role of miRNAs in PAH is expanding quickly, and it is increasingly difficult to identify which miRNAs have the highest translational potential. This review summarizes the current knowledge of miRNA expression in PAH, discusses the challenges in miRNA analysis and interpretation, and highlights 4 promising miRNAs in this field (miR-29, miR-124, miR-140, and miR-204).

Effect of Increased Lactate Dehydrogenase A Activity and Aerobic Glycolysis on the Proinflammatory Profile of Autoimmune CD8+ T Cells in Rheumatoid Arthritis.

OBJECTIVE: CD8+ T cells contribute to rheumatoid arthritis (RA) by releasing proinflammatory and cytolytic mediators, even in a challenging hypoxic and nutrient-poor microenvironment such as the synovial membrane. This study was undertaken to explore the mechanisms through which CD8+ T cells meet their metabolic demands in the blood and synovial membrane of patients with RA. METHODS: Purified blood CD8+ T cells from patients with RA, patients with psoriatic arthritis (PsA), and patients with spondyloarthritis (SpA), as well as healthy control subjects, and CD8+ T cells from RA synovial membrane were stimulated in medium containing 13 C-labeled metabolic substrates in the presence or absence of metabolic inhibitors, under conditions of normoxia or hypoxia. The production of metabolic intermediates was quantified by 1 H-nuclear magnetic resonance. The expression of metabolic enzymes, transcription factors, and immune effector molecules was assessed at both the messenger RNA (mRNA) and protein levels. CD8+ T cell functional studies were performed. RESULTS: RA blood CD8+ T cells met their metabolic demands through aerobic glycolysis, production of uniformly 13 C-enriched lactate in the RA blood (2.6 to 3.7-fold higher than in patients with SpA, patients with PsA, and healthy controls; P < 0.01), and induction of glutaminolysis. Overexpression of Warburg effect-linked enzymes in all RA CD8+ T cell subsets maintained this metabolic profile, conferring to the cells the capacity to proliferate under hypoxia and low-glucose conditions. In all RA CD8+ T cell subsets, lactate dehydrogenase A (LDHA) was overexpressed at the mRNA level (P < 0.03 versus controls; n = 6 per group) and protein level (P < 0.05 versus controls; n = 17 RA patients, n = 9 controls). In RA blood, inhibition of LDHA with FX11 led to reductions in lipogenesis, migration and proliferation of CD8+ T cells, and CD8+ T cell effector functions, while production of reactive oxygen species was increased by 1.5-fold (P < 0.03 versus controls). Following inhibition of LDHA with FX11, RA CD8+ T cells lost their capacity to induce healthy B cells to develop a proinflammatory phenotype. Similar metabolic alterations were observed in RA CD8+ T cells from the synovial membrane. CONCLUSION: Remodeling glucose and glutamine metabolism in RA CD8+ T cells by targeting LDHA activity can reduce the deleterious inflammatory and cytolytic contributions of these cells to the development of autoimmunity.

Comparación de los métodos de Willems I y Willems II en la estimación forense de la edad en niños venezolanos / Comparison of the Willems I and Willems II methods for forensic age estimation in Venezuelan children

INTRODUCCIÓN: El presente trabajo tuvo como objetivo comparar los métodos de Willems et al. (WI y WII) en la estimación forense de la edad en niños venezolanos. MATERIAL Y MÉTODOS: Esta investigación fue de tipo retrospectiva y observacional. Se emplearon 516 ortopantomografías de individuos de ambos sexos (6-18 años), se asignaron los estadios de maduración dental descritos por Demirjian et al. en los 7 dientes inferiores izquierdos, para luego determinar la edad dental (ED) mediante WI y WII. Se calculó la diferencia de media entre la edad cronológica (EC) y la ED estimada mediante una prueba «t» de Student para muestras relacionadas. RESULTADOS: Para el total de la muestra se observó la subestimación de la edad por ambos métodos (0,17+/-1,75 años), siendo la diferencia EC-ED estadísticamente significativa (p = 0,04). Considerando todos los grupos de edad, WII mostró la menor diferencia EC-ED (0,05 años+/-1,40) en el sexo femenino, mientras que para el masculino se encontró menor diferencia para WI (0,04+/-0,54 años). En una submuestra hasta los 16 años se evidenció una subestimación de la edad en el sexo femenino para WI (0,03+/-1,23 años) y una sobreestimación para WII (-0,18+/-1,24 años), en el sexo masculino ambos métodos sobreestimaron la edad (W1=-0,21+/-1,17; WII=-0,06+/-1,20 años). CONCLUSIONES: La precisión de los métodos varió de acuerdo al sexo y grupo de edad, sin embargo, ambos resultaron aplicables a la muestra estudiada

INTRODUCTION: The purpose of this study was to compare the methods of Willems et al., (WI and WII) in estimating the forensic age in Venezuelan children. MATERIAL AND METHODS: A retrospective and observational study was performed on 516 orthopantomographs of individuals of both genders (6-18 years) in order to evaluate the stages of dental maturation described by Demirjian et al. Using the dental age (DA) in the seven left inferior teeth, DA was calculated using the WI and WII methods. The mean difference between the chronological age (CA) and DA was calculated using a paired-sample t-Test. RESULTS: The under-estimation of age by both methods (0.17+/-1.75 years) was observed, with the CA-DA difference being statistically significant (P=.04). Considering all age groups, WII showed the smallest difference between CA-DA (0.05+/-1.40 years) in girls, while a smaller difference was found for WI (0.04+/-0.54 years)for boys. In a subsample up to the age of 16, there was an under-estimation of age in females for WI (0.03+/-1.23 years) and an over-estimation for WII (-0.18+/-1.24 years). In the males both methods over-estimated the age (W1=-0.21+/-1.17 years, WII=-0.06+/-1.20 years). CONCLUSIONS: The accuracy of the methods varied according to gender and age group: however, both were applicable to the studied sample

Dealing with wicked problems in socio-ecological systems affected by industrial disasters: A framework for collaborative and adaptive governance.

This paper puts forward an analytical framework for collaborative and adaptive governance in dealing with so-called "wicked problems" in socio-ecological systems that are affected by industrial disasters. Wicked problems are dilemmas in social and political planning that resist clear definitions and predetermined solutions. An industrial disaster can be transformed into a wicked problem when organizations face institutional complexity, and multiple interests emerge during the damage-recovery phase whenever the available information is confusing. Mitigation actions are associated with incomplete knowledge of the various interests involved and with different perspectives on values. In this context, approaches to public, collaborative, and adaptive governance can help show how to proceed in the face of industrial disasters that turn into wicked problems. These governance regimes operate at multiple levels, consider interdependencies, integrate adjacent policies, promote innovation and social learning, and recognize that solutions are not unique but require continuous adjustments. In this paper, we draw up an analytical framework to enable transformative adaptations to be made to industrial disaster-recovery processes. The proposed framework has some applications for improving the handling of industrial disasters and therefore has relevance for those studying and managing disaster relief and resilience-planning.

A bioinformatic analysis identifies circadian expression of splicing factors and time-dependent alternative splicing events in the HD-MY-Z cell line.

The circadian clock regulates key cellular processes and its dysregulation is associated to several pathologies including cancer. Although the transcriptional regulation of gene expression by the clock machinery is well described, the role of the clock in the regulation of post-transcriptional processes, including splicing, remains poorly understood. In the present work, we investigated the putative interplay between the circadian clock and splicing in a cancer context. For this, we applied a computational pipeline to identify oscillating genes and alternatively spliced transcripts in time-course high-throughput data sets from normal cells and tissues, and cancer cell lines. We investigated the temporal phenotype of clock-controlled genes and splicing factors, and evaluated their impact in alternative splice patterns in the Hodgkin Lymphoma cell line HD-MY-Z. Our data points to a connection between clock-controlled genes and splicing factors, which correlates with temporal alternative splicing in several genes in the HD-MY-Z cell line. These include the genes DPYD, SS18, VIPR1 and IRF4, involved in metabolism, cell cycle, apoptosis and proliferation. Our results highlight a role for the clock as a temporal regulator of alternative splicing, which may impact malignancy in this cellular model.

The Interplay between Colon Cancer Cells and Tumour-Associated Stromal Cells Impacts the Biological Clock and Enhances Malignant Phenotypes.

Cancer cells interrelate with the bordering host microenvironment that encompasses the extracellular matrix and a nontumour cellular component comprising fibroblasts and immune-competent cells. The tumour microenvironment modulates cancer onset and progression, but the molecular factors managing this interaction are not fully understood. Malignant transformation of a benign tumour is among the first crucial events in colorectal carcinogenesis. The role of tumour stroma fibroblasts is well-described in cancer, but less well-characterized in benign tumours. In the current work we utilized fibroblasts isolated from tubulovillous adenoma, which has high risk for malignant transformation, to study the interaction between benign tumour stroma and the circadian clock machinery. We explored the role of the biological clock in this interplay taking advantage of an experimental model, represented by the co-culture of colon cancer cells with normal fibroblasts or tumour-associated fibroblasts, isolated from human colorectal tumour specimens. When co-cultured with tumour-associated fibroblasts, colon cancer cells showed alterations in their circadian and metabolic parameters, with decreased apoptosis, increased colon cancer cell viability, and increased resistance to chemotherapeutic agents. In conclusion, the interactions among colon cancer cells and tumour-associated fibroblasts affect the molecular clockwork and seem to aggravate malignant cell phenotypes, suggesting a detrimental effect of this interplay on cancer dynamics.

Temporal Splicing Switches in Elements of the TNF-Pathway Identified by Computational Analysis of Transcriptome Data for Human Cell Lines.

Alternative splicing plays an important role in numerous cellular processes and aberrant splice decisions are associated with cancer. Although some studies point to a regulation of alternative splicing and its effector mechanisms in a time-dependent manner, the extent and consequences of such a regulation remains poorly understood. In the present work, we investigated the time-dependent production of isoforms in two Hodgkin lymphoma cell lines of different progression stages (HD-MY-Z, stage IIIb and L-1236, stage IV) compared to a B lymphoblastoid cell line (LCL-HO) with a focus on tumour necrosis factor (TNF) pathway-related elements. For this, we used newly generated time-course RNA-sequencing data from the mentioned cell lines and applied a computational pipeline to identify genes with isoform-switching behaviour in time. We analysed the temporal profiles of the identified events and evaluated in detail the potential functional implications of alterations in isoform expression for the selected top-switching genes. Our data indicate that elements within the TNF pathway undergo a time-dependent variation in isoform production with a putative impact on cell migration, proliferation and apoptosis. These include the genes TRAF1, TNFRSF12A and NFKB2. Our results point to a role of temporal alternative splicing in isoform production, which may alter the outcome of the TNF pathway and impact on tumorigenesis.

Influencia de la edad, sexo y dentición en índices radiomorfométricos mandibulares de una población adulta venezolana / Influence of age, sex and dental status in mandible radiomorphometric indices of a Venezuelan adult sample

Objetivo: Evaluar la influencia de la edad, el sexo y el estado dental, sobre índices radiomorfométricos de la mandíbula, obtenidos en panorámicas de adultos venezolanos. Métodos: La muestra estuvo constituida por 156 radiografías panorámicas digitales de individuos de ambos sexos (78 hombres y 78 mujeres), con edades entre los 20 y 81 años, donde se midieron mediante el software ImageJ, los siguientes índices: EC=espesor de la cortical mandibular, ARM= altura región mentoniana, GRA= grado de reabsorción de la cresta alveolar, IPM= índice panorámico mandibular, IAG= índice antegonial, IG=índice gonial, AMR=altura máxima de la rama, AG=ángulo gonial. Se obtuvieron estadísticos descriptivos, correlaciones de Spearman y diferencias de medias mediante test de Mann-Whitney. Resultados: Se observó una tendencia a la disminución de las medias obtenidas para los índices en los grupos de mayor edad, para ambos sexos, asimismo, se evidenciaron diferencias estadísticamente significativas entre hombres y mujeres, con excepción de EC e IAG. Al comparar los valores observados para los sexos por grupos de edad, se evidenciaron diferencias significativas en los grupos de mayor edad (p<0,001). Se verificó una correlación negativa entre los índices y la edad cronológica, siendo estadísticamente significativa en ARM, GRA, IPM, IAG e IG. En ambos sexos el estado de la dentición mostró una correlación positiva y significativa con GRA. Conclusiones: Los valores de los índices disminuyeron con la edad y mostraron dimorfismo sexual, lo que fue evidente a partir de los 50 años, el estado dental se relacionó significativamente con el GRA. Palabras clave: Adulto; Mandíbula; Panorámica; Radiografía.

Objective: To evaluate the influence of age, sex and dental status on radiomorphometric indexes of the mandible, obtained from panoramic radiographs of a Venezuelan adults. Methods: The sample consisted of 156 digital panoramic radiographs of individuals of both sexes (78 men and 78 women), with ages between 20 and 81 years old, where the following indexes were measured by ImageJ software: MC=thickness of the mandibu-lar cortex, MRH= mental region height, ARR=degree of alveolar ridge reabsorption, PMI=panoramic mandibular index, AGI=antegonial index, GI=gonial index, MRH=-maximum height of the ramus, GA=gonial angle. Descriptive statistics, Spearman correlations and differences in means were obtained using the Mann-Whitney test. Results:There was a tendency to measurement values reduction for the indices obtained from age groups, for both sexes, as well as statistically significant differences between men and wo-men, with the exception of MC and AGI. When comparing the observed values for the sexes by age groups, significant differences were found in the older age groups (p<0.001). There was a negative correlation between the indexes and the chronological age, being statistically significant in MRH, ARR, PMI, AGI and GI. In both sexes the dentition status showed a positive and significant correlation with ARR. Conclusions: The values of the indexes decreased with age and showed sexual dimorphism, which was evident after 50 years, the dental status was significantly related to the ARR. Keywords: Mandible; Panoramic; Radiography; Adult.

The reciprocal interplay between TNFα and the circadian clock impacts on cell proliferation and migration in Hodgkin lymphoma cells.

A bidirectional interaction between the circadian network and effector mechanisms of immunity brings on a proper working of both systems. In the present study, we used Hodgkin lymphoma (HL) as an experimental model for a type of cancer involving cells of the immune system. We identified this cancer type among haematological malignancies has having a strong differential expression of core-clock elements. Taking advantage of bioinformatics analyses and experimental procedures carried out in III- and IV-stage HL cells, and lymphoblastoid B cells, we explored this interplay and bear out diverse interacting partners of both systems. In particular, we assembled a wide-ranging network of clock-immune-related genes and pinpointed TNF as a crucial intermediary player. A robust circadian clock hallmarked III-stage lymphoma cells, differently from IV-stage HL cells, which do not harbour a properly functioning clockwork. TNF and circadian gene modulation impacted on clock genes expression and triggered phenotypic changes in lymphoma cells, suggesting a crucial involvement of core-clock elements and TNF in the physiopathological mechanisms hastening malignancy. Our results move forward our understanding of the putative role of the core-clock and TNF in the pathobiology of Hodgkin lymphoma, and highlight their influence in cellular proliferation and migration in lymphatic cancers.

Construção de um índice de desenvolvimento sustentável e análise espacial das desigualdades nos municípios cearenses / Construcción de un índice de desarrollo sostenible y análisis espacial de las desigualdades en los municipios del estado de Ceará / Building a sustainable development index and spacial assessment of municipalities inequalities in the state of Ceará

Resumo O artigo apresenta a análise espacial da sustentabilidade nos municípios cearenses a partir da construção de um índice de desenvolvimento sustentável, embasado em dimensões social, ambiental, econômica e institucional. Os índices de sustentabilidade forçam instituições e governos a questionarem seus padrões e atuam como "forças motrizes" para a elaboração de políticas públicas. A validade dos índices de sustentabilidade depende da definição do conjunto de indicadores e das técnicas adotadas para definição dos ponderadores e agregação dos indicadores. A pesquisa é de natureza quantitativa, desenvolvida por meio de dados secundários, e adotou análise fatorial confirmatória para a construção do índice de desenvolvimento sustentável (IDS) e modelagem econométrica espacial para representação das desigualdades no mapa dos municípios cearenses. Os resultados da pesquisa revelam um ajuste regular do índice de desenvolvimento sustentável. O IDS permite uma visualização geográfica e identificação de associação espacial dos indicadores de desenvolvimento sustentável, e apresenta-se como uma ferramenta de suporte à definição de políticas públicas. A pesquisa revelou que áreas mais urbanas do estado do Ceará apresentam um melhor índice de desenvolvimento sustentável, e confirmou a fragilidade das políticas públicas em promover o equilíbrio regional.

Resumen El artículo presenta el análisis espacial de la sostenibilidad en los municipios cearenses a partir de la construcción de un índice de desarrollo sostenible, basado en dimensiones social, ambiental, económica e institucional. Los índices de sostenibilidad obligan a instituciones y gobiernos a cuestionar sus patrones y actúan como "fuerzas motrices" para la elaboración de políticas públicas. La validez de los índices de sostenibilidad depende de la definición del conjunto de indicadores y de las técnicas adoptadas para definir los ponderadores y la agregación de los indicadores. La investigación es de naturaleza cuantitativa, desarrollada por medio de datos secundarios, y adoptó análisis factorial confirmatorio para la construcción del índice de desarrollo sostenible (IDS) y modelado econométrico espacial para representación de las desigualdades en el mapa de los municipios cearenses. Los resultados de la investigación revelan un ajuste regular del índice de desarrollo sostenible. El IDS permite una visualización geográfica e identificación de asociación espacial de los indicadores de desarrollo sostenible, y se presenta como una herramienta de apoyo a la definición de políticas públicas. La investigación reveló que áreas más urbanas del estado de Ceará presentan un mejor índice de desarrollo sostenible, y confirmó la fragilidad de las políticas públicas en promover el equilibrio regional

Abstract The paper builds a sustainable development index based on environmental, social, economic and institutional dimensions and presents a spatial assessment of municipalitties inequalities in the State of Ceará. Sustainability indices force institutions and governments to question their standards and act as "driving forces" for public policy-making. The validity of sustainability indexes is heavily dependent on how their components are weighted and aggregated. The research is quantitative developed through secondary data acquired in public agency databases of the country (Brazil) and federal unit (State of Ceara). Data analysis included confirmatory factor analysis for the construction of general sustainability indexes, descriptive analysis of these indices and spatial econometric modeling to represent on the map of Ceará municipalities. The survey results reveal a regular adjustment of the sustainable development index. The model allows a geographical view of sustainable development indicators and presents with a tool in the definition of public policies for social equity, environmental and economic. The analysis of social, economic, environmental, institutional and general indices revealed that the most urban areas of the State of Ceará have a greater sustainable development and confirms the fragility of public policies in promoting regional balance.

The Ink4a/Arf locus operates as a regulator of the circadian clock modulating RAS activity.

The mammalian circadian clock and the cell cycle are two major biological oscillators whose coupling influences cell fate decisions. In the present study, we use a model-driven experimental approach to investigate the interplay between clock and cell cycle components and the dysregulatory effects of RAS on this coupled system. In particular, we focus on the Ink4a/Arf locus as one of the bridging clock-cell cycle elements. Upon perturbations by the rat sarcoma viral oncogene (RAS), differential effects on the circadian phenotype were observed in wild-type and Ink4a/Arf knock-out mouse embryonic fibroblasts (MEFs), which could be reproduced by our modelling simulations and correlated with opposing cell cycle fate decisions. Interestingly, the observed changes can be attributed to in silico phase shifts in the expression of core-clock elements. A genome-wide analysis revealed a set of differentially expressed genes that form an intricate network with the circadian system with enriched pathways involved in opposing cell cycle phenotypes. In addition, a machine learning approach complemented by cell cycle analysis classified the observed cell cycle fate decisions as dependent on Ink4a/Arf and the oncogene RAS and highlighted a putative fine-tuning role of Bmal1 as an elicitor of such processes, ultimately resulting in increased cell proliferation in the Ink4a/Arf knock-out scenario. This indicates that the dysregulation of the core-clock might work as an enhancer of RAS-mediated regulation of the cell cycle. Our combined in silico and in vitro approach highlights the important role of the circadian clock as an Ink4a/Arf-dependent modulator of oncogene-induced cell fate decisions, reinforcing its function as a tumour-suppressor and the close interplay between the clock and the cell cycle network.

Critical elements for eco-retrofitting a conventional industrial park: Social barriers to be overcome.

This paper aims to explore critical elements for eco-retrofitting a conventional industrial park, based on a survey of companies and institutions located in Brazil. The study investigates social barriers to be overcome in promotion of opportunities for waste exchange. Our results indicate that values, trust behaviour, waste cognitive domain and environment engagement are necessary for the creation of an eco-industrial park. Similar values of benevolence and universalism are essential for company engagement to eco-retrofit. Low levels of trust behaviour combining with limited waste cognitive domain prevent firms from establishing agreement on waste exchange initiatives. The findings lend support to the view that social barriers are pre-requisites to engagement among firms in establishing technological and logistical solutions. Serious attention needs to be given to these social barriers because they are not easily overcome in the social and economic context of developing countries.

Circadian systems biology: When time matters.

The circadian clock is a powerful endogenous timing system, which allows organisms to fine-tune their physiology and behaviour to the geophysical time. The interplay of a distinct set of core-clock genes and proteins generates oscillations in expression of output target genes which temporally regulate numerous molecular and cellular processes. The study of the circadian timing at the organismal as well as at the cellular level outlines the field of chronobiology, which has been highly interdisciplinary ever since its origins. The development of high-throughput approaches enables the study of the clock at a systems level. In addition to experimental approaches, computational clock models exist which allow the analysis of rhythmic properties of the clock network. Such mathematical models aid mechanistic understanding and can be used to predict outcomes of distinct perturbations in clock components, thereby generating new hypotheses regarding the putative function of particular clock genes. Perturbations in the circadian timing system are linked to numerous molecular dysfunctions and may result in severe pathologies including cancer. A comprehensive knowledge regarding the mechanistic of the circadian system is crucial to develop new procedures to investigate pathologies associated with a deregulated clock. In this manuscript we review the combination of experimental methodologies, bioinformatics and theoretical models that have been essential to explore this remarkable timing-system. Such an integrative and interdisciplinary approach may provide new strategies with regard to chronotherapeutic treatment and new insights concerning the restoration of the circadian timing in clock-associated diseases.

Assembly of a comprehensive regulatory network for the mammalian circadian clock: a bioinformatics approach.

By regulating the timing of cellular processes, the circadian clock provides a way to adapt physiology and behaviour to the geophysical time. In mammals, a light-entrainable master clock located in the suprachiasmatic nucleus (SCN) controls peripheral clocks that are present in virtually every body cell. Defective circadian timing is associated with several pathologies such as cancer and metabolic and sleep disorders. To better understand the circadian regulation of cellular processes, we developed a bioinformatics pipeline encompassing the analysis of high-throughput data sets and the exploitation of published knowledge by text-mining. We identified 118 novel potential clock-regulated genes and integrated them into an existing high-quality circadian network, generating the to-date most comprehensive network of circadian regulated genes (NCRG). To validate particular elements in our network, we assessed publicly available ChIP-seq data for BMAL1, REV-ERBα/ß and RORα/γ proteins and found strong evidence for circadian regulation of Elavl1, Nme1, Dhx6, Med1 and Rbbp7 all of which are involved in the regulation of tumourigenesis. Furthermore, we identified Ncl and Ddx6, as targets of RORγ and REV-ERBα, ß, respectively. Most interestingly, these genes were also reported to be involved in miRNA regulation; in particular, NCL regulates several miRNAs, all involved in cancer aggressiveness. Thus, NCL represents a novel potential link via which the circadian clock, and specifically RORγ, regulates the expression of miRNAs, with particular consequences in breast cancer progression. Our findings bring us one step forward towards a mechanistic understanding of mammalian circadian regulation, and provide further evidence of the influence of circadian deregulation in cancer.