Spatial distribution and temporal evolution of DRONPA-fused SNAP25 clusters in adrenal chromaffin cells

Abstract Sub-diffraction imaging of plasma membrane localized proteins, such as the SNARE (SolubleNSFAttachment Protein Receptor) proteins involved in exocytosis, in fixed cells have resulted in images with high spatial resolution, at the expense of dynamical information. Here, we have imaged localized fluorescence bursts of DRONPA-fused SNAP-25 molecules in live chromaffin cells by Total Internal Reflection Fluorescence (TIRF) imaging. We find that this method allows tracking protein cluster dynamics over relatively long times (∼20 min.), partly due to the diffusion into the TIRF field of fresh molecules, making possible the simultaneous identification of cluster size, location and temporal evolution. The results indicate that the DRONPA-fused SNAP-25 clusters display rich dynamics, going from staying constant to disappearing and reappearing in specific cluster domains within minutes.</jats:p

Paisagem e Literatura: análises geoliterárias no poema Morte e Vida Severina de João Cabral de Melo Neto

Literatura e Geografia caminham lado a lado na materialização do ficcional para o real, dentro de um processo mimético. Assim, este trabalho tem por objetivo trazer observações feitas a partir da leitura do poema Morte e Vida Severina, escrito por João Cabral de Melo Neto, entre os anos de 1954 e 1955, e publicado em 1956. Este poema conta a história de Severino, homem que sai do Sertão em busca de uma melhor qualidade de vida. A metodologia utiliza-se de uma revisão bibliográfica em que apontamos como principais fontes teóricas de consulta: Bachelard (1986), Monteiro (2002), Feitosa (2010), Pinheiro Neto (2012) e Alves (2018).Literatura y Geografía caminan lado a lado en la materialización de lo ficcional a lo real, dentro de un proceso mimético. Así, este trabajo tiene por objetivo traer observaciones hechas a partir del poema Morte e Vida Severina, escrito por João Cabral de Melo Neto, entre los años 1954 y 1955, y publicado en 1956. Este poema cuenta la historia de Severino, hombre que sale del Sertão en busca de una mejor calidad de vida. La metodología usa de una revisión bibliográfica en la que apuntamos como principales fuentes teóricas de consulta: Bachelard (1986), Monteiro (2002), Feitosa (2010), Pinheiro Neto (2012) e Alves (2018).Literature and Geography take together in the materialization of the fictional to the real, within a mimetic process. Thus, this paper aims to bring observations made from the poem Morte e Vida Severina, written by João Cabral de Melo Neto, between the years 1954 and 1955, and published in 1956. That tells the story of Severino, a man who leaves the hinterland in search of a better quality of life. The methodology uses a bibliographical review in which we point out as main theoretical sources of consultation: Bachelard (1986), Monteiro (2002), Feitosa (2010), Pinheiro Neto (2012) e Alves (2018)

NODULAÇÃO DE LEGUMINOSAS COM POTENCIAL DE USO NA RECUPERAÇÃO DE ÁREAS DEGRADADAS EM CAMPO RUPESTRE QUARTZÍTICO

The objective of this work was to evaluate the composition and structure of leguminous species in vegetal formation of rocky fields in Serra do Espinhaço Meridional, as well as to verify the diversity of nitrogen-fixing bacteria of the nodular species. The floristic survey was carried out in a reserve area belonging to the Anglo American mining company, based in the municipality of Conceição do Mato Dentro - MG. The region is located on the eastern edge of the Serra do Espinhaço Meridional and has a Cwa climate. To carry out the floristic survey, 25 fixed plots (10×10 m) were installed for the shrub-tree compartment and 60 fixed plots (1 × 2 m) for the herb-subshrub compartment. Absolute and relative density, absolute and relative frequency were estimated for both compartments. After the previous floristic survey, 40 root nodules of the species of Calliandra fasciculata and 23 of Moldenhawera emarginata were collected. Afterwards, bacteria were isolated from the nodules from the species C. fasciculata and M. emarginata. The cultural characteristics of the isolates were evaluated and from the data, it was possible to group the isolates through a cultural dendrogram using the Jaccard coefficient, using the MVSP (Multivariate Statistical Package) software. Three leguminous species were recorded in both compartments, totaling 1192 individuals distributed among C. fasciculata (with 1080 ind.), M. emarginata (with 86 ind.) and Periandra mediterranea (with 26 ind.). However, for the species P. mediterranea, no nodulation was observed in the field. The species C. fasciculata was responsible for 92.62% of the total number of individuals. From the morphological characteristics successfully evaluated, of the 23 isolates of C. fasciculata, 6 cultural groups were obtained. This result suggests high morphophysiological diversity of nitrogen-fixing bacteria nodulating the species in question, in a quartzite rock field environment. Due to the importance of this species, its use in recovery projects is recommended, especially in environments similar to the one in this study.Este trabalho teve o objetivo de avaliar a composição e estrutura de espécies leguminosas em formação vegetal de campo rupestre na Serra do Espinhaço Meridional, assim como verificar a diversidade de bactérias fixadoras de nitrogênio das espécies nodulíferas. O levantamento florístico foi realizado em uma área de Reserva Legal da empresa Anglo American no município de Conceição do Mato Dentro, Minas Gerais. A região está inserida na Borda Leste da Serra do Espinhaço Meridional e apresenta clima Cwa. Instalou-se 25 parcelas fixas (10 × 10 m) para a comunidade arbustivo-arbóreo e 60 parcelas fixas (1 × 2 m) para a comunidade herbáceo-subarbustivo no levantamento florístico. Foram estimadas a densidade absoluta e relativa, frequência absoluta e relativa para ambas as comunidades. Após o levantamento florístico prévio, realizou-se a coleta de 40 nódulos da raiz das espécies de Calliandra fasciculata e 23 de Moldenhawera emarginata. Em seguida, realizou-se o isolamento das bactérias dos nódulos provenientes das espécies C. fasciculata e M. emarginata. Foram avaliadas as características culturais dos isolados e a partir dos dados, foi possível agrupar os isolados por meio de um dendrograma cultural com a utilização do coeficiente de Jaccard, pelo software MVSP (Multivariate Stastitical Package). Três espécies leguminosas foram registradas em ambos as comunidades, totalizando 1.192 indivíduos distribuídos dentre C. fasciculata (com 1080 ind.), M. emarginata (com 86 ind.) e Periandra mediterranea (com 26 ind.). Contudo, na espécie P. mediterranea não foi constatada nodulação em campo. A espécie C. fasciculata foi responsável por 92,62% do total de indivíduos. A partir das características morfológicas avaliadas, dos 23 isolados de C. fasciculata, obteve-se 6 grupos culturais. Este resultado sugere alta diversidade morfofisiológica de bactérias fixadoras de nitrogênio nodulando essa espécie em ambiente de campo rupestre quartzítico. Devido à importância desta espécie, recomenda-se seu uso em projetos de recuperação, especialmente em ambientes semelhantes ao deste estudo. Palavras-chave: endemismo; Fabaceae; nitrogênio; nodulação.   Nodulating legumes with potential for use in the recovery of degraded areas in quartzite rocky fields   ABSTRACT: The objective of this work was to evaluate the composition and structure of leguminous species in the vegetal formation of rocky fields in Serra do Espinhaço Meridional, as well as to verify the diversity of nitrogen-fixing bacteria of the nodular species. The floristic survey was carried out in a reserve area based in the municipality of Conceição do Mato Dentro, Minas Gerais. The region is located on the eastern edge of the Serra do Espinhaço Meridional and has a Cwa climate. Twenty-five fixed plots (10 × 10 m) were installed for the shrub-tree compartment and 60 fixed plots (1 × 2 m) for the herb-subshrub compartment to carry out the floristic survey. Absolute and relative density, absolute and relative frequency were estimated for both compartments. After the previous floristic survey, 40 root nodules of the species of Calliandra fasciculata and 23 of Moldenhawera emarginata were collected. Afterward, bacteria were isolated from the nodules of C. fasciculata and M. emarginata. The cultural characteristics of the isolates were evaluated and from the data, it was possible to group the isolates through a cultural dendrogram using the Jaccard coefficient, using the MVSP (Multivariate Statistical Package) software. Three leguminous species were recorded in both compartments, totaling 1192 individuals distributed among C. fasciculata (with 1080 ind.), M. emarginata (with 86 ind.) and Periandra mediterranea (with 26 ind.). However, for the species P. mediterranea, no nodulation was observed in the field. The species C. fasciculata was responsible for 92.62% of the total number of individuals. From the morphological characteristics successfully evaluated, of the 23 isolates of C. fasciculata, 6 cultural groups were obtained. This result suggests high morphophysiological diversity of nitrogen-fixing bacteria nodulating this species in a quartzite rock field environment. Due to the importance of this species, its use in recovery projects is recommended, especially in environments similar to the one in this study. Keywords: endemism; Fabaceae; nitrogen; nodulation

Influence of daily usage times on patients’ compliance during at-home bleaching : a randomized clinical trial

The effectiveness of at-home dental bleaching treatments depends on the time that bleaching products are in contact with the teeth surface and, consequently, on the adequate use of associated custom acetate trays. Objective: This randomized single-blinded trial aimed to analyze if the daily usage time of these products influences the patient’s compliance behavior when submitted to monitored at-home dental bleaching. Secondary outcomes were color change and tooth sensitivity. Methodology: Sixty-six volunteers were randomly distributed into three groups (n=22): patients were instructed to use the trays for 2 (G2), 4 (G4), and 8 (G8) hours daily. The daily dental bleaching compliance behavior was measured using a microsensor inserted into the trays. Subjective and objective color evaluation assessments were adopted at baseline (T0), one (T1), two (T2), and three weeks (T3) after the beginning of the bleaching treatment, as well as two weeks after the treatment (T4). Tooth sensitivity was analyzed using the VAS scale, ranging from T1 to T4. Results: G2 showed a greater degree of cooperation than G8 and cooperation was inversely proportional to the recommended usage time. Significantly higher color change was observed in the upper arch for G8 when compared to G2 in subjective analysis, from T1 to T4. There were no statistical differences between the groups in objective analysis. Conclusion: Shorter recommended usage time of the bleaching product may improve the patient's compliance with at-home dental bleaching treatments. However, increased daily usage time may promote better subjective color change. Bleaching sensitivity was more significant in the first week for a longer time of use

Heavy Metal Poisoning in a Cockatiel (Nymphicus hollandicus)

Background: In recent decades the demand for unconventional pets has been relatively increasing, a situation that increasingly causes veterinarians to encounter these animals in medical and surgical practice. Of these animals, the birds stand out. Animals of the order Psittaciform are known as very curious and active creatures that have the tendency to chew objectsin their environment. Among the several occurrences that lead this animal to attend the veterinary clinic, we highlight the poisoning by heavy metals, especially lead poisoning (Pb) and zinc (Zn). The objective of this work was to report a case of heavy metal intoxication in cockatiel (Nymphicus hollandicus).Case: A cockatiel was taken to the veterinarian with a history of apathy, motor incoordination, exacerbated water consumption and regurgitation. Complete anamnesis was instrumental in directing suspected heavy metal intoxication. Radiopaque particles were visualized through radiographic examination, suggesting heavy metal intoxication. The diagnosis wasconcluded through complementary examinations since the clinical symptoms are nonspecific. The treatment was intended to provide emergency intervention, avoid further absorption, use of antidotes, provide supportive measures and provide guidance to the owner. It can be concluded that the diagnosis and treatment were successful.Discussion: Metal poisoning can kill birds. The veterinarian should always seek the literature in order to perform the best support and treatment. For this, detailed history and detailed medical history must be taken into account, since the time of ingestion and the type of metal interfere with the therapeutic conduct. The use of imaging tests such as x-rays andultrasound are essential to assist the clinician, especially in cases where the tutor does not know whether or not the animal has ingested an object. In the radiographic examination, the heavy metal has the characteristic of having high radiopacity, which was evidenced in the case in question. Radiographic positions should be considered in order to avoid false negatives. In the literature, the treatment of chelation therapy is prioritized to remove the circulating heavy metal and thus act on the cause of the problem. In the case in question calcium EDTA was used intramuscularly, which showed clinical improvement in the animal after the second application. Calcium EDTA binds to metals and facilitates their transport and excretion. The use of fluid therapy is necessary as a supportive treatment to prevent kidney damage, since heavy metals are highlyharmful to nephrons. Especially in cases where the animal stops feeding and ingesting water. The use of antibiotics is essential because in many cases the animal, in addition to not feeding, becomes prone to infections due to metal toxicity, therefore, prophylactic use is essential for a better prognosis. In the case in question, the use of enrofloxacin was effective,as reported in the literature. It is recommended that the diagnosis of serum lead and zinc dosage should be made, however, due to the difficulties of obtaining the samples, and since other metals may also cause intoxication, in the case in question the treatment was started without these results. According to the results obtained in this work, the treatment described in the literature is effective and can be performed immediately to save animal life without subsequent sequelae.Keywords: bird, lead, zinc, calcium EDTA

K- and L-shell theoretical fluorescence yields for the Fe isonuclear sequence

Funding Information: This research was funded in part by FCT (Portugal) under research center grant UID/FIS/04559/2020 (LIBPhys). This work was also funded through the project PTDC/FIS-AQM/31969/2017, “Ultra-high-accuracy X-ray spectroscopy of transition metal oxides and rare earths”. F.G. acknowledges support from FCT, Portugal through contract UI/BD/151000/2021 . J. M and J.P.S acknowledge the support of EMPIR, Germany , under Contract No. 20FUN04 PrimA-LTD. The EMPIR initiative is co-funded by the European Union’s Horizon 2020 research and innovation programme and the EMPIR, Germany participating States. Part of this work has been carried out under the High Performance Computing Chair - a R&D infrastructure (based at the University of Évora; PI: M. Avillez), endorsed by Hewlett Packard Enterprise (HPE), and involving a consortium of higher education institutions (University of Algarve, University of Évora, NOVA University Lisbon, and University of Porto), research centres (CIAC, CIDEHUS, CHRC), enterprises (HPE, ANIET, ASSIMAGRA, Cluster Portugal Mineral Resources, DECSIS, FastCompChem, GeoSense, GEOtek, Health Tech, Starkdata), and public/private organizations (Alentejo Tourism-ERT, KIPT Colab). Funding Information: This research was funded in part by FCT (Portugal) under research center grant UID/FIS/04559/2020 (LIBPhys). This work was also funded through the project PTDC/FIS-AQM/31969/2017, “Ultra-high-accuracy X-ray spectroscopy of transition metal oxides and rare earths”. F.G. acknowledges support from FCT, Portugal through contract UI/BD/151000/2021. J. M and J.P.S acknowledge the support of EMPIR, Germany, under Contract No. 20FUN04 PrimA-LTD. The EMPIR initiative is co-funded by the European Union's Horizon 2020 research and innovation programme and the EMPIR, Germany participating States. Part of this work has been carried out under the High Performance Computing Chair - a R&D infrastructure (based at the University of Évora; PI: M. Avillez), endorsed by Hewlett Packard Enterprise (HPE), and involving a consortium of higher education institutions (University of Algarve, University of Évora, NOVA University Lisbon, and University of Porto), research centres (CIAC, CIDEHUS, CHRC), enterprises (HPE, ANIET, ASSIMAGRA, Cluster Portugal Mineral Resources, DECSIS, FastCompChem, GeoSense, GEOtek, Health Tech, Starkdata), and public/private organizations (Alentejo Tourism-ERT, KIPT Colab). Publisher Copyright: © 2022 The Author(s)In this work, we present K- and L- shell fluorescence yield values of the full isonuclear sequence of Fe ions, using a state-of-the-art multiconfiguration Dirac–Fock approach. These results may be of importance for spectral fitting and plasma modeling, both in laboratory and astrophysical studies, where Fe is an important benchmark element. The K-shell fluorescence yields were found to be very similar up to the removal of 14 electrons.publishersversionpublishe

Antioxidant Activity and Stable Free Radicals in Robusta Green Coffee Genotypes/ Atividade antioxidante e Radicais Estáveis Livres em Genótipos de Café Verde Robusta

This study reports on physicochemical properties (total soluble solids, titratable acidity plus caffeine, trigonelline, 5-CQA, and total phenols contents) of five genotypes of Robusta coffee beans; Bamburral, Beira Rio, Clementino, Coringa, and Pirata. Green bean of Clementino presents the highest concentration of total soluble solids in dry basis (18.0%) and the lowest titratable acidity (154.0 mL-NaOH). Moreover, green beans of Clementino and Pirata show intermediate values of caffeine and trigonelline contents while presenting the highest yield index ofstable free radical formation after roasting, respectively equal to 26 and 23 (electron paramagnetic resonance-EPR data), and the highest content of 5-CQA (around 42 mg/g). Green beans of Bamburral, Beira Rio and Coringa show the highest concentration of total phenols (53 - 56 mg/g), meaning the highest antioxidant activities (IC50 test) but reduced yield index of formation of stable free radicals after roasting, respectively equal to 13.0, 5.9, and 13.0 (EPR data)

Gut Microbiota Diversity and C-Reactive Protein Are Predictors of Disease Severity in COVID-19 Patients

Funding Information: We acknowledge support from the NOVA Medical School of Universidade NOVA de Lisboa, CINTESIS, and CHRC. Funding. This study was sponsored by the Funda??o para a Ci?ncia e a Tecnologia (FCT, project no. 268_596883842), BIOCODEX, and CINTESIS (reference UIDB/4255/2020). The funders had no role in study design, data collection, data analysis, data interpretation, or manuscript writing. Publisher Copyright: © Copyright © 2021 Moreira-Rosário, Marques, Pinheiro, Araújo, Ribeiro, Rocha, Mota, Pestana, Ribeiro, Pereira, de Sousa, Pereira-Leal, de Sousa, Morais, Teixeira, Rocha, Silvestre, Príncipe, Gatta, Amado, Santos, Maltez, Boquinhas, de Sousa, Germano, Sarmento, Granja, Póvoa, Faria and Calhau.The risk factors for coronavirus disease 2019 (COVID-19) severity are still poorly understood. Considering the pivotal role of the gut microbiota on host immune and inflammatory functions, we investigated the association between changes in the gut microbiota composition and the clinical severity of COVID-19. We conducted a multicenter cross-sectional study prospectively enrolling 115 COVID-19 patients categorized according to: (1) the WHO Clinical Progression Scale—mild, 19 (16.5%); moderate, 37 (32.2%); or severe, 59 (51.3%), and (2) the location of recovery from COVID-19—ambulatory, 14 (household isolation, 12.2%); hospitalized in ward, 40 (34.8%); or hospitalized in the intensive care unit, 61 (53.0%). Gut microbiota analysis was performed through 16S rRNA gene sequencing, and the data obtained were further related to the clinical parameters of COVID-19 patients. The risk factors for COVID-19 severity were identified by univariate and multivariable logistic regression models. In comparison to mild COVID-19 patients, the gut microbiota of moderate and severe patients have: (a) lower Firmicutes/Bacteroidetes ratio; (b) higher abundance of Proteobacteria; and (c) lower abundance of beneficial butyrate-producing bacteria such as the genera Roseburia and Lachnospira. Multivariable regression analysis showed that the Shannon diversity index [odds ratio (OR) = 2.85, 95% CI = 1.09–7.41, p = 0.032) and C-reactive protein (OR = 3.45, 95% CI = 1.33–8.91, p = 0.011) are risk factors for severe COVID-19 (a score of 6 or higher in the WHO Clinical Progression Scale). In conclusion, our results demonstrated that hospitalized patients with moderate and severe COVID-19 have microbial signatures of gut dysbiosis; for the first time, the gut microbiota diversity is pointed out as a prognostic biomarker of COVID-19 severity.publishersversionpublishe