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1.
BMC Vet Res ; 20(1): 275, 2024 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-38918814

RESUMO

Transgene silencing provides a significant challenge in animal model production via gene engineering using viral vectors or transposons. Selecting an appropriate strategy, contingent upon the species is crucial to circumvent transgene silencing, necessitating long-term observation of in vivo gene expression. This study employed the PiggyBac transposon to create a GFP rat model to address transgene silencing in rats. Surprisingly, transgene silencing occurred while using the CAG promoter, contrary to conventional understanding, whereas the Ef1α promoter prevented silencing. GFP expression remained stable through over five generations, confirming efficacy of the Ef1α promoter for long-term protein expression in rats. Additionally, GFP expression was consistently maintained at the cellular level in various cellular sources produced from the GFP rats, thereby validating the in vitro GFP expression of GFP rats. Whole-genome sequencing identified a stable integration site in Akap1 between exons 1 and 2, mitigating sequence-independent mechanism-mediated transgene silencing. This study established an efficient method for producing transgenic rat models using PiggyBac transposon. Our GFP rats represent the first model to exhibit prolonged expression of foreign genes over five generations, with implications for future research in gene-engineered rat models.


Assuntos
Elementos de DNA Transponíveis , Proteínas de Fluorescência Verde , Ratos Transgênicos , Animais , Elementos de DNA Transponíveis/genética , Proteínas de Fluorescência Verde/genética , Ratos , Técnicas de Transferência de Genes/veterinária , Transgenes , Masculino , Inativação Gênica , Feminino , Regiões Promotoras Genéticas
2.
Int J Mol Sci ; 23(18)2022 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-36142294

RESUMO

Lycopene epsilon-cyclase (LcyE) is a key enzyme in the carotenoid biosynthetic pathway of higher plants. Using the CRSPR/Cas9 and the geminiviral replicon, we optimized a method for targeted mutagenesis and golden SNP replacement of the LcyE gene in rice. We have exploited the geminiviral replicon amplification as a means to provide a large amount of donor template for the repair of a CRISPR-Cas-induced DNA double-strand break (DSB) in the target gene via homology-directed repair (HDR). Mutagenesis experiments performed on the Donggin variety achieved precise modification of the LcyE loci with an efficiency of up to 90%. In HDR experiments, our target was the LcyE allele (LcyE-H523L) derived from anther culture containing a golden SNP replacement. The phenotype of the homologous recombination (HR) mutant obtained through the geminiviral replicon-based template delivery system was tangerine color, and the frequency was 1.32% of the transformed calli. In addition, the total carotenoid content of the LcyEsg2-HDR1 and LcyEsg2-HDR2 lines was 6.8-9.6 times higher than that of the wild-type (WT) calli, respectively. The reactive oxygen species content was lower in the LcyEsg2-HDR1 and LcyEsg2-HDR2 lines. These results indicate that efficient HDR can be achieved in the golden SNP replacement using a single and modular configuration applicable to different rice targets and other crops. This work demonstrates the potential to replace all genes with elite alleles within one generation and greatly expands our ability to improve agriculturally important traits.


Assuntos
Edição de Genes , Oryza , Sistemas CRISPR-Cas , Carotenoides , DNA , Edição de Genes/métodos , Liases Intramoleculares , Oryza/genética , Espécies Reativas de Oxigênio , Replicon/genética
3.
BMC Vet Res ; 17(1): 44, 2021 Jan 22.
Artigo em Inglês | MEDLINE | ID: mdl-33482811

RESUMO

BACKGROUND: Murine is the most abundantly used as laboratory animal models. There has been a tremendous amount of research including; their evolution, growth, physiology, disease modeling as well as genomic mapping. Rats and mice are the most widely used among them. Although both rats and mice fall under the same category still both are different a lot too. As regarding in vitro maturation and development mouse studies are well established as compared to rats which still lies in the early phase of development. So, we tried to figure out rat oocytes in vitro maturation and their developmental potential by performing 3 experiments i.e. superovulation, in vitro Maturation as simple culture (COC's only), and COC's & cumulus cells co-culture, which later further developed using parthenogenetic activation after IVM. Female Sprague Dawley rat 3-4 week used for these studies, we hyper-stimulated their ovaries using PMSG and hCG 150 IU/kg each. After that, we collected ovaries via dissection and retrieved oocytes. We matured them in TCM 199 supplemented with FSH, Estrogen, EGF, and Pyruvate. After maturation, we activated them using two types of activators i.e. Ethanol 7%, Ionomycin. After that, we saw and compared their developmental potential in vitro. RESULTS: Oocytes matured in COC's and Cumulus cell monolayer co-culture (59% ± 4*) showed significantly more even growth and extrusion of the first polar body as compared to the COC's only culture (53.8 ± 7%*). While oocytes activated using Ionomycin showed more promising development until 8 cells/blastocyst level compared to ethanol 7%. CONCLUSION: we concluded that COC's and cumulus monolayer co-culture is better than COC's only culture. Cumulus monolayer provides extra aid in the absorption of nutrients and supplements thus providing a better environment for oocytes growth. Also, we concluded that matured oocytes showed more developmental capacity after activation via ionomycin compared to ethanol.


Assuntos
Técnicas de Maturação in Vitro de Oócitos/veterinária , Oócitos/fisiologia , Animais , Técnicas de Cocultura/métodos , Técnicas de Cocultura/veterinária , Meios de Cultura , Células do Cúmulo/citologia , Células do Cúmulo/fisiologia , Etanol/farmacologia , Feminino , Técnicas de Maturação in Vitro de Oócitos/métodos , Ionomicina/farmacologia , Oócitos/citologia , Oócitos/efeitos dos fármacos , Partenogênese , Ratos Sprague-Dawley
4.
Emerg Infect Dis ; 26(10): 2465-2468, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32673193

RESUMO

We analyzed reports for 59,073 contacts of 5,706 coronavirus disease (COVID-19) index patients reported in South Korea during January 20-March 27, 2020. Of 10,592 household contacts, 11.8% had COVID-19. Of 48,481 nonhousehold contacts, 1.9% had COVID-19. Use of personal protective measures and social distancing reduces the likelihood of transmission.


Assuntos
Busca de Comunicante/estatística & dados numéricos , Infecções por Coronavirus/epidemiologia , Surtos de Doenças , Pneumonia Viral/epidemiologia , Adolescente , Adulto , Distribuição por Idade , Fatores Etários , Idoso , Idoso de 80 Anos ou mais , Betacoronavirus , COVID-19 , Criança , Pré-Escolar , Infecções por Coronavirus/transmissão , Características da Família , Humanos , Lactente , Recém-Nascido , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/transmissão , República da Coreia/epidemiologia , SARS-CoV-2 , Adulto Jovem
5.
PLoS Genet ; 13(9): e1006981, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28892488

RESUMO

The filamentous fungus Chromocrea spinulosa (Trichoderma spinulosum) exhibits both self-fertile (homothallic) and self-sterile (heterothallic) sexual reproductive behavior. Self-fertile strains produce progeny cohorts that are 50% homothallic, 50% heterothallic. Heterothallic progeny can mate only with homothallic strains, and progeny also segregate 50% homothallic, 50% heterothallic. Sequencing of the mating type (MAT) region of homothallic and heterothallic strains revealed that both carry an intact MAT1-1 locus with three MAT1-1 genes (MAT1-1-1, MAT1-1-2, MAT1-1-3), as previously described for the Sordariomycete group of filamentous fungi. Homothallic strains, however, have a second version of MAT with the MAT1-2 locus genetically linked to MAT1-1. In this version, the MAT1-1-1 open reading frame is split into a large and small fragment and the truncated ends are bordered by 115bp direct repeats (DR). The MAT1-2-1 gene and additional sequences are inserted between the repeats. To understand the mechanism whereby C. spinulosa can exhibit both homothallic and heterothallic behavior, we utilized molecular manipulation to delete one of the DRs from a homothallic strain and insert MAT1-2 into a heterothallic strain. Mating assays indicated that: i) the DRs are key to homothallic behavior, ii) looping out of MAT1-2-1 via intra-molecular homologous recombination between the DRs in self-fertile strains results in two nuclear types in an individual (one carrying both MAT1-1 and MAT1-2 and one carrying MAT1-1 only), iii) self-fertility is achieved by inter-nuclear recognition between these two nuclear types before meiosis, iv) the two types of nuclei are in unequal proportion, v) having both an intact MAT1-1-1 and MAT1-2-1 gene in a single nucleus is not sufficient for self-fertility, and vi) the large truncated MAT1-1-1 fragment is expressed. Comparisons with MAT regions of Trichoderma reesei and Trichoderma virens suggest that several crossovers between misaligned parental MAT chromosomes may have led to the MAT architecture of homothallic C. spinulosa.


Assuntos
Proteínas Fúngicas/genética , Genes Fúngicos Tipo Acasalamento/genética , Reprodução/genética , Trichoderma/genética , Núcleo Celular/genética , Citoplasma/genética , Fertilidade/genética , Meiose/genética , Filogenia , Sequências Repetitivas de Ácido Nucleico/genética , Trichoderma/crescimento & desenvolvimento
6.
J Korean Med Sci ; 35(31): e288, 2020 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-32776726

RESUMO

BACKGROUND: In February 2020, a coronavirus disease 2019 (COVID-19) outbreak was reported in fitness centers in Cheonan, Korea. METHODS: From February 24 to March 13, an epidemiological investigation was conducted on the fitness center outbreak. All those who were screened were tested for severe acute respiratory syndrome coronavirus-2 (SARS CoV-2) using real-time reverse transcriptase polymerase chain reaction. Contacts were traced and self-isolated for 14 days. We determined the epidemiological characteristics of confirmed cases of SARS-CoV-2 infection, and estimated the time-dependent reproduction number to assess the transmission dynamics of the infection. RESULTS: A total of 116 cases were confirmed, and 1,687 contacts were traced. The source cases were 8 Zumba instructors who led aerobics classes in 10 fitness centers, and had the largest average number of contacts. A total of 57 Zumba class participants, 37 of their family members, and 14 other contacts were confirmed as cases. The attack rate was 7.3%. The contacts at Zumba classes and homes had a higher attack rate than other contacts. The mean serial interval (± standard deviation) were estimated to be 5.2 (± 3.8) days. The time-dependent reproduction number was estimated to be 6.1 at the beginning of the outbreak, but it dropped to less than 1, 2 days after the epidemiological investigation was launched. CONCLUSION: The results suggest that the COVID-19 outbreak was effectively contained with rigorous contact tracing, isolating, and testing in combination with social distancing without a lock-down.


Assuntos
Infecções por Coronavirus/epidemiologia , Pneumonia Viral/epidemiologia , Adulto , Betacoronavirus/genética , Betacoronavirus/isolamento & purificação , COVID-19 , Busca de Comunicante , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Surtos de Doenças , Feminino , Academias de Ginástica , Humanos , Masculino , Pessoa de Meia-Idade , Pandemias , Pneumonia Viral/diagnóstico , Pneumonia Viral/virologia , Quarentena , RNA Viral/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , República da Coreia/epidemiologia , SARS-CoV-2
7.
PLoS Pathog ; 13(10): e1006670, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29073267

RESUMO

Fusarium fujikuroi causes bakanae ("foolish seedling") disease of rice which is characterized by hyper-elongation of seedlings resulting from production of gibberellic acids (GAs) by the fungus. This plant pathogen is also known for production of harmful mycotoxins, such as fusarins, fusaric acid, apicidin F and beauvericin. Recently, we generated the first de novo genome sequence of F. fujikuroi strain IMI 58289 combined with extensive transcriptional, epigenetic, proteomic and chemical product analyses. GA production was shown to provide a selective advantage during infection of the preferred host plant rice. Here, we provide genome sequences of eight additional F. fujikuroi isolates from distant geographic regions. The isolates differ in the size of chromosomes, most likely due to variability of subtelomeric regions, the type of asexual spores (microconidia and/or macroconidia), and the number and expression of secondary metabolite gene clusters. Whilst most of the isolates caused the typical bakanae symptoms, one isolate, B14, caused stunting and early withering of infected seedlings. In contrast to the other isolates, B14 produced no GAs but high amounts of fumonisins during infection on rice. Furthermore, it differed from the other isolates by the presence of three additional polyketide synthase (PKS) genes (PKS40, PKS43, PKS51) and the absence of the F. fujikuroi-specific apicidin F (NRPS31) gene cluster. Analysis of additional field isolates confirmed the strong correlation between the pathotype (bakanae or stunting/withering), and the ability to produce either GAs or fumonisins. Deletion of the fumonisin and fusaric acid-specific PKS genes in B14 reduced the stunting/withering symptoms, whereas deletion of the PKS51 gene resulted in elevated symptom development. Phylogenetic analyses revealed two subclades of F. fujikuroi strains according to their pathotype and secondary metabolite profiles.


Assuntos
Fusarium/genética , Fusarium/patogenicidade , Oryza/microbiologia , Doenças das Plantas/microbiologia , Proteínas Fúngicas/biossíntese , Fusariose/genética , Fusarium/metabolismo , Genes Fúngicos/genética , Filogenia , Virulência
8.
PLoS Genet ; 11(9): e1005486, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26334536

RESUMO

Fusarium graminearum, the causal agent of Fusarium head blight in cereal crops, produces sexual progeny (ascospore) as an important overwintering and dissemination strategy for completing the disease cycle. This homothallic ascomycetous species does not require a partner for sexual mating; instead, it carries two opposite mating-type (MAT) loci in a single nucleus to control sexual development. To gain a comprehensive understanding of the regulation of sexual development in F. graminearum, we used in-depth and high-throughput analyses to examine the target genes controlled transcriptionally by two-linked MAT loci (MAT1-1, MAT1-2). We hybridized a genome-wide microarray with total RNAs from F. graminearum mutants that lacked each MAT locus individually or together, and overexpressed MAT1-2-1, as well as their wild-type progenitor, at an early stage of sexual development. A comparison of the gene expression levels revealed a total of 1,245 differentially expressed genes (DEGs) among all of the mutants examined. Among these, genes involved in metabolism, cell wall organization, cellular response to stimuli, cell adhesion, fertilization, development, chromatin silencing, and signal transduction, were significantly enriched. Protein binding microarray analysis revealed the presence of putative core DNA binding sequences (ATTAAT or ATTGTT) for the HMG (high mobility group)-box motif in the MAT1-2-1 protein. Targeted deletion of 106 DEGs revealed 25 genes that were specifically required for sexual development, most of which were regulated transcriptionally by both the MAT1-1 and MAT1-2 loci. Taken together with the expression patterns of key target genes, we propose a regulatory pathway for MAT-mediated sexual development, in which both MAT loci may be activated by several environmental cues via chromatin remodeling and/or signaling pathways, and then control the expression of at least 1,245 target genes during sexual development via regulatory cascades and/or networks involving several downstream transcription factors and a putative RNA interference pathway.


Assuntos
Grão Comestível/microbiologia , Fusarium/crescimento & desenvolvimento , Genes Fúngicos , Meios de Cultura , Grão Comestível/genética , Fusarium/genética , Fusarium/fisiologia , Regulação da Expressão Gênica de Plantas , Transcrição Gênica
9.
J Phys Ther Sci ; 29(2): 290-294, 2017 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28265160

RESUMO

[Purpose] The purpose of this study was to investigate the effects of radiofrequency (RF), electroacupuncture (EA), and low-level laser therapy (LLLT), which are used in physical therapies, on facial wrinkles and moisture. [Subjects and Methods] A total of 30 female participants aged 30-55 years participated in this study and the results will be used as a reference for further development of skin physical therapy. Thirty adult females were assigned to an LLLT (n=10), EA (n=10), or RF group (n=10). The intervention was performed in two 15-minute sessions per week for six weeks. Subjects' skin tone and pigmentation were observed before and after the intervention. [Results] Treatment of the under eye area showed that wrinkles were significantly decreased on both sides after RF, EA, and LLLT. Treatment of the eye rims indicated that wrinkles significantly decreased on the right side after RF, EA, and LLLT. [Conclusion] The application of LLLT, EA, and RF had positive effects on wrinkle and moisture content of adult women's faces.

10.
J Phys Ther Sci ; 28(5): 1407-11, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-27313340

RESUMO

[Purpose] In this study, the effects of low-level laser therapy (LLLT), electroacupuncture (EA), and radiofrequency (RF), which are used in physical therapy, on the pigmentation and skin tone of adult women's faces were investigated to provide basic data for skin interventions. [Subjects and Methods] Thirty adult females were assigned to either an LLLT group (n=10), an EA group (n=10), or an RF group (n=10). The intervention was performed in two 15-minute sessions per week for six weeks. Subjects' skin tone and pigmentation were observed before and after the intervention. [Results] The EA group showed significant reductions in pigmentation in the left and right eye rims, as well as in the left cheek. The RF group showed significant post-intervention reductions in pigmentation under the left eye, as well as in the left and right eye rims and the left cheek. The LLLT group showed significant increases in skin tone in the forehead and both eye rims. The RF group showed significant increases in skin tone under both eyes. [Conclusion] The application of LLLT, EA, and RF had positive effects on pigmentation and skin tone of adult women's faces.

11.
Gynecol Obstet Invest ; 78(4): 259-65, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25277375

RESUMO

BACKGROUND: Normal fetal development requires adequate folate levels during pregnancy. Although folate metabolic enzymes have important roles in the maintenance of normal fetal development, the location of folate metabolic enzymes, methionine synthase (MTR) and 5,10-methylenetetrahydrofolate reductase (MTHFR), has not been previously examined. METHODS: We investigated the expression of MTR and MTHFR in human term placenta obtained from normal and pregnancy-induced hypertension (PIH) patients. RESULTS: MTR is expressed in the villous syncytiotrophoblast and MTHFR is expressed in the extravillous trophoblast. There was no difference in the quantity and location of these enzymes between control and PIH patients. CONCLUSION: These results suggest that MTR in the villous trophoblast participates in the metabolism of homocysteine by using folate, and MTHFR in the extravillous trophoblast is associated with extratrophoblast invasion.


Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/análise , Hipertensão Induzida pela Gravidez/enzimologia , Metilenotetra-Hidrofolato Redutase (NADPH2)/análise , Placenta/enzimologia , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/metabolismo , Western Blotting , Feminino , Ácido Fólico/metabolismo , Idade Gestacional , Homocisteína/metabolismo , Humanos , Imuno-Histoquímica , Metilenotetra-Hidrofolato Redutase (NADPH2)/genética , Gravidez , RNA Mensageiro/análise , Trofoblastos/enzimologia , Trofoblastos/fisiologia
12.
Vaccine ; 42(7): 1440-1444, 2024 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-38365479

RESUMO

South Korea experienced a low prevalence of SARS-CoV-2 until the emergence of the omicron in early 2022, triggering a major community epidemic. To evaluate effectiveness of NVX-CoV2373 and BNT162b2 vaccines in Korean population, we conducted an observational study utilizing individual-level case data on laboratory-confirmed SARS-CoV-2 infection, along with vaccination record. A total of 47,078 recipients of NVX-CoV2373 vaccine and 7,561 recipients of BNT162b2 vaccine were eligible for the study. Thirty days post-second doses, COVID-19 rates were 7.9% (595 out of 7561) of NVX-CoV2373 recipients and 8.6 % (647 out of 7561) of BNT162b2 recipients experienced COVID-19. NVX-CoV2373 rates increased to 9.8 % and 11.2 % at 60 and 90 days, while BNT162b2 rates were 10.5 % and 11.3 % at the same intervals. The 22-weeks risk ratios for recipients of the NVX-CoV2373 vaccine as compared with recipients of the BNT162b2 vaccine were 1.11 (95 % CI, 0.99 to 1.25) for laboratory-confirmed SARS-CoV-2 infection. Continued monitoring is essential to evaluate the duration of protection across different vaccine platforms and schedules.


Assuntos
Vacinas contra COVID-19 , COVID-19 , Adulto , Humanos , COVID-19/epidemiologia , COVID-19/prevenção & controle , Vacina BNT162 , SARS-CoV-2 , Infecções Irruptivas , Vacinação , República da Coreia/epidemiologia
13.
Microbiology (Reading) ; 158(Pt 7): 1723-1733, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22516221

RESUMO

The velvet genes are conserved in ascomycetous fungi and function as global regulators of differentiation and secondary metabolism. Here, we characterized one of the velvet genes, designated FgVelB, in the plant-pathogenic fungus Fusarium graminearum, which causes fusarium head blight in cereals and produces mycotoxins within plants. FgVelB-deleted (ΔFgVelB) strains produced fewer aerial mycelia with less pigmentation than those of the wild-type (WT) during vegetative growth. Under sexual development conditions, the ΔFgVelB strains produced no fruiting bodies but retained male fertility, and conidiation was threefold higher compared with the WT strain. Production of trichothecene and zearalenone was dramatically reduced compared with the WT strain. In addition, the ΔFgVelB strains were incapable of colonizing host plant tissues. Transcript analyses revealed that FgVelB was highly expressed during the sexual development stage, and may be regulated by a mitogen-activated protein kinase cascade. Microarray analysis showed that FgVelB affects regulatory pathways mediated by the mating-type loci and a G-protein alpha subunit, as well as primary and secondary metabolism. These results suggest that FgVelB has diverse biological functions, probably by acting as a member of a possible velvet protein complex, although identification of the FgVelB-FgVeA complex and the determination of its roles require further investigation.


Assuntos
Proteínas Fúngicas/metabolismo , Fusarium/fisiologia , Regulação Fúngica da Expressão Gênica , Micotoxinas/biossíntese , Doenças das Plantas/microbiologia , Recombinação Genética , Fatores de Virulência/biossíntese , Grão Comestível/microbiologia , Proteínas Fúngicas/genética , Fusarium/genética , Fusarium/metabolismo , Fusarium/patogenicidade , Deleção de Genes , Perfilação da Expressão Gênica , Análise em Microsséries , Micélio/crescimento & desenvolvimento , Pigmentos Biológicos/metabolismo , Esporos Fúngicos/crescimento & desenvolvimento , Virulência
14.
Curr Genet ; 58(3): 179-89, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22531843

RESUMO

Protein-protein interactions play important roles in controlling many cellular events. To date, several techniques have been developed for detection of protein-protein interactions in living cells, among which split luciferase complementation has been applied in animal and plant cells. Here, we examined whether the split luciferase assay could be used in filamentous ascomycetes, such as Gibberella zeae and Cochliobolus heterostrophus. The coding sequences of two strongly interacting proteins (the F-box protein, FBP1, and its partner SKP1) in G. zeae, under the control of the cryparin promoter from Cryphonectria parasitica, were translationally fused to the C- and N-terminal fragments of firefly luciferase (luc), respectively. Each fusion product inserted into a fungal transforming vector carrying the gene for resistance to either geneticin or hygromycin B, was transformed into both fungi. We detected complementation of split luciferase proteins driven by interaction of the two fungal proteins with a high luminescence intensity-to-background ratio only in the fungal transformants expressing both N-luc and C-luc fusion constructs. Using this system, we also confirmed a novel protein interaction between transcription factors, GzMCM1 and FST12 in G. zeae, which could hardly be proven by the yeast two-hybrid method. This is the first study demonstrating that monitoring of split luciferase complementation is a sensitive and efficient method of studying in vivo protein-protein interactions in filamentous ascomycetes.


Assuntos
Ascomicetos/metabolismo , Proteínas Fúngicas/metabolismo , Luciferases de Vaga-Lume/genética , Mapeamento de Interação de Proteínas/métodos , Ascomicetos/genética , Frutose-Bifosfatase/genética , Frutose-Bifosfatase/metabolismo , Proteínas Fúngicas/genética , Ordem dos Genes , Teste de Complementação Genética , Luciferases de Vaga-Lume/metabolismo , Plasmídeos , Ligação Proteica , Proteínas Quinases Associadas a Fase S/genética , Proteínas Quinases Associadas a Fase S/metabolismo , Transformação Genética
15.
Plants (Basel) ; 12(1)2022 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-36616184

RESUMO

We generated an orange-colored (OC) rice callus line by targeted mutagenesis of the orange gene (OsOr) using the CRISPR-Cas9 system. The OC line accumulated more lutein, ß-carotene, and two ß-carotene isomers compared to the WT callus line. We also analyzed the expression levels of carotenoid biosynthesis genes by qRT-PCR. Among the genes encoding carotenoid metabolic pathway enzymes, the number of transcripts of the PSY2, PSY3, PDS, ZDS and ß-LCY genes were higher in the OC line than in the WT line. In contrast, transcription of the ε-LCY gene was downregulated in the OC line compared to the WT line. In addition, we detected increases in the transcript levels of two genes involved in carotenoid oxidation in the OC lines. The developed OC lines also showed increased tolerance to salt stress. Collectively, these findings indicate that targeted mutagenesis of the OsOr gene via CRISPR/Cas9-mediated genome editing results in ß-carotene accumulation in rice calli. Accordingly, we believe that this type of genome-editing technology could represent an effective alternative approach for enhancing the ß-carotene content of plants.

16.
Korean J Hepatol ; 17(1): 19-26, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21494074

RESUMO

BACKGROUND/AIMS: Abdominal ultrasonography is useful for the detection and diagnosis of nonalcoholic fatty liver disease (NAFLD). The aims of this study were to establish a predictive model for the selection of subjects for abdominal ultrasonography for the diagnosis of NAFLD and to assess validity of the model. METHODS: The subjects included 901 people who visited the health examination center of the Busan Medical Center. We conducted multiple logistic regression analyses of potential risk factors to identify independent risk factors for NAFLD, and developed an index system. RESULTS: Four independent risk factors were identified. The index system was developed by assigning 1 clinical scoring point to approximately 0.7 logistic regression coefficients to each factor as follows: alanine aminotransferase/aspartate aminotransferase ratio >1.5 (odds ratio [OR], 2.22; 95% confidence interval [CI], 1.21-4.07; P=0.010), 1 point; γ-glutamyl transpeptidase >50 (OR, 2.15; 95% CI, 1.13-4.07; P=0.019), 1 point; triglyceride >150 mg/dL (OR, 1.92; 95% CI, 1.14-3.24; P=0.015), 1 point; 23 kg/m(2)≤BMI⟨25 kg/m(2) (OR, 3.68; 95% CI, 2.05-6.63; P<0.001), 2 points; and BMI 25 kg/m(2) (OR, 7.65; 95% CI, 4.29-13.62; P<0.001), 3 points. The area under the receiver operating characteristics curve was 0.797 (95% CI, 0.751-0.842), and when 3 points was used as a cut-off value, the sensitivity and specificity were 71.7% and 75.9%, respectively. CONCLUSIONS: NAFLD can be predicted through the clinical application of the index system established herein. If abdominal ultrasonography is used for high-risk patients, NAFLD will be diagnosed and managed in its early stage.


Assuntos
Fígado Gorduroso/diagnóstico , Adulto , Idoso , Alanina Transaminase/sangue , Área Sob a Curva , Aspartato Aminotransferases/sangue , Índice de Massa Corporal , Fígado Gorduroso/diagnóstico por imagem , Feminino , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica , Razão de Chances , Valor Preditivo dos Testes , Fatores de Risco , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Ultrassonografia , gama-Glutamiltransferase/sangue
17.
Vaccines (Basel) ; 9(4)2021 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-33920117

RESUMO

Increased awareness of adverse events following immunization (AEFI) can disrupt vaccination programs. In South Korea, a report of alleged influenza vaccine-related deaths attracted significant media attention in 2020. We retrieved the vaccination coverage and AEFI data to determine their association with media coverage. Between 2015 and 2019, the vaccination coverage rate ranged between 80.5% and 83.3%; however, the vaccination coverage rate declined significantly from 2020 to 2021 to 73.6% (p < 0.0001). During the 43rd week of 2020, following a large amount of media coverage on vaccine safety issues, the number of cases with AEFI reached 60. Between 2015 and 2020, the mortality rate ratios for influenza vaccines and non-vaccines ranged between 0.1296 (95% confidence interval (CI), 0.1262-0.1331, p < 0.0001) and 0.1608 (95% CI, 0.1572-0.1644, p < 0.0001). Vaccine safety surveillance should be continued in conjunction with investigation and transparent risk communication to maintain public trust in vaccines and vaccinations.

18.
Sci Rep ; 10(1): 21257, 2020 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-33277567

RESUMO

In vitro plant regeneration involves a two-step practice of callus formation and de novo organogenesis. During callus formation, cellular competence for tissue regeneration is acquired, but it is elusive what molecular processes and genetic factors are involved in establishing cellular pluripotency. To explore the mechanisms underlying pluripotency acquisition during callus formation in monocot plants, we performed a transcriptomic analysis on the pluripotent and non-pluripotent rice calli using RNA-seq. We obtained a dataset of differentially expressed genes (DEGs), which accounts for molecular processes underpinning pluripotency acquisition and maintenance. Core regulators establishing root stem cell niche were implicated in pluripotency acquisition in rice callus, as observed in Arabidopsis. In addition, KEGG analysis showed that photosynthetic process and sugar and amino acid metabolism were substantially suppressed in pluripotent calli, whereas lipid and antioxidant metabolism were overrepresented in up-regulated DEGs. We also constructed a putative coexpression network related to cellular pluripotency in rice and proposed potential candidates conferring pluripotency in rice callus. Overall, our transcriptome-based analysis can be a powerful resource for the elucidation of the molecular mechanisms establishing cellular pluripotency in rice callus.


Assuntos
Oryza/genética , Sementes/metabolismo , Regulação da Expressão Gênica de Plantas/genética , RNA-Seq , Nicho de Células-Tronco/fisiologia , Transcriptoma/genética
19.
20.
Sci Rep ; 10(1): 11148, 2020 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-32636400

RESUMO

Thyroid hormone (TH) has long been believed to play a minor role in male reproduction. However, evidences from experimental model of thyrotoxicosis or hypothyroidism suggests its role in spermatogenesis. Cellular action of TH requires membrane transport via specific transporters such as monocarboxylate transporter 8 (MCT8). SLC16A2 (encodes for MCT8) inactivating mutation in humans can lead to Allan-Herndon Dudley-syndrome, a X-linked psychomotor and growth retardation. These patients present cryptorchidism which suggests a role of MCT8 during spermatogenesis. In this study, we found that Mct8 is highly expressed during early postnatal development and decreases its expression in the adulthood of testis of wild-type male rats. Histological analysis revealed that spermatogonia largely lacks MCT8 expression while spermatocytes and maturing spermatids highly express MCT8. To further understand the role of Mct8 during spermatogenesis, we generated Slc16a2 (encodes MCT8) knockout rats using CRISPR/Cas9. Serum THs (T3 and T4) level were significantly altered in Slc16a2 knockout rats when compared to wild-type littermates during early to late postnatal development. Unlike Slc16a2 knockout mice, Slc16a2 knockout rats showed growth delay during early to late postnatal development. In adult Slc16a2 knockout rats, we observed reduced sperm motility and viability. Collectively, our data unveil a functional involvement of MCT8 in spermatogenesis, underscoring the importance of TH signaling and action during spermatogenesis.


Assuntos
Transportadores de Ácidos Monocarboxílicos/fisiologia , Espermatozoides/crescimento & desenvolvimento , Testículo/crescimento & desenvolvimento , Animais , Proteína 9 Associada à CRISPR , Sistemas CRISPR-Cas , Edição de Genes/métodos , Técnicas de Silenciamento de Genes/métodos , Masculino , Transportadores de Ácidos Monocarboxílicos/genética , Transportadores de Ácidos Monocarboxílicos/metabolismo , Ratos , Ratos Sprague-Dawley , Espermatogênese/genética , Espermatogênese/fisiologia , Espermatozoides/fisiologia , Testículo/metabolismo , Glândula Tireoide/metabolismo , Glândula Tireoide/fisiologia
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