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Bats are important reservoirs for viruses of public health and veterinary concern. Virus studies in Australian bats usually target the families Paramyxoviridae, Coronaviridae and Rhabdoviridae, with little known about their overall virome composition. We used metatranscriptomic sequencing to characterise the faecal virome of grey-headed flying foxes from three colonies in urban/suburban locations from two Australian states. We identified viruses from three mammalian-infecting (Coronaviridae, Caliciviridae, Retroviridae) and one possible mammalian-infecting (Birnaviridae) family. Of particular interest were a novel bat betacoronavirus (subgenus Nobecovirus) and a novel bat sapovirus (Caliciviridae), the first identified in Australian bats, as well as a potentially exogenous retrovirus. The novel betacoronavirus was detected in two sampling locations 1,375 km apart and falls in a viral lineage likely with a long association with bats. This study highlights the utility of unbiased sequencing of faecal samples for identifying novel viruses and revealing broad-scale patterns of virus ecology and evolution.

Immunosuppressive therapy is standard for the treatment of inflammatory diseases and for minimizing rejection in transplant patients. However, immunosuppressant drugs are associated with an increased risk of certain cancers. In particular, melanoma is an immunogenic tumor and as such, is strongly influenced by the immune system. We performed this literature review to summarize the effects of commonly used immunomodulating agents on melanoma development, recurrence and progression. We outline the mechanism of action of each drug and discuss the available evidence on its influence on melanoma. Based on existing literature, we recommend avoiding the following agents in patients with a history of invasive melanoma: cyclosporine, sirolimus, natalizumab, IL-6 inhibitors, cyclophosphamide, methotrexate and the tumor necrosis factor-alpha inhibitors infliximab and etanercept. If there are no viable alternative agents, we recommend for these patients to see a dermatologist every 6 months for a thorough skin examination.

Supplementary Information, Figure 1, Table 1 from A Laminin-Collagen Complex Drives Human Epidermal Carcinogenesis through Phosphoinositol-3-Kinase Activation

Ticks harbour a high diversity of viruses, bacteria and protozoa. The soft tick Carios vespertilionis (Argasidae) is a common ectoparasite of bats in the Palearctic region and is suspected to be vector and reservoir of viruses and other microbial species in bat populations, some of which may act as zoonotic agents for human disease. The Soprano pipistrelle (Pipistrellus pygmaeus, Vespertilionidae) is widely distributed in Europe, where it can be found inside or close to human habitation. We used meta-transcriptomic sequencing to determine the RNA virome and common microbiota in blood-fed C. vespertilionis ticks collected from a Soprano pipistrelle bat roosting site in south-central Sweden. Our analyses identified 16 viruses from 11 virus families, of which 15 viruses were novel. For the first time in Sweden we identified Issuk-Kul virus, a zoonotic arthropod-borne virus previously associated with outbreaks of acute febrile illness in humans. Probable bat-associated and tick-borne viruses were classified within the families Nairoviridae, Caliciviridae and Hepeviridae, while other invertebrate-associated viruses included members of the Dicistroviridae, Iflaviridae, Nodaviridae, Partitiviridae, Permutotetraviridae, Polycipiviridae and Solemoviridae. Similarly, we found abundant bacteria in C. vespertilionis, including genera with known tick-borne bacteria, such as Coxiella spp. and Rickettsia spp. These findings demonstrate the remarkable diversity of RNA viruses and bacteria present in C. vespertilionis and highlight the importance of bat-associated ectoparasite surveillance as an effective and non-invasive means to track viruses and bacteria circulating in bats and ticks.

More than 70 bat species are found in mainland Australia, including five species of megabat from a single genus (family Pteropodidae) and more than 65 species representing six families of microbats. The conservation status of these animals varies from least concern to endangered. Research directed at evaluating the impact of microorganisms on bat health has been generally restricted to surveillance for specific pathogens. While most of the current bat virome studies focus on sampling apparently healthy individuals, little is known about the infectome of diseased bats. We performed traditional diagnostic techniques and metatranscriptomic sequencing on tissue samples from 43 individual bats, comprising three flying fox and two microbat species experiencing a range of disease syndromes, including mass mortality, neurological signs, pneumonia and skin lesions. We identified reads from four pathogenic bacteria and two pathogenic fungi, includingPseudomonas aeruginosain lung samples from flying foxes with peracute pneumonia, and with dermatitis. Of note, we identified the recently discovered Hervey pteropid gammaretrovirus, with evidence of replication consistent with an exogenous virus, in a bat with lymphoid leukemia. In addition, one novel picornavirus, at least three novel astroviruses and bat pegiviruses were identified. We suggest that the most likely cause of peracute lung disease wasPseudomonas aeruginosa, while we suspect Hervey pteropid gammaretrovirus was associated with lymphoid leukemia. It is possible that any of the novel astroviruses could have contributed to the presentation of skin lesions in individual microbats. This study highlights the importance of studying the role of microorganisms in bat health and conservation.IMPORTANCEBats have been implicated as reservoir hosts for zoonotic disease of concern, however, the burden of microorganism including viruses on bat health and disease is understudied. Here we incorporated veterinary diagnostics and RNA sequencing to identify the presence of microbes and viruses with possible pathogenic status in Australian bats with varying disease presentations. These techniques were able to effectively identify and describe several pathogenic species of bacteria and fungi in addition to known and novel viruses. This study emphasises the importance of screening pathogens in cases of bat mortality for the conservation of this diverse order.

The Andean Cordilleras of Colombia, especially the Cordillera Occidental, are among the areas with the highest diversity of Pristimantis frogs in the world. Within the Cordillera Occidental, the Se...

Arthropods harbor a largely undocumented diversity of RNA viruses. Some arthropods, like mosquitoes, can transmit viruses to vertebrates but are themselves parasitized by other arthropod species, such as mites. Very little is known about the viruses of these ectoparasites and how they move through the host-parasite relationship. To address this, we determined the virome of both mosquitoes and the mites that feed on them. The mosquito

MAPK-pathway up-regulation is responsible for over 40% of human cancer cases. Finding effective therapeutic targets for melanoma therapy continues to be a challenge due to drug resistance. Using a computational and experimental pipeline, we discovered the nuclear enriched long non-coding RNA (lncRNA) TRASH, which is induced upon MAPK-pathway-activation in melanoma. LncRNA hold essential regulatory functions in many cancer types. TRASH-targeting Antisense Oligonucleotides (TRASH-ASOs) greatly reduced cell-growth of melanoma cells in culture and systemic TRASH-ASO treatment significantly inhibited tumor growth of melanoma cell-line and patient-derived tumor xenografts without apparent side effects. We found that TRASH is essential for protein stability of the MAPK-pathway-regulating and apoptosis-inhibiting oncogene hnRNPA2/B1. TRASH knockdown induced apoptosis by down-regulating anti-apoptotic kinase activity and pro-survival signaling pathways. Compared to the well-studied oncogenic lncRNA MALAT1, the unique feature of TRASH is that it governs tumor cell-survival through maintaining activity of anti-apoptotic kinases and pro-survival signaling pathways in melanoma cells. TRASH-ASO treatment can bypass MEK-inhibitor (MEKi) resistance, and unlike MEKi, it does not induce early-onset treatment resistance. Furthermore, TRASH-ASO and MEKi combinations confer synergistic effects in melanoma treatment. Our findings show that TRASH-ASO treatment presents promising features for long lasting RNA-targeting melanoma therapy.

Arthropods harbor a largely undocumented diversity of RNA viruses. Some arthropods, like mosquitoes, can transmit viruses to vertebrates but are themselves parasitized by other arthropod species, such as mites. Very little is known about the viruses of these ectoparasites and how they move through the host–parasite relationship. To address this, we determined the virome of both mosquitoes and the mites that feed on them. The mosquito Aedes communis is an abundant and widely distributed species in Sweden, in northern Europe. These dipterans are commonly parasitized by water mite larvae (Trombidiformes: Mideopsidae) that are hypothesized to impose negative selection pressures on the mosquito by reducing fitness. In turn, viruses are dual-host agents in the mosquito–mite interaction. We determined the RNA virus diversity of mite-free and mite-detached mosquitoes, as well as their parasitic mites, using meta-transcriptomic sequencing. Our results revealed an extensive RNA virus diversity in both mites and mosquitoes, including thirty-seven putative novel RNA viruses that cover a wide taxonomic range. Notably, a high proportion of viruses (20/37) were shared between mites and mosquitoes, while a limited number of viruses were present in a single host. Comparisons of virus composition and abundance suggest potential virus transfer between mosquitoes and mites during their symbiotic interaction. These findings shed light on virome diversity and ecology in the context of arthropod host–parasite–virus relationships.

The endosymbiont bacteria of the genus Wolbachia are associated with multiple mutualistic effects on insect biology, including nutritional and antiviral properties. Members of the genus Wolbachia naturally occur in fly species of the genus Drosophila, providing an operational model host for studying how virome composition may be affected by its presence. Drosophila simulans populations can carry a variety of strains of members of the genus Wolbachia , with the wAu strain associated with strong antiviral protection under experimental conditions. We used D. simulans sampled from the Perth Hills, Western Australia, to investigate the potential virus protective effect of the wAu strain of Wolbachia on individual wild-caught flies. Our data revealed no appreciable variation in virus composition and abundance between individuals infected or uninfected with Wolbachia associated with the presence or absence of wAu. However, it remains unclear whether wAu might affect viral infection and host survival by increasing tolerance rather than inducing complete resistance. These data also provide new insights into the natural virome diversity of D. simulans. Despite the small number of individuals sampled, we identified a repertoire of RNA viruses, including nora virus, galbut virus, thika virus and La Jolla virus, that have been identified in other species of the genus Drosophila. Chaq virus-like sequences associated with galbut virus were also detected. In addition, we identified five novel viruses from the families Reoviridae, Tombusviridae, Mitoviridae and Bunyaviridae. Overall, this study highlights the complex interaction between Wolbachia and RNA virus infections and provides a baseline description of the natural virome of D. simulans.

The endosymbiont bacterium Wolbachia is associated with multiple mutualistic effects on insect biology, including nutritional and antiviral properties. Wolbachia naturally occurs in Drosophila fly species, providing an operational model host to study how virome composition may be impacted by its presence. Drosophila simulans populations can carry a variety of Wolbachia strains. In particular, the wAu strain of Wolbachia has been associated with strong antiviral protection under experimental conditions. We used D. simulans sampled from the Perth Hills, Western Australia, to investigate the potential virus protective effect of the wAu strain on individual wild-caught flies. Our data revealed no appreciable variation in virus composition and abundance between Wolbachia infected/uninfected individuals associated with the presence/absence of wAu. However, it remains unclear whether wAu might impact viral infection and host survival by increasing tolerance rather than inducing complete resistance. These data also provide new insights into the natural virome diversity of D. simulans. Despite the small number of individuals sampled, we identified a repertoire of RNA viruses, including Nora virus, Galbut virus, Chaq virus, Thika virus and La Jolla virus, that have been identified in other Drosophila species. In addition, we identified five novel viruses from the families Reoviridae, Tombusviridae, Mitoviridae and Bunyaviridae. Overall, this study highlights the complex interaction between Wolbachia and RNA virus infections and provides a baseline description of the natural virome of D. simulans.

The Serrania de los Paraguas is a mountainous region situated in the central part of the Cordillera Occidental of Colombia. It rises some 2500 m above the wet forest and supports a high concentration of species richness and endemism for plants and terrestrial vertebrates (amphibians, birds, and mammals). In the case of amphibians, 61 named species occurring in The Serrania de los Paraguas are recognized in the literature, which are mostly the works of J.D. Lynch and P. Ruiz-Carranza in the 1990s. During a 9-d survey in the wet season (19–27 July 2019) on the Reserva Natural Comunitaria Cerro El Ingles (one of the hills of the Serrania de los Paraguas) on the frontier between Choco and Valle del Cauca departments, we discovered two new species of the genus Pristimantis. Both species share a divided palmar tubercle as well as a distinctive color pattern on the groin, of which both are distinctive within Pristimantis. The first new species is distinguished by its small size and having a distinctive carmine red color pattern on groin, uniform lateral fringes on fingers (no crenulated), palmar tubercle deeply bifid (or divided), advertisement call composed of a short single note distinctly pulsed, and absence of nuptial pads as well as hyperdistal tubercles, inner tarsal tubercle and fold, and partial fusion of Toe IV and V. The second new species is distinguished by having dermal ridges on scapular region, conical tubercle on upper eyelid and heel, palmar tubercle divided, differentiated tympanic membrane, males with vocal slits, orange spots on groin, advertisement call composed of distinctly pulsed 6–8 notes, and absence of dorsolateral folds as well as cranial crest, hyperdistal tubercles, and toe webbing. Considering these new species, the number of amphibians inhabiting the Serrania de los Paraguas has increased to 63 species. It is important to point out that within this outstanding species richness of amphibians, there are 10 endemic species and 17 threatened species according to the International Union for Conservation of Nature Red List (7 Critically Endangered, 5 Endangered, and 5 Vulnerable). Likewise, it is noteworthy that 29 terraranan species can be found in the Reserva Natural Comunitaria Cerro El Ingles. Thus, based on the species richness, endemism, diversity at higher taxonomic levels, and the number of threatened species, it seems reasonable to say that the Serrania de los Paraguas is a priority site for conservation of amphibians in Colombia.

Revealing the determinants of virome composition is central to placing disease emergence in a broader evolutionary context. Fish are the most species-rich group of vertebrates and so provide an ideal model system to study the factors that shape virome compositions and their evolution. We characterised the viromes of 19 wild-caught species of marine fish using total RNA sequencing (meta-transcriptomics) combined with analyses of sequence and protein structural homology to identify divergent viruses that often evade characterisation. From this, we identified 25 new vertebrate-associated viruses and a further 22 viruses likely associated with fish diet or their microbiomes. The vertebrate-associated viruses identified here included the first fish virus in the Matonaviridae (single-strand, negative-sense RNA virus). Other viruses fell within the Astroviridae, Picornaviridae, Arenaviridae, Reoviridae, Hepadnaviridae, Paramyxoviridae, Rhabdoviridae, Hantaviridae, Filoviridae and Flaviviridae and were sometimes phylogenetically distinct from known fish viruses. We also show how key metrics of virome composition – viral richness, abundance and diversity – can be analysed along with host ecological and biological factors as a means to understand virus ecology. Accordingly, these data suggest that that the vertebrate-associated viromes of the fish sampled here are predominantly shaped by the phylogenetic history (i.e. taxonomic order) of their hosts, along with several biological factors including water temperature, habitat depth, community diversity and swimming behaviour. No such correlations were found for viruses associated with porifera, molluscs, arthropods, fungi and algae, that are unlikely to replicate in fish hosts. Overall, these data indicate that fish harbour particularly large and complex viromes and the vast majority of fish viromes are undescribed.

Revealing the determinants of virome composition is central to placing disease emergence in a broader evolutionary context. Fish are the most species-rich group of vertebrates and so provide an ideal model system to study the factors that shape virome compositions and their evolution. We characterized the viromes of nineteen wild-caught species of marine fish using total RNA sequencing (meta-transcriptomics) combined with analyses of sequence and protein structural homology to identify divergent viruses that often evade characterization. From this, we identified twenty-five new vertebrate-associated viruses and a further twenty-two viruses likely associated with fish diet or their microbiomes. The vertebrate-associated viruses identified here included the first fish virus in the

The Red fox (Vulpes vulpes) has established large populations in Australia’s urban and rural areas since its introduction following European settlement. The cryptic and highly adaptable nature of foxes allows them to invade cities and live among humans whilst remaining largely unnoticed. Urban living and access to anthropogenic food resources also influence fox ecology. Urban foxes grow larger, live at higher densities, and are more social than their rural counterparts. These ecological changes in urban red foxes are likely to impact the pathogens that they harbour, and foxes could pose a disease risk to humans and other species that share these urban spaces. To investigate this possibility, we used a meta-transcriptomic approach to characterise the virome of urban and rural foxes across the Greater Sydney region in Australia. Urban and rural foxes differed significantly in virome composition, with rural foxes harbouring a greater abundance of viruses compared to their urban counterparts. We identified ten potentially novel vertebrate-associated viruses in both urban and rural foxes, some of which are related to viruses associated with disease in domestic species and humans. These included members of the Astroviridae, Picobirnaviridae, Hepeviridae, and Picornaviridae as well as rabbit haemorrhagic disease virus-2. This study sheds light on the viruses carried by urban and rural foxes and emphasises the need for greater genomic surveillance of foxes and other invasive species at the human–wildlife interface.

Tilapia lake virus (TiLV) has caused mass mortalities in farmed and wild tilapia with serious economic and ecological consequences. Until recently, this virus was the sole member of the Amnoonviridae, a family within the order Articulavirales comprising segmented negative-sense RNA viruses. We sought to identify additional viruses within the Amnoonviridae through total RNA sequencing (meta-transcriptomics) and data mining of published transcriptomes. Accordingly, we sampled marine fish species from both Australia and China and discovered several segments of two new viruses within the Amnoonviridae, tentatively called Flavolineata virus and Piscibus virus, respectively. In addition, by mining vertebrate transcriptome data, we identified nine additional virus transcripts matching to multiple genomic segments of TiLV in both marine and freshwater fish. These new viruses retained sequence conservation with the distantly related Orthomyxoviridae in the RdRp subunit PB1, but formed a distinct and diverse phylogenetic group. These data suggest that the Amnoonviridae have a broad host range within fish and that greater animal sampling will identify additional divergent members of the Articulavirales.

The Red fox (Vulpes vulpes) has established large populations in Australia’s urban and rural areas since its introduction following European settlement. Foxes’ cryptic and highly adaptable nature allows them to invade cities and live among humans while remaining largely unnoticed. Urban living and access to anthropogenic food resources also influences fox ecology. Urban foxes grow larger, live at higher densities and are more social than their rural counterparts. These ecological changes in urban red foxes are likely to impact the pathogens that they harbour, and foxes could pose a disease risk to humans and other species that share these urban spaces. To assess this possibility, we used a meta-transcriptomic approach to characterise the viromes of urban and rural foxes across the Greater Sydney region in Australia. Urban and rural foxes differed significantly in virome composition, with rural foxes harbouring a greater abundance of viruses compared to their urban counterparts. In contrast, urban fox viromes comprised a greater diversity of viruses compared to rural foxes. We identified nine potentially novel vertebrate-associated viruses in both urban and rural foxes, some of which are related to viruses associated with disease in domestic species and humans. These included members of theAstroviridae, Picobirnaviridae, HepeviridaeandPicornaviridaeas well as rabbit haemorrhagic disease virus-2 (RHDV2). This study sheds light on the viruses carried by urban and rural foxes and emphasises the need for greater genomic surveillance of foxes and other invasive species at the human-wildlife interface.ImportanceUrbanisation of wild environments is increasing as human populations continue to expand. Remnant pockets of natural environments and other green spaces in urban landscapes provide invasive wildlife such as red foxes with refuges within urban areas, where they thrive on the food resources provisioned by humans. Close contact between humans, domestic species and foxes likely increases the risk of novel pathogen emergence. Indeed, the vast majority of emerging infectious diseases in humans originate in wild animals. Here, we explored potential differences in viromes between urban fox invaders and their rural counterparts. Viromes of foxes and their ectoparasites comprise a diversity of viruses including those from theAstroviridae, Picobirnaviridae, Hepeviridae, CaliciviridaeandPicornaviridae. Microbial surveillance in foxes and other urban wildlife is vital for monitoring viral emergence and for the prevention of infectious diseases.

Targeted therapy has become a cornerstone for the treatment of melanoma patients. Targeting NRAS function is particularly challenging. To date, only single MEK inhibitor treatment was able to show minimal clinical efficacy. The discovery that co-targeting of MEK and CDK4,6 has antitumor activity created excitement for patients and clinicians; however, it is largely unknown if only NRAS mutant patients might benefit from MEK/CDK4,6 blockade. In this study we investigate response patterns of NRAS, BRAF mutant and ‘wild type’ melanoma cells in vitro and in vivo when challenged with inhibitors of MEK, CDK4,6 and the combination of both. Data revealed, that in vitro growth response patterns of cells treated with the MEK/CDK4,6 combination correspond to in vivo efficacy of MEK/CDK4,6 co-targeting in melanoma xenograft models. Strikingly, this was consistently observed in NRAS and BRAF mutant, as well as in ‘wild type’ melanoma cells. Additionally, cells displaying elevated p-Rb levels after single MEK inhibition, showed more effective growth reduction with MEK/CDK4,6 co-targeting compared to single MEK inhibitor treatment in vivo. Findings indicate that combined MEK/CDK4,6 inhibition could offer an effectively therapeutic modality in a subset of BRAF and NRAS mutant, as well as ‘wild type’ melanoma patients.

Background Wildlife species carry a remarkable diversity of trypanosomes. The detection of trypanosome infection in native Australian fauna is central to understanding their diversity and host-parasite associations. The implementation of total RNA sequencing (meta-transcriptomics) in trypanosome surveillance and diagnosis provides a powerful methodological approach to better understand the host species distribution of this important group of parasites. Methods We implemented a meta-transcriptomic approach to detect trypanosomes in a variety of tissues (brain, liver, lung, skin, gonads) sampled from native Australian wildlife, comprising four marsupials (koala, Phascolarctos cinereus; southern brown bandicoot, Isoodon obesulus; swamp wallaby, Wallabia bicolor; bare-nosed wombat, Vombatus ursinus), one bird (regent honeyeater, Anthochaera phrygia) and one amphibian (eastern dwarf tree frog, Litoria fallax). Samples corresponded to both clinically healthy and diseased individuals. Sequencing reads were de novo assembled into contigs and annotated. The evolutionary relationships among the trypanosomatid sequences identified were determined through phylogenetic analysis of 18S rRNA sequences. Results We detected trypanosome sequences in all six species of vertebrates sampled, with positive samples in multiple organs and tissues confirmed by PCR. Phylogenetic analysis indicated that the trypanosomes infecting marsupials were related to those previously detected in placental and marsupial mammals, while the trypanosome in the regent honeyeater grouped with avian trypanosomes. In contrast, we provide the first evidence for a trypanosome in the eastern dwarf tree frog that was phylogenetically distinct from those described in other amphibians. Conclusions To our knowledge, this is the first meta-transcriptomic analysis of trypanosomes in native Australian wildlife, expanding the known genetic diversity of these important parasites. We demonstrated that RNA sequencing is sufficiently sensitive to detect low numbers of Trypanosoma transcripts and from diverse hosts and tissues types, thereby representing an effective means to detect trypanosomes that are divergent in genome sequence.

The largest outbreak of yellow fever of the 21st century in the Americas began in 2016, with intense circulation in the southeastern states of Brazil, particularly in sylvatic environments near densely populated areas including the metropolitan region of São Paulo city (MRSP) during 2017–2018. Herein, we describe the origin and molecular epidemiology of yellow fever virus (YFV) during this outbreak inferred from 36 full genome sequences taken from individuals who died following infection with zoonotic YFV. Our analysis revealed that these deaths were due to three genetic variants of sylvatic YFV that belong the South American I genotype and that were related to viruses previously isolated in 2017 from other locations in Brazil (Minas Gerais, Espírito Santo, Bahia and Rio de Janeiro states). Each variant represented an independent virus introduction into the MRSP. Phylogeographic and geopositioning analyses suggested that the virus moved around the peri-urban area without detectable human-to-human transmission, and towards the Atlantic rain forest causing human spill-over in nearby cities, yet in the absence of sustained viral transmission in the urban environment.

Actualmente los profesores que realizan investigacion y que desarrollan la practica docente, se enfrentan al problema de validar la confiabilidad de un instrumento de medicion, que ayude a verificar y tener certeza sobre los temas que investigan y los desempenos de los alumnos. El metodo para validar el instrumento fue el de consistencia interna alfa de Cronbach. El articulo tiene como objetivo determinar la fiabilidad de un instrumento de medicion de satisfaccion del alumno en la aplicacion de un test, usado como estrategia de medicion en tercer grado de basica primaria en Cucuta, Colombia. Investigacion cuantitativa, descriptiva, con muestra de 1000 estudiantes de basica primaria en instituciones ubicadas en zona vulnerable. Se utilizo software SPSS version 21 para analisis e interpretacion estadistica. Esta ofrece un analisis de fiabilidad y como mejorar la probabilidad de exito del instrumento al aumentar la confiabilidad del mismo. Conclusion: el instrumento es fiable para instituciones ubicadas en zona de alta vulnerabilidad por el conflicto armado. Palabras claves Confiabilidad, instrumento de medicion, consistencia interna, alfa de Cronbach, interpretacion estadistica. Abstract Currently teachers who conduct research and who develop teaching practice, face the problem of validating the reliability of a measurement instrument, which helps to verify and be certain about the topics they investigate and the performances of the students. The most commonly used method to validate an instrument is Cronbach's alpha internal consistency, which is used in this work. This article shows a practical example to determine the reliability of a measurement instrument of student satisfaction in the application of a test, used as a measurement strategy in the third grade of elementary school at Colegio Andres Bello in Cucuta, Colombia. SPSS software version 21 and its statistical interpretation are used. This paper offers an analysis of reliability and how to improve the probability of success of the instrument by increasing the reliability of it. Keywords: Reliability, measurement instrument, internal consistency, Cronbach's alpha, statistical interpretation. Resumo Atualmente professores que conduzem pesquisas e que desenvolvem praticas de ensino, enfrentam o problema de validar a confiabilidade de um instrumento de medicao, o que ajuda a verificar e ter certeza dos topicos que investigam e dos desempenhos dos alunos. O metodo mais utilizado para validar um instrumento e a consistencia interna alfa de Cronbach, que e usada neste trabalho. Este artigo mostra um exemplo pratico para determinar a confiabilidade de um instrumento de medicao de satisfacao dos alunos na aplicacao de um teste, utilizado como estrategia de medicao na terceira serie da escola primaria no Colegio Andres Bello em Cucuta, Colombia. A versao de software SPSS 21 e sua interpretacao estatistica sao usadas. Este artigo oferece uma analise de confiabilidade e como melhorar a probabilidade de sucesso do instrumento, aumentando a confiabilidade do mesmo. Palavras-chave: Confiabilidade, instrumento de medicao, consistencia interna, alfa de Cronbach, interpretacao estatistica

In melanoma, mutant and thereby constantly active neuroblastoma rat sarcoma (NRAS) affects 15-20% of tumors, contributing to tumor initiation, growth, invasion, and metastasis. Recent therapeutic approaches aim to mimic RAS extinction by interfering with critical signaling pathways downstream of the mutant protein. This study investigates the phosphoproteome of primary human melanocytes bearing mutations in the two hot spots of NRAS, NRAS(G12) and NRAS(Q61). Stable isotope labeling by amino acids in cell culture followed by mass spectrometry identified 14,155 spectra of 3,371 unique phosphopeptides mapping to 1,159 proteins (false discovery rate < 2%). Data revealed pronounced PI3K/AKT signaling in NRAS(G12V) mutant cells and pronounced mitogen-activated protein kinase (MAPK) signaling in NRAS(Q61L) variants. Computer-based prediction models for kinases involved, revealed that CK2α is significantly overrepresented in primary human melanocytes bearing NRAS(Q61L) mutations. Similar differences were found in human NRAS(Q61) mutant melanoma cell lines that were also more sensitive to pharmacologic CK2α inhibition compared with NRAS(G12) mutant cells. Furthermore, CK2α levels were pronounced in patient samples of NRAS(Q61) mutant melanoma at the mRNA and protein level. The preclinical findings of this study reveal that codon 12 and 61 mutant NRAS cells have distinct signaling characteristics that could allow for the development of more effective, mutation-specific treatment modalities.

Melanoma is one of the leading cancers worldwide, distinguished for its malignancy and low survival rates. Although the poor outcome could improve with an early diagnosis and a good monitoring of the disease, current melanoma biomarkers display several limitations which make them useless. Interestingly, long-noncoding RNAs are secreted into the bloodstream inside exosomes by a wide range of malignant cells, and several of them have been validated as promising circulating molecular signatures of other tumors, but not melanoma. In this review we propose to explore the booming field of long-noncoding RNAs in order to find potential candidates to be tested as novel melanoma biomarkers, with the ultimate goal of improving melanoma detection, diagnosis and prognosis.

Up to 18% of melanomas harbor mutations in the neuroblastoma rat-sarcoma homolog (NRAS). Yet, decades of research aimed to interfere with oncogenic RAS signaling have been largely disappointing and have not resulted in meaningful clinical outputs. Recent advances in disease modeling, structural biology, and an improved understanding of RAS cycling as well as RAS signaling networks have renewed hope for developing strategies to selectively block hyperactive RAS function. This review discusses direct and indirect blocking of activated RAS with a focus on current and potential future therapeutic approaches for NRAS mutant melanoma.

The aim of this study was to use a 3-D finite element method (FEM) to compare the sliding resistance of 0.019"x0.025" stainless steel conventional archwires versus 0.019"x0.025" stainless steel beveled archwires in active (In-Ovation® Dentsply) and passive (SmartClip®, 3M) self-ligating brackets with 0.022" x 0.028" slots. A model was designed for each kind of bracket-archwire system and the following parameters were introduced in the models: friction coefficient calculated for stainless steel bracket-wire: 0.7 pm; Poisson ratio for stainless steel wire: 0.3, and elastic module: 205 GPa for bracket and 190 GPa for archwire. Static structural analysis was applied for homogeneous, linear and isotropic properties considering contacts between wire and bracket as frictional. The results indicate that the beveled archwire generates less stress than the rectangular wire in SmartClip® and In-Ovation® brackets. Comparing brackets, SmartClip® generated less stress than InOvation®. It is concluded that beveled rectangular arch wires provide the advantage of reduced sliding resistance, which is better in some clinical situations to improve orthodontic mechanics.El objetivo de este artículo fue comparar con el método tridimensional de elementos finitos (MEF) el comportamiento de la resistencia al deslizamiento expresado en esfuerzos de arcos de acero inoxidable 0.019"x0.025" convencionales y arcos de acero inoxidable con bisel de 0.019"x0.025" en brackets de autoligado slot 0.022" x 0.028" activo (InOvation® "R" Dentsply) y pasivo (SmartClip® 3M). Se diseñó un modelo de los brackets de autoligado In-Ovation® "R" Dentsply, SmartClip® 3My de los arcos de acero inoxidable convencionales y arcos de acero inoxidable con bisel, el análisis se calculó con el coeficiente de fricción para el bracket de acero inoxidable con el del arco de acero inoxidable: p 0.7. La relación de Poisson 0.3 para el arco de acero inoxidable. El módulo de elasticidad del bracket: 205 GPa y del arco: 190 GPa. La aplicación del método se realizó para un análisis estructural estático con condiciones de material homogéneas, lineales e isotrópicas y con contactos de tipo fricción entre el arco y el bracket. Se observó que el arco biselado generó menos esfuerzo que el arco rectangular en el bracket SmartClip® y en el bracket In-Ovation® "R", siendo el SmartClip® en el que se generó menor esfuerzo. Se encontró que existe una ventaja en el uso de los arcos rectangulares con bisel ya que presentan menor resistencia al deslizamiento optimizando en algunas situaciones clínicas las mecánicas ortodónticas.

Analiza cómo la aplicación del marketing social en el pueblo de Antioquía (Perú) contribuyó en la promoción, aceptación y modificación del comportamiento voluntario de las personas, logrando mejorar su realidad socioeconómica. En este contexto, observamos que las sociedades se esfuerzan constantemente por conseguir y sostener su bienestar, y así poder mejorar la calidad de vida de los individuos que la integran. Dichos esfuerzos son de incumbencia del marketing social, ya que este pretende influir en el comportamiento de las personas generando un cambio positivo.

BACKGROUND Dengue virus type 4 (DENV-4) was first reported in Brazil in 1982 and since then no more cases were detected again in Brazil until 2010, when the virus was reintroduced. Over the following years, the virus spread to several Brazilian states and resulted in about 1,400,000 dengue cases, in 2013. The largest number of cases were documented in the Southeast macro-region. OBJECTIVES To determine the phylogeography of DENV-4 Genotype IIB strains isolated during the epidemics in 2012-2013 in São Paulo, Brazil, we aimed to contextualise the contribution of viruses sampled in different localities across the overall movement of DENV-4 in Brazil. METHODS Based on the envelope gene sequences retrieved from GenBank, we employed a Bayesian phylogeographic approach to assess the spatiotemporal dynamics of DENV-4 Genotype IIB in São Paulo, Brazil. FINDINGS The dispersal dynamics of DENV-4 Genotype IIB in Brazil indicated Rio de Janeiro and Mato Grosso states as the most likely routes toward São Paulo before the 2012-2013 outbreak. Likewise, Guarujá and São José do Rio Preto facilitated viral spread and transmission to other localities in the South and Southeast macro-regions in Brazil. CONCLUSIONS The spread pattern of DENV-4 Genotype IIB strains across the country supports two independent introductions of the virus in São Paulo in a short period of time. Furthermore, São Paulo appears to have played a pivotal role in the dissemination of DENV-4 to other locations in Brazil.

Despite major advances in targeted melanoma therapies, drug resistance limits their efficacy. Long noncoding RNAs (lncRNAs) are transcriptome elements that do not encode proteins but are important regulatory molecules. LncRNAs have been implicated in cancer development and response to different therapeutics and are thus potential treatment targets; however, the majority of their functions and molecular interactions remain unexplored. In this study, we identify a novel cytoplasmic intergenic lincRNA (MIRAT), which is upregulated following prolonged MAPK inhibition in NRAS mutant melanoma and modulates MAPK signaling by binding to the MEK scaffold protein IQGAP1. Collectively, our results present MIRAT’s direct modulatory effect on the MAPK pathway and highlight the relevance of cytoplasmic lncRNAs as potential targets in drug resistant cancer.