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1.
Front Mol Biosci ; 11: 1427352, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39176391

RESUMO

Asthma comprises one of the most common chronic inflammatory conditions, yet still lacks effective diagnostic markers and treatment targets. To gain deeper insights, we comprehensively analyzed microarray datasets of airway epithelial samples from asthmatic patients and healthy subjects in the Gene Expression Omnibus database using three machine learning algorithms. Our investigation identified a pivotal gene, STEAP4. The expression of STEAP4 in patients with allergic asthma was found to be reduced. Furthermore, it was found to negatively correlate with the severity of the disease and was subsequently validated in asthmatic mice in this study. A ROC analysis of STEAP4 showed the AUC value was greater than 0.75. Functional enrichment analysis of STEAP4 indicated a strong correlation with IL-17, steroid hormone biosynthesis, and ferroptosis signaling pathways. Subsequently, intercellular communication analysis was performed using single-cell RNA sequencing data obtained from airway epithelial cells. The results revealed that samples exhibiting low levels of STEAP4 expression had a richer MIF signaling pathway in comparison to samples with high STEAP4 expression. Through both in vitro and in vivo experiments, we further confirmed the overexpression of STEAP4 in airway epithelial cells resulted in decreased expression of MIF, which in turn caused a decrease in the levels of the cytokines IL-33, IL-25, and IL-4; In contrast, when the STEAP4 was suppressed in airway epithelial cells, there was an upregulation of MIF expression, resulting in elevated levels of the cytokines IL-33, IL-25, and IL-4. These findings suggest that STEAP4 in the airway epithelium reduces allergic asthma Th2-type inflammatory reactions by inhibiting the MIF signaling pathway.

2.
Taiwan J Obstet Gynecol ; 60(6): 1066-1071, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34794739

RESUMO

OBJECTIVE: To evaluate the detection rate (DR) by prenatal cell-free DNA test for pathogenic copy number variations (CNVs)>2 Mb among pregnancies with fetal ultrasound abnormalities. MATERIALS AND METHODS: This was a retrospective study on 29 pregnant women with fetuses diagnosed as microdeletion/microduplication syndromes by prenatal chromosome microarray analysis (CMA). Cell-free DNA from the maternal plasma was sequenced on the NextSeq CN500 sequencer. The quality standard of unique map reads in a single sample was greater than 10 M and only gains and losses of more than 2 Mb were reported. RESULTS: A total of 24 CNVs were identified by cell-free DNA test among the 21 fetuses with pathogenic CNVs identified by prenatal CMA, including 20 consistent CNVs and 4 inconsistent CNVs. Overall, the DR of cell-free DNA test for pathogenic CNVs >2 Mb was 69%. Microdeletions or microduplications at 22q11.2 were the most common CNVs, with a DR of 4/5 (80%) and 3/4 (75%) respectively. CONCLUSION: Cell-free DNA test exhibited a moderate DR for pathogenic CNVs >2 Mb among fetuses with ultrasound abnormalities. Cell-free DNA test could provide an opportunity for early screening before the appearance of abnormalities on fetal ultrasound, while further clinical data and cost-effectiveness assessment are needed.


Assuntos
Transtornos Cromossômicos/diagnóstico , Variações do Número de Cópias de DNA/genética , DNA/sangue , Testes Genéticos/métodos , Análise em Microsséries/métodos , Diagnóstico Pré-Natal/métodos , Adulto , Ácidos Nucleicos Livres/genética , Aberrações Cromossômicas , Transtornos Cromossômicos/genética , Feminino , Humanos , Gravidez , Estudos Retrospectivos , Análise de Sequência de DNA
3.
Taiwan J Obstet Gynecol ; 60(2): 232-237, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33678321

RESUMO

OBJECTIVE: To present the experience on prenatal features of 17q12 microdeletion and microduplication syndromes. MATERIALS AND METHODS: Prenatal chromosomal microarray analysis (CMA) were conducted between January 2015 and December 2018 at a single Chinese tertiary medical centre. Information of cases identified with 17q12 microdeletion or microduplication syndromes were retrospectively collected. Foetal ultrasonographic findings were reviewed, and other information about the gestation week at diagnosis, inheritance and pregnancy outcomes were also included. RESULTS: Ten pregnancies with 17q12 microdeletion and 4 with 17q12 microduplication were identified. The copy number variation (CNV) sizes were 1.39-1.94 Mb in the deleted cases and 1.42-1.48 Mb in the duplicated cases, respectively. All the duplicated and deleted regions included HNF1B and LHX1 genes. Most individuals with 17q12 deletion presented kidney anomalies (9/10), with renal hyperechogenicity being the most common finding (7/10). Fetuses with 17q12 duplication presented a wide phenotypic spectrum, including "double bubble" sign, structural anomalies of the heart and growth anomalies. CONCLUSIONS: Our experience further demonstrated the high correlation between 17q12 microdeletion and renal anomalies especially hyperechogenic kidneys. Structural anomalies of the heart were newly identified phenotypes of 17q12 duplication during prenatal period. Besides, growth anomalies and duodenal atresia might be associated with the duplication.


Assuntos
Deleção Cromossômica , Duplicação Cromossômica , Cromossomos Humanos Par 17/genética , Anormalidades Congênitas/embriologia , Anormalidades Congênitas/genética , Adulto , Anormalidades Congênitas/diagnóstico , Variações do Número de Cópias de DNA , Feminino , Cardiopatias Congênitas/diagnóstico , Cardiopatias Congênitas/embriologia , Fator 1-beta Nuclear de Hepatócito/genética , Humanos , Rim/anormalidades , Rim/embriologia , Proteínas com Homeodomínio LIM/genética , Análise em Microsséries , Fenótipo , Gravidez , Diagnóstico Pré-Natal , Estudos Retrospectivos , Síndrome , Fatores de Transcrição/genética
4.
Taiwan J Obstet Gynecol ; 58(2): 251-254, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30910148

RESUMO

OBJECTIVE: To investigate the clinical value of chromosomal microarray analysis (CMA) in the prenatal diagnosis of genetic abnormalities in fetal isolated mild ventriculomegaly. MATERIALS AND METHODS: This retrospective study reviewed 101 fetuses with isolated mild ventriculomegaly who had undergone invasive prenatal diagnosis at our hospital. CMA was performed in all cases to detect chromosomal aneuploidy as well as copy number variations (CNVs) that are too small to be detected by conventional karyotyping. Real time quantitative PCR (qPCR) or multiplex ligation dependent probe amplification (MLPA) was used to confirm all fetal CNVs <400 Kb. RESULTS: Except for three cases of chromosomal aneuploidy, CMA revealed pathogenic copy number variations (CNVs) in 3.0% (3/101) of the fetuses; these cases demonstrated involvement in the chromosomal regions 15q11.2, 1q21.1 and Xq27.3q28. Furthermore, we detected three likely pathogenic (3.0%) and two variants of uncertain significance (2.0%) among 101 fetuses diagnosed as isolated mild ventriculomegaly on ultrasound examination. CONCLUSION: Our study suggests that CNVs could aid in the risk assessment and genetic counseling in fetuses with isolated ventriculomegaly.


Assuntos
Variações do Número de Cópias de DNA/genética , Doenças Fetais/diagnóstico , Hidrocefalia/diagnóstico , Análise em Microsséries , Diagnóstico Pré-Natal/métodos , Aneuploidia , Feminino , Doenças Fetais/genética , Idade Gestacional , Humanos , Hidrocefalia/embriologia , Hidrocefalia/genética , Masculino , Gravidez , Estudos Retrospectivos , Medição de Risco
5.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 30(1): 45-8, 2013 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-23450478

RESUMO

OBJECTIVE: To detect potential mutations for probands from families affected with Duchenne/Becker muscular dystrophy (DMD/BMD), and to carry out prenatal diagnosis through identification of female carriers. METHODS: A total of 43 DMD/BMD families were recruited. Multiplex PCR was used to analyze 18 exons within hotspots for DMD gene deletions. Multiplex ligation-dependent probe amplification (MLPA) was used to detect potential deletions and duplications of DMD gene for 43 patients and 36 females from 32 families. Prenatal diagnosis was performed for 27 families. RESULTS: Deletional mutations were detected in 26 patients with multiplex PCR. In addition, MLPA has detected 3 deletions and 6 duplicational mutations, and the ranges of mutations were all determined. Among 36 female members, 18 were determined as carriers of deletional mutations, 10 were excluded as mutation carriers. The status of remaining 8 could not be determined. For prenatal diagnosis, 3 out of 18 male fetuses were diagnosed as patients and 1 female fetus was identified as carrier. CONCLUSION: MLPA is an accurate and reliable method for detecting deletional/duplicational mutations of DMD gene as well as for prenatal diagnosis and detection of female carriers.


Assuntos
Distrofina/genética , Distrofia Muscular de Duchenne/diagnóstico , Distrofia Muscular de Duchenne/genética , Adolescente , Criança , Pré-Escolar , Feminino , Heterozigoto , Humanos , Lactente , Masculino , Reação em Cadeia da Polimerase Multiplex , Mutação , Linhagem , Gravidez , Diagnóstico Pré-Natal
6.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 28(6): 625-9, 2011 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-22161092

RESUMO

OBJECTIVE: To provide genetic diagnosis and counseling for a 2-year-old girl with typical Rett syndrome through analyzing the methyl-CpG binding protein 2 (MECP2) gene. METHODS: Potential mutation of the MECP2 gene was screened by DNA sequencing and multiplex ligation-dependent probe amplification (MLPA) analysis of members of the family as well as normal controls. Lymphocyte culture for karyotype analysis was carried out for the patient to exclude chromosomal abnormalities. RESULTS: The karyotype of the girl was normal. No variation of the MECP2 gene was detected in the patient by direct sequencing. A heterozygosis variation, c.1072G>A in exon 4 of the MECP2 gene was detected in a normal female control, which was not found in other controls. The son and daughter of the female control were respectively heterozygous and homozygous carriers of the same mutation. By MLPA analysis, a heterozygosis deletion of exon 3 and part of exon 4 was detected in the patient. cDNA amplification and sequencing confirmed the presence of a 1176 bp deletion (c.27-1202del1176). The same deletion was not detected in the parents. CONCLUSION: A large deletion in MECP2 gene was detected with MLPA in a patient featuring typical Rett syndrome. The same deletion was missed by sequencing analysis. With cDNA sequencing, the breakage point of the mutation can be mapped precisely.


Assuntos
Proteína 2 de Ligação a Metil-CpG/genética , Síndrome de Rett/genética , Sequência de Bases , Pré-Escolar , Éxons , Feminino , Testes Genéticos , Genótipo , Humanos , Cariotipagem , Mutação
7.
Chin Med J (Engl) ; 124(19): 3054-7, 2011 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22040554

RESUMO

BACKGROUND: Multiple osteochondromas (MO), an inherited autosomal dominant disorder, is characterized by the presence of multiple exostoses on the long bones. MO is caused by mutations in the EXT1 or EXT2 genes which encode glycosyltransferases implicated in heparin sulfate biosynthesis. METHODS: In this study, efforts were made to identify the underlying disease-causing mutations in patients from two MO families in China. RESULTS: Two novel EXT1 gene mutations were identified and no mutation was found in EXT2 gene. The mutation c.497T > A in exon 1 of the EXT1 gene was cosegregated with the disease phenotype in family 1 and formed a stop codon at amino acid site 166. The fetus of the proband was diagnosed negative. In family 2, the mutation c.1430-1431delCC in exon 6 of the EXT1 gene would cause frameshift and introduce a premature stop codon after the reading frame being open for 42 amino acids. The fetus of this family inherited this mutation from the father. CONCLUSIONS: Mutation analysis of two MO families in this study demonstrates its further application in MO genetic counseling and prenatal diagnosis.


Assuntos
Exostose Múltipla Hereditária/genética , Mutação , N-Acetilglucosaminiltransferases/genética , Adulto , Povo Asiático/genética , Pré-Escolar , Feminino , Humanos , Masculino
8.
Zhonghua Yi Xue Za Zhi ; 89(25): 1753-6, 2009 Jul 07.
Artigo em Chinês | MEDLINE | ID: mdl-19862979

RESUMO

OBJECTIVE: To analyze the pathogenic mutation of sporadic patients in Duchenne/ Becker muscular dystrophy (DMD/BMD) families and to perform prenatal diagnosis, identify the female carriers and evaluate the ratio of de novo mutation in these pedigrees. METHODS: A total of 30 sporadic DMD/BMD families were included. Traditional multiplex PCR was used to detect the 18 exons in the deletion hot area of DMD gene. MLPA was used to detect the deletion or duplication mutations of DMD gene for 30 patients and 28 females from 23 families. Prenatal diagnosis was performed in 19 families. RESULTS: Deletion mutation was detected in 19 patients through mPCR. Twenty-one deletions and 3 duplication mutations were detected by MLPA. Among 21 mothers, 10 mothers were carriers of deletion mutation and two duplication mutation carriers. The other 9 mothers were non-carriers, the mutations in these families were de novo and the ratio was 37.5%. Among seven sisters, five were carriers and two non-carriers. In the prenatal diagnosis families, 2 of 8 female fetuses were carriers and 5 of 12 male fetus patients. CONCLUSION: The MLPA technique has proved to be an accurate and reliable tool not only for the deletion and duplication mutations of DMD patients, but also for the prenatal diagnosis and the female carriers in these families. Prenatal diagnosis;


Assuntos
Análise Mutacional de DNA/métodos , Deleção de Genes , Distrofia Muscular de Duchenne/genética , Diagnóstico Pré-Natal , Criança , Pré-Escolar , Éxons , Feminino , Frequência do Gene , Heterozigoto , Humanos , Masculino , Mutação , Técnicas de Amplificação de Ácido Nucleico , Linhagem , Reação em Cadeia da Polimerase/métodos , Gravidez
9.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 25(4): 421-3, 2008 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-18683141

RESUMO

OBJECTIVE: To detect the mutation of the SEDL gene in an X-linked spondyloepiphyseal dysplasia tarda (SEDL) family. METHODS: Two patients and three females of the X-SEDL family were detected using reverse transcriptase PCR (RT-PCR) and sequence analysis. RESULTS: A G209A mutation of SEDL gene was detected in the cDNA sequences of the patients, which was confirmed by sequence analysis of the exon 4 of the SEDL gene. The daughter of the proband was a carrier of the mutation. CONCLUSION: Since the SEDL gene is relatively small, sequence analysis of cDNA of the SEDL gene was possible after extraction of total RNA followed by RT-PCR. Mutations in the open reading frame can be detected y by cDNA sequencing. It was relatively more rapid and direct than amplifying and detecting the exons one by one.


Assuntos
Cromossomos Humanos X , Análise Mutacional de DNA , Doenças Genéticas Ligadas ao Cromossomo X/genética , Osteocondrodisplasias/genética , DNA Complementar/análise , Feminino , Ligação Genética , Humanos , Masculino , Linhagem , Deleção de Sequência
10.
Zhonghua Yi Xue Za Zhi ; 88(46): 3246-9, 2008 Dec 16.
Artigo em Chinês | MEDLINE | ID: mdl-19159546

RESUMO

OBJECTIVE: To analyze the pathogenic mutation of an X-linked ichthyosis (XLI) family, and identify the genetic diagnosis of three probable female carriers in this family. To evaluate the availability of different detect methods for steroid sulfatase (STS) gene mutation. METHODS: Peripheral blood samples were collected from the family, including the proband, proband's mother, younger sister, and younger female cousin, and 10 males and 10 females as controls. Ordinary PCR was used to detect whether there was STS gene deletion in the male proband. Then, multiplex quantitative fluorescent PCR (QF-PCR) was used to detect the STS gene in the proband and his 3 female family members. Fluorescence in situ hybridization (FISH) was used to authenticate the results of multiplex QF-PCR method. RESULTS: No amplified product of the exons 1-10 of STS gene deletion was detected by ordinary PCR in the proband. The proband's mother was diagnosed as a carrier, but his sister and cousin were diagnosed as normal females by multiplex QF-PCR. FISH confirmed the results of multiplex QF-PCR. CONCLUSION: Both multiplex QF-PCR and FISH are effective to detect the complete deletion mutation of STS gene and identify the female carrier, and multiplex QF-PCR is more convenient and automatic compared with FISH.


Assuntos
Ictiose Ligada ao Cromossomo X/diagnóstico , Ictiose Ligada ao Cromossomo X/genética , Reação em Cadeia da Polimerase/métodos , Adulto , Éxons , Feminino , Humanos , Hibridização in Situ Fluorescente , Masculino , Linhagem , Gravidez , Esteril-Sulfatase/genética
11.
Zhonghua Yi Xue Za Zhi ; 87(16): 1123-5, 2007 Apr 24.
Artigo em Chinês | MEDLINE | ID: mdl-17672996

RESUMO

OBJECTIVE: To investigate the genotype of oculocutaneous albinism type II (OCA2) and perform prenatal gene diagnosis for OCA2. METHODS: Peripheral blood samples were collected from a 9-year-old girl with OCA and her parents, the mother being pregnant. PCR, automatic sequence analysis and denaturing high performance liquid chromatography (DHPLC) were used to analyze the TYR gene and P gene so as to screen the OCA genes. Amniocentesis was conducted when the mother was 20-week pregnant and the amniotic cells underwent screening of the 2 specific mutations. Peripheral blood samples were collected from 100 healthy persons without phenotype of OCA to undergo genetic analysis. RESULTS: The TYR gene of the proband did not show any mutation, and 2 new mutations were found in her P gene: p. N476D (c. 1426 A>G) and p.Y827H (c.2479T>C). Her father and mother were heterozygote of Y827H and N476D respectively. Based on these findings the amniotic cells underwent sequencing of enlarged fragments of the exons 14 and 24 containing the mutation sites. The result showed that the fetus only got the maternal N476D mutation and didn't get the paternal Y827H mutation. So the fetus was predicted to be a carrier of OCA2 with normal appearance. The baby was born normal later as predicted. None of these 2 mutations was found in the 100 healthy persons. CONCLUSION: This is a success of prenatal gene diagnosis of OCA2. Two novel mutations of the P gene related to OCA have been discovered.


Assuntos
Albinismo Oculocutâneo/diagnóstico , Albinismo Oculocutâneo/genética , Mutação , Diagnóstico Pré-Natal/métodos , Adulto , Albinismo Oculocutâneo/embriologia , Sequência de Aminoácidos , Sequência de Bases , Criança , Cromatografia Líquida de Alta Pressão/métodos , Análise Mutacional de DNA , Feminino , Genótipo , Humanos , Masculino , Proteínas de Membrana Transportadoras/genética , Dados de Sequência Molecular , Mães , Homologia de Sequência de Aminoácidos
12.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 23(6): 614-7, 2006 Dec.
Artigo em Chinês | MEDLINE | ID: mdl-17160937

RESUMO

OBJECTIVE: To investigate gene mutations of a consanguineous family with two oculocutaneous albinism (OCA) patients. METHODS: Genomic DNA was prepared from peripheral leukocytes. All of the exons and flanking introns of P gene and TYR gene were PCR-direct-sequenced. Hha I restriction fragment length polymorphism in codon 787 of the P gene was studied in the family and 102 unrelated normal Chinese individuals. RESULTS: Although no mutations were found in TYR gene, a missense mutation A787T was found in P gene. Two patients of the family were both homozygous for A787T. Their parents and brother were heterozygous for the mutation. The mutation was not observed among 102 normally pigmented subjects. CONCLUSION: The A787T mutation is not a common polymorphism among normal Chinese and it seems most likely to be a pathological OCA2 mutation. This is the first report on the study of gene diagnosis in Chinese OCA2 patients.


Assuntos
Albinismo Oculocutâneo/genética , Proteínas de Membrana Transportadoras/genética , Mutação de Sentido Incorreto , Adulto , Albinismo Oculocutâneo/etnologia , Sequência de Aminoácidos , Povo Asiático/genética , Sequência de Bases , China , Códon , Consanguinidade , Análise Mutacional de DNA , Éxons , Feminino , Homozigoto , Humanos , Íntrons , Masculino , Dados de Sequência Molecular , Monofenol Mono-Oxigenase/genética , Linhagem , Homologia de Sequência do Ácido Nucleico
13.
Yi Chuan ; 28(9): 1149-52, 2006 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-16963427

RESUMO

Oculocutaneous albinism (OCA) is a complex genetic disease with great clinical heterogeneity. Four different types of OCA have been reported to date (OCA1, OCA2, OCA3, and OCA4). OCA4 was firstly reported in a Turkish OCA patient. The gene responsible for OCA4 is the human homologue of the mouse underwhite (uw) gene, which encodes the mem-brane-associated transporter protein (MATP). MATP gene is located on chromosome 5p13.3 and is divided into 7 exons and 6 introns. MATP gene is transcriptionally modulated by MITF, and encodes a protein of 530 amino acids. There are at least 18 pathologic mutations and 8 non-pathologic polymorphisms have been found.


Assuntos
Albinismo Oculocutâneo/genética , Albinismo Oculocutâneo/patologia , Albinismo Oculocutâneo/classificação , Albinismo Oculocutâneo/metabolismo , Animais , Humanos , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Mutação , Polimorfismo Genético
14.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 23(3): 280-2, 2006 Jun.
Artigo em Chinês | MEDLINE | ID: mdl-16767664

RESUMO

OBJECTIVE: Mutation analysis and prenatal gene diagnosis for the mutated tyrosinase (TYR) gene in two families with oculocutaneous albinism type I (OCA1). METHODS: To define the fetus genotypes and gene mutation sites, the PCR and sequencing techniques were applied to amplify and analyze the regions of exon, exon-intron and promoter of TYR gene in probands and their parents of 2 families. RESULTS: The patient or proband of family 1 showed as a compound heterozygote with mutants R278X and 929insC. However, the fetus did not get any one of the two mutations, and so was with a normal genotype and phenotype. The parents of proband in family 2 were heterozygous with IVS4+ 3A>T or G253E respectively, but their fetus was heterozygous only with IVS4+3A>T but without G253E, and so was a carrier as his father. CONCLUSION: In the mainland of China, the prenatal gene diagnosis of OCA1 is reported for the first time.


Assuntos
Albinismo Oculocutâneo/diagnóstico , Albinismo Oculocutâneo/genética , Diagnóstico Pré-Natal/métodos , Pré-Escolar , Saúde da Família , Feminino , Humanos , Masculino , Monofenol Mono-Oxigenase/genética , Mutação , Linhagem , Reação em Cadeia da Polimerase , Gravidez
15.
Yi Chuan ; 27(6): 984-8, 2005 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-16378950

RESUMO

Oculocutaneous albinism typeII(OCA2), the most common type of albinism, is an autosomal recessive disorder. It is caused by mutations in the P gene, which is located on chromosome 15q11.1-q12 and divided into 24 exons and 23 introns. P gene codes for 838-amino-acid integral membrane protein with 12 putative transmembrane domains, but the exact function is not clear yet. There are at least 60 pathologic mutations and 43 non-pathologic polymorphisms have been found. Pathologic mutations include missense mutations, nonsense mutations, frameshift mutations and splice-sit mutations. But unlike TYR gene, most of P gene mutations are located on the C-terminal and don' t cluster in defined regions. It is difficult to define the pathologic mutations since many non-pathologic polymorphisms also lie in exons. Some non-pathologic missense mutations may be associated with phenotypic variation in normally pigmented individuals and need to further study.


Assuntos
Albinismo Oculocutâneo/genética , Proteínas de Membrana Transportadoras/genética , Mutação , Polimorfismo Genético , Albinismo Oculocutâneo/classificação , Cromossomos Humanos Par 15 , Humanos
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