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1.
Ophthalmic Plast Reconstr Surg ; 39(6): 617-620, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37450648

RESUMO

PURPOSE: To perform a radiological analysis of the relation of the anterior ethmoidal foramen (AEF) to the cranial base and olfactory fossa (OF). METHODS: Retrospective analysis of computed tomography coronal scans of 35 orbits of 35 patients with Graves orbitopathy. Following a standard multiplanar reconstruction of the orbit, the authors measured the vertical and the horizontal distances of the AEF to the cranial base and to the lateral border of the OF, respectively. The height of the OF was measured at the level of the AEF and at the position of Keros classification. The presence of supraorbital ethmoid cells (SOEC) was also recorded. RESULTS: The vertical distance of the AEF to the cranial base ranged from 0.1 to 7.3 mm with a mean = 2.5 mm (SD = 2.17). In 47.5% of the orbits the foramen was adjacent (<1 mm) to the cranial base. The horizontal distance of the AEF to the lateral border of the OF ranged from 2.3 to 9.5 mm (mean = 5.4 mm, SD = 1.98). The presence of an SOEC (n = 12) increased significantly both the distance from the AEF to the cranial base and to the lateral border of the OF. The height of the OF at the level of AEF is not correlated with the depth of the OF at the level of Keros classification. CONCLUSIONS: In the absence of supraorbital pneumatization, the AEF location is a dangerous landmark for the upper limit of the transconjunctival medial wall decompression.


Assuntos
Oftalmopatia de Graves , Humanos , Estudos Retrospectivos , Base do Crânio/diagnóstico por imagem , Osso Etmoide/cirurgia , Órbita/diagnóstico por imagem , Órbita/cirurgia
2.
Orbit ; : 1-9, 2023 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-37942625

RESUMO

PURPOSE: To review the literature on the location of the anterior ethmoidal foramen (AEF) and trauma during transconjunctival medial wall decompression. METHODS: A comprehensive literature search was conducted using the PubMed, Embase, and Scopus databases, combining the terms "olfactory fossa" and "fovea ethmoidalis" with "trauma," "cerebrospinal fluid leak," "pneumocephalus," "orbital decompression," and "anterior ethmoidal artery" (AEA). All cases of cranial base trauma during medial orbital decompression and the anatomical studies on the location of the AEF and the course of the AEA were reviewed. RESULTS: Ninety-four articles were identified, of which 37 were related to the AEF, 41 reported the course of the AEA, and 16 to reported cases of cranial base trauma. Out of these cases, 10 were related to transconjunctival medial orbital decompression, affecting 11 patients. Most AEFs are situated at the frontoethmoidal suture, but up to 38.15% of AEFs are located above the suture on the frontal bone. Most AEFs are adjacent to the roof of the ethmoidal sinus. The distance of the AEF to the cranial base increases in the presence of supraorbital ethmoidal cells (SOEC). CONCLUSIONS: The position of the AEF is variable and should not be considered a safe landmark for all patients.

3.
Int Ophthalmol ; 43(4): 1363-1367, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36149617

RESUMO

PURPOSE: To measure the changes of the eyebrow's contour after frontalis muscle (FM) contraction. METHODS: Two consecutive pictures of the eyes and frontal region of 36 volunteers (15 men and 21 women) were obtained with the forehead relaxed and upon maximal frontalis muscle contraction. Bézier lines representing the brows' contours, obtained with the ImageJ software, were graphically sampled with a resolution of 0.025 mm. The contours of the relaxed and elevated brows were compared regarding the position of the contour peak (CP), the degree of elevation of brow central point, and the medial and lateral areas outlined between the brow contours before and after frontalis contraction. The asymmetry of the brow contour after FM contraction was calculated by the Naeije formula as (Medial-Lateral)/(Medial + Lateral). A difference of 10% or less between the lateral and medial areas was taken as a cutoff value for symmetrical brow elevation. RESULTS: In 62 (86.1%) of the eyes, the location of the brow's CP was lateral to the brow's midpoint. When the brows were raised, the CP of 60% of the brows was displaced medially and 40% laterally. The motion of the CP with medial displacement (mean = 3.7 mm) was significantly larger than those displaced laterally (mean = 1.9 mm) decreasing the number of brows with lateral peaks from 62 (86.1%) to 54 (75%). No difference was observed between the mean elevation of the central point of the brow for males (7.9 mm (3.2 SD) and females (7.4 mm, 3.0 SD). In 58.3% of the sample, the positional change of the medial and lateral sectors of the brows was symmetrical, and among the 41.7% asymmetrical cases, the elevation of the medial side was higher than the lateral in 66.7% of the brows. CONCLUSIONS: The brow changes during FM contraction concur well with the anatomic data of the FM shape. The contour of most brows is displaced toward the medial direction when the brow is elevated. The pattern of brow shape change is an indication of the FM anatomy and must be carefully analyzed before any procedure involving the FM, such as chemodenervation injections or frontalis slings for blepharoptosis correction.


Assuntos
Blefaroplastia , Blefaroptose , Masculino , Humanos , Feminino , Sobrancelhas , Blefaroptose/cirurgia , Blefaroplastia/métodos , Contração Muscular , Olho
4.
Orbit ; : 1-4, 2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35670333

RESUMO

Two patients with thyroid eye disease underwent inferomedial orbital decompression complicated by a cerebrospinal fluid (CSF) leak. One of the cases developed a postoperative pneumocephalus resolved with conservative measures. In the second case, the CSF leak was managed intraoperatively. In both patients, a computed tomography (CT) scan revealed a pronounced slope of the lateral lamella of the cribriform plate, forming an obtuse angle with its lateral bony extension. When this anatomical disposition of the olfactory fossa (OF) is present, the course of the anterior ethmoidal artery (AEA) is usually embedded in the skull base, and its foramen should not be taken as the upper limit of the transconjunctival ethmoidectomy. The shape and relative height of the olfactory fossa and fovea ethmoidalis, and the course of the AEA should always be assessed before transconjunctival medial decompressions.

5.
Physiol Mol Biol Plants ; 28(8): 1607-1624, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-36389096

RESUMO

Water deficit is a major constraint for crops of economic importance in almost all agricultural regions. However, plants have an active defense system to adapt to these adverse conditions, acting in the reprogramming of gene expression responsible for encoding microRNAs (miRNAs). These miRNAs promote the regulation to the target gene expression by the post-transcriptional (PTGS) and transcriptional gene silencing (TGS), modulating several pathways including defense response to water deficit. The broader knowledge of the miRNA expression profile and its regulatory networks in response to water deficit can provide evidence for the development of new biotechnological tools for genetic improvement of several important crops. In this study, we used Setaria viridis accession A10.1 as a C4 model plant to widely investigate the miRNA expression profile in early responses to different levels of water deficit. Ecophysiological studies in Setaria viridis under water deficit and after rewatering demonstrated a drought tolerant accession, capable of a rapid recovery from the stress. Deep small RNA sequencing and degradome studies were performed in plants submitted to drought to identify differentially expressed miRNA genes and their predicted targets, using in silico analysis. Our findings showed that several miRNAs were differentially modulated in response to distinctive levels of water deficit and after rewatering. The predicted mRNA targets mainly corresponded to genes related to cell wall remodeling, antioxidant system and drought-related transcription factors, indicating that these genes are rapidly regulated in early responses to drought stress. The implications of these modulations are extensively discussed, and higher-effect miRNAs are suggested as major players for potential use in genetic engineering to improve drought tolerance in economically important crops, such as sugarcane, maize, and sorghum. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01226-z.

6.
BMC Plant Biol ; 21(1): 300, 2021 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-34187360

RESUMO

BACKGROUND: A major limiting factor for plant growth is the aluminum (Al) toxicity in acidic soils, especially in tropical regions. The exclusion of Al from the root apex through root exudation of organic acids such as malate and citrate is one of the most ubiquitous tolerance mechanisms in the plant kingdom. Two families of anion channels that confer Al tolerance are well described in the literature, ALMT and MATE family. RESULTS: In this study, sugarcane plants constitutively overexpressing the Sorghum bicolor MATE gene (SbMATE) showed improved tolerance to Al when compared to non-transgenic (NT) plants, characterized by sustained root growth and exclusion of aluminum from the root apex based on the result obtained with hematoxylin staining. In addition, genome-wide analysis of the recently released sugarcane genome identified 11 ALMT genes and molecular studies showed potential new targets for aluminum tolerance. CONCLUSIONS: Our results indicate that the transgenic plants overexpressing the Sorghum bicolor MATE has an improved tolerance to Al. The expression profile of ALMT genes revels potential candidate genes to be used has an alternative for agricultural expansion in Brazil and other areas with aluminum toxicity in poor and acid soils.


Assuntos
Alumínio/metabolismo , Proteínas de Transporte de Ânions/genética , Genes de Plantas/genética , Proteínas de Plantas/genética , Saccharum/genética , Alumínio/toxicidade , Proteínas de Transporte de Ânions/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas/fisiologia , Estudo de Associação Genômica Ampla , Filogenia , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas , Saccharum/metabolismo , Sorghum/genética , Sorghum/metabolismo , Transcriptoma
7.
Ophthalmic Plast Reconstr Surg ; 37(6): e206-e208, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34366384

RESUMO

The association of C-antineutrophil cytoplasmic antibody (ANCA) vasculitis and IgG4 positivity is a new condition not well described in clinical terms. The authors examined a 28-year-old man with a previous diagnosis of eosinophilic granulomatosis with polyangiitis, formerly known as Churg-Strauss disease, who presented with bilateral orbital inflammation. Magnetic resonance imaging revealed diffuse orbital infiltration and enlargement of the major divisions of the trigeminal nerve. Biopsy of the orbital contents showed necrotizing granulomatous vasculitis and a high number of IgG4-positive plasma cells (IgG4/IgG = 60%).


Assuntos
Síndrome de Churg-Strauss , Granulomatose com Poliangiite , Adulto , Anticorpos Anticitoplasma de Neutrófilos , Síndrome de Churg-Strauss/complicações , Síndrome de Churg-Strauss/diagnóstico , Granulomatose com Poliangiite/complicações , Granulomatose com Poliangiite/diagnóstico , Humanos , Imunoglobulina G , Masculino , Plasmócitos
8.
Int Ophthalmol ; 41(5): 1929-1947, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33517506

RESUMO

PURPOSE: To systematically review the literature on the deep lateral orbital decompression (DLD). METHODS: The authors searched the MEDLINE, Lilac, Scopus, and EMBASE databases for all articles in English, Spanish, and French that used as keywords the terms orbital decompression and lateral wall. Two articles in German were also included. Data retrieved included the number of patients and orbits operated, types of the approach employed, exophthalmometric and horizontal eye position changes, and complications. The 95% confidence intervals (CI) of the mean Hertel changes induced by the surgery were calculated from series with 15 or more data. RESULTS: Of the 204 publications initially retrieved, 131 were included. Detailed surgical techniques were analyzed from 59 articles representing 4559 procedures of 2705 patients. In 45.8% of the reports, the orbits were decompressed ab-interno. Ab-externo and rim-off techniques were used in 25.4% and 28.8% of the orbits, respectively. Mean and 95% CI intervals of Hertel changes, pooled from 15 articles, indicate that the effect of the surgery is not related to the technique and ranges from 2.5 to 4.5 mm. The rate of new onset of diplopia varied from zero to 8.6%. Several complications have been reported including dry eye, oscillopsia, temporal howling, lateral rectus damage, and bleeding. Unilateral amaurosis and subdural hematoma have been described in only one patients each. CONCLUSIONS: The low rate of new-onset diplopia is the main benefit of DLD. Prospective studies are needed to compare the rate of complications induced by the 3 main surgical techniques used.


Assuntos
Oftalmopatia de Graves , Descompressão Cirúrgica , Oftalmopatia de Graves/cirurgia , Humanos , Órbita/cirurgia , Estudos Prospectivos , Estudos Retrospectivos
9.
Oral Dis ; 25(6): 1545-1554, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31077633

RESUMO

OBJECTIVES: Saliva metabolome is a promising diagnostic tool concerning oral and systemic diseases. We aimed at establishing a suitable protocol for saliva collection and gauging the relative impacts of gender, dentition stage, and caries on the saliva metabolome of a small children cohort. SUBJECTS AND METHODS: A nuclear magnetic resonance-based metabolomics cross-sectional study of children saliva (n = 38) compared the effects of: (a) stimulation and unstimulation conditions, and (b) collection through passive drool and using an absorbing device. Multivariate and univariate statistical analyses were applied to evaluate such effects and those related to gender, dentition stage and caries. RESULTS: No significant differences were found between unstimulated and stimulated saliva, and the former was used for subsequent studies. Swab collection induced significant changes in sample composition, indicating passive drool as preferential. The impacts of gender and dentition stage were not significant compared to that of caries, which induced variations in the levels of 21 metabolites. These comprised amino acids and monosaccharides observed for the first time to our knowledge regarding children caries, suggesting protein hydrolysis and deglycosylation. CONCLUSIONS: Unstimulated passive drool saliva metabolome may carry a caries signature.


Assuntos
Cárie Dentária , Metabolômica , Saúde Bucal , Saliva , Criança , Estudos Transversais , Humanos , Espectroscopia de Ressonância Magnética
10.
New Phytol ; 218(1): 81-93, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29315591

RESUMO

Feruloylation of arabinoxylan (AX) in grass cell walls is a key determinant of recalcitrance to enzyme attack, making it a target for improvement of grass crops, and of interest in grass evolution. Definitive evidence on the genes responsible is lacking so we studied a candidate gene that we identified within the BAHD acyl-CoA transferase family. We used RNA interference (RNAi) silencing of orthologs in the model grasses Setaria viridis (SvBAHD01) and Brachypodium distachyon (BdBAHD01) and determined effects on AX feruloylation. Silencing of SvBAHD01 in Setaria resulted in a c. 60% decrease in AX feruloylation in stems consistently across four generations. Silencing of BdBAHD01 in Brachypodium stems decreased feruloylation much less, possibly due to higher expression of functionally redundant genes. Setaria SvBAHD01 RNAi plants showed: no decrease in total lignin, approximately doubled arabinose acylated by p-coumarate, changes in two-dimensional NMR spectra of unfractionated cell walls consistent with biochemical estimates, no effect on total biomass production and an increase in biomass saccharification efficiency of 40-60%. We provide the first strong evidence for a key role of the BAHD01 gene in AX feruloylation and demonstrate that it is a promising target for improvement of grass crops for biofuel, biorefining and animal nutrition applications.


Assuntos
Biomassa , Parede Celular/metabolismo , Coenzima A-Transferases/genética , Ácidos Cumáricos/metabolismo , Genes de Plantas , Setaria (Planta)/enzimologia , Setaria (Planta)/genética , Supressão Genética , Ácidos/metabolismo , Brachypodium/genética , Metabolismo dos Carboidratos , Coenzima A-Transferases/metabolismo , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Hidrólise , Lignina/metabolismo , Espectroscopia de Ressonância Magnética , Tamanho do Órgão , Filogenia , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas , Sementes/anatomia & histologia , Sementes/crescimento & desenvolvimento , Transcriptoma/genética , Xilanos/metabolismo
11.
Biomacromolecules ; 17(6): 1985-97, 2016 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-27203709

RESUMO

Gradients of physical and chemical cues are characteristic of specific tissue microenvironments and contribute toward morphogenesis and tissue regeneration upon injury. Recent advances on microfluidics and hydrogel manipulation raised the possibility of generating biomimetic biomaterials enriched with bioactive factors and encapsulating cells following designs specifically tailored for a target application. The novelty of this work relies on the combination of methacrylated gellan gum (MeGG) with platelet lysate (PL), aiming to generate novel advanced 3D PL-enriched photo-cross-linkable hydrogels and overcoming the lack of adhesion sites provided by the native MeGG hydrogels. This combination takes advantage of the availability, enriched growth factor composition, and potential autologous application of PL while simultaneously preserving the ability provided by MeGG to tailor mechanical properties, protein release kinetics, and shape of the construct according to the desired goal. Incorporation of PL in the hydrogels significantly improved cellular adhesion and viability in the constructs. The use of microfluidic tools allowed the design of a fiber-like hydrogel incorporating a gradient of PL along the length of the fiber. These spatial protein gradients led to the viability and cell number gradients caused by maintenance of human umbilical vein endothelial cells (HUVECs) survival in the fibers toward the PL-enriched sections in comparison with the nonloaded MeGG sections of the fibers. Altogether, we propose a proof of concept strategy to design a PL gradient biomaterial with potential in tissue engineering approaches and analysis of cell-microenvironment interactions.


Assuntos
Materiais Biomiméticos , Plaquetas/química , Microambiente Celular , Hidrogéis , Células-Tronco/citologia , Engenharia Tecidual , Acroleína/análogos & derivados , Acroleína/química , Tecido Adiposo/citologia , Materiais Biomiméticos/síntese química , Materiais Biomiméticos/química , Adesão Celular , Sobrevivência Celular , Células Endoteliais da Veia Umbilical Humana , Humanos , Hidrogéis/síntese química , Hidrogéis/química , Microfluídica , Polissacarídeos Bacterianos/química , Propriedades de Superfície , Alicerces Teciduais/química
13.
Appl Microbiol Biotechnol ; 97(3): 1161-73, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22446793

RESUMO

Carboxylesterase 2 (CES2), the main carboxylesterase expressed in human intestine, is an increasingly important enzyme in anti-cancer combined therapies for the treatment of different pathologies like colon adenocarcinoma and malignant glioma. The production of human recombinant CES2, in human embryonic kidney cells (HEK-293T cells) using serum-free media, is herein described. CES2 secretion to the media was achieved by the simple addition of an in-frame C-terminal 10× histidine tag (CES2-10xHis) without the need of addition of extra N-terminal signalling sequences or the mutation or deletion of the C-terminal HTEL motif responsible for retaining the protein in the lumen of endoplasmic reticulum. This secretion allowed a fourfold increase in CES2 production. The characterization of human recombinant CES2 showed that this protein exists in other active and inactive forms than the described 60 kDa monomer.


Assuntos
Carboxilesterase/metabolismo , Multimerização Proteica , Carboxilesterase/química , Carboxilesterase/genética , Técnicas de Cultura de Células/métodos , Linhagem Celular , Meios de Cultura Livres de Soro/química , Humanos , Peso Molecular , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
14.
Saudi J Ophthalmol ; 35(3): 193-197, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-35601849

RESUMO

The IgG4-related disease (IgG4-RD) is a systemic condition defined as a fibro-inflammatory disorder, characterized by the occurrence of tumor-like lesions in multiple organs including the eye adnexa. The main diagnostic criterion is based on histopathological findings, especially on the IgG4+/IgG+ plasma cell ratio. In this article, we reviewed the literature of non-IgG4-RD orbital conditions with IgG4 positivity. There were 20 reports of inflammatory non-IgG4-RD orbital lesions and 14 reports of orbital lymphoid proliferations with significant IgG4 positivity. The role of plasma cells IgG4 in the pathogenesis of non-IgG4-RD is not clear. Considering the large spectrum of diseases caused by a variety of different etiopathogenic mechanisms, we think that the common denominator of IgG4+ in these conditions might be related to the peculiar properties of down regulation of immune response of the IgG4 and not to a specific link to IgG4-RD.

15.
Plant Biotechnol (Tokyo) ; 38(2): 227-238, 2021 Jun 25.
Artigo em Inglês | MEDLINE | ID: mdl-34393601

RESUMO

The CRISPR/Cas9 system has been used for genome editing in several organisms, including higher plants. This system induces site-specific mutations in the genome based on the nucleotide sequence of engineered guide RNAs. The complex genomes of C4 grasses makes genome editing a challenge in key grass crops like maize (Zea mays), sorghum (Sorghum bicolor), Brachiaria spp., switchgrass (Panicum virgatum), and sugarcane (Saccharum spp.). Setaria viridis is a diploid C4 grass widely used as a model for these C4 crop plants. Here, an optimized CRISPR/Cas9 binary vector that exploits the non-homologous end joining (NHEJ) system was used to knockout a green fluorescent protein (gfp) transgene in S. viridis accession A10.1. Transformation of embryogenic callus by A. tumefaciens generated ten glufosinate-ammonium resistant transgenic events. In the T0 generation, 60% of the events were biallelic mutants in the gfp transgene with no detectable accumulation of GFP protein and without insertions or deletions in predicted off-target sites. The gfp mutations generated by CRISPR/Cas9 were stable and displayed Mendelian segregation in the T1 generation. Altogether, the system described here is a highly efficient genome editing system for S. viridis, an important model plant for functional genomics studies in C4 grasses. Also, this system is a potential tool for improvement of agronomic traits in C4 crop plants with complex genomes.

16.
Obes Surg ; 30(8): 3093-3098, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32415633

RESUMO

BACKGROUND: Roux-en-Y gastric bypass (RYGB) is among the most performed bariatric surgery techniques. One known complication of RYGB surgery is food intolerance, which may limit the intake of protein. OBJECTIVE: To investigate the relationship of food intolerance after RYGB surgery with masticatory efficiency, chewing time and cycles, and consumption of protein and red meat. METHODS: A case-control study in subjects with and without food intolerance (regurgitation and/or vomiting more than once a week) aged over 18 years old who had undergone RYGB more than 2 years prior, with an absence of no more than 2 dental units and normal oral motor system evaluation. Masticatory efficiency was evaluated by the granulometry of red meat chewed by the study subject according to a predefined protocol using a sieving technique and classified from very poor to excellent. Protein and red meat consumption were evaluated by usual food recall and a 3-day dietary diary. RESULTS: The study population consisted of 24 cases (37.7 ± 7.57 years old, 79.2% females) and 68 controls (38.0 ± 8.75 years old, 61.8% females). There was a statistically significant association (p = 0.001 by the ranksum test) between food intolerance and masticatory efficiency, with 58.3% of cases and 23.5% of controls showing very poor masticatory efficiency. No evidence was found of an association of food intolerance with chewing time, chewing cycles, low protein or red meat consumption. CONCLUSION: Masticatory inefficiency is a contributing factor to food intolerance after RYGB, regardless of time and chewing cycles. No relationship was found between food intolerance and consumption of red meat and protein.


Assuntos
Derivação Gástrica , Obesidade Mórbida , Adulto , Estudos de Casos e Controles , Proteínas Alimentares , Feminino , Intolerância Alimentar , Derivação Gástrica/efeitos adversos , Humanos , Masculino , Pessoa de Meia-Idade , Obesidade , Obesidade Mórbida/cirurgia
17.
Dent Mater J ; 38(3): 411-417, 2019 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-30867347

RESUMO

To evaluate the mechanical properties (MP) and antimicrobial activity of a glass ionomer cement (GIC) incorporated with an antimicrobial agent from the aerial parts of Dioscorea altissima. MP were: syneresis & imbibition; solubility; elasticity module; surface microhardness and fluoride release, and the anti-Streptococcus mutans (Smut) activity was accessed by microdilution broth assay and 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) analysis. Syneresis & imbibition did not change over time for both groups, but EG showed lower values at days 7th and 30th. The control group (CG)'s and the experimental group (EG)'s weights were similar before the solubility assay, but after seven days, CG exhibited weight gain in comparison to EG. The elasticity module had no significant differences between groups. The CG showed lower surface microhardness compared to the EG. CG and EG had similar behaviors regarding fluoride release. EG showed diminished Streptococcus mutans count in comparison to CG. EG showed similar or better mechanical properties and an improvement in the antibacterial activity than the original cement.


Assuntos
Cimentos de Ionômeros de Vidro , Streptococcus mutans , Fluoretos , Teste de Materiais , Extratos Vegetais
18.
Methods Mol Biol ; 1864: 49-65, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30415328

RESUMO

Setaria viridis is an emerging model for C4 species, and it is an important model to validate some genes for further C4 crop transformation, such as sugarcane, maize, and wheat. Here, we describe two protocols for stable transformation of S. viridis mediated by Agrobacterium tumefaciens with three different reporter genes and two selectable markers. Routine transformation efficiency reaching 29% was achieved using embryogenic callus in S. viridis (accession A10.1). Alternatively, we developed a transformation method by floral dip with 0.6% efficiency. The developed protocols could be useful for genetic and genomics studies of important food-feed-fiber-fuel C4 crops.


Assuntos
Agrobacterium tumefaciens/genética , Plantas Geneticamente Modificadas/genética , Setaria (Planta)/genética , Transformação Bacteriana/genética , Produtos Agrícolas/genética , Genes Reporter/genética , Genômica/métodos , Proteínas de Plantas/genética , Regiões Promotoras Genéticas/genética , Zea mays/genética
19.
Biotechnol J ; 14(8): e1800570, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31106962

RESUMO

Currently, marketed influenza vaccines are only efficient against homologous viruses, thus requiring a seasonal update based on circulating subtypes. This constant reformulation adds several challenges to manufacturing, particularly in purification due to the variation of the physicochemical properties of the vaccine product. A universal platform approach capable of handling such variation is therefore of utmost importance. In this work, a filtration-based approach is explored to purify influenza virus-like particles. Switching from adsorptive separation to size-based purification allows overcoming the differences in retention observed for different influenza strains. The proposed process employs a cascade of ultrafiltration and diafiltration steps, followed by a sterile filtration step. Different process parameters are assessed in terms of product recovery and impurities' removal. Membrane chemistry, pore size, operation modes, critical flux, transmembrane pressure, and permeate control strategies are evaluated. After membrane selection and parameter optimization, concentration factors and diafiltration volumes are also defined. By optimizing the filtration mode of operation, it is possible to achieve product recoveries of approximately 80%. Overall, the process time is decreased by 30%, its scalability is improved, and the costs are reduced due to the removal of chromatography and associated buffer consumptions, cleaning, and its validation steps.


Assuntos
Biotecnologia/métodos , Vírus da Influenza A , Ultrafiltração/métodos , Vírion/isolamento & purificação , Animais , Linhagem Celular , Vacinas contra Influenza , Membranas Artificiais , Esterilização , Ultrafiltração/instrumentação , Vacinas de Partículas Semelhantes a Vírus
20.
Sci Rep ; 9(1): 4028, 2019 03 11.
Artigo em Inglês | MEDLINE | ID: mdl-30858491

RESUMO

Abscisic acid (ABA) is an essential phytohormone that regulates growth, development and adaptation of plants to environmental stresses. In Arabidopsis and other higher plants, ABA signal transduction involves three core components namely PYR/PYL/RCAR ABA receptors (PYLs), type 2C protein phosphatases (PP2Cs) and class III SNF-1-related protein kinase 2 (SnRK2s). In the present study, we reported the identification and characterization of the core ABA signaling components in Setaria viridis, an emerging model plant for cereals and feedstock crops presenting C4 metabolism, leading to the identification of eight PYL (SvPYL1 to 8), twelve PP2C (SvPP2C1 to 12) and eleven SnRK2 (SvSnRK2.1 through SvSnRK2.11) genes. In order to study the expression profiles of these genes, two different S. viridis accessions (A10.1 and Ast-1) were submitted to drought, salinity and cold stresses, in addition to application of exogenous ABA. Differential gene expression profiles were observed in each treatment and plant genotype, demonstrating variations of ABA stress responses within the same species. These differential responses to stresses were also assessed by physiological measurements such as photosynthesis, stomatal conductance and transpiration rate. This study allows a detailed analysis of gene expression of the core ABA signaling components in Setaria viridis submitted to different treatments and provides suitable targets for genetic engineering of C4 plants aiming tolerance to abiotic stresses.


Assuntos
Ácido Abscísico/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/metabolismo , Setaria (Planta)/metabolismo , Adaptação Fisiológica , Resposta ao Choque Frio/fisiologia , Secas , Fotossíntese/fisiologia , Transpiração Vegetal/fisiologia , Estresse Salino/fisiologia , Setaria (Planta)/genética
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