Avaliação dos níveis de autofagia e morfometria nuclear no carcinoma epidermoide de esôfago

O câncer de esôfago é a sexta causa de morte relacionada a neoplasias malignas no mundo. A taxa de sobrevida em 5 anos é inferior a 20 %, após o diagnóstico. O conhecimento da biologia básica e o desenvolvimento de novos marcadores diagnósticos e prognósticos podem contribuir para o manejo clínico desses tumores. A autofagia é um mecanismo fisiológico envolvido na degradação de organelas e proteínas disfuncionais ou envelhecidas. Perturbações na autofagia têm sido associadas à carcinogênese. Em nosso estudo: (a) comparamos os níveis de três indicadores de autofagia, SQSTM1, MAP1LC3B e BECN1 no epitélio esofágico não neoplásico e em amostras de carcinoma epidermoide de esôfago (ESCC); (b) analisamos a influência da idade, sexo e tabagismo no nível desses indicadores; (c) avaliamos a morfometria nuclear e sua correlação com a autofagia; e (d) examinamos a influência da autofagia na taxa de sobrevida e no estadiamento do ESCC. Para isso, selecionamos 32 biópsias de esôfago sem alterações associadas a neoplasia (grupo controle) e 53 amostras de ESCC. O material foi imunomarcado para SQSTM1, MAP1LC3B e BECN1, seguido da quantificação dos níveis proteicos e geração de um Índice Autofágico para cada amostra. Também foi realizada a análise morfométrica nuclear. A influência da autofagia nas taxas de sobrevida dos pacientes com ESCC foi estimada usando o banco de dados TCGA. Observamos um aumento nos níveis dos três indicadores de autofagia nas amostras de ESCC comparadas com o grupo controle. O AutoIndex, que acreditamos melhor representar o estado autofágico, comparado aos indicadores isolados, demonstrou diferença ainda maior entre os grupos. Encontramos uma redução na área nuclear no ESCC em comparação às amostras controle. Essa redução se correlacionou negativamente com o AutoIndex, sugerindo que alterações nucleares e na autofagia ocorrem concomitantemente ao longo da carcinogênese do ESCC. Utilizando o TCGA identificamos níveis elevados de MAP1LC3A/B, SQSTM1, Atg4A e Atg12 associados a um pior prognóstico e estadiamento avançado, sugerindo que a autofagia no ESCC contribua para a agressividade tumoral. Ainda usando o TCGA verificamos que a combinação de genes de autofagia dois a dois apresenta uma diferença ainda maior nas curvas de Kaplan-Meier entre pacientes com níveis elevados e baixos de autofagia. Ao final, a partir dos nossos resultados e dados da literatura, levantamos a hipótese de que quanto maior a diferença, para mais ou menos, da intensidade da autofagia em um tumor em relação ao tecido não neoplásico, pior o prognóstico. Em conclusão, observamos um aumento da autofagia no ESCC, e sugerimos que a combinação de indicadores deste mecanismo, mais do que o uso de marcadores isolados, tem potencial para uso na avaliação prognóstica no ESCC.Esophageal cancer is the sixth leading cause of deaths by malignant neoplasias in the word. Less than 20 % of patients survive 5 years after diagnosis, so the knowledge of basic tumor biology as well as the development of new prognostic markers may contribute to the management of these tumors. Autophagy is a physiological mechanism involved in the degradation of aged or dysfunctional organelles and proteins. Disturbs in autophagy have been associated with the carcinogenesis. Here, we: (a) compared the levels of three autophagy markers, SQSTM1, MAP1LC3B and BECN1 in non-neoplastic esophagus and in Esophagus Squamous Cell Carcinoma (ESCC) samples; (b) assessed the influence of age, gender and smoking in the level of these markers; (c) evaluate nuclear morphometry and its correlation with autophagy; and (d) examined the influence of autophagy genes expression in the survival and staging of ESCC. We select 32 non-neoplastic esophageal samples (control group) and 53 ESCC samples, which were immunostained to SQSTM1, MAP1LC3B and BECN1, followed by the quantification of these proteins levels and the generation of an Autophagic Index (AutoIndex) to each sample. Nuclear morphometry was assessed by the Nuclear Morphometric Analysis. The influence of autophagy in the overall survival time of ESCC patients was estimated using the TCGA database. We found that the three autophagy markers were significantly increased in ESCC samples in comparison to non-neoplastic samples. The AutoIndex, which we believe that better represents the autophagic status of a given sample, then single markers, showed an even higher difference between groups. We found a reduction in nuclear area in ESCC in comparison to controls. This reduction correlated negatively with AutoIndex, suggesting these alterations in the structure of the chromatin and autophagy occur concomitantly along the carcinogenesis of ESCC. Using the TCGA data bank we found that high levels of MAP1LC3 A/B, SQSTM1, Atg4A and Atg12 gene expression were associated with a poor prognosis and advanced staging, suggesting that autophagy contributes to tumor aggressiveness. The combination of autophagy gene markers two by two showed an even larger gap in Kaplan-Meier curves between patients with low and high levels of autophagy. Based on our data and those from the literature we raised the hypothesis that the greater the difference of autophagy in tumours compared to non-neoplastic tissue, the poorer the prognosis, regardless the alteration is up or down regulated. In conclusion, we found that an increase in autophagy may be involved in the progression of ESCC, and that the combination, more than a single marker analysis, has the potential to be used as a prognostic index in ESCC

Combining functional traits and phylogeny to disentangling Amazonian butterfly assemblages on anthropogenic gradients

Environmental gradients consist of sequential changes in the physical and structural characteristics of a region. These allow us to follow species responses and tolerances under different habitat conditions. Among them, forest fragmentation and succession comprise the most common examples of forest gradients, where organismal responses require distinct morphological, physiological, and behavioral adaptations. However, environmental changes can impose ecological and evolutionary constraints that act on species traits, as well as on local species assemblies through their phylogenetic history. In this study, we evaluated the differences in species distribution and composition on fruit-feeding butterfly assemblages along forest fragmentation and succession gradients. We combine functional and phylogenetic methods for determining butterfly assemblages, and inferred species resistance and resilience according to habitat changes in tropical forests. We used a database of 471 fruit-feeding butterflies of 60 species sampled from different environments in the central Amazon rainforest. A total of 13 functional traits were measured, and a phylogenetic tree was obtained for the sampled species. The trait–environment relationship was analyzed along both forest fragmentation and succession gradients, controlling for phylogenetic signal on species distribution and functional composition when necessary. Several traits presented phylogenetic signal, and phylogeny was also driving butterfly species distribution along the successional gradient. After controlling for phylogeny, individual characteristics related to flight speed (thoracic weight) and antipredatory strategies (camouflage) increased in early-successional forests, with large butterflies (body length) prevailing in primary forests. No clear functional and phylogenetic pattern was identified for the fragmentation gradient. Our results are consistent with the idea that butterflies may be employing distinct functional strategies to attenuate habitat change effects. Larger butterflies, with lower dispersal ability, are preferentially susceptible to local extinctions in the early-successional environments, mainly when forested habitat and its resources become spatially restricted. In addition, several anti-predatory strategies related to conspicuous colors may be losing their functionality in open areas, where not being distinctive against the background becomes the primary defense against predation

Butterflies (Lepidoptera: Papilionoidea and Hesperioidea) from Swamp forests and Restinga forests at the southern Brazilian Coastal Plain

This paper presents a butterfly species list of Swamp and Resting forests in the Coastal Plain of Rio Grande do Sul and Santa Catarina States aiming to contribute to the knowledge of butterflies for these environments in the Atlantic Forest. Data compilation was obtained through inventories carried out in the years 2005 and 2011 with two sampling protocols (bait traps and butterfly nets). After 10.920 trap-hours and 360 net-hours 225 species of butterflies were recorded belonging to six families and 19 subfamilies. Twenty-five species are new records for the Rio Grande do Sul State and 35 species are new records for the Atlantic Forest in Rio Grande do Sul, with six Nymphalidae, ten Hesperiidae, twelve Lycaenidae and seven Riodinidae. The results obtained in the present study are fundamental for the knowledge and conservation of the taxa studied and to their associated habitats.Este trabalho teve como objetivo elaborar uma lista de espécies de borboletas de Matas Paludosas e de Restingas da Planície Costeira do Rio Grande do Sul e de Santa Catarina contribuindo para o conhecimento da fauna de borboletas da Mata Atlântica. Foram compilados dados obtidos com amostragens realizadas através de duas técnicas de coleta (armadilha com isca atrativa e rede entomológica) nos anos de 2005 e 2011. Com um esforço amostral de 10.920 horas com armadilhas e de 360 horas-rede foram encontradas 225 espécies de borboletas nos dois ambientes, distribuídas em seis famílias e 19 subfamílias. Vinte e cinco espécies são registros novos ainda não publicados para o Rio Grande do Sul e 35 espécies são novos registros para a Mata Atlântica do Rio Grande do Sul, sendo seis Nymphalidae, dez Hesperiidae, doze Lycaenidae e sete Riodinidae. Os resultados gerados no presente estudo são fundamentais para o conhecimento e conservação dos táxons estudados bem como dos ambientes aos quais estão associados.181190Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Avaliação dos níveis de autofagia e morfometria nuclear no carcinoma epidermoide de esôfago

O câncer de esôfago é a sexta causa de morte relacionada a neoplasias malignas no mundo. A taxa de sobrevida em 5 anos é inferior a 20 %, após o diagnóstico. O conhecimento da biologia básica e o desenvolvimento de novos marcadores diagnósticos e prognósticos podem contribuir para o manejo clínico desses tumores. A autofagia é um mecanismo fisiológico envolvido na degradação de organelas e proteínas disfuncionais ou envelhecidas. Perturbações na autofagia têm sido associadas à carcinogênese. Em nosso estudo: (a) comparamos os níveis de três indicadores de autofagia, SQSTM1, MAP1LC3B e BECN1 no epitélio esofágico não neoplásico e em amostras de carcinoma epidermoide de esôfago (ESCC); (b) analisamos a influência da idade, sexo e tabagismo no nível desses indicadores; (c) avaliamos a morfometria nuclear e sua correlação com a autofagia; e (d) examinamos a influência da autofagia na taxa de sobrevida e no estadiamento do ESCC. Para isso, selecionamos 32 biópsias de esôfago sem alterações associadas a neoplasia (grupo controle) e 53 amostras de ESCC. O material foi imunomarcado para SQSTM1, MAP1LC3B e BECN1, seguido da quantificação dos níveis proteicos e geração de um Índice Autofágico para cada amostra. Também foi realizada a análise morfométrica nuclear. A influência da autofagia nas taxas de sobrevida dos pacientes com ESCC foi estimada usando o banco de dados TCGA. Observamos um aumento nos níveis dos três indicadores de autofagia nas amostras de ESCC comparadas com o grupo controle. O AutoIndex, que acreditamos melhor representar o estado autofágico, comparado aos indicadores isolados, demonstrou diferença ainda maior entre os grupos. Encontramos uma redução na área nuclear no ESCC em comparação às amostras controle. Essa redução se correlacionou negativamente com o AutoIndex, sugerindo que alterações nucleares e na autofagia ocorrem concomitantemente ao longo da carcinogênese do ESCC. Utilizando o TCGA identificamos níveis elevados de MAP1LC3A/B, SQSTM1, Atg4A e Atg12 associados a um pior prognóstico e estadiamento avançado, sugerindo que a autofagia no ESCC contribua para a agressividade tumoral. Ainda usando o TCGA verificamos que a combinação de genes de autofagia dois a dois apresenta uma diferença ainda maior nas curvas de Kaplan-Meier entre pacientes com níveis elevados e baixos de autofagia. Ao final, a partir dos nossos resultados e dados da literatura, levantamos a hipótese de que quanto maior a diferença, para mais ou menos, da intensidade da autofagia em um tumor em relação ao tecido não neoplásico, pior o prognóstico. Em conclusão, observamos um aumento da autofagia no ESCC, e sugerimos que a combinação de indicadores deste mecanismo, mais do que o uso de marcadores isolados, tem potencial para uso na avaliação prognóstica no ESCC.Esophageal cancer is the sixth leading cause of deaths by malignant neoplasias in the word. Less than 20 % of patients survive 5 years after diagnosis, so the knowledge of basic tumor biology as well as the development of new prognostic markers may contribute to the management of these tumors. Autophagy is a physiological mechanism involved in the degradation of aged or dysfunctional organelles and proteins. Disturbs in autophagy have been associated with the carcinogenesis. Here, we: (a) compared the levels of three autophagy markers, SQSTM1, MAP1LC3B and BECN1 in non-neoplastic esophagus and in Esophagus Squamous Cell Carcinoma (ESCC) samples; (b) assessed the influence of age, gender and smoking in the level of these markers; (c) evaluate nuclear morphometry and its correlation with autophagy; and (d) examined the influence of autophagy genes expression in the survival and staging of ESCC. We select 32 non-neoplastic esophageal samples (control group) and 53 ESCC samples, which were immunostained to SQSTM1, MAP1LC3B and BECN1, followed by the quantification of these proteins levels and the generation of an Autophagic Index (AutoIndex) to each sample. Nuclear morphometry was assessed by the Nuclear Morphometric Analysis. The influence of autophagy in the overall survival time of ESCC patients was estimated using the TCGA database. We found that the three autophagy markers were significantly increased in ESCC samples in comparison to non-neoplastic samples. The AutoIndex, which we believe that better represents the autophagic status of a given sample, then single markers, showed an even higher difference between groups. We found a reduction in nuclear area in ESCC in comparison to controls. This reduction correlated negatively with AutoIndex, suggesting these alterations in the structure of the chromatin and autophagy occur concomitantly along the carcinogenesis of ESCC. Using the TCGA data bank we found that high levels of MAP1LC3 A/B, SQSTM1, Atg4A and Atg12 gene expression were associated with a poor prognosis and advanced staging, suggesting that autophagy contributes to tumor aggressiveness. The combination of autophagy gene markers two by two showed an even larger gap in Kaplan-Meier curves between patients with low and high levels of autophagy. Based on our data and those from the literature we raised the hypothesis that the greater the difference of autophagy in tumours compared to non-neoplastic tissue, the poorer the prognosis, regardless the alteration is up or down regulated. In conclusion, we found that an increase in autophagy may be involved in the progression of ESCC, and that the combination, more than a single marker analysis, has the potential to be used as a prognostic index in ESCC

Stichelia pelotensis (Lepidoptera, Riodinidae): conservation, notes, and rediscovery of an endangered butterfly from southern Brazil

AbstractStichelia pelotensis (Lepidoptera, Riodinidae) is an endemic and threatened butterfly from the Pampa biome in southern Brazil, and has not been recorded in its type locality in the last 56 years. Recently, a population was found in two sites from extreme south Brazil, Pelotas, Rio Grande do Sul state. These records are an important find given the conservation status of S. pelotensis, since all the information gathered is new and involve the natural history of this species. The information obtained is useful for the management, monitoring and conservation priorities of this species and its associated habitats, since its known distribution is restricted to a narrow area in the Rio Grande do Sul Coastal Plain inside this threatened biome in southern Brazil

First Record of Aricoris cinericia (Stichel, 1910) (Riodinidae) from Brazil and Update on Its Geographical Distribution

Se da a conocer a Aricoris cinericia por vez primera para Brasil, así como se mencionan otros registros en Argentina, conducta y distribución geográfica.Fil: Russo Siewert, Ricardo. Universidade Federal Do Parana; BrasilFil: Agra Iserhard, Cristiano. Universidade Federal de Pelotas; BrasilFil: Núñez Bustos, Ezequiel Osvaldo. Fundación de Historia Natural Felix de Azara; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Piccoli Romanowski, Helena. Universidade Federal do Rio Grande do Sul; Brasi

Distribution patterns of riodinid butterflies (Lepidoptera: Riodinidae) from southern Brazil

Background: The aim of this study was to synthesize the knowledge of Riodinidae butterflies (Lepidoptera: Papilionoidea) in Rio Grande do Sul state (RS), southern Brazil, evaluating the role of climatic, topographic, and vegetational variables on the observed patterns of occurrence and distribution of these butterflies in the Pampa and Atlantic Forest biomes. The records of riodinid butterflies in RS were collected from published studies and the examination of museum collections in Brazil. Results: A total of 97 taxa of Riodinidae were recorded, distributed in 92 municipalities. The NMDS analysis and the Constrained Analysis of Principal Coordinates grouped the municipalities according to the phytogeographic regions and biomes - Pampa and Atlantic Forest domains - in which the species records were made. Distance from the ocean, precipitation and temperature were the environmental variables which most contributed to explain the distribution patterns of these butterflies. The multivariate Mantel correlogram suggests that over short distances, the composition of species shows significant levels of spatial autocorrelation, and as geographic distance increases, these levels tend to present negative values. Conclusions: The results suggest that the observed distribution pattern of Riodinidae in the different biomes and phytogeographic regions in the extreme southern Brazil could be explained by climatic, environmental variables and geographic distance