Landscape resistance index aiming at functional forest connectivity.

Resistance models may quantify the ability of the landscape to impede species' movement and represent suitable habitats. Moreover, the performance of resistance models parameterized by land-use/land cover attributes evidence that the integrity of the environments subject to urban sprawl is poorly understood. In this sense, the study assumed we could identify the forest functional connectivity in a landscape considering the disparity in the landscape mosaic. In this context, we sought to develop a landscape resistance index through structural equation modeling (SEM), supported by the criteria of heat emission, biomass, and anthropogenic barriers, obtained by remote sensing, called observed variables. The landscape studied in the Green Belt Biosphere Reserve of São Paulo has significant remnants of the Atlantic Forest, a biodiversity hotspot. However, our results indicated criteria variability in the landscape modeled through the SEM, obtaining a significant adjustment of the landscape resistance index, with comparative fit index (CFI) of 1.00 and root mean square error of approximation (RMSEA) of 0.00. The index reflects the resistance levels of the land use/land cover, expressed by the class interval, ranging from 0% (1.73) to 100% (493.88), with the highest values associated with the anthropized uses and forest isolation. Thus, our index based on environmental attributes reflects the structure of functional forest connectivity and offers a framework to design forest corridors across landscapes.

An innovative application of event structure analysis (ESA).

This paper presents an innovative application of event structure analysis (ESA). The key improvements incorporated on the method are: (i) a robust system for coding events; (ii) the use of causal process tracing tests for inferring necessary connections; (iii) the combination of ESA with network analyses. Finally, we propose five types of analysis for event network models (i.e., critical elements, critical associations, critical connections, critical specific happenings, and critical antecedents) and exemplify some of them in a causal case study about the process of capability construction for open innovation management in an Industrial Electronic Manufacturer.•ESA can be combined with process-tracing tests to ground counterfactual causal inferences.•ESA can be combined with network analysis to explore quantitative patterns in event structures.•ESA is an outstanding method to conduct process research in management and engineering.

Functional and Structural Characterization of ClC-1 and Nav1.4 Channels Resulting from CLCN1 and SCN4A Mutations Identified Alone and Coexisting in Myotonic Patients.

Non-dystrophic myotonias have been linked to loss-of-function mutations in the ClC-1 chloride channel or gain-of-function mutations in the Nav1.4 sodium channel. Here, we describe a family with members diagnosed with Thomsen's disease. One novel mutation (p.W322*) in CLCN1 and one undescribed mutation (p.R1463H) in SCN4A are segregating in this family. The CLCN1-p.W322* was also found in an unrelated family, in compound heterozygosity with the known CLCN1-p.G355R mutation. One reported mutation, SCN4A-p.T1313M, was found in a third family. Both CLCN1 mutations exhibited loss-of-function: CLCN1-p.W322* probably leads to a non-viable truncated protein; for CLCN1-p.G355R, we predict structural damage, triggering important steric clashes. The SCN4A-p.R1463H produced a positive shift in the steady-state inactivation increasing window currents and a faster recovery from inactivation. These gain-of-function effects are probably due to a disruption of interaction R1463-D1356, which destabilizes the voltage sensor domain (VSD) IV and increases the flexibility of the S4-S5 linker. Finally, modelling suggested that the p.T1313M induces a strong decrease in protein flexibility on the III-IV linker. This study demonstrates that CLCN1-p.W322* and SCN4A-p.R1463H mutations can act alone or in combination as inducers of myotonia. Their co-segregation highlights the necessity for carrying out deep genetic analysis to provide accurate genetic counseling and management of patients.

Effect of setting data collection parameters on the reliability of a circular dichroism spectrum.

Circular dichroism (CD) spectroscopy is a well-established biophysical technique used to investigate the structure of molecules. The analysis of a protein CD spectrum depends on the quality of the original CD data, which can be affected by the sample purity, background absorption of the additives/solvent/buffer, the choice of the parameters used for data collection, etc. In this paper, the CD spectrum of myoglobin was used as a model to exploit how variations on each data collection parameter could affect the final protein CD spectrum and, the subsequent effect of them on the quantitative analysis of protein secondary structure. Bioinformatics analysis carried out with SESCA package and PDBMD2CD server predicted a theoretical myoglobin CD spectrum, and a Monte Carlo-like model was implemented to estimate the uncertainty in secondary structure predictions performed with CDSSTR, Selcon 3 and ContinLL algorithms. An inappropriate choice of data collection parameters can lead to a misinterpretation of the CD data in terms of the protein structural content.

The role of a socio-integrated recycling system in implementing a circular economy - The case of Belo Horizonte, Brazil.

Waste pickers (WPs) are considered a strong suggestion to become practical mediators of the circular economy (CE) in emerging economies. This new recommendation intends to strengthen WPs' role in household solid waste management while supporting the establishment of CE. Municipalities often do not recognize WPs as service providers and frequently discriminate against them. In such a challenging situation, could a socio-integrated recycling system with integrated WPs be a robust strategy to boost a CE? Belo Horizonte is a learning platform to answer this research question because this Brazilian city has a long-term commitment to social integration. The work applies the combination of participatory observation, multi-year material flow analysis (MFA), and structural agent analysis (SAA) to identify allocative resources, legitimation, and cultural values that are fundamental to operationalizing CE. The MFA results show a significant increase in waste generation, but not more than 4% of recyclable waste generated could be collected as input for WP cooperatives. The number of WPs registered in cooperatives, the market price of recyclables, and regulatory legislation for packaging products are classified as barriers for the successful extension of a socio-integrated recycling system identified in the SAA. This study suggests that knowing the target group (e.g., city hall and industries) brings opportunities for WPs to disclose niches (based on a small network of agents with expectations and visions) and can potentially create socio-technical regimes to implement a conscious and sustainable CE.

aeBlue Chromoprotein Color is Temperature Dependent.

BACKGROUND: Marine sessile organisms display a color palette that is the result of the expression of fluorescent and non-fluorescent proteins. Fluorescent proteins have uncovered transcriptional regulation, subcellular localization of proteins, and the fate of cells during development. Chromoproteins have received less attention until recent years as bioreporters. Here, we studied the properties of aeBlue, a a 25.91 kDa protein from the anemone Actinia equina. OBJECTIVE: To assess the properties of aeBlue chromoprotein under different physicochemical conditions. METHODS: In this article, during the purification of aeBlue we uncovered that it suffered a color shift when frozen. We studied the color shift by different temperature incubation and physicochemical conditions and light spectroscopy. To assess the possible structural changes in the protein, circular dichroism analysis, size exclusion chromatography and native PAGE was performed. RESULTS: We uncover that aeBlue chromoprotein, when expressed from a synthetic construct in Escherichia coli, showed a temperature dependent color shift. Protein purified at 4 °C by metal affinity chromatography exhibited a pinkish color and shifts back at higher temperatures to its intense blue color. Circular dichroism analysis revealed that the structure in the pink form of the protein has reduced secondary structure at 4 °C, but at 35 °C and higher, the structure shifts to a native conformation and Far UV- vis CD spectra revealed the shift in an aromatic residue of the chromophore. Also, the chromophore retains its properties in a wide range of conditions (pH, denaturants, reducing and oxidants agents). Quaternary structure is also maintained as a tetrameric conformation as shown by native gel and size exclusion chromatography. CONCLUSION: Our results suggest that the chromophore position in aeBlue is shifted from its native position rendering the pink color and the process to return it to its native blue conformation is temperature dependent.

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2).

BACKGROUND: Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors. METHODS: The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed. RESULTS: V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (α/ß)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4. CONCLUSION: The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2)

Background:Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors.Methods:The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed.Results:V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (α/β)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4.Conclusion:The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.(AU)

4-Hydroxy-pyran-2-one and 3-hydroxy-N-methyl-2-oxindole derivatives of Salinispora arenicola from Brazilian marine sediments.

Three new 3-hydroxy-N-methyl-2-oxindole (1 and 2) and 4-hydroxy-pyran-2-one (3) derivatives, along with the known 3-hydroxy-N-methyl-2-oxindole (4) and 6-methoxy-N-methylisatin (5) were isolated from a marine Salinispora arenicola strain from sediments of the St. Peter and St. Paul Archipelago, Brazil. The structures of the new compounds were elucidated by a combination of spectroscopic (1D and 2D NMR and HR-ESIMS) data, including single-crystal X-ray diffraction analysis for 2 and 3. Compounds 1 to 5 were assayed for their antimicrobial properties, but only 4 and 5 were active against Enterococcus faecalis with MIC value of 15.6 µg/mL.

A novel single amino acid deletion impairs fibronectin function and causes familial glomerulopathy with fibronectin deposits: case report of a family.

BACKGROUND: Glomerulopathy with fibronectin deposits is an autosomal dominant disease associated with proteinuria, hematuria, hypertension and renal function decline. Forty percent of the cases are caused by mutations in FN1, the gene that encodes fibronectin. CASE PRESENTATION: This report describes two cases of Glomerulopathy with fibronectin deposits, involving a 47-year-old father and a 14-year-old son. The renal biopsies showed glomeruli with endocapillary hypercellularity and large amounts of mesangial and subendothelial eosinophilic deposits. Immunohistochemistry for fibronectin was markedly positive. Whole exome sequencing identified a novel FN1 mutation that leads to an amino-acid deletion in both patients (Ile1988del), a variant that required primary amino-acid sequence analysis for assessment of pathogenicity. Our primary sequence analyses revealed that Ile1988 is very highly conserved among relative sequences and is positioned in a C-terminal FN3 domain containing heparin- and fibulin-1-binding sites. This mutation was predicted as deleterious and molecular mechanics simulations support that it can change the tertiary structure and affect the complex folding and its molecular functionality. CONCLUSION: The current report not only documents the occurrence of two GFND cases in an affected family and deeply characterizes its anatomopathological features but also identifies a novel pathogenic mutation in FN1, analyzes its structural and functional implications, and supports its pathogenicity.

Rethinking medicalization: discursive positions of children and their caregivers on the diagnosis and treatment of ADHD in Chile / Repensando la medicalización: posiciones discursivas de niños y de sus cuidadores sobre el diagnóstico y tratamiento del TDAH en Chile

Abstract The debates around the diagnosis and pharmacological treatment of Attention Deficit and Hyperactivity Disorder (ADHD) have traditionally been approached from the perspective of the "medicalization processes" of children's behaviour. However, this perspective tends to overlook the meanings of diagnosis and treatment of ADHD for children and their caregivers. The purpose of this article is to describe the discursive positions of children and their caregivers on the diagnosis and treatment of ADHD. In-depth interviews were conducted with seven Chilean children and their caregivers. The material was analysed following the procedures of the discourse structure analysis. A discursive structure was identified that configures four emerging realities: the myth of origin of the child's behaviour and learning problems; the ambivalences in/of medicalization; the process of identity (dis)stabilization under diagnosis and treatment; and the subversion of medicalization. It is observed that the subjective experience of the diagnosis and treatment of ADHD is not homogeneous, since different discursive positions, family and institutional understandings that enter into conflict cross it. The experiences of ADHD are shaped by discursive structures that condition the meanings of this experience. The medicalization process is not univocal, but can take different forms and have consequences on children's experiences and social trajectories.

Resumen Los debates en torno al diagnóstico y tratamiento farmacológico del Trastorno de Déficit Atencional e Hiperactividad (TDAH) han sido tradicionalmente abordados desde la perspectiva de los "procesos de medicalización" del comportamiento infantil. Sin embargo, esta perspectiva tiende a pasar por alto los sentidos y significaciones del diagnóstico y tratamiento del TDAH para los niños y sus cuidadores. El objetivo de este artículo es describir las posiciones discursivas de niños y de sus cuidadores sobre el diagnóstico y tratamiento farmacológico del TDAH. Se realizaron entrevistas en profundidad a siete niños chilenos y a sus cuidadores. El material fue analizado siguiendo los procedimientos del análisis estructural del discurso. Se identificó una estructura discursiva que configura cuatro realidades emergentes: el mito de origen de los problemas de comportamiento y de aprendizaje del niño; las ambivalencias en/de la medicalización; el proceso de (des)estabilización identitaria bajo el diagnóstico y tratamiento; y la subversión de la medicalización. Se observa que la experiencia subjetiva del diagnóstico y tratamiento del TDAH no es homogénea, dado que se encuentra atravesada por distintas posiciones discursivas, comprensiones familiares e institucionales que entran en conflicto. Las experiencias del diagnóstico y tratamiento del TDAH se encuentran modeladas por estructuras discursivas que condicionan las posibilidades de dar sentido a dicha experiencia. El proceso de medicalización no es unívoco, sino que puede asumir formas diferentes y tener consecuencias diversas sobre las experiencias y trayectorias sociales de niños y niñas.

Heterologous expression and mutagenesis of recombinant Vespa affinis hyaluronidase protein (rVesA2)

Background:Crude venom of the banded tiger waspVespa affinis contains a variety of enzymes including hyaluronidases, commonly known as spreading factors.Methods:The cDNA cloning, sequence analysis and structural modelling of V. affinis venom hyaluronidase (VesA2) were herein described. Moreover, heterologous expression and mutagenesis of rVesA2 were performed.Results:V. affinis venom hyaluronidase full sequence is composed of 331 amino acids, with four predicted N-glycosylation sites. It was classified into the glycoside hydrolase family 56. The homology modelling exhibited a central core (α/β)7 composed of Asp107 and Glu109, acting as the catalytic residues. The recombinant protein was successfully expressed in E. coli with hyaluronidase activity. A recombinant mutant type with the double point mutation, Asp107Asn and Glu109Gln, completely lost this activity. The hyaluronidase from crude venom exhibited activity from pH 2 to 7. The recombinant wild type showed its maximal activity at pH 2 but decreased rapidly to nearly zero at pH 3 and was completely lost at pH 4.Conclusion:The recombinant wild-type protein showed its maximal activity at pH 2, more acidic pH than that found in the crude venom. The glycosylation was predicted to be responsible for the pH optimum and thermal stability of the enzymes activity.

Response to comment on 'Hydrogen bonds in crystalline d-alanine: diffraction and spectroscopic evidence for differences between enantiomers'.

A response is given to comments by Bürgi & Macchi [IUCrJ (2018), 5, 654-657] about Belo et al. [IUCrJ (2018), 5, 6-12.].

Hydrogen bonds in crystalline d-alanine: diffraction and spectroscopic evidence for differences between enantiomers.

Enantiomeric amino acids have specific physiological functions in complex biological systems. Systematic studies focusing on the solid-state properties of d-amino acids are, however, still limited. To shed light on this field, structural and spectroscopic studies of d-alanine using neutron powder diffraction, polarized Raman scattering and ab initio calculations of harmonic vibrational frequencies were carried out. Clear changes in the number of vibrational modes are observed as a function of temperature, which can be directly connected to variations of the N-D bond lengths. These results reveal dissimilarities in the structural properties of d-alanine compared with l-alanine.

Phase quantification of antihypertensive drugs - Chlorthalidone, Hydrochlorothiazide, Losartan and combinations, Losartan/Chlorthalidone and Losartan/Hydrochlorothiazide - by the Rietveld method.

The identification and quantification of crystalline phases of antihypertensive drugs - Losartan potassium (LOS-K), Hydrochlorothiazide (HCTZ) and Chlorthalidone (CTD) were carried out by means of X-ray powder diffraction data and the Rietveld method. Quantitative phase analyses of Losartan potassium/Chlorthalidone (LOS-K/CTD) and Losartan potassium/Hydrochlorothiazide (LOS-K/HCTZ) combinations were also evaluated. The results indicated that for diuretics (HCTZ and CTD) only one crystalline phase was found in samples, and for LOS-K the crystal structure showed similarity between the Bragg peaks to the phase described as monoclinic and space group P21/c. After one year storage, the orthorhombic one was also observed in this sample.

NMR elucidation and crystal structure analysis of 1-hydroxy-3, 6-dimethoxy-8-methyl-9h-xanthen-9-one (lichexanthone) isolated from Vismia baccifera (Guttiferae) / Elucidación por RMN y análisis de la estructura cristalina de 1-hidroxi-3,6-dimetox1-8-methil-9h-xanten-9-ona (lichexanthone)

In the present investigation the structural analysis of 1-hydroxy-3,6-dimethoxy-8-methyl-9h-xanthen-9-one (lichexanthone) isolated from Vismia baccifera var. dealbata collected in Mérida-Venezuela, was established by NMR (1H and 13C), mass spectrometry and single crystal X-ray diffraction. Lichexanthone crystallizes in the monoclinic system, space group P21/c (No 14) with unit cell parameters a = 11.6405(5) Å; b = 7.5444(3) Å; c = 15.2341(6) Å; = 102.280(1)°; V = 1307.26(9) Å3; Z = 4. The structure refinement converged to R = 0.0397, wR2 = 0.1076, S = 1.04. Lichexanthone had been isolated before from Parmotrema sp and Ruprechtia tangarana (Polygonaceae). However, to the best of our knowledge, this is the first report of this compound obtained from V. baccifera var. dealbata (Guttiferae).

En la presente investigación el análisis estructural de 1-hidroxi-3,6-dimetoxi-8-metil-9h-xanten-9-ona (lichexanthone) aislada de Vismia baccifera var. dealbata colectada en Mérida-Venezuela, fue determinado por RMN (1H y 13C), espectrometría de masas y difracción de rayos X. La lichexanthona cristaliza en un sistema monoclínico con un grupo espacial P21/c (No 14) y parámetros de celda de a = 11.6405 (5) Å; b = 7.5444 (3) Å; c = 15.2341 (6) Å; = 102.280(1)°; V = 1307.26(9) Å3; Z = 4. El refinamiento de la estructura convergió a los valores de R = 0.0397, wR2 = 0.1076, S = 1.04. La lichexanthona ha sido aislada de Parmotrema sp y Ruprechtia tangarana (Polygonaceae). Sin embargo, para nuestro conocimiento, esta es la primera vez que se reporta el aislamiento de este compuesto en la especie V. baccifera var. dealbata (Guttiferae).

A estrutura da representação social da morte na interface com as religiosidades em equipes multiprofissionais de saúde / The structure of social representation of death in the interface with the religiosities of healthcare professionals groups

O estudo objetivou acessar a estrutura da representação social da morte em equipes multiprofissionais de saúde na interface com a religiosidade, no que esta interfere nas significações atribuídas à morte por esta população em específico. Utilizou-se um instrumento com questões abertas e fechadas sobre a morte e o morrer, bem como sobre a vivência religiosa singular dos participantes. Protocolos escritos foram capturados em uma amostra de 80 profissionais (49 médicos, 13 psicólogas, 18 enfermeiras) pertencentes a 10 equipes multiprofissionais de saúde. Estes foram analisados através de Análise de Conteúdo e de Análise de Estrutura de Similaridade (SSA) e interpretados pela Teoria das Representações Sociais. Os dados revelaram uma interferência expressiva das crenças religiosas na estruturação da representação da Morte.

The study aims to investigate the structure of social representation of death in a Healthcare professional group establishing an interface with religiosity and how the latter interferes in the meaning attributed to death by this specific population. Open and closed questions on death or the act of dying, and on the singular religious experience of the participants were applied. Written protocols were collected from a sample of 80 professionals (49 physicians, 13 psychologists, and 18 nurses), which were analyzed according to content analysis and non-metric multidimensional analysis, SSA (Similarity Structure Analysis), and interpreted according to the theory of Social Representations, revealing a significant interference of religious believes in the structure of death representation.

The Star STING server: a multiplatform environment for protein structure analysis

Star STING is the latest version of the STING suite of programs and corresponding database. We report on five important aspects of this package that have acquired some new characteristics, designed to add key advantages to the whole suite: 1) availability for most popular platforms and browsers, 2) introduction of the STING_DB quality assessment, 3) improvement in algorithms for calculation of three STING parameters, 4) introduction of five new STING modules, and 5) expansion of the existing modules. Star STING is freely accessible at: http://sms.cbi.cnptia.embrapa.br/SMS/, http://trantor.bioc.columbia.edu/SMS, http://www.es.embnet.org/SMS/, http://gibk26.bse.kyutech.ac.jp/SMS/ and http://www.ar.embnet.org/SMS.