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Omics analyses in citrus reveal a possible role of RNA translation pathways and Unfolded Protein Response regulators in the tolerance to combined drought, high irradiance, and heat stress

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Horticulture Research 2023年第7期10卷 165-175页

作者： Damián Balfagón Sara IZandalinas Tadeu dos Reis de Oliveira Claudete Santa-Catarina Aurelio Gómez-Cadenas departamento de Biología Bioquímica y Ciencias NaturalesAv.Sos Baynat s/n.Universitat Jaume I46520 Castellóde la PlanaSpain Laboratório de biologia Celular e tecidual(LBCT) Centro de Biociências E Biotecnologia(CBB)Universidade Estadual Do Norte Fluminense Darcy Ribeiro(UENF)Av.Alberto Lamego 2000Campos Dos GoytacazesRJ28013-602Brazil

environmental changes derived from global warming and human activities increase the intensity and frequency of stressful conditions for plants.Multiple abiotic factors acting simultaneously enhance stress pressure and drastically reduce plant growth,yield,and survival.Stress combination causes a specific stress situation that induces a particular plant response different to the sum of responses to the individual stresses.Here,by comparing transcriptomic and proteomic profiles to different abiotic stress combinations in two citrus genotypes,Carrizo citrange(Citrus sinensis×Poncirus trifoliata)and Cleopatra mandarin(Citrus reshni),with contrasting tolerance to different abiotic stresses,we revealed key responses to the triple combination of heat stress,high irradiance and drought.The specific transcriptomic response to this stress combination in Carrizo was directed to regulate RNA metabolic pathways and translation processes,potentially conferring an advantage with respect to Cleopatra.In addition,we found endoplasmic reticulum stress response as common to all individual and combined stress conditions in both genotypes and identified the accumulation of specific groups of heat shock proteins(HSPs),such as small HSPs and HSP70s,and regulators of the unfolded protein response,BiP2 and PDIL2-2,as possible factors involved in citrus tolerance to triple stress combination.Taken together,our findings provide new insights into the acclimation process of citrus plants to multiple stress combination,necessary for increasing crop tolerance to the changing climatic conditions.

关键词： citrus Citrus stress

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effect of dietary supplementation with olive and sunflower oils on lipid profile and liver histology in rats fed high cholesterol diet

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Asian Pacific Journal of Tropical Medicine 2017年第6期10卷 609-613页

作者： Sandra Mara Pimentel Duavy Gerson Javier Torres Salazar Gerlania de Oliveira Leite Assis ecker Nilda Vargas Barbosa departamento de Química Universidade Regional do Cariri(URCA)Campus Pimenta departamento de bioquímica e biologia Molecular Programa de Pós-Graduacao em Ciências Biológicas:Bioquímica ToxicológicaUniversidade Federal de Santa Maria(UFSM)Campus Universitário-Camobi departamento de Farmacologia Programa de Pós-Graduacao em FarmacologiaUniversidade Federal de Santa Maria(UFSM)Campus Universitário-Camobi

Objective: To compare the effects of high-monounsaturated(MUFA) and polyunsaturated fatty acids(PUFA) against the metabolic disorders elicited by a high-cholesterol diet(HC) in rats. Methods: Using in vivo dietary manipulation, rats were fed with different diets containing 4% soybean oil(cholesterol free diet) and 1% HC containing 12% olive oil(HC+OO) enriched with MUFA and 12% sunflower oil(HC+SO) enriched with PUFA for 60 d. Serum lipid levels and hepatic steatosis were evaluated after the treatment period. Results: Comparatively, rats treated with HC+OO diet experienced a decrease in the serum LDL-C, VLDL-C and CT levels compared to those fed with HC+SO diet(Perwise, HC+OO provoked significant microvesicular steatosis situated in the hepatic acinar zone 1. Conclusions: HC+OO diet has high absorption velocity in the acinar zone 1 of liver compared to the HC+SO diet. Based on this, the reduction of the LDL-C, VLDL-C and CT serum levels in the animals treated with HC+OO diet can be caused by the delay in the FA release to the blood.

关键词： Cholesterol diets Olive oil Sunflower oil Serum lipids Liver steatosis

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electrooxidation of sulfanilamide and its voltammetric determination in pharmaceutical formulation,human urine and serum on glassy carbon electrode

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Journal of Pharmaceutical Analysis 2018年第1期8卷 55-59页

作者： Bruno R.L.Ferraz Tiago Guimaraes demetrius Profeti Luciene P.R.Profeti departamento de biologia Universidade Federal do Espírito SantoAlegre 29.500-000ESBrazil departamento de Química e Física Universidade Federal do Espírito SantoAlegre 29.500-000ESBrazil

For the first time, sulfanilamide(SFD) was determined in otologic solution, human urine and serum by electroanalytical techniques on glassy carbon electrode(GCe). The cyclic voltammetry(CV) experiments showed an irreversible oxidation peak at t 1.06 V in 0.1 mol/L BRBS(p H ? 2.0) at 50 m V/s. Different voltammetric scan rates(from 10 to 250 m V/s) suggested that the oxidation of SFD on the GCe was a diffusioncontrolled process. Square-wave voltammetry(SWV) method under optimized conditions showed a linear response to SFD from 5.0 to 74.7 μmol/L(R ? 0.999) with detection and quantification limits of 0.92 and3.10 μmol/L, respectively. The developed SWV method showed better results for detection limit and linear range than the chronoamperometry method. It has been successfully applied to determine SFD concentration in pharmaceutical formulation, human urine and serum samples with recovery close to 100%.

关键词： Square-wave voltammetry Sulfanilamide Glassy carbon electrode Biological fluids

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Remodeling of the cell wall as a drought-tolerance mechanism of a soybean genotype revealed by global gene expression analysis

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aBIOTeCH 2021年第1期2卷 14-31页

作者： Flaviane Silva Coutinho Juliano Mendonça Rodrigues Lucas Leal Lima Rosilene Oliveira Mesquita Paola Avelar Carpinetti João Paulo Batista Machado Camilo elber Vital Pedro Marcus Vidigal Maria eduarda Soares Ramos Mariana Rocha Maximiano Angela Mehta Maria Goreti Almeida Oliveira elizabeth Pacheco Batista Fontes Humberto Josuéde Oliveira Ramos departamento de bioquímica e biologia Molecular Universidade Federal de ViçsaViçsaMinas GeraisBrazil departamento de Fitotecnia Universidade Federal do CearáFortalezaCearáBrazil Núcleo de Análise de Biomoléculas(NuBioMol) Universidade Federal de ViçsaViçsaMinas GeraisBrazil Instituto de Ciéncias Agrárias Universidade Federal de ViçsaFlorestalMinas GeraisBrazil embrapa Recursos Genéticos e Biotecnologia BrasíliaDFBrazil Universidade Federal de Juiz de Fora Juiz de ForaMinas GeraisBrazil

Drought stress is major abiotic stress that affects soybean production.Therefore,it is widely desirable that soybean becomes more tolerant to stress.To provide insights into regulatory mechanisms of the stress response,we compared the global gene expression profiles from leaves of two soybean genotypes that display different responses to water-deficit(BR 16 and embrapa 48,drought-sensitive and droughttolerant,respectively).After the RNA-seq analysis,a total of 5335 down-regulated and 3170 up-regulated genes were identified in the BR16.On the other hand,the number of genes differentially expressed was markedly lower in the embrapa 48,355 up-regulated and 471 down-regulated genes.However,induction and expression of protein kinases and transcription factors indicated signaling cascades involved in the drought tolerance.Overall,the results suggest that the metabolism of pectin is differently modulated in response to drought stress and may play a role in the soybean defense mechanism against drought.This occurs via an increase of the cell wall plasticity and crosslink,which contributed to a higher hydraulic conductance(Kf)and relative water content(RWC%).The drought-tolerance mechanism of the embrapa 48 genotype involves remodeling of the cell wall and increase of the hydraulic conductance to the maintenance of cell turgor and metabolic processes,resulting in the highest leaf RWC,photosynthetic rate(A),transpiration(e)and carboxylation(A/Ci).Thus,we concluded that the cell wall adjustment under drought is important for a more efficient water use which promoted a more active photosynthetic metabolism,maintaining higher plant growth under drought stress.

关键词： Gene expression RNA-seq Molecular physiology Water-use efficiency

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Changes in cellular proliferation and plasma products are associated with liver failure

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World Journal of Hepatology 2016年第32期8卷 1370-1383页

作者： Juliana Gil Melgaco Frederico Marianetti Soriani Pedro Henrique Ferreira Sucupira Leonardo Assaf Pinheiro Yasmine Rangel Vieira Jaqueline Mendes de Oliveira Lia Laura Lewis-Ximenez Cristina Carvalho Vianna Araújo Lúcio Filgueiras Pacheco-Moreira Gustavo Batista Menezes Oswaldo Goncalves Cruz Claudia Lamarca Vitral Marcelo Alves Pinto departamento de Microbiologia e Parasitologia Universidade Federal Fluminense Instituto Oswaldo Cruz/Fiocruz Laboratório de Desenvolvimento Tecnológico em Virologia departamento de biologia Geral Instituto de Ciencias BiologicasUniversidade Federal de Minas Gerais Instituto Oswaldo Cruz/Fiocruz Ambulatório de Hepatites Virais Hospital Federal de Bonsucesso Laboratório de Imunofarmacologia Departamento de Bioquímica e ImunologiaInstituto de Ciencias BiológicasUniversidade Federal de Minas Gerais Fundacao Oswaldo Cruz Programa de Computacao Científica

AIM To study the differences in immune response and cytokine profile between acute liver failure and selflimited acute hepatitis.MeTHODS Forty-six patients with self-limited acute hepatitis(AH), sixteen patients with acute liver failure(ALF), and twenty-two healthy subjects were involved in this study. The inflammatory and anti-inflammatory products in plasma samples were quantified using commercial enzyme-linked immunoassays and quantitative real-time PCR. The cellular immune responses were measured by proliferation assay using flow cytometry. The groups were divided into viral- and non-viral-induced selflimited AH and ALF. Thus, we worked with five groups: Hepatitis A virus(HAV)-induced self-limited acute hepatitis(HAV-AH), HAV-induced ALF(HAV-ALF), nonviral-induced self-limited acute hepatitis(non-viral AH), non-viral-induced acute liver failure(non-viral ALF), and healthy subjects(HC). Comparisons among HAV and non-viral-induced AH and ALF were performed.ReSULTS The levels of mitochondrial DNA(mt DNA) and the cytokines investigated [interleukin(IL)-6, IL-8, IL-10, interferon gamma, and tumor necrosis factor] were significantly increased in ALF patients, independently of etiology(P ere the best markers associated with ALF [mt DNA: OR = 320.5(95%CI: 14.42-7123.33), P death [mt DNA: OR = 12.1(95%CI: 2.57-57.07), P = 0.002; and IL-10: OR = 8.01(95%CI: 1.26-50.97), P = 0.027]. In the cellular proliferation assay, NK^(bright), NKT and regulatory T cells(TReg) predominated in virusspecific stimulation in HAV-induced ALF patients with an anergic behavior in the cellular response to mitotic stimulation. Therefore, in non-viral-induced ALF, anergic behavior of activated T cells was not observed after mitotic stimulation, as expected and as described by the literature. CONCLUSION mt DNA and IL-10 may be predictors of ALF and death. TReg cells are involved in immunological disturbance in HAV-induced ALF.

关键词： Acute liver failure Cytokines Mitochondrial DNA Cellular immune response Hepatitis A virus

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Nanovesicles Are Secreted during Pollen Germination and Pollen Tube Growth： A Possible Role in Fertilization

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Molecular Plant 2014年第3期7卷 573-577页

作者： Noela Prado Juan de Dios Alche Juan Casado-Vela Salvador Mas Mayte Villalba Rosalia Rodriguez eva Batanero departamento de bioquimica y biologia Molecular Facultad de Qu[mica Universidad Complutense de Madrid 28040 Madrid Spain departamento de bioquimica Biologia Celular y Molecular de Plantas Estacion Experimental de1Zaidin (CSIC) E-18080 Granada Spain Spanish National Research Council (CSIC) Spanish National Centre for Biotechnoiogy (CNB) Darwin 3 Cantoblanco 28049 Madrid Spain

dear editor, During recent decades, a novel mechanism of secretion has been identified in a wide range of mammalian cells. It involves the release of bioactive membrane nanovesicles （30-100 nm）, termed exosomes, upon the fusion of multivesicular bodies with the plasma membrane （Thery et al., 2009）. exosomes are implicated in diverse functions, such as scavenging of archaic proteins, intercellular messengers delivering cell-specific signals, and vehicles for transmissible pathogens. exosomes have also been described in other organisms such as bacte- ria, Drosophila, and fungi.

关键词： OLIVe POLLeN ARABIDOPSIS eXPReSSION PROTeINS ALLeRGeN PeCTIN

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The spectrum of building block conformers sustains the biophysical properties of clinically-oriented self-assembling protein nanoparticles

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Science China Materials 2022年第6期65卷 1662-1670页

作者： eric Voltà-Durán Julieta M Sánchez Hèctor-López-Laguna eloi Parladé Laura Sánchez-García Alejandro Sánchez-Chardi Ario de Marco Ugutz Unzueta esther Vázquez Antonio Villaverde Institut de Biotecnologia i de Biomedicina Universitat Autònoma de BarcelonaBellaterra 08193BarcelonaSpain departament de Genètica i de Microbiologia Universitat Autònoma de BarcelonaBellaterra 08193BarcelonaSpain CIBeR de Bioingeniería Biomateriales y Nanomedicina(CIBeR-BBN) Bellaterra 08193BarcelonaSpain Instituto de Investigaciones Biológicas y Tecnológicas(IIBYT)(CONICeT-Universidad Nacional de Córdoba) ICTA&Cátedra de Química BiológicaDepartamento de QuímicaFCEFyNUNC.Av.Velez Sarsfield 1611X 5016GCA CórdobaArgentina Servei de Microscòpia Universitat Autònoma de BarcelonaBellaterra 08193BarcelonaSpain departament de biologia evolutiva Ecologia i Ciències AmbientalsFacultat de BiologiaUniversitat de Barcelona08028 BarcelonaSpain Laboratory for environmental and Life Sciences University of Nova GoricaNova Gorica 5000Slovenia Biomedical Research Institute Sant Pau(IIB Sant Pau) Sant Antoni Ma Claret 16708025 BarcelonaSpain

Histidine-rich peptides confer self-assembling properties to recombinant proteins through the supramolecular coordination with divalent cations.This fact allows the cost-effective,large-scale generation of microscopic and macroscopic protein materials with intriguing biomedical properties.Among such materials,resulting from the simple bioproduction of protein building blocks,homomeric nanoparticles are of special value as multivalent interactors and drug carriers.Interestingly,we have here identified that the assembly of a given His-tagged protein might render distinguishable categories of self-assembling protein nanoparticles.This fact has been scrutinized through the nanobody-containing fusion proteins eM1-GFP-H6 and A3C8-GFP-H6,whose biosynthesis results in two distinguishable populations of building blocks.In one of them,the assembling and disassembling is controllable by cations.However,a second population immediately self-assembles upon purification through a non-regulatable pathway,rendering larger nanoparticles with specific biological properties.The structural analyses of both model proteins and nanoparticles revealed important conformational variability in the building blocks.This fact renders different structural and functional categories of the final soft materials resulting from the participation of energetically unstable intermediates in the oligomerization process.These data illustrate the complexity of the Hismediated protein assembling in recombinant proteins but they also offer clues for a better design and refinement of protein-based nanomedicines,which,resulting from biological fabrication,show an architectonic flexibility unusual among biomaterials.

关键词： recombinant proteins self-assembling protein materials nanoparticles biomaterials

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Three-dimensional models of antigens with serodiagnostic potential for leprosy:An in silico study

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World Journal of Clinical Infectious Diseases 2023年第1期13卷 1-10页

作者： Bianca Luiza Melo de Assis Rafaela Viana Vieira Ian Theodoro Rudenco Gomes Palma Matheus Bertolini Coutinho Juliana de Moura Gabrielle Caroline Peiter Kádima Nayara Teixeira Campus Toledo Universidade Federal do ParanáToledo 85.919-899ParanáBrazil departamento de Patologia Básica Universidade Federal do Paraná-Setor de Ciências BiológicasCuritiba 81.531-980ParanáBrazil Programa Multicêntrico de Pós-graduação em bioquímica e biologia Molecular-Setor Palotina Universidade Federal do ParanáPalotina 85.950-000ParanáBrazil

BACKGROUND Leprosy is a disease caused by Mycobacterium leprae(M.leprae),an intracellular pathogen that has tropism and affects skin and nervous system cells.The disease has two forms of presentation:Paucibacillary and multibacillary,with different clinical and immunological manifestations.Unlike what occurs in the multibacillary form,the diagnostic tests for the paucibacillary form are nonspecific and not very sensitive,allowing the existence of infected individuals without treatment,which contributes to the spread of the pathogen in the population.To mitigate this contamination,more sensitive diagnostic tests capable of detecting paucibacillary patients are needed.AIM To predict the three-dimensional structure models of M.leprae antigens with serodiagnostic potential for leprosy.MeTHODS In this in silico study,satisfactory templates were selected in the Protein Data Bank(PDB)using Basic Local Alignment Search Tool to predict the structural templates of ML2038,ML0286,ML0050,and 85B antigens by comparative modeling.The templates were selected according to general criteria such as sequence identity,coverage,X-ray resolution,Global Model Quality estimate value and phylogenetic relationship;Clustal X 2.1 software was used in this analysis.Molecular modeling was completed using the software Modeller 9v13.Visualization of the models was made using ViewerLite 4.2 and PyMol software,and analysis of the quality of the predicted models was performed using the QMeAN score and Z-score.Finally,the three-dimensional moels were validated using the MolProbity and Verify 3D platforms.ReSULTS The three-dimensional structure models of ML2038,ML0286,ML0050,and 85B antigens of M.leprae were predicted using the templates PDB:3UOI(90.51%identity),PDB:3eKL(87.46%identity),PDB:3FAV(40.00%identity),and PDB:1F0N(85.21%identity),respectively.The QMeAN and Z-score values indicated the good quality of the structure models.These data refer to the monomeric units of antigens,since some of these antigens have quaternary structure.The validation

关键词： Antigens Leprosy diagnosis Mycobacterium leprae Molecular modelling Serological test In silico study

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Myostatin gene promoter:structure,conservation and importance as a target for muscle modulation

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Journal of Animal Science and Biotechnology 2019年第2期10卷 255-273页

作者： Carla Vermeulen Carvalho Grade Carolina Stefano Mantovani Lúcia elvira Alvares Universidade Federal da Integracao Latino-Americana UNILAInstituto Latino-Americano de Ciências da Vida e da NaturezaAvenida Tarquínio Joslin dos Santos1000Foz do IguacuPR CEP 85870-901Brazil departamento de bioquímica e biologia tecidual Universidade Estadual de Campinas-UNICAMPRua Monteiro Lobato255CampinasSP CEP 13083-862Brazil.

Myostatin(MSTN) is one of the key factors regulating myogenesis. Because of its role as a negative regulator of muscle mass deposition, much interest has been given to its protein and, in recent years, several studies have analysed MSTN gene regulation. This review discusses the MSTN gene promoter, focusing on its structure in several animal species, both vertebrate and invertebrate. We report the important binding sites considering their degree of phylogenetic conservation and roles they play in the promoter activity. Finally, we discuss recent studies focusing on MSTN gene regulation via promoter manipulation and the potential applications they have both in medicine and agriculture.

关键词： CAAT box e-box Gene promoter Myogenesis Myostatin SNP Transcription factors TATA box

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“Stop Ne(c)king around”: How interactomics contributes to functionally characterize Nek family kinases

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World Journal of Biological Chemistry 2014年第2期5卷 141-160页

作者： Gabriela Vaz Meirelles Arina Marina Perez edmárcia elisa de Souza Ferna Luisa Basei Priscila Ferreira Papa Talita Diniz Melo Hanchuk Vanessa Bomfim Cardoso Jrg Kobarg Laboratório Nacional de Biociências Centro Nacional de Pesquisa em Energia e Materiais Campinas SP 13084-971Brazil departamento de bioquímica-Programa de Pós-gradua o em biologia Funcional e Molecular Instituto de Biologia Universidade Estadual de Campinas Campinas SP 13084-971 Brasil departamento de Genética Evolu o e Bioagentes-Programa de Pós-gradua o em Genética e Biologia Molecular Instituto de Biologia Universidade Estadual de Campinas1 Campinas SP 13084-97 Brasil

Aside from Polo and Aurora, a third but less studied kinase family involved in mitosis regulation is the never in mitosis-gene A(NIMA)-related kinases(Neks). The founding member of this family is the sole member NIMA of Aspergillus nidulans, which is crucial for the initiation of mitosis in that organism. All 11 human Neks have been functionally assigned to one of the three core functions established for this family in mammals:(1) centrioles/mitosis;(2) primary ciliary function/ciliopathies; and(3) DNA damage response(DDR). Recent findings, especially on Nek 1 and 8, showed however, that several Neks participate in parallel in at least two of these contexts: primary ciliary function and DDR. In the core section of this in-depth review, we report the current detailed functional knowledge on each of the 11 Neks. In the discussion, we return to the cross-connections among Neks and point out how our and other groups' functional and interactomics studies revealed that most Neks interact with protein partners associated with two if not all three of the functional contexts. We then raise the hypothesis that Neks may be the connecting regulatory elements that allow the cell to fine tune and synchronize the cellular events associated with these three core functions. The new and exciting findings on the Nek family open new perspectives and should allow the Neks to finally claim the attention they deserve in the field of kinases and cell cycle biology.

关键词： Cell cycle Mitosis DNA damage response Protein interactions Kinases

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