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1.
Prep Biochem Biotechnol ; : 1-13, 2023 Oct 10.
Artigo em Inglês | MEDLINE | ID: mdl-37814816

RESUMO

Collagenolytic proteases produced by Aspergillus heteromorphus URM0269 were extracted using a PEG/sulfate aqueous two-phase system (ATPS). A 23 factorial design was performed to analyze the independent variables: PEG molar mass (MPEG), PEG concentration (CPEG), and sulfate concentration (Csulf). The extracted proteases were also evaluated for their optimum pH and stability at different pH levels (4.0 - 11.0) after 20 h of incubation. Collagen was extracted from mutton snapper (Lutjanus analis) skin using acetic acid (0.5 mol L-1). The enzyme was preferentially partitioned to the PEG-rich phase (K > 1), whose highest purification factor and recovery (PF = 6.256 and Y = 404.432%) were obtained under specific conditions: MPEG 8000 g.mol-1, CPEG 30%, Csulf 10%. The ATPS extraction provided an enzymatic activity range of pH 7.0 - 11.0, exhibiting greater stability compared to the crude extract. Approximately 80% of protease activity was maintained after 20 hours of incubation at all analyzed pH levels, except pH 11.0. Collagen extraction from L. analis skin yielded 8.056%, and both crude extract samples and ATPS-derived samples successfully hydrolyzed the extracted collagen, reaching peak hydrolysis after 36 hours of treatment. These findings demonstrate the feasibility of extracting highly purified and active proteases capable of hydrolyzing L. analis collagen.

2.
Fam Pract ; 39(6): 1017-1023, 2022 11 22.
Artigo em Inglês | MEDLINE | ID: mdl-35477768

RESUMO

BACKGROUND: It was verified the level of religiosity and spirituality, and symptoms of depression, stress, and anxiety of caregivers of children and adolescents with type 1 diabetes, and its interference in glycaemic control. METHODS: Socio-economic and demographic data were collected from caregivers of 59 children and adolescents with type 1 diabetes and obtained dosages of fasting glucose (FG); postprandial glycaemia (PPG); fructosamine (FRUTO); and HbA1c, as well as the glycaemic variability-∆HbA1c. Levels of religiosity were obtained by the DUREL scale; the use of religious/spiritual coping was verified by the SRCOPE-Brief scale. Symptoms of depression, anxiety, and stress were analysed by the DASS-21 scale. The correlations between the variables were analysed by the Pearson coefficient, with significance at 5% level. RESULTS: Inverse correlations were observed between caregivers' schooling with PPG (r = -0.30; P = 0.002) and FRUTO (r = -0.34; P = 0.008) and between family income and FRUTO (r = -0.37; P = 0.004). Direct, moderate correlations were observed between negative religious/spiritual coping (NSRCOPE) with symptoms of depression (r = 0.588; P < 0.0001), stress (r = 0.500; P < 0.0001), and anxiety (r = 0.551; P < 0.0001). CONCLUSION: The direct association between NSRCOPE with symptoms of depression, stress, and anxiety was the highlighted item in the present study emphasizing the need for greater attention to the emotional health of informal caregivers of children and adolescents with DM1. It is necessary to reflect on the religious/spiritual support especially for the main caregiver of children and adolescents with DM1.


Assuntos
Cuidadores , Diabetes Mellitus Tipo 1 , Criança , Adolescente , Humanos , Cuidadores/psicologia , Depressão/psicologia , Adaptação Psicológica , Ansiedade/psicologia
3.
Genet Mol Biol ; 43(4): e20190418, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33174976

RESUMO

Peanut is a crop of the Kayabi tribe, inhabiting the Xingu Indigenous Park, Brazil. Morphological analysis of Xingu accessions showed variation exceeding that described for cultivated peanuts. This raised questions as to the origin of the Xingu accessions: are they derived from different species, or is their diversity a result of different evolutionary and selection processes? To answer these questions, cytogenetic and genotyping analyses were conducted. The karyotypes of Xingu accessions analyzed are very similar to each other, to an A. hypogaea subsp. fastigiata accession and to the wild allotetraploid A. monticola. The accessions share the number and general morphology of the chromosomes; DAPI+ bands; 5S and 45S rDNA loci distribution and a high genomic affinity with A. duranensis and A. ipaënsis genomic probes. However, the number of CMA3+ bands differs from those determined for A. hypogaea and A. monticola, which are also different from each other. SNP genotyping grouped all Arachis allotetraploids into four taxonomic groups: Xingu accessions were closer to A. monticola and A. hypogaea subsp. hypogaea. Our data suggests that the morphological diversity within these accessions is not associated with a different origin and can be attributed to morphological plasticity and different selection by the Indian tribes.

4.
BMC Plant Biol ; 18(1): 159, 2018 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-30081841

RESUMO

BACKGROUND: The Root-Knot Nematode (RKN), Meloidogyne arenaria, significantly reduces peanut grain quality and yield worldwide. Whilst the cultivated species has low levels of resistance to RKN and other pests and diseases, peanut wild relatives (Arachis spp.) show rich genetic diversity and harbor high levels of resistance to many pathogens and environmental constraints. Comparative transcriptome analysis can be applied to identify candidate resistance genes. RESULTS: Transcriptome analysis during the early stages of RKN infection of two peanut wild relatives, the highly RKN resistant Arachis stenosperma and the moderately susceptible A. duranensis, revealed genes related to plant immunity with contrasting expression profiles. These included genes involved in hormone signaling and secondary metabolites production and also members of the NBS-LRR class of plant disease resistance (R) genes. From 345 NBS-LRRs identified in A.duranensis reference genome, 52 were differentially expressed between inoculated and control samples, with the majority occurring in physical clusters unevenly distributed on eight chromosomes with preferential tandem duplication. The majority of these NBS-LRR genes showed contrasting expression behaviour between A. duranensis and A. stenosperma, particularly at 6 days after nematode inoculation, coinciding with the onset of the Hypersensitive Response in the resistant species. The physical clustering of some of these NBS-LRR genes correlated with their expression patterns in the contrasting genotypes. Four NBS-LRR genes exclusively expressed in A. stenosperma are located within clusters on chromosome Aradu. A09, which harbors a QTL for RKN resistance, suggesting a functional role for their physical arrangement and their potential involvement in this defense response. CONCLUSION: The identification of functional novel R genes in wild Arachis species responsible for triggering effective defense cascades can contribute to the crop genetic improvement and enhance peanut resilience to RKN.


Assuntos
Arachis/metabolismo , Resistência à Doença/genética , Genes de Plantas/genética , Raízes de Plantas/metabolismo , Tylenchoidea , Animais , Arachis/genética , Arachis/parasitologia , Perfilação da Expressão Gênica , Genes de Plantas/fisiologia , Filogenia , Doenças das Plantas/imunologia , Doenças das Plantas/parasitologia , Raízes de Plantas/genética , Raízes de Plantas/parasitologia , Reação em Cadeia da Polimerase em Tempo Real , Transcriptoma
5.
Plant Mol Biol ; 94(1-2): 79-96, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28243841

RESUMO

Expansins are plant cell wall-loosening proteins involved in adaptive responses to environmental stimuli and various developmental processes. The first genome-wide analysis of the expansin superfamily in the Arachis genus identified 40 members in A. duranensis and 44 in A. ipaënsis, the wild progenitors of cultivated peanut (A. hypogaea). These expansins were further characterized regarding their subfamily classification, distribution along the genomes, duplication events, molecular structure, and phylogeny. A RNA-seq expression analysis in different Arachis species showed that the majority of these expansins are modulated in response to diverse stresses such as water deficit, root-knot nematode (RKN) infection, and UV exposure, with an expansin-like B gene (AraEXLB8) displaying a highly distinct stress-responsive expression profile. Further analysis of the AraEXLB8 coding sequences showed high conservation across the Arachis genotypes, with eight haplotypes identified. The modulation of AraEXLB8 expression in response to the aforementioned stresses was confirmed by qRT-PCR analysis in distinct Arachis genotypes, whilst in situ hybridization revealed transcripts in different root tissues according to the stress imposed. The overexpression of AraEXLB8 in soybean (Glycine max) composite plants remarkably decreased the number of galls in transformed hairy roots inoculated with RKN. This study improves the current understanding of the molecular evolution, divergence, and gene expression of expansins in Arachis, and provides molecular and functional insights into the role of expansin-like B, the less-studied plant expansin subfamily.


Assuntos
Arachis/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Família Multigênica/fisiologia , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Parede Celular/fisiologia , Estudo de Associação Genômica Ampla , Filogenia , Células Vegetais/fisiologia , Doenças das Plantas/microbiologia , Raios Ultravioleta , Água
6.
Am J Bot ; 104(3): 379-388, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-28341626

RESUMO

PREMISE OF THE STUDY: Several species of Arachis have been cultivated for their edible seeds, historically and to the present day. The diploid species that have a history of cultivation show relatively small signatures of domestication. In contrast, the tetraploid species A. hypogaea evolved into highly domesticated forms and became a major world crop, the cultivated peanut. It seems likely that allotetraploidization (hybridity and/or tetraploidization) in some way enhanced attractiveness for cultivation. Here we investigate this using six different hybridization and tetraploidization events, from distinct Arachis diploid species, including one event derived from the same wild species that originated peanut. METHODS: Twenty-six anatomical, morphological, and physiological traits were examined in the induced allotetraploid plants and compared with their wild diploid parents. KEY RESULTS: Nineteen traits were transgressive (showed strong response to hybridization and chromosome duplication): allotetraploids had larger leaves, stomata and epidermal cells than did their diploid parents. In addition, allotetraploids produced more photosynthetic pigments. These traits have the same trend across the different hybrid combinations, suggesting that the changes are more likely due to ploidy rather than hybridity. In contrast, seed dimensions and seed mass did not significantly change in response to hybridization or tetraploidization. CONCLUSIONS: We suggest that the original allotetraploid that gave rise to cultivated peanut may have been attractive because of an increase in plant size, different transpiration characteristics, higher photosynthetic capacity, or other characteristics, but contrary to accepted knowledge, increased seed size was unlikely to have been important in the initial domestication.


Assuntos
Arachis/genética , Domesticação , Genoma de Planta/genética , Fotossíntese , Arachis/anatomia & histologia , Arachis/crescimento & desenvolvimento , Arachis/fisiologia , Produtos Agrícolas , Diploide , Genótipo , Hibridização Genética , Fenótipo , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/fisiologia , Poliploidia , Sementes/anatomia & histologia , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/fisiologia , Tetraploidia
7.
Plant Biotechnol J ; 13(7): 884-92, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25572960

RESUMO

There is an urgent need to provide effective anti-HIV microbicides to resource-poor areas worldwide. Some of the most promising microbicide candidates are biotherapeutics targeting viral entry. To provide biotherapeutics to poorer areas, it is vital to reduce the cost. Here, we report the production of biologically active recombinant cyanovirin-N (rCV-N), an antiviral protein, in genetically engineered soya bean seeds. Pure, biologically active rCV-N was isolated with a yield of 350 µg/g of dry seed weight. The observed amino acid sequence of rCV-N matched the expected sequence of native CV-N, as did the mass of rCV-N (11 009 Da). Purified rCV-N from soya is active in anti-HIV assays with an EC50 of 0.82-2.7 nM (compared to 0.45-1.8 nM for E. coli-produced CV-N). Standard industrial processing of soya bean seeds to harvest soya bean oil does not diminish the antiviral activity of recovered rCV-N, allowing the use of industrial soya bean processing to generate both soya bean oil and a recombinant protein for anti-HIV microbicide development.


Assuntos
Proteínas de Bactérias/biossíntese , Proteínas de Transporte/biossíntese , Glycine max/genética , Engenharia de Proteínas , Sementes/genética , Fármacos Anti-HIV , Proteínas de Bactérias/genética , Proteínas de Transporte/genética , Sementes/metabolismo , Glycine max/metabolismo
8.
Plants (Basel) ; 13(15)2024 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-39124136

RESUMO

Mosses are an early lineage of the plant kingdom, with around 13,000 species. Although an important part of biodiversity, providing crucial ecosystem services, many species are threatened with extinction. However, only circa 300 species have so far had their extinction risk evaluated globally for the IUCN Red List. Functional traits are known to help predict the extinction risk of species in other plant groups. In this study, a matrix of 15 functional traits was produced for 723 moss species from around the world to evaluate the potential of such predictability. Binary generalized linear models showed that monoicous species were more likely to be threatened than dioicous species, and the presence of a sporophyte (sexual reproduction), vegetative reproduction and an erect (straight) capsule instead of a pendent (immersed) one lowers the risk of species extinction. A longer capsule, seta and stem length, as well as broader substrate breadth, are indicative of species with a lower risk of extinction. The best-performing models fitted with few traits were able to predict extinction risks of species with good accuracy. These models applied to Data Deficient (DD) species proved how useful they may be to speed up the IUCN Red List assessment process while reducing the number of listed DD species, by selecting species most in need of a full, detailed assessment. Some traits tested in this study are a novelty in conservation research on mosses, opening new possibilities for future studies. The traits studied and the models presented here are a significant contribution to the knowledge of mosses at risk of extinction and will help to improve conservation efforts.

9.
Codas ; 35(2): e20210262, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37098939

RESUMO

PURPOSE: To analyze the effects of lingual frenotomy on the breastfeeding of infants, based on the electrical activity of the masseter and suprahyoid muscles and assessment of the breastfeeding. METHODS: Observational study developed between October 2017 and June 2018 with a sample of 20 newborns and infants who attended a dental clinic and were diagnosed with ankyloglossia. Another 20 were excluded for meeting some of the following exclusion criteria: babies more than 6 months old, who were not on exclusive or mixed breastfeeding, who had other clinical impairments that interfered with breastfeeding, who had other foods introduced into their diet, who had neurological changes and/or craniofacial deformities, and/or who did not finish all the stages of the study. Breastfeeding was assessed with the UNICEF Breastfeeding Assessment and Observation Protocol, while the muscle electrical activity was assessed with the Electrical Activity Assessment Protocol for the Masseter and Suprahyoid Muscles in Newborns During Breastfeeding. The same speech-language-hearing therapist conducted the two assessments both before the conventional frenotomy and 7 days after it. RESULTS: The signs suggestive of breastfeeding difficulties changed 7 days after the surgery, with a p-value ≤ 0.002 for general observation of the mother, position of the infant, latch, and sucking. The maximum voluntary contraction of the masseter was the only integral parameter with a difference, as the electrical activity had decreased. CONCLUSION: Behaviors favorable to breastfeeding increased 7 days after the frenotomy in all the breastfeeding assessment categories, whereas the electrical activity of the masseter decreased.


Assuntos
Anquiloglossia , Aleitamento Materno , Lactente , Feminino , Recém-Nascido , Humanos , Freio Lingual/cirurgia , Freio Lingual/anormalidades , Anquiloglossia/cirurgia , Mães , Músculos do Pescoço , Resultado do Tratamento
10.
Front Public Health ; 11: 1248121, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38026344

RESUMO

Background: To effectively combat the rising incidence of syphilis, the Brazilian Ministry of Health (MoH) created a National Rapid Response to Syphilis with actions aimed at bolstering epidemiological surveillance of acquired, congenital syphilis, and syphilis during pregnancy complemented with communication activities to raise population awareness and to increase uptake of testing that targeted mass media outlets from November 2018 to March 2019 throughout Brazil, and mainly areas with high rates of syphilis. This study analyzes the volume and quality of online news content on syphilis in Brazil between 2015 and 2019 and examines its effect on testing. Methods: The collection and processing of online news were automated by means of a proprietary digital health ecosystem established for the study. We applied text data mining techniques to online news to extract patterns from categories of text. The presence and combination of such categories in collected texts determined the quality of news that were analyzed to classify them as high-, medium-and low-quality news. We examined the correlation between the quality of news and the volume of syphilis testing using Spearman's Rank Correlation Coefficient. Results: 1,049 web pages were collected using a Google Search API, of which 630 were categorized as earned media. We observed a steady increase in the number of news on syphilis in 2015 (n = 18), 2016 (n = 26), and 2017 (n = 42), with a substantial rise in the number of news in 2018 (n = 107) and 2019 (n = 437), although the relative proportion of high-quality news remained consistently high (77.6 and 70.5% respectively) and in line with similar years. We found a correlation between news quality and syphilis testing performed in primary health care with an increase of 82.32, 78.13, and 73.20%, respectively, in the three types of treponemal tests used to confirm an infection. Conclusion: Effective communication strategies that lead to dissemination of high quality of information are important to increase uptake of public health policy actions.


Assuntos
Sífilis Congênita , Sífilis , Feminino , Humanos , Gravidez , Brasil/epidemiologia , Saúde Pública , Sífilis/epidemiologia , Sífilis Congênita/epidemiologia
12.
Int J Paediatr Dent ; 22(4): 286-91, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22092596

RESUMO

BACKGROUND: In dentistry, clinical practice is directed towards attitudes that promote oral health and the paediatricians occupy a privileged position in this process. AIM: To assess the knowledge and attitudes of paediatricians in relation to the oral health of their patients. DESIGN: A cross-sectional study was carried out at the Institute of Integrative Medicine Professor Fernando Figueira, Recife, Brazil. A total of 182 paediatricians participated by filling out a questionnaire. RESULTS: A total of 63.9% believed the first visit to the dentist should occur before the child completes 1 year of life. Moreover, 67.8% considered their knowledge on oral health to be insufficient. Approximately 78% of the paediatricians diagnosed caries through an analysis of cavities. Only 29.9% always recommended fluoride dentifrice. The term 'fluorosis' was unknown by 48.3% of the respondents. Concerning pacifiers, 32.6% did not allow it and 66.9% did not either recommend it or restrict it. A total of 83.4% classified the oral health content in their medical education as either nonexistent or deficient; this figure remained high (72.4%) in relation to residency. CONCLUSIONS: It is important to develop oral health information programmes to paediatricians. Information on oral health should be included in medical curricula and residency.


Assuntos
Atitude do Pessoal de Saúde , Educação Médica , Saúde Bucal , Pediatria/educação , Médicos/psicologia , Adulto , Idoso , Brasil , Cariostáticos/uso terapêutico , Competência Clínica , Estudos Transversais , Currículo , Assistência Odontológica para Crianças , Cárie Dentária/diagnóstico , Dentifrícios/uso terapêutico , Feminino , Fluoretos/uso terapêutico , Fluorose Dentária/diagnóstico , Educação em Saúde Bucal , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Chupetas , Encaminhamento e Consulta , Adulto Jovem
13.
Plants (Basel) ; 11(24)2022 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-36559595

RESUMO

The association of both cell-surface PRRs (Pattern Recognition Receptors) and intracellular receptor NLRs (Nucleotide-Binding Leucine-Rich Repeat) in engineered plants have the potential to activate strong defenses against a broad range of pathogens. Here, we describe the identification, characterization, and in planta functional analysis of a novel truncated NLR (TNx) gene from the wild species Arachis stenosperma (AsTIR19), with a protein structure lacking the C-terminal LRR (Leucine Rich Repeat) domain involved in pathogen perception. Overexpression of AsTIR19 in tobacco plants led to a significant reduction in infection caused by Sclerotinia sclerotiorum, with a further reduction in pyramid lines containing an expansin-like B gene (AdEXLB8) potentially involved in defense priming. Transcription analysis of tobacco transgenic lines revealed induction of hormone defense pathways (SA; JA-ET) and PRs (Pathogenesis-Related proteins) production. The strong upregulation of the respiratory burst oxidase homolog D (RbohD) gene in the pyramid lines suggests its central role in mediating immune responses in plants co-expressing the two transgenes, with reactive oxygen species (ROS) production enhanced by AdEXLB8 cues leading to stronger defense response. Here, we demonstrate that the association of potential priming elicitors and truncated NLRs can produce a synergistic effect on fungal resistance, constituting a promising strategy for improved, non-specific resistance to plant pathogens.

14.
Artigo em Inglês | MEDLINE | ID: mdl-36554926

RESUMO

BACKGROUND: The "Syphilis No!" campaign the Brazilian Ministry of Health (MoH) launched between November 2018 and March 2019, brought forward the concept "Test, Treat and Cure" to remind the population of the importance of syphilis prevention. In this context, this study aims to analyze the similarity of syphilis online news to comprehend how public health communication interventions influence media coverage of the syphilis issue. METHODS: This paper presented a computational approach to assess the effectiveness of communication actions on a public health problem. Data were collected between January 2015 and December 2019 and processed using the Hermes ecosystem, which utilizes text mining and machine learning algorithms to cluster similar content. RESULTS: Hermes identified 1049 google-indexed web pages containing the term 'syphilis' in Brazil. Of these, 619 were categorized as news stories. In total, 157 were grouped into clusters of at least two similar news items and a single cluster with 462 news classified as "single" for not featuring similar news items. From these, 19 clusters were identified in the pre-campaign period, 23 during the campaign, and 115 in the post-campaign. CONCLUSIONS: The findings presented in this study show that the volume of syphilis-related news reports has increased in recent years and gained popularity after the SNP started, having been boosted during the campaign and escalating even after its completion.


Assuntos
Ecossistema , Saúde Pública , Comunicação , Brasil/epidemiologia , Meios de Comunicação de Massa
15.
Sci Rep ; 12(1): 6371, 2022 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-35430625

RESUMO

COVID-19-related in-hospital mortality has been reported at 30.7-47.3% in Brazil, however studies assessing exclusively private hospitals are lacking. This is important because of significant differences existing between the Brazilian private and public healthcare systems. We aimed to determine the COVID-19-related in-hospital mortality and associated risk factors in a Brazilian private network from March/2020 to March/2021. Data were extracted from institutional database and analyzed using Cox regression model. Length of hospitalization and death-related factors were modeled based on available independent variables. In total, 38,937 COVID-19 patients were hospitalized of whom 3058 (7.8%) died. Admission to the intensive care unit occurred in 62.5% of cases, and 11.5% and 3.8% required mechanical ventilation (MV) and renal replacement therapy (RRT), respectively. In the adjusted model, age ≥ 61 years-old, comorbidities, and the need for MV and/or RRT were significantly associated with increased mortality (p < 0.05). Obesity and hypertension were associated with the need for MV and RRT (p < 0.05).


Assuntos
COVID-19 , Brasil/epidemiologia , COVID-19/epidemiologia , Estudos de Coortes , Atenção à Saúde , Mortalidade Hospitalar , Hospitalização , Humanos , Unidades de Terapia Intensiva , Pessoa de Meia-Idade , Respiração Artificial/efeitos adversos , Estudos Retrospectivos , Fatores de Risco , SARS-CoV-2
16.
Transgenic Res ; 20(4): 841-55, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21069460

RESUMO

The seed-based production of recombinant proteins is an efficient strategy to achieve the accumulation, correct folding, and increased stability of these recombinant proteins. Among potential plant molecular farming systems, soybean [Glycine max (L.) Merrill] is a viable option for the production of recombinant proteins due to its high protein content, known regulatory sequences, efficient gene transfer protocols, and a scalable production system under greenhouse conditions. We report here the expression and stable accumulation of human coagulation factor IX (hFIX) in transgenic soybean seeds. A biolistic process was utilised to co-introduce a plasmid carrying the hFIX gene under the transcriptional control of the α' subunit of a ß-conglycinin seed-specific promoter and an α-Coixin signal peptide in soybean embryonic axes from mature seeds. The 56-kDa hFIX protein was expressed in the transgenic seeds at levels of up to 0.23% (0.8 g kg(-1) seed) of the total soluble seed protein as determined by an enzyme-linked immunosorbent assay (ELISA) and western blot. Ultrastructural immunocytochemistry assays indicated that the recombinant hFIX in seed cotyledonary cells was efficiently directed to protein storage vacuoles. Mass spectrometry characterisation confirmed the presence of the hFIX recombinant protein sequence. Protein extracts from transgenic seeds showed a blood-clotting activity of up to 1.4% of normal plasma. Our results demonstrate the correct processing and stable accumulation of functional hFIX in soybean seeds stored for 6 years under room temperature conditions (22 ± 2°C).


Assuntos
Fator IX/metabolismo , Glycine max/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Proteínas Recombinantes/metabolismo , Sequência de Aminoácidos , Antígenos de Plantas/genética , Coagulação Sanguínea/efeitos dos fármacos , Fator IX/genética , Fator IX/farmacologia , Globulinas/genética , Humanos , Dados de Sequência Molecular , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Regiões Promotoras Genéticas , Sinais Direcionadores de Proteínas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Proteínas de Armazenamento de Sementes/genética , Sementes/genética , Sementes/metabolismo , Proteínas de Soja/genética , Glycine max/genética
17.
Transgenic Res ; 20(4): 811-26, 2011 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-21069461

RESUMO

We produced human growth hormone (hGH), a protein that stimulates growth and cell reproduction, in genetically engineered soybean [Glycine max (L.) Merrill] seeds. Utilising the alpha prime (α') subunit of ß-conglycinin tissue-specific promoter from soybean and the α-Coixin signal peptide from Coix lacryma-jobi, we obtained transgenic soybean lines that expressed the mature form of hGH in their seeds. Expression levels of bioactive hGH up to 2.9% of the total soluble seed protein content (corresponding to approximately 9 g kg(-1)) were measured in mature dry soybean seeds. The results of ultrastructural immunocytochemistry assays indicated that the recombinant hGH in seed cotyledonary cells was efficiently directed to protein storage vacuoles. Specific bioassays demonstrated that the hGH expressed in the soybean seeds was fully active. The recombinant hGH protein sequence was confirmed by mass spectrometry characterisation. These results demonstrate that the utilisation of tissue-specific regulatory sequences is an attractive and viable option for achieving high-yield production of recombinant proteins in stable transgenic soybean seeds.


Assuntos
Glycine max/genética , Hormônio do Crescimento Humano/biossíntese , Plantas Geneticamente Modificadas/genética , Proteínas Recombinantes/biossíntese , Sementes/genética , Sequência de Aminoácidos , Antígenos de Plantas/genética , Globulinas/genética , Hormônio do Crescimento Humano/genética , Humanos , Dados de Sequência Molecular , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/metabolismo , Regiões Promotoras Genéticas , Sinais Direcionadores de Proteínas/genética , Proteínas Recombinantes/genética , Proteínas de Armazenamento de Sementes/genética , Sementes/metabolismo , Proteínas de Soja/genética , Glycine max/metabolismo , Vacúolos/metabolismo
18.
Oral Health Prev Dent ; 9(4): 329-37, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22238731

RESUMO

PURPOSE: The aim of the present investigation was to determine the degree of anxiety and fear in patients treated at the dental school clinics of the Universidade Federal de Pernambuco (Brazil) regarding the different types of treatment offered. MATERIALS AND METHODS: The sample was made up of 400 patients from both genders over 18 years of age treated between July and October 2007. Data collection was carried out through the administration of validated questionnaires by a calibrated researcher. The Modified Dental Anxiety Scale (MDAS) and Gatchel's Scale were used to evaluate the degree of anxiety and fear among the patients. RESULTS: There was a 23.0% prevalence of dental anxiety; 9.5% were very anxious, 13.5% were anxious and 77.0% were not anxious. Women were more anxious than men (20.7% and 11.3%, respectively, P = 0.995). The age group that exhibited the highest prevalence of anxiety was 30 to 39 years (29.3%). Regarding fear, 13.5% felt extreme fear; 30.5% felt moderate fear and 56% felt no fear. Schooling was the only significant socioeconomic factor for both anxiety and fear. In the association between fear and anxiety, 38.9% of the patients with extreme fear were classified as very anxious. CONCLUSION: The majority of individuals were classified as non-anxious and without fear regarding dental procedures.


Assuntos
Ansiedade ao Tratamento Odontológico/epidemiologia , Clínicas Odontológicas , Adolescente , Adulto , Fatores Etários , Brasil/epidemiologia , Distribuição de Qui-Quadrado , Restauração Dentária Permanente/psicologia , Escolaridade , Feminino , Humanos , Masculino , Escala de Ansiedade Manifesta , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Bucais/psicologia , Prevalência , Tratamento do Canal Radicular/psicologia , Fatores Sexuais , Inquéritos e Questionários , Adulto Jovem
19.
G3 (Bethesda) ; 11(11)2021 10 19.
Artigo em Inglês | MEDLINE | ID: mdl-34510200

RESUMO

Polyploidy is considered a driving force in plant evolution and domestication. Although in the genus Arachis, several diploid species were traditionally cultivated for their seeds, only the allotetraploid peanut Arachis hypogaea became the successful, widely spread legume crop. This suggests that polyploidy has given selective advantage for domestication of peanut. Here, we study induced allotetraploid (neopolyploid) lineages obtained from crosses between the peanut's progenitor species, Arachis ipaënsis and Arachis duranensis, at earlier and later generations. We observed plant morphology, seed dimensions, and genome structure using cytogenetics (FISH and GISH) and SNP genotyping. The neopolyploid lineages show more variable fertility and seed morphology than their progenitors and cultivated peanut. They also showed sexual and somatic genome instability, evidenced by changes of number of detectable 45S rDNA sites, and extensive homoeologous recombination indicated by mosaic patterns of chromosomes and changes in dosage of SNP alleles derived from the diploid species. Genome instability was not randomly distributed across the genome: the more syntenic chromosomes, the higher homoeologous recombination. Instability levels are higher than observed on peanut lines, therefore it is likely that more unstable lines tend to perish. We conclude that early stages of the origin and domestication of the allotetraploid peanut involved two genetic bottlenecks: the first, common to most allotetraploids, is composed of the rare hybridization and polyploidization events, followed by sexual reproductive isolation from its wild diploid relatives. Here, we suggest a second bottleneck: the survival of the only very few lineages that had stronger mechanisms for limiting genomic instability.


Assuntos
Arachis , Fabaceae , Arachis/genética , Fabaceae/genética , Genoma de Planta , Humanos , Poliploidia , Sintenia
20.
Sci Rep ; 11(1): 11097, 2021 05 27.
Artigo em Inglês | MEDLINE | ID: mdl-34045561

RESUMO

Nematodes and drought are major constraints in tropical agriculture and often occur simultaneously. Plant responses to these stresses are complex and require crosstalk between biotic and abiotic signaling pathways. In this study, we explored the transcriptome data of wild Arachis species subjected to drought (A-metaDEG) and the root-knot nematode Meloidogyne arenaria (B-metaDEG) via meta-analysis, to identify core-stress responsive genes to each individual and concurrent stresses in these species. Transcriptome analysis of a nematode/drought bioassay (cross-stress) showed that the set of stress responsive DEGs to concurrent stress is distinct from those resulting from overlapping A- and B-metaDEGs, indicating a specialized and unique response to combined stresses in wild Arachis. Whilst individual biotic and abiotic stresses elicit hormone-responsive genes, most notably in the jasmonic and abscisic acid pathways, combined stresses seem to trigger mainly the ethylene hormone pathway. The overexpression of a cross-stress tolerance candidate gene identified here, an endochitinase-encoding gene (AsECHI) from Arachis stenosperma, reduced up to 30% of M. incognita infection and increased post-drought recovery in Arabidopsis plants submitted to both stresses. The elucidation of the network of cross-stress responsive genes in Arachis contributes to better understanding the complex regulation of biotic and abiotic responses in plants facilitating more adequate crop breeding for combined stress tolerance.


Assuntos
Arachis/genética , Arachis/parasitologia , Secas , Estresse Fisiológico/fisiologia , Tylenchoidea , Animais , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/genética , Transcriptoma
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