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1.
Microsc Microanal ; 29(6): 2149-2160, 2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-37967302

RESUMO

Mammalian oocyte maturation relies on mitochondrial ATP production, but this can lead to damaging reactive oxygen species (ROS). SIRT3, a mitochondrial sirtuin, plays a critical role in regulating mitochondrial redox balance in mouse oocytes under stress; however, its specific roles in porcine oocytes remain unclear. In this study, we utilized the SIRT3 inhibitor 3-TYP to investigate SIRT3's importance in porcine oocyte maturation. Our findings revealed that SIRT3 is expressed in porcine oocytes and its inhibition leads to maturation failure. This was evident through reduced polar body extrusion, arrested cell cycle, as well as disrupted spindle organization and actin distribution. Furthermore, SIRT3 inhibition resulted in a decrease in mitochondrial DNA copy numbers, disruption of mitochondrial membrane potential, and reduced ATP levels, all indicating impaired mitochondrial function in porcine oocytes. Additionally, the primary source of damaged mitochondria was associated with decreased levels of deacetylated superoxide dismutase 2 (SOD2) after SIRT3 inhibition, which led to ROS accumulation and oxidative stress-induced apoptosis. Taken together, our results suggest that SIRT3 regulates the levels of deacetylated SOD2 to maintain redox balance and preserve mitochondrial function during porcine oocyte maturation, with potential implications for improving pig reproduction.


Assuntos
Doenças Mitocondriais , Sirtuína 3 , Camundongos , Animais , Suínos , Espécies Reativas de Oxigênio , Sirtuína 3/genética , Sirtuína 3/metabolismo , Estresse Oxidativo , Oócitos/metabolismo , Trifosfato de Adenosina/metabolismo , Doenças Mitocondriais/metabolismo , Mamíferos/metabolismo
2.
Hum Reprod ; 38(8): 1628-1642, 2023 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-37218343

RESUMO

STUDY QUESTION: Can multiple-site low-pass genome sequencing (GS) of products of conception (POCs) improve the detection of genetic abnormalities, especially heterogeneously distributed mosaicism and homogeneously distributed mosaicism in first-trimester miscarriage? SUMMARY ANSWER: Multiple-site sampling combined with low-pass GS significantly increased genetic diagnostic yield (77.0%, 127/165) of first-trimester miscarriages, with mosaicisms accounting for 17.0% (28/165), especially heterogeneously distributed mosaicisms (75%, 21/28) that are currently underappreciated. WHAT IS KNOWN ALREADY: Aneuploidies are well known to cause first-trimester miscarriage, which are detectable by conventional karyotyping and next-generation sequencing (NGS) on a single-site sampling basis. However, there are limited studies demonstrating the implications of mosaic genetic abnormalities in first-trimester miscarriages, especially when genetic heterogeneity is present in POCs. STUDY DESIGN, SIZE, DURATION: This is a cross-sectional cohort study carried out at a university-affiliated public hospital. One hundred seventy-four patients diagnosed with first-trimester miscarriage from December 2018 to November 2021 were offered ultrasound-guided manual vacuum aspiration (USG-MVA) treatment. Products of conception were subjected to multiple-site low-pass GS for the detection of chromosomal imbalances. PARTICIPANTS/MATERIALS, SETTING, METHODS: For each POC, multiple sites of villi (three sites on average) were biopsied for low-pass GS. Samples with maternal cell contamination (MCC) and polyploidy were excluded based on the quantitative fluorescence polymerase chain reaction (QF-PCR) results. The spectrum of chromosomal abnormalities, including mosaicism (heterogeneously distributed and homogeneously distributed) and constitutional abnormalities was investigated. Chromosomal microarray analysis and additional DNA fingerprinting were used for validation and MCC exclusion. A cross-platform comparison between conventional karyotyping and our multiple-site approach was also performed. MAIN RESULTS AND THE ROLE OF CHANCE: One hundred sixty-five POCs (corresponding to 490 DNA samples) were subjected to low-pass GS. Genetic abnormalities were detected in 77.0% (127/165) of POCs by our novel approach. Specifically, 17.0% (28/165) of cases had either heterogeneously distributed mosaicism (12.7%, 21/165) or homogeneously distributed mosaicism (6.1%, 10/165) (three cases had both types of mosaicism). The remaining 60.0% (99/165) of cases had constitutional abnormalities. In addition, in the 71 cases with karyotyping performed in parallel, 26.8% (19/71) of the results could be revised by our approach. LIMITATIONS, REASONS FOR CAUTION: Lack of a normal gestational week-matched cohort might hinder the establishment of a causative link between mosaicisms and first-trimester miscarriage. WIDER IMPLICATIONS OF THE FINDINGS: Low-pass GS with multiple-site sampling increased the detection of chromosomal mosaicisms in first-trimester miscarriage POCs. This innovative multiple-site low-pass GS approach enabled the novel discovery of heterogeneously distributed mosaicism, which was prevalent in first-trimester miscarriage POCs and frequently observed in preimplantation embryos, but is currently unappreciated by conventional single-site cytogenetic investigations. STUDY FUNDING/COMPETING INTEREST(S): This work was supported partly by Research Grant Council Collaborative Research Fund (C4062-21GF to K.W.C), Science and Technology Projects in Guangzhou (202102010005 to K.W.C), Guangdong-Hong Kong Technology Cooperation Funding Scheme (TCFS), Innovation and Technology Fund (GHP/117/19GD to K.W.C), HKOG Direct Grant (2019.050 to J.P.W.C), and Hong Kong Health and Medical Research Fund (05160406 to J.P.W.C). The authors have no competing interests to declare. TRIAL REGISTRATION NUMBER: N/A.


Assuntos
Aborto Espontâneo , Gravidez , Feminino , Humanos , Aborto Espontâneo/genética , Primeiro Trimestre da Gravidez , Mosaicismo , Estudos Transversais , Projetos Piloto
3.
Brain Behav ; 13(6): e3013, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37072935

RESUMO

INTRODUCTION: Regulation of brain-derived neurotrophic factor (BDNF) in the basal forebrain ameliorates sleep deprivation-induced fear memory impairments in rodents. Antisense oligonucleotides (ASOs) targeting ATXN2 was a potential therapy for spinocerebellar ataxia, whose pathogenic mechanism associates with reduced BDNF expression. We tested the hypothesis that ASO7 targeting ATXN2 could affect BDNF levels in mouse basal forebrain and ameliorate sleep deprivation-induced fear memory impairments. METHODS: Adult male C57BL/6 mice were used to evaluate the effects of ASO7 targeting ATXN2 microinjected into the bilateral basal forebrain (1 µg, 0.5 µL, each side) on spatial memory, fear memory and sleep deprivation-induced fear memory impairments. Spatial memory and fear memory were detected by the Morris water maze and step-down inhibitory avoidance test, respectively. Immunohistochemistry, RT-PCR, and Western blot were used to evaluate the changes of levels of BDNF, ATXN2, and postsynaptic density 95 (PSD95) protein as well as ATXN2 mRNA. The morphological changes in neurons in the hippocampal CA1 region were detected by HE staining and Nissl staining. RESULTS: ASO7 targeting ATXN2 microinjected into the basal forebrain could suppress ATXN2 mRNA and protein expression for more than 1 month and enhance spatial memory but not fear memory in mice. BDNF mRNA and protein expression in basal forebrain and hippocampus was increased by ASO7. Moreover, PSD95 expression and synapse formation were increased in the hippocampus. Furthermore, ASO7 microinjected into the basal forebrain increased BDNF and PSD95 protein expression in the basal forebrain of sleep-deprived mice and counteracted sleep deprivation-induced fear memory impairments. CONCLUSION: ASOs targeting ATXN2 may provide effective interventions for sleep deprivation-induced cognitive impairments.


Assuntos
Prosencéfalo Basal , Privação do Sono , Camundongos , Masculino , Animais , Privação do Sono/complicações , Privação do Sono/metabolismo , Fator Neurotrófico Derivado do Encéfalo/genética , Memória Espacial , Prosencéfalo Basal/metabolismo , Oligonucleotídeos Antissenso/farmacologia , Oligonucleotídeos Antissenso/metabolismo , Camundongos Endogâmicos C57BL , Transtornos da Memória/etiologia , Transtornos da Memória/complicações , RNA Mensageiro/metabolismo
4.
Plant Sci ; 324: 111446, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36041562

RESUMO

Heading date is crucial for rice reproduction and the geographical expansion of cultivation. We fine-mapped qHD5 and identified LOC_Os05g03040, a gene that encodes an AP2 transcription factor, as the candidate gene of qHD5 in our previous study. In this article, using two near-isogenic lines NIL(BG1) and NIL(XLJ), which were derived from the progeny of the cross between BigGrain1 (BG1) and Xiaolijing (XLJ), we verified that LOC_Os05g03040 represses heading date in rice through genetic complementation and CRISPR/Cas9 gene-editing experiments. Complementary results showed that qHD5 is a semi-dominant gene and that the qHD5XLJ and qHD5BG1 alleles are both functional. The homozygous mutant line generated from knocking out qHD5XLJ in NIL(XLJ) headed earlier than NIL(XLJ) under both short-day and long-day conditions. In addition, the homozygous mutant line of qHD5BG1 in NIL(BG1) also headed slightly earlier than NIL(BG1). All of these results show that qHD5 represses the heading date in rice. Transient expression showed that the qHD5 protein localizes to the nucleus. Transactivation activity assays showed that the C-terminus is the critical site that affects self-activation in qHD5XLJ. qRT-PCR analysis revealed that qHD5 represses flowering by down-regulating Ehd2. qHD5 may have been selected during indica rice domestication.


Assuntos
Oryza , Alelos , Mapeamento Cromossômico , Regulação da Expressão Gênica de Plantas , Oryza/metabolismo , Locos de Características Quantitativas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
5.
Ying Yong Sheng Tai Xue Bao ; 33(8): 2260-2270, 2022 Aug.
Artigo em Chinês | MEDLINE | ID: mdl-36043835

RESUMO

With the continuous growth of vehicle per capita in recent years, tire wear particles (TWP) produced by tire wear on roads have been widely found in various environmental media, whose environmental behavior and effects have attracted extensive attention. After being produced, these microsized and nanosized TWP will drift into the atmosphere or enter the surrounding soil, rivers, and even into the ocean with rain and runoff. The existence of TWP significantly affects the composition of organic materials in the environment. Meanwhile, the release of heavy metals and organic additives from TWP will be harmful to organisms and human health. We summarized the sources and characteristics of TWP, their migration and diffusion in water, atmosphere, and soil environment. By analyzing the environmental impacts and ecological risks of TWP pollution, we proposed the key questions urgently to be solved and prevention measures.


Assuntos
Metais Pesados , Poluentes Químicos da Água , Monitoramento Ambiental , Humanos , Microplásticos , Plásticos , Solo , Poluentes Químicos da Água/análise
6.
J Clin Anesth ; 75: 110408, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34237489

RESUMO

STUDY OBJECTIVE: To develop and validate a simple delirium-predicting scoring system in patients undergoing major abdominal surgery by incorporating preoperative risk factors and intraoperative surgical Apgar score (SAS). DESIGN: Observational retrospective cohort study. SETTING: A tertiary general hospital in China. PATIENTS: 1055 patients who received major abdominal surgery from January 2015 to December 2019. MEASUREMENTS: We collected data on preoperative and intraoperative variables, and postoperative delirium. A risk scoring system for postoperative delirium in patients after major open abdominal surgery was developed and validated based on traditional logistic regression model. The elastic net algorithm was further developed and evaluated. MAIN RESULTS: The incidence of postoperative delirium was 17.8% (188/1055) in these patients. They were randomly divided into the development (n = 713) and validation (n = 342) cohorts. Both the logistic regression model and the elastic net regression model identified that advanced age, arrythmia, hypoalbuminemia, coagulation dysfunction, mental illness or cognitive impairments and low surgical Apgar score are related with increased risk of postoperative delirium. The elastic net algorithm has an area under the receiver operating characteristic curve (AUROC) of 0.842 and 0.822 in the development and validation cohorts, respectively. A prognostic score was calculated using the following formula: Prognostic score = Age classification (0 to 3 points) + arrythmia + 2 * hypoalbuminemia + 2 * coagulation dysfunction + 4 * mental illness or cognitive impairments + (10-surgical Apgar score). The 22-point risk scoring system had good discrimination and calibration with an AUROC of 0.823 and 0.834, and a non-significant Hosmer-Lemeshow test P = 0.317 and P = 0.853 in the development and validation cohorts, respectively. The bootstrapping internal verification method (R = 1000) yielded a C-index of 0.822 (95% CI: 0.759-0.857). CONCLUSION: The prognostic scoring system, which used both preoperative risk factors and surgical Apgar score, serves as a good first step toward a clinically useful predictive model for postoperative delirium in patients undergoing major open abdominal surgery.


Assuntos
Delírio , Abdome/cirurgia , Índice de Apgar , Delírio/diagnóstico , Delírio/epidemiologia , Delírio/etiologia , Humanos , Recém-Nascido , Complicações Pós-Operatórias/diagnóstico , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Estudos Retrospectivos , Fatores de Risco
7.
Front Plant Sci ; 12: 804444, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-35126429

RESUMO

Grain size is a key constituent of grain weight and appearance in rice. However, insufficient attention has been paid to the small-effect quantitative trait loci (QTLs) on the grain size. In the present study, residual heterozygous populations were developed for mapping two genetically linked small-effect QTLs for grain size. After the genotyping and the phenotyping of five successive generations, qGS7.1 was dissected into three QTLs and two were selected for further analysis. The qTGW7.2a was finally mapped into a 21.10 kb interval containing four annotated candidate genes. Transcript levels assay showed that the expression of the candidates LOC_Os07g39490 and the LOC_Os07g39500 were significantly reduced in the NIL-qTGW7.2aBG1 . The cytological observation indicated that qTGW7.2a regulated the grain width through controlling the cell expansion. Using the same strategy, qTGW7.2b was fine-mapped into a 52.71 kb interval containing eight annotated candidate genes, showing a significant effect on the grain length and width with opposite allelic directions, but little on the grain weight. Our study provides new genetic resources for yield improvement and for fine-tuning of grain size in rice.

8.
Theor Appl Genet ; 134(2): 453-471, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33089345

RESUMO

Key message Rice male fertility gene Baymax1, isolated through map-based cloning, encodes a MYB transcription factor and is essential for rice tapetum and microspore development.Abstract The mining and characterization of male fertility gene will provide theoretical and material basis for future rice production. In Arabidopsis, the development of male organ (namely anther), usually involves the coordination between MYB (v-myb avian myeloblastosis viral oncogene homolog) and bHLH (basic helix-loop-helix) members. However, the role of MYB proteins in rice anther development remains poorly understood. In this study, we isolated and characterized a male sterile mutant (with normal vegetative growth) of Baymax1 (BM1), which encodes a MYB protein. The bm1 mutant exhibited slightly lagging meiosis, aborted transition of the tapetum to a secretory type, premature tapetal degeneration, and abnormal pollen exine formation, leading to ultimately lacks of visible pollens in the mature white anthers. Map-based cloning, complementation and targeted mutagenesis using CRISPR/Cas9 technology demonstrated that the mutated LOC_Os04g39470 is the causal gene in bm1. BM1 is preferentially expressed in rice anthers from stage 5 to stage 10. Phylogenetic analysis indicated that rice BM1 and its homologs in millet, maize, rape, cabbage, and pigeonpea are evolutionarily conserved. BM1 can physically interacts with bHLH protein TIP2, EAT1, and PHD (plant homeodomain)-finger member TIP3, respectively. Moreover, BM1 affects the expression of several known genes related to tapetum and microspore development. Collectively, our results suggest that BM1 is one of key regulators for rice male fertility and may serve as a potential target for rice male-sterile line breeding and hybrid seed production.


Assuntos
Regulação da Expressão Gênica de Plantas , Oryza/fisiologia , Fenótipo , Infertilidade das Plantas , Proteínas de Plantas/metabolismo , Pólen/química , Proteínas Proto-Oncogênicas c-myb/metabolismo , Mutação , Oryza/genética , Filogenia , Melhoramento Vegetal/métodos , Proteínas de Plantas/genética , Proteínas Proto-Oncogênicas c-myb/genética
9.
Genes (Basel) ; 11(9)2020 08 21.
Artigo em Inglês | MEDLINE | ID: mdl-32825792

RESUMO

Chromosomal mosaicism is at high occurrence in early developmental-stage embryos, but much lower in those at prenatal stage. Recent studies provided evidence on the viability of mosaic embryos by reporting pregnancy outcomes. Expanded research is warranted to evaluate its clinical significance. This is a multi-center prospective cohort study on 137 mosaic, 476 euploid and 835 non-preimplantation genetic testing (non-PGT) embryos from three in vitro fertilization (IVF) providers of three countries in Asia, applying the same preimplantation genetic testing for aneuploidies (PGT-A) reporting criteria. Mosaic embryo transfers (METs) resulted in a significantly lower clinical pregnancy rate (40.1% versus 59.0% versus 48.4%), lower ongoing/live birth rate (27.1% versus 47.0% versus 35.1%) and higher miscarriage rate (33.3% versus 20.5% versus 27.4%) than euploid and non-PGT transfers, respectively. Pregnancy losses after METs were different between embryos carrying numerical and segmental chromosomal abnormalities (p = 0.04). Our meta-analysis concluded that METs gave rise to pregnancies but were associated with a reduced ongoing/live birth rate and a higher miscarriage rate. All 37 MET live births were confirmed viable, among which 8 completed prenatal genetic testing with normal results. Longitudinal investigation on one MET pregnancy evidenced the aneuploidy depletion hypothesis. This is the first multi-center prospective study reporting a full MET pregnancy outcome with complementary information from prenatal genetic testing as compared to euploid and non-PGT cohorts.


Assuntos
Aborto Espontâneo , Aneuploidia , Implantação do Embrião , Transferência Embrionária/métodos , Mosaicismo/embriologia , Taxa de Gravidez , Adulto , Blastocisto , Estudos de Casos e Controles , Feminino , Humanos , Nascido Vivo , Gravidez , Resultado da Gravidez , Estudos Prospectivos
10.
Insects ; 10(9)2019 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-31484469

RESUMO

Intrapuparial development is a special pattern of metamorphosis in cyclorrhaphous flies, in which the pupa forms in an opaque, barrel-like puparium. This has been well studied in forensic insects for age estimations. In this study, the intrapuparial development of a quarantine agricultural pest, Bactrocera dorsalis (Hendel), was studied under a constant temperature of 27 ± 1 °C and 70 ± 5% relative humidity. Results showed that intrapuparial development could be divided into five stages: Larval-pupal apolysis, cryptocephalic pupa, phanerocephalic pupa, pharate adult, and emergent adult. It lays a morphology-based foundation for molecular mechanism studies and enhances the understanding of the physiological basis for changes in intrapuparial development. More importantly, the chronology of intrapuparial development can be used to predict the emergence time of tephritid flies, indicating when to spray insecticides to control these phytophagous agricultural pests. This may be an effective approach to reduce the use of insecticides and slow down the evolution of insecticidal resistance.

11.
ACS Omega ; 4(7): 11493-11499, 2019 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-31460254

RESUMO

A three-dimensional supramolecular framework containing both left- and right-handed helixes, {[Zn(TTPA)Cl2]·1.63H2O} n (1), has been converted to a novel octanuclear cluster containing a metal-organic framework, [Zn8(ptptp)6Cl2](ClO4)2·4CH3OH·2H2O (1b), induced by ligand rearrangement during a solvothermal process. The luminescent properties of 1b indicate that the material can act as a selective probe toward Pb2+ and Cr2O7 2-.

12.
Artigo em Inglês | MEDLINE | ID: mdl-31326828

RESUMO

Olfaction contributes to many crucial behaviors in insects, such as foraging, locating hosts, mating, and avoiding predators. In the first step of the olfaction process in insects, odorant-binding proteins (OBPs) bind with the odorants and transport hydrophobic odorants. OBPs are also believed to accelerate the termination of the odorant response. The oriental fruit fly, Bactrocera dorsalis, is one of the most destructive fruit-eating pests, causing enormous economic losses to the fruit and vegetable industry worldwide. However, information relating to the number, diversity, and expression patterns of OBPs still remains fragmented in this insect pest. Here, we attempted to identify the OBPs in B. dorsalis using genomic and transcriptomic information. In this study, we expanded the repository of B. dorsalis OBPs to 49. Phylogenetic analysis of BdorOBPs with other species revealed that these proteins grouped into four subfamilies. Furthermore, we determined the expression profiles in six body parts (namely, the legs, wings, antenna, cuticles of the head, thorax, and abdomen) and five internal tissues (namely, the fat body, midgut, Malpighian tubule, testis, and ovary). The results indicated that 21 BdorOBPs showed high expression levels in the antenna, legs, and head cuticles and may thus perform olfactory functions, which corroborates previous evidence. Two BdorOBPs were specifically expressed in the abdomen cuticles. Nineteen OBPs were highly expressed in the fat body, while four OBPs were highly expressed in the reproductive organs. This indicated that they may have physiological roles other than in chemoreception. In summary, our results contribute to the knowledge base of insect OBPs and provide a foundation for the further study of the molecular mechanisms of chemoreception in B. dorsalis.


Assuntos
Dípteros/genética , Perfilação da Expressão Gênica , Genoma de Inseto , Receptores Odorantes/genética , Animais , Transcrição Gênica
13.
J Assist Reprod Genet ; 36(8): 1609-1621, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31292818

RESUMO

PURPOSE: This study was to evaluate if spent culture media (SCM) of embryos could be used as a non-invasive tool to achieve aneuploidy screening. Ploidy calls, as well as concordance rates between PGT-A results from trophectoderm (TE) and SCM, were compared. Clinical outcomes of single euploid transfers were also evaluated. METHODS: The study was conducted from March 2017 to June 2018 in a university-based ART center. SCM of day 3 to the day(s) of TE biopsy of all biopsied blastocysts were collected for testing. PGT-A results of SCM were compared with the standard results of TE, with clinical relevance and outcomes examined. RESULTS: NiPGT-A using SCM gave a sensitivity of 81.6%, specificity of 48.3%, positive predictive value of 82.6%, and negative predictive value of 46.7% in ploidy calling. The concordance rates for autosomes and sex determination were 62.1% and 82.4%, respectively. There were 14 single embryo transfer cycles of euploids as determined by TE biopsy. Clinical outcomes not only confirmed 3 false positive results from SCM but also reflected the true ploidy status of the transferred embryo in one case. If ploidy calls were dichotomized without mosaic embryos, the sensitivity and NPV would increase to 91.0% and 66.7% (p = 0.60 and p = 0.25), respectively. CONCLUSIONS: Cell-free DNA found in SCM could provide ploidy information of an embryo as in PGT-A from its TE. Given its potential to reflect the comprehensive chromosomal profile of the whole embryo, more research based on clinical outcomes is required to determine if SCM could be a reliable selection tool in PGT-A.


Assuntos
Aneuploidia , Meios de Cultura/metabolismo , Fertilização in vitro , Testes Genéticos/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Diagnóstico Pré-Implantação/métodos , Trofoblastos/metabolismo , Técnicas de Cultura Embrionária , Feminino , Humanos , Recuperação de Oócitos , Indução da Ovulação , Gravidez , Estudos Prospectivos , Trofoblastos/citologia
14.
Insects ; 10(7)2019 Jun 27.
Artigo em Inglês | MEDLINE | ID: mdl-31252564

RESUMO

Glutamine synthetase (GS) is a key enzyme in glutamine synthesis and is associated with multiple physiological processes in insects, such as embryonic development, heat shock response, and fecundity regulation. However, little is known about the influence of GS on female fecundity in the oriental fruit fly, Bactrocera dorsalis. Based on the cloning of BdGSs, mitochondrial BdGSm and cytoplasmic BdGSc, we determined their expressions in the tissues of adult B. dorsalis. BdGSm was highly expressed in the fat body, while BdGSc was highly expressed in the head and midgut. Gene silencing by RNA interference against two BdGSs isoforms suppressed target gene expression at the transcriptional level, leading to a reduced ovarian size and lower egg production. The specific inhibitor L-methionine S-sulfoximine suppressed enzyme activity, but only the gene expression of BdGSm was suppressed. A similar phenotype of delayed ovarian development occurred in the inhibitor bioassay. Significantly lower expression of vitellogenin and vitellogenin receptor was observed when GS enzyme activity was suppressed. These data illustrate the effects of two GS genes on adult fecundity by regulating vitellogenin synthesis in different ways.

15.
Int J Mol Sci ; 20(9)2019 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-31035645

RESUMO

Pyrimidine nucleotides are important metabolites that are building blocks of nucleic acids, which participate in various aspects of plant development. Only a few genes involved in pyrimidine metabolism have been identified in rice and the majority of their functions remain unclear. In this study, we used a map-based cloning strategy to isolate a UMPK gene in rice, encoding the UMP kinase that phosphorylates UMP to form UDP, from a recessive mutant with pale-green leaves. In the mutant, UDP content always decreased, while UTP content fluctuated with the development of leaves. Mutation of UMPK reduced chlorophyll contents and decreased photosynthetic capacity. In the mutant, transcription of plastid-encoded RNA polymerase-dependent genes, including psaA, psbB, psbC and petB, was significantly reduced, whereas transcription of nuclear-encoded RNA polymerase-dependent genes, including rpoA, rpoB, rpoC1, and rpl23, was elevated. The expression of UMPK was significantly induced by various stresses, including cold, heat, and drought. Increased sensitivity to cold stress was observed in the mutant, based on the survival rate and malondialdehyde content. High accumulation of hydrogen peroxide was found in the mutant, which was enhanced by cold treatment. Our results indicate that the UMP kinase gene plays important roles in regulating chloroplast development and stress response in rice.


Assuntos
Cloroplastos/fisiologia , Resposta ao Choque Frio , Núcleosídeo-Fosfato Quinase/metabolismo , Oryza/fisiologia , Desenvolvimento Vegetal , Clonagem Molecular , Resposta ao Choque Frio/genética , Regulação da Expressão Gênica de Plantas , Mutação , Núcleosídeo-Fosfato Quinase/genética , Fenótipo , Desenvolvimento Vegetal/genética , Plastídeos/genética , Transcrição Gênica
16.
Insects ; 10(3)2019 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-30893923

RESUMO

Peptidoglycans (PGNs) are major bacterial components recognized by the immune systems of insects and mammals. PGN recognition proteins (PGRPs) are widely distributed and highly conserved in vertebrates and invertebrates. PGRPs are a family of pattern recognition receptors that recognize peptidoglycan and regulate immune responses. In this study, we cloned two PGRP genes (BdPGRP-SA and BdPGRP-SD) from Bactrocera dorsalis (Hendel), which encode 192 and 196 amino acid residues, respectively. Both genes were highly expressed in adults, especially in the fat body and midgut. These two genes were up-regulated when challenged by the immune triggers, PGN-EB (Escherichia coli O111:B4) and PGN-SA (Staphylococcus aureus). The suppression of transcriptional expression of either gene by RNA interference (RNAi) resulted in increased sensitivities to Gram-negative E. coli and Gram-positive S. aureus PGNs. Suppression of BdPGRP-SA and -SD expression by RNAi resulted in weak expressions of four antimicrobial peptides (AMPs) upon injected with E. coli or S. aureus. BdPGRP-SA and -SD are involved in recognizing both Gram-negative and Gram-positive bacteria independently to activate the downstream AMP's response to bacterial infection.

17.
Artigo em Inglês | MEDLINE | ID: mdl-30909163

RESUMO

The oriental fruit fly, Bactrocera dorsalis, is one of the most destructive pests worldwide. The frequent use of chemical insecticides has led B. dorsalis to develop resistance to many insecticides in recent decades. New high-throughput-sequenced transcriptomes, as well as genomes, have revealed a large number of reference genes for functional target identification. Here, we performed digital gene expression profiling of ovary and testis of B. dorsalis adults. Various genes were identified to be highly expressed in B. dorsalis ovary. The genes encoding components of eggshell, vitelline membrane proteins (Vmps) and chorion-related proteins, were identified to be tissue-specifically expressed in ovary. Five cytochrome P450 genes were also identified to be highly expressed in ovary. Three of them were ecdysone synthesis pathway genes indicating the ovary as a potential synthesis site of female. The up-regulated expression of Vmps by exogenous 20-hydroxyecdysone implied the hormonal regulation of eggshell formation during ovarian development. Many other genes with potential functions in ovarian development were also identified, including vitellogenin receptor, insulin receptor, NASP protein, and odorant binding protein. These findings should promote our understanding of the regulation of vitellogenesis and eggshell formation and enable exploration of potentially novel pest control targets.


Assuntos
Ecdisterona/metabolismo , Proteínas do Ovo/genética , Proteínas de Insetos/genética , Tephritidae/genética , Transcriptoma , Animais , Sistema Enzimático do Citocromo P-450/genética , Feminino , Perfilação da Expressão Gênica , Ovário/metabolismo , Tephritidae/metabolismo
18.
Biomed Res Int ; 2018: 8518631, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29707579

RESUMO

BACKGROUND: Persistent postural-perceptual dizziness (PPPD) is a common vestibular disorder. This study was conducted to assess whether the addition of cognitive behavior therapy (CBT) could significantly improve the efficacy and acceptability of sertraline in treating PPPD. METHODS: PPPD patients were recruited and randomly assigned to control and experiment groups. Patients in both groups received sertraline 50-200 mg/day, and only patients in the experiment group received CBT (twice a week, one hour per time). The treatment was continued for eight weeks. At baseline, week 2, week 4, and week 8, the 25-item Dizziness Handicap Inventory (DHI), Hamilton Anxiety Rating Scale (HARS), and Hamilton Depression Rating Scale (HDRS) were used to assess the self-perceived handicapping effects caused by PPPD, anxiety, and depressive symptoms, respectively. The dose of sertraline used and the adverse events in both groups were recorded and analyzed. RESULTS: In total, 91 PPPD patients were randomly assigned to the control group (n = 45) and experiment group (n = 46). After eight weeks of treatment, the average DHI scores, HDRS scores, and HARS scores were significantly decreased in both groups. But compared to the control group, the experiment group had significantly lower average DHI score, HDRS score, and HARS score at weeks 4 and 8. Moreover, the dose of sertraline used in the experiment group was significantly lower than that in the control group, and adverse events occurred more frequently in the control group than in the experiment group (48.9% versus 26.1%, p = 0.025). CONCLUSION: These results demonstrated that the addition of CBT could significantly improve the efficacy and acceptability of sertraline in treating PPPD and reduce the dose of sertraline used.


Assuntos
Terapia Cognitivo-Comportamental , Tontura , Equilíbrio Postural , Sertralina/administração & dosagem , Doenças Vestibulares , Adolescente , Adulto , Tontura/fisiopatologia , Tontura/terapia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sertralina/efeitos adversos , Doenças Vestibulares/fisiopatologia , Doenças Vestibulares/terapia
19.
Surg Innov ; 25(3): 230-235, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29671380

RESUMO

BACKGROUND AND OBJECTIVE: Transilluminated powered phlebectomy using the TriVex system is a new procedure for minimally invasive varicose vein surgery. We used the TriVex system for the removal of benign superficial subcutaneous soft tissue masses, and we also achieved satisfactory clinical effects. The objective of this study was to explore the feasibility, benefits, and technique-related complications of using the TriVex system for the removal of benign superficial masses as a novel and minimally invasive surgical technique. METHOD: Between January 2010 and August 2016, 452 patients with benign superficial soft tissue masses underwent surgical removal of masses using the TriVex II system in our department. The surgical complications, postoperative cosmetic outcome, and personal satisfaction were reviewed at a mean follow-up of 24 months. RESULTS: The TriVex procedure was successfully completed in all patients using only local tumescent anesthesia without any technical problem and serious complications. The mean operation duration was 8.5 minutes, and the average blood loss was 5 mL. Surgical complications were observed in 69 cases (15.3%) of subcutaneous ecchymosis, 34 cases (7.5%) of skin perforation, 14 cases (3.1%) of subcutaneous hematoma, 13 cases (2.9%) of wound infection, and 7 cases (1.5%) of induration of operation area. Eleven patients (2.4%) showed recurrence during the follow-up and were cured by reoperation by the TriVex system. Overall, the majority of patients (N = 436, 96.5%) were very satisfied or satisfied with the outcome. CONCLUSION: The TriVex procedure for the removal of benign superficial masses is simple, safe, and effective with advantages of short operation time, small incisions, and good cosmetic outcome, which is predicted as a new minimally invasive surgery of superficial masses.


Assuntos
Procedimentos Cirúrgicos Minimamente Invasivos , Procedimentos Cirúrgicos Robóticos , Neoplasias de Tecidos Moles/cirurgia , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Cirúrgicos Minimamente Invasivos/efeitos adversos , Procedimentos Cirúrgicos Minimamente Invasivos/métodos , Procedimentos Cirúrgicos Minimamente Invasivos/estatística & dados numéricos , Complicações Pós-Operatórias/epidemiologia , Procedimentos Cirúrgicos Robóticos/efeitos adversos , Procedimentos Cirúrgicos Robóticos/métodos , Procedimentos Cirúrgicos Robóticos/estatística & dados numéricos , Adulto Jovem
20.
Biomed Pharmacother ; 99: 1-8, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29324307

RESUMO

BACKGROUND: Cell based therapy has been heralded as a novel, promising therapeutic strategy for cardiovascular diseases including pulmonary arterial hypertension (PAH). However, the low survival rate after transplantation due to cell death via anoikis is a major obstacle in stem cell therapy. Cells adhesion via Integrin alpha5beta1 (ITGA5B1) has a tendency to exert higher maximum forces. The present study aimed to evaluate the potential protective effect of ITGA5B1 on rat bone marrow mesenchymal stem cells (rBMSCs) from anoikis. METHODS: Mononuclear cells were isolated by density gradient centrifugation with Ficoll, and rBMSCs cell surface markers were evaluated by flow cytometry. Osteogenic and adipocyte differentiation was determined by Alizarin Red S and Oil Red O staining respectively. The expression of Integrin A5 (ITGA5), Integrin B1 (ITGB1), eNOS and actived-caspase-3 mRNA or protein was confirmed by qPCR and western-blot. Cell adhesion, cell viability, anoikis and the migration of rBMSCs were also evaluated. Nitric oxide (NO) production was detected by the greiss assay. RESULTS: Co-infected with Integrin A5 and B1 lentivirus to rBMSCs increased ITGA5 and ITGB1 mRNA and protein expression. ITGA5B1 enhanced the cell adhesion, cell viability, cell migration and NO production but reduced the cell anoikis in rBMSCs/ITGA5B1 groups. CONCLUSION: Transduction of rat rBMSCs with ITGA5B1 lentivirus could prevent cell anoikis and increase NO production.


Assuntos
Anoikis , Integrina alfa5beta1/metabolismo , Células-Tronco Mesenquimais/metabolismo , Óxido Nítrico/biossíntese , Animais , Adesão Celular , Movimento Celular , Sobrevivência Celular , Células Cultivadas , Masculino , Células-Tronco Mesenquimais/citologia , Ratos , Transdução Genética , Cicatrização
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