Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear un derstanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5–7 vast areas of the tropics remain understudied.8–11 In the American tropics, Amazonia stands out as the world’s most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepre sented in biodiversity databases.13–15 To worsen this situation, human-induced modifications16,17 may elim inate pieces of the Amazon’s biodiversity puzzle before we can use them to understand how ecological com munities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple or ganism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region’s vulnerability to environmental change. 15%–18% of the most ne glected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lostinfo:eu-repo/semantics/publishedVersio

Pervasive gaps in Amazonian ecological research

Biodiversity loss is one of the main challenges of our time,1,2 and attempts to address it require a clear understanding of how ecological communities respond to environmental change across time and space.3,4 While the increasing availability of global databases on ecological communities has advanced our knowledge of biodiversity sensitivity to environmental changes,5,6,7 vast areas of the tropics remain understudied.8,9,10,11 In the American tropics, Amazonia stands out as the world's most diverse rainforest and the primary source of Neotropical biodiversity,12 but it remains among the least known forests in America and is often underrepresented in biodiversity databases.13,14,15 To worsen this situation, human-induced modifications16,17 may eliminate pieces of the Amazon's biodiversity puzzle before we can use them to understand how ecological communities are responding. To increase generalization and applicability of biodiversity knowledge,18,19 it is thus crucial to reduce biases in ecological research, particularly in regions projected to face the most pronounced environmental changes. We integrate ecological community metadata of 7,694 sampling sites for multiple organism groups in a machine learning model framework to map the research probability across the Brazilian Amazonia, while identifying the region's vulnerability to environmental change. 15%–18% of the most neglected areas in ecological research are expected to experience severe climate or land use changes by 2050. This means that unless we take immediate action, we will not be able to establish their current status, much less monitor how it is changing and what is being lost

Analysis of the M93 olfactory receptor expression in heterologous systems

O sistema olfatório de mamífero pode discriminar milhares de odores presentes no meio ambiente. Aproximadamente 1000 diferentes receptores olfatórios (ORs) são expressos no epitélio olfatório (OE) do nariz, Os ORs detectam os odores e transmitem os sinais resultantes para o bulbo olfatório (OB) no cérebro. Os ORs pertencem a super família dos receptores acoplados a proteína G (GPCR) e apresentam sete domínios transmembrânicos putativos. Por razões desconhecidas, os ORs são retidos no retículo endoplasmático quando expressos em linhagens de células de mamíferos heterólogas. Provavelmente, proteínas acessórias sejam requeridas para o endereçamento dos Ors para a superficie celular. No presente estudo, utilizamos o OR M93 para estudar os mecanismos de expressão de um ORo A dissertação teve como objetivos específicos: (l) construção de um vetor para expressão do OR M93 em fusão com GFP em levedura e análise de sua localização celular; (2) identificar proteínas expressas no epitélio olfatório de camundongo que interajam com os ORs. A análise por microscopia de fluorescência revelou que a expressão do OR M93 fusionado a GFP demonstrou um padrão de fluorescência que sugere a retenção do OR M93 no retículo endoplasmático. Nós utilizamos o sistema de duplo híbrido em levedura para varrer uma biblioteca de cDNA de epitélio olfatório de camundongo com uma isca correspondente à região N-terminal do OR M93. Quatro proteínas candidatas foram identificadas: HLA-B associado ao transcrito 3 (BAT-3/ Scythe), superfamília transmembrana 4 (membro CD82), superfamília transmembrana 4 (membro OAP-I) e sindecan (membro SDC2) (\"GenBank accession numbers\": BC026647, D14883, BC0430n e BC047144). A análise da hibridação in situ destas proteínas, revelou que a proteína OAP-1 é a melhor candidata a interação com OR M93. Dessa maneira, nós indicamos a proteína OAP-1 como possível proteína candidata a auxiliar o OR a ser expresso de maneira funcional em sistemas heterólogos.The mammalian olfactory system can discrim inate thousands of odorants present in the environrnent. Approximately 1000 different olfactory receptors (ORs) are expressed in the olfactory epithelium (OE) of the nose. The ORs detect odorants and transmit the resulting signals to the olfactory bulb (OB) of the brain. ORs belong to the G-protein-coupled receptor (GPCR) super family and have seven putative transmembrane domains. For unknown reasons, the ORs are retained in the endoplasmatic reticulum when expressed in heterologous mammalian cell lines. Probably accessory proteins are required for the sorting of the ORs to the cell surface. In the present work, we used the OR M93 to study the mechanisms of OR expression. Our goals were to (1) construct an expression vector for OR M93 in fusion with GFP in yeast and (2) to identify proteins expressed in the mouse OE that interact with ORs. The analysis by fluorescence microscopy suggested that OR M93 in fusion with GFP was retained in the endoplasmic reticulum (ER) of yeast. We used the yeast two-hybrid system to screen a mouse OE cDNA library with a bait corresponding to the N-terminal region ofthe üR M93. Four potential candidates were identified: HLA-B associated transcript 3 (BAT-3/Scythe), transmembrane 4 superfamily (CD82 member), transmembrane 4 superfamily (TSPN-3 member) and syndecan (SDC2). In situ hybridization analysis suggests that OAP-l protein represents the best candidate for interaction with OR M93. We suggest the OAP-l protein could be an accessory protein required for the sorting of the ORs to the cell surface in heterologous cell lines

Analysis of the M93 olfactory receptor expression in heterologous systems

O sistema olfatório de mamífero pode discriminar milhares de odores presentes no meio ambiente. Aproximadamente 1000 diferentes receptores olfatórios (ORs) são expressos no epitélio olfatório (OE) do nariz, Os ORs detectam os odores e transmitem os sinais resultantes para o bulbo olfatório (OB) no cérebro. Os ORs pertencem a super família dos receptores acoplados a proteína G (GPCR) e apresentam sete domínios transmembrânicos putativos. Por razões desconhecidas, os ORs são retidos no retículo endoplasmático quando expressos em linhagens de células de mamíferos heterólogas. Provavelmente, proteínas acessórias sejam requeridas para o endereçamento dos Ors para a superficie celular. No presente estudo, utilizamos o OR M93 para estudar os mecanismos de expressão de um ORo A dissertação teve como objetivos específicos: (l) construção de um vetor para expressão do OR M93 em fusão com GFP em levedura e análise de sua localização celular; (2) identificar proteínas expressas no epitélio olfatório de camundongo que interajam com os ORs. A análise por microscopia de fluorescência revelou que a expressão do OR M93 fusionado a GFP demonstrou um padrão de fluorescência que sugere a retenção do OR M93 no retículo endoplasmático. Nós utilizamos o sistema de duplo híbrido em levedura para varrer uma biblioteca de cDNA de epitélio olfatório de camundongo com uma isca correspondente à região N-terminal do OR M93. Quatro proteínas candidatas foram identificadas: HLA-B associado ao transcrito 3 (BAT-3/ Scythe), superfamília transmembrana 4 (membro CD82), superfamília transmembrana 4 (membro OAP-I) e sindecan (membro SDC2) (\"GenBank accession numbers\": BC026647, D14883, BC0430n e BC047144). A análise da hibridação in situ destas proteínas, revelou que a proteína OAP-1 é a melhor candidata a interação com OR M93. Dessa maneira, nós indicamos a proteína OAP-1 como possível proteína candidata a auxiliar o OR a ser expresso de maneira funcional em sistemas heterólogos.The mammalian olfactory system can discrim inate thousands of odorants present in the environrnent. Approximately 1000 different olfactory receptors (ORs) are expressed in the olfactory epithelium (OE) of the nose. The ORs detect odorants and transmit the resulting signals to the olfactory bulb (OB) of the brain. ORs belong to the G-protein-coupled receptor (GPCR) super family and have seven putative transmembrane domains. For unknown reasons, the ORs are retained in the endoplasmatic reticulum when expressed in heterologous mammalian cell lines. Probably accessory proteins are required for the sorting of the ORs to the cell surface. In the present work, we used the OR M93 to study the mechanisms of OR expression. Our goals were to (1) construct an expression vector for OR M93 in fusion with GFP in yeast and (2) to identify proteins expressed in the mouse OE that interact with ORs. The analysis by fluorescence microscopy suggested that OR M93 in fusion with GFP was retained in the endoplasmic reticulum (ER) of yeast. We used the yeast two-hybrid system to screen a mouse OE cDNA library with a bait corresponding to the N-terminal region ofthe üR M93. Four potential candidates were identified: HLA-B associated transcript 3 (BAT-3/Scythe), transmembrane 4 superfamily (CD82 member), transmembrane 4 superfamily (TSPN-3 member) and syndecan (SDC2). In situ hybridization analysis suggests that OAP-l protein represents the best candidate for interaction with OR M93. We suggest the OAP-l protein could be an accessory protein required for the sorting of the ORs to the cell surface in heterologous cell lines

Study of cell division in Bacillus subtilis by fluorescence microscopy

A divisão celular em B. subtilis inicia-se pela formação de um complexo multiprotéico, o divisomo, no sítio onde a bactéria irá se dividir. FtsZ é a primeira proteína a se localizar no futuro sitio de divisão, formando uma estrutura em anel (anel Z) que se estende por toda a circunferência da célula. O anel Z funciona como um arcabouço responsável por recrutar outras quinze proteínas de divisão que irão participar da montagem do divisomo. Nesta tese, utilizamos abordagens quantitativas e qualitativas de microscopia de fluorescência vital para estudarmos duas questões ainda não esclarecidas sobre o funcionamento do divisomo. A primeira delas é como o divisomo é montado. Para estudarmos a montagem do divisomo nós realizamos ensaios de co-localização entre o anel Z (FtsZ-mCherry) e as proteínas ZapA, EzrA, FtsW, FtsL, YpsB , DivIVA, e MinC fusionadas a GFP. Quanto maior a freqüência de co-localização entre FtsZ e outra proteína de divisão, mais inicial é a participação da proteína na formação do divisomo. Portanto, a medida da freqüência de co-localização entre o anel Z e as proteínas componentes do divisomo permite que se deduza uma cinética da montagem deste complexo. Estes ensaios demonstraram uma freqüência de co-localização de 97,33% para ZapA; 98,31% para EzrA; 83,90% para FtsW; 78,43% para FtsL; 50% para YpsB; 41,7% para DivIVA e 31,64% para MinC. Estes resultados sugerem que o divisomo seja formado em três etapas. ZapA e EzrA se associam ao divisomo imediatamente após a formação do anel Z, em seguida FtsW e FtsL são recrutados para o divisomo, e por último YpsB, DivIVA, MinC associam-se ao divisomo. A segunda questão que investigamos nesta tese foi o mecanismo da mudança de posição do divisomo que ocorre durante a esporulação em B. subtilis. Na fase de esporulação a célula divide-se assimetricamente, com a formação do septo próxima a um dos pólos. Durante o crescimento vegetativo a divisão não ocorre próxima aos pólos por causa da ação das proteínas MinC, MinD e DivIVA, importantes reguladores espaciais da divisão. MinCD e DivIVA são inibidores da formação do anel Z que durante o crescimento vegetativo se localizam nos pólos das células.. Uma hipótese para explicar o uso dos sítios polares para a divisão durante a esporulação seria que as proteínas MinCD e DivIVA seriam removidas dos pólos celulares. Para testarmos esta hipótese, estudamos a localização das proteínas MinCD e DivIVA durante a esporulação. Nossos resultados demonstraram que MinCD e DivIVA se re-localizam e saem dos pólos celulares durante a esporulação. Porém esta dinâmica ocorre após a formação do anel Z assimétrico, sugerindo que o anel Z seja insensível a estes inibidores durante a esporulação. Por ensaios genéticos em B. subtilis demonstramos que a proteína SpoIIE, conhecida como provável proteína responsável por promover a formação do septo assimétrico, seja capaz de contrapor a ação de MinC no início da esporulação. Dessa maneira nós propomos um novo modelo de mudança da divisão simétrica para assimétrica durante a esporulação, diferentemente da simples saída do complexo MinCD dos pólos como é proposto na literatura.Bacillus subtilis division begins through the formation of a multiprotein complex, the divisome, at the site of division. FtsZ is the earliest known protein to localize to the future division site where the protein forms a ring-like structure (Z-ring) that extends around the circumference of the cell. The Z-ring functions as a scaffold and recruits about fifteen other division proteins that compose the divisome. In this work, we used quantitative and qualitative methods of vital fluorescence microscopy to study two questions that have not been elucidated about the divisome dynamics. The first is how divisome is assembled. To address that problem, we made co-localization between Z-ring (FtsZ-mCherry) and proteins ZapA, EzrA, FtsW, FtsL, YpsB, DivIVA, and MinC fused to GFP. Higher is the match between GFP fusions to Z-ring, earlier is the assembly of division proteins to divisome. Therefore, the co-localization frequency between Z ring and divisome proteins will allow us to deduce the assemble kinetics of the divisome. This assays showed a co-localization frequency of 97,33% for ZapA; 98,31% for EzrA; 83,90 for FtsW; 78,43% for FtsL; 50% for YpsB; 41,7% for DivIVA and 31,64% for MinC. This data suggests that the divisome does not assemble in two but in three steps. ZapA and EzrA assemble into the divisome immediately after Z ring formation, secondly FtsW and FtsL were recruited to the divisome, and finally YpsB, DivIVA, MinC associated with the divisome. The second question that we investigated in this work is the mechanism responsible for change the divisome position that occurs during sporulation in B. subtilis. In sporulation the cell divides asymmetrically, with a septum formation near poles. During vegetative grown the divisiome does not occur near poles because of MinC, MinD and DivIVA action, relevant for spatial regulation of division. MinCD and DivIVA are inhibitors of Z ring formation that during vegetative growth are located at poles. A hypothesis to explain the use of polar sites for division during sporulation would be that MinCD and DivIVA would be removed from cellular poles. To test this hypothesis, we studied the location of MinCD and DivIVA proteins during sporulation. Our results demonstrated that MinCD and DivIVA re-localize and leave to cell poles during sporulation. However this process occurs after asymmetric Z ring formation, suggesting that Z ring would be unresponsive to this inhibitors during sporulation. Through genetics assays in B. subtilis we demonstrated that SpoIIE protein, known to probably play a role in asymmetric septum formation, would be able to contrapose MinC action during early sporulation. Therefore, we propose a novel model for change the symmetric to asymmetric division during sporulation, unlike the release of MinCD from pole proposed in the literature

Cytological Characterization of YpsB, a Novel Component of the Bacillus subtilis Divisome▿ †

Cell division in bacteria is carried out by an elaborate molecular machine composed of more than a dozen proteins and known as the divisome. Here we describe the characterization of a new divisome protein in Bacillus subtilis called YpsB. Sequence comparisons and phylogentic analysis demonstrated that YpsB is a paralog of the division site selection protein DivIVA. YpsB is present in several gram-positive bacteria and likely originated from the duplication of a DivIVA-like gene in the last common ancestor of bacteria of the orders Bacillales and Lactobacillales. We used green fluorescent protein microscopy to determine that YpsB localizes to the divisome. Similarly to that for DivIVA, the recruitment of YpsB to the divisome requires late division proteins and occurs significantly after Z-ring formation. In contrast to DivIVA, however, YpsB is not retained at the newly formed cell poles after septation. Deletion analysis suggests that the N terminus of YpsB is required to target the protein to the divisome. The high similarity between the N termini of YpsB and DivIVA suggests that the same region is involved in the targeting of DivIVA. YpsB is not essential for septum formation and does not appear to play a role in septum positioning. However, a ypsB deletion has a synthetic effect when combined with a mutation in the cell division gene ftsA. Thus, we conclude that YpsB is a novel B. subtilis cell division protein whose function has diverged from that of its paralog DivIVA

Re-introduction of dengue virus serotype 2 in the state of Rio de Janeiro after almost a decade of epidemiological silence.

The Asian/American genotype of dengue virus serotype 2 (DENV-2) has been introduced in Brazil through the state of Rio de Janeiro around 1990, and since then it has been spreading and evolving, leading to several waves of dengue epidemics throughout the country that cause a major public health problem. Of particular interest has been the epidemic of 2008, whose highest impact was evidenced in the state of Rio de Janeiro, with a higher number of severe cases and mortality rate, compared to previous outbreaks. Interestingly, no circulation of DENV-2 was witnessed in this region during the preceding 9-year period. By early 2010, phylogenetic analysis of the 2008 epidemic strain revealed that the outbreak was caused by a new viral lineage of the Asian/American genotype, which was pointed as responsible for the outbreak severity as well. The same scenario is repeating in 2019 in this state; however, only a few cases have been detected yet. To provide information that helps to the understanding of DENV-2 dynamics in the state of Rio de Janeiro, and thereafter contribute to public health control and prevention actions, we employed phylogenetic studies combined with temporal and dynamics geographical features to determine the origin of the current viral strain. To this effect, we analyzed a region of 1626 nucleotides entailing the Envelope/NS1 viral genes. Our study reveals that the current strain belongs to the same lineage that caused the 2008 outbreak, however, it is phylogenetically distant from any Brazilian strain identified so far. Indeed, it seemed to be originated in Puerto Rico around 2002 and has been introduced into the state in late 2018. Taking into account that no DENV-2 case was reported over the last decade in the state (representing a whole susceptible children generation), and the fact that a new viral strain may be causing current dengue infections, these results will be influential in strengthening dengue surveillance and disease control, mitigating the potential epidemiological consequences of virus spread

Early Evidence for Zika Virus Circulation among Aedes aegypti Mosquitoes, Rio de Janeiro, Brazil

During 2014–2016, we conducted mosquito-based Zika virus surveillance in Rio de Janeiro, Brazil. Results suggest that Zika virus was probably introduced into the area during May–November 2013 via multiple in-country sources. Furthermore, our results strengthen the hypothesis that Zika virus in the Americas originated in Brazil during October 2012–May 2013