Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 10.112
Filtrar
1.
J Exp Med ; 217(11)2020 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-32692348

RESUMO

The emergence of SARS-CoV-2 and the ensuing explosive epidemic of COVID-19 disease has generated a need for assays to rapidly and conveniently measure the antiviral activity of SARS-CoV-2-specific antibodies. Here, we describe a collection of approaches based on SARS-CoV-2 spike-pseudotyped, single-cycle, replication-defective human immunodeficiency virus type-1 (HIV-1), and vesicular stomatitis virus (VSV), as well as a replication-competent VSV/SARS-CoV-2 chimeric virus. While each surrogate virus exhibited subtle differences in the sensitivity with which neutralizing activity was detected, the neutralizing activity of both convalescent plasma and human monoclonal antibodies measured using each virus correlated quantitatively with neutralizing activity measured using an authentic SARS-CoV-2 neutralization assay. The assays described herein are adaptable to high throughput and are useful tools in the evaluation of serologic immunity conferred by vaccination or prior SARS-CoV-2 infection, as well as the potency of convalescent plasma or human monoclonal antibodies.


Assuntos
Anticorpos Neutralizantes/análise , Anticorpos Antivirais/análise , Betacoronavirus/imunologia , Infecções por Coronavirus/imunologia , Imunoensaio/métodos , Pneumonia Viral/imunologia , Animais , Anticorpos Monoclonais/imunologia , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Betacoronavirus/genética , Linhagem Celular , Quimera/genética , Quimera/imunologia , Chlorocebus aethiops , Infecções por Coronavirus/virologia , Células HEK293 , HIV-1/genética , HIV-1/imunologia , Humanos , Testes de Neutralização/métodos , Pandemias , Pneumonia Viral/virologia , Recombinação Genética , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/imunologia , Células Vero , Vírus da Estomatite Vesicular Indiana/genética , Vírus da Estomatite Vesicular Indiana/imunologia
2.
Science ; 368(6492): 731-736, 2020 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-32409469

RESUMO

The establishment of reproductive barriers between populations can fuel the evolution of new species. A genetic framework for this process posits that "incompatible" interactions between genes can evolve that result in reduced survival or reproduction in hybrids. However, progress has been slow in identifying individual genes that underlie hybrid incompatibilities. We used a combination of approaches to map the genes that drive the development of an incompatibility that causes melanoma in swordtail fish hybrids. One of the genes involved in this incompatibility also causes melanoma in hybrids between distantly related species. Moreover, this melanoma reduces survival in the wild, likely because of progressive degradation of the fin. This work identifies genes underlying a vertebrate hybrid incompatibility and provides a glimpse into the action of these genes in natural hybrid populations.


Assuntos
Ciprinodontiformes/genética , Doenças dos Peixes/genética , Proteínas de Peixes/genética , Hibridização Genética , Melanoma/genética , Melanoma/virologia , Receptores Proteína Tirosina Quinases/genética , Alelos , Nadadeiras de Animais/patologia , Animais , Quimera , Loci Gênicos , Estudo de Associação Genômica Ampla
3.
Gene ; 752: 144791, 2020 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-32439378

RESUMO

Prkaca consists of the catalytic subunit alpha protein kinase A (PKA), which is involved in many cellular processes. In this study, the cDNA and genomic sequences of prkaca in tilapia hybrids (Oreochromis mossambicus × Oreochromis hornorum) were cloned and analysed. The results showed the prkaca gene consists of 11 exons and 10 introns, and its protein contains 351 amino acid residues and is clustered with Oreochromis niloticus, Maylandia zebra and Haplochromis burtoni first in a phylogenetic tree. Amino acid alignment indicates that prkaca shares the highest identity (100%) to Oreochromis niloticus, Maylandia zebra and Haplochromis burtoni. Two CpG islands of prkaca were found by MethPrimer software, and 32 CG sites were found in the proximal promoter. The methylation level of prkaca in the hybrids (0.31%) was significantly lower than that of their parents (0.94% and 3.43%) in kidney tissue (P < 0.05). The gene expression levels and DNA methylation levels of prkaca in muscle and kidney tissues of the tilapia hybrids were detected by quantitative real-time PCR and bisulfite sequencing PCR and showed a negative correlation under saline-alkali stress. The results of this research demonstrated that DNA methylation levels and prkaca mRNA expression levels were inversely correlated under saline-alkali stress, implying that heterosis is likely accompanied by DNA methylation alterations. This research provides new clues for further investigations of DNA methylation and heterosis in hybrid fish.


Assuntos
Subunidades Catalíticas da Proteína Quinase Dependente de AMP Cíclico/genética , Osmorregulação/genética , Tilápia/genética , Sequência de Aminoácidos/genética , Animais , Quimera/genética , China , Subunidades Catalíticas da Proteína Quinase Dependente de AMP Cíclico/metabolismo , Metilação de DNA/genética , DNA Complementar/genética , Feminino , Genômica , Vigor Híbrido/genética , Masculino , Filogenia , Regiões Promotoras Genéticas/genética , Equilíbrio Hidroeletrolítico/genética
4.
J Parasitol ; 106(2): 316-322, 2020 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-32330281

RESUMO

Fascioliasis is a zoonotic infection of humans and, more commonly, ruminants. It is caused by 2 liver fluke species, Fasciola hepatica and Fasciola gigantica, which differ in size. The traditional morphological methods used to distinguish the 2 species can be unreliable, particularly in the presence of hybrids between the 2 species. The development of advanced molecular methods has allowed for more definitive identification of Fasciola species, including their hybrids. Hybrids are of concern, as it is thought that they could acquire advantageous traits such as increased pathogenicity and host range. In 2013, we collected flukes from Fasciola-positive cattle, sheep, and goats slaughtered in 4 Chadian abattoirs. DNA from 27 flukes was extracted, amplified, and analyzed to identify species using the ITS1+2 locus. Twenty-six of the 27 flukes were identified as F. gigantica, while the remaining fluke showed heterozygosity at all variable sites that distinguish F. hepatica and F. gigantica. Cloning and sequencing of both alleles confirmed the presence of 1 F. hepatica and 1 F. gigantica allele. To our knowledge, this is the first unambiguous, molecular demonstration of the presence of such a hybrid in a bovine in sub-Saharan Africa.


Assuntos
Doenças dos Bovinos/parasitologia , Quimera/genética , Fasciola hepatica/genética , Fasciolíase/veterinária , Matadouros , Animais , Bovinos , Chade , Quimera/classificação , Sequência Consenso , Fasciola/classificação , Fasciola/genética , Fasciola/isolamento & purificação , Fasciola hepatica/classificação , Fasciola hepatica/isolamento & purificação , Fasciolíase/parasitologia , Feminino , Inspeção de Alimentos , Doenças das Cabras/parasitologia , Cabras , Polimorfismo de Nucleotídeo Único/genética , Ovinos , Doenças dos Ovinos/parasitologia
5.
PLoS One ; 15(4): e0230422, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32271764

RESUMO

The inability of beef cattle to maintain full term pregnancies has become an economic concern for the beef industry. Herd management and nutritional improvements have alleviated environmental impacts on embryonic and fetal loss, yet additional gains can be made through genomic selection. The objectives of this study were to identify loci and gene-sets in crossbred beef heifers associated with the number of services required to become pregnant (TBRD) and heifer conception rate at first service (HCR1). Heifers (n = 709) from a commercial beef operation underwent one round of artificial insemination, before exposure to bulls for natural service for 50 days. Pregnancy and time of conception was determined by ultrasound 35 days after the breeding season. Heifers were genotyped using the GeneSeek (Lincoln, NE) Bovine GGP50K BeadChip prior to genome-wide association analyses (GWAA) conducted using an EIGENSTRAT-like model to identify loci associated (P < 1 × 10-5) with TBRD and HCR1. One locus was associated (P = 8.97 × 10-6) with TBRD on BTA19 and included the positional candidate gene ASIC2, which is differentially expressed in the endometrium of fertility classified heifers, and the positional candidate gene, SPACA3. Gene-set enrichment analyses using SNP (GSEA-SNP) data, was performed and identified one gene-set, oxidoreductase activity, acting on paired donors, with incorporation or reduction of molecular oxygen as enriched (NES = 3.15) with TBRD and contained nine leading edge genes that contributed to the enrichment of the gene set. The enriched gene-set is involved in catalyzing oxidation-reduction reactions, which have been associated with oxidative stressors impacting pregnancy success. No loci were associated nor gene-sets enriched with HCR1. Identification of loci, positional candidate genes, gene-sets and leading edge genes enriched for fertility facilitate genomic selection that allows producers to select for reproductively superior cattle, reduce costs associated with infertility, and increase percent calf crop.


Assuntos
Bovinos/genética , Loci Gênicos , Hibridização Genética/genética , Taxa de Gravidez , Prenhez , Reprodução/genética , Animais , Cruzamento , Quimera/genética , Cruzamentos Genéticos , Feminino , Fertilidade/genética , Fertilização/genética , Estudos de Associação Genética/veterinária , Técnicas de Genotipagem , Masculino , Polimorfismo de Nucleotídeo Único , Gravidez , Prenhez/genética
6.
PLoS Negl Trop Dis ; 14(4): e0007143, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32310945

RESUMO

Parasites of the genus Leishmania (Kinetoplastida: Trypanosomatidae) cause widespread and devastating human diseases. Visceral leishmaniasis due to Leishmania donovani is endemic in Ethiopia where it has also been responsible for major epidemics. The presence of hybrid genotypes has been widely reported in surveys of natural populations, genetic variation reported in a number of Leishmania species, and the extant capacity for genetic exchange demonstrated in laboratory experiments. However, patterns of recombination and the evolutionary history of admixture that produced these hybrid populations remain unclear. Here, we use whole-genome sequence data to investigate Ethiopian L. donovani isolates previously characterized as hybrids by microsatellite and multi-locus sequencing. To date there is only one previous study on a natural population of Leishmania hybrids based on whole-genome sequences. We propose that these hybrids originate from recombination between two different lineages of Ethiopian L. donovani occurring in the same region. Patterns of inheritance are more complex than previously reported with multiple, apparently independent, origins from similar parents that include backcrossing with parental types. Analysis indicates that hybrids are representative of at least three different histories. Furthermore, isolates were highly polysomic at the level of chromosomes with differences between parasites recovered from a recrudescent infection from a previously treated individual. The results demonstrate that recombination is a significant feature of natural populations and contributes to the growing body of data that shows how recombination, and gene flow, shape natural populations of Leishmania.


Assuntos
Quimera , Leishmania donovani/genética , Leishmaniose Visceral/parasitologia , Etiópia , Genótipo , Humanos , Recombinação Genética , Sequenciamento Completo do Genoma
7.
Science ; 368(6487): 181-186, 2020 04 10.
Artigo em Inglês | MEDLINE | ID: mdl-32273467

RESUMO

Embryonic development is a complex process that is unamenable to direct observation. In this study, we implanted a window to the mouse uterus to visualize the developing embryo from embryonic day 9.5 to birth. This removable intravital window allowed manipulation and high-resolution imaging. In live mouse embryos, we observed transient neurotransmission and early vascularization of neural crest cell (NCC)-derived perivascular cells in the brain, autophagy in the retina, viral gene delivery, and chemical diffusion through the placenta. We combined the imaging window with in utero electroporation to label and track cell division and movement within embryos and observed that clusters of mouse NCC-derived cells expanded in interspecies chimeras, whereas adjacent human donor NCC-derived cells shrank. This technique can be combined with various tissue manipulation and microscopy methods to study the processes of development at unprecedented spatiotemporal resolution.


Assuntos
Embrião de Mamíferos/citologia , Embrião de Mamíferos/fisiologia , Desenvolvimento Embrionário , Microscopia Intravital/métodos , Crista Neural , Animais , Encéfalo/embriologia , Encéfalo/fisiologia , Divisão Celular , Movimento Celular , Quimera/embriologia , Quimera/fisiologia , Eletroporação , Feminino , Técnicas de Transferência de Genes , Camundongos , Camundongos Transgênicos , Neovascularização Fisiológica , Crista Neural/irrigação sanguínea , Crista Neural/citologia , Crista Neural/embriologia , Placenta/fisiologia , Gravidez , Retina/embriologia , Retina/fisiologia , Transmissão Sináptica , Útero
8.
Nat Commun ; 11(1): 1577, 2020 03 27.
Artigo em Inglês | MEDLINE | ID: mdl-32221280

RESUMO

Microglia, the brain-resident macrophages, exhibit highly dynamic functions in neurodevelopment and neurodegeneration. Human microglia possess unique features as compared to mouse microglia, but our understanding of human microglial functions is largely limited by an inability to obtain human microglia under homeostatic states. Here, we develop a human pluripotent stem cell (hPSC)-based microglial chimeric mouse brain model by transplanting hPSC-derived primitive macrophage progenitors into neonatal mouse brains. Single-cell RNA-sequencing of the microglial chimeric mouse brains reveals that xenografted hPSC-derived microglia largely retain human microglial identity, as they exhibit signature gene expression patterns consistent with physiological human microglia and recapitulate heterogeneity of adult human microglia. Importantly, the engrafted hPSC-derived microglia exhibit dynamic response to cuprizone-induced demyelination and species-specific transcriptomic differences in the expression of neurological disease-risk genes in microglia. This model will serve as a tool to study the role of human microglia in brain development and degeneration.


Assuntos
Encéfalo/citologia , Diferenciação Celular , Quimera/fisiologia , Células-Tronco Pluripotentes Induzidas/citologia , Microglia/citologia , Animais , Linhagem Celular , Cuprizona , Doenças Desmielinizantes/patologia , Feminino , Humanos , Imageamento Tridimensional , Camundongos , Microglia/transplante , RNA-Seq , Análise de Célula Única , Transcriptoma/genética
9.
PLoS One ; 15(3): e0230138, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32176708

RESUMO

Surface-expressed colonization factors and their subunits are promising candidates for inclusion into a multivalent vaccine targeting enterotoxigenic Escherichia coli (ETEC), a leading cause of acute bacterial diarrhea in developing regions. However, soluble antigens are often poorly immunogenic in the absence of an adjuvant. We show here that the serum immune response to CfaE, the adhesin of the ETEC colonization factor CFA/I, can be enhanced in BALB/c mice by immunization with a chimeric antigen containing CfaE and pentameric cholera toxin B subunit (CTB) of cholera toxin from Vibrio cholerae. We constructed this antigen by replacing the coding sequence for the A1 domain of the cholera toxin A subunit (CTA) with the sequence of donor strand complemented CfaE (dscCfaE) within the cholera toxin operon, resulting in a dscCfaE-CTA2 fusion. After expression, via non-covalent interactions between CTA2 and CTB, the fusion and CTB polypeptides assemble into a complex containing a single dscCfaE-CTA2 protein bound to pentameric CTB (dscCfaE-CTA2/CTB). This holotoxin-like chimera retained the GM1 ganglioside binding activity of CTB, as well as the ability of CfaE to mediate the agglutination of bovine red blood cells when adsorbed to polystyrene beads. When administered intranasally to mice, the presence of CTB in the chimera significantly increased the serum immune response to CfaE compared to dscCfaE alone, stimulating a response similar to that obtained with a matched admixture of dscCfaE and CTB. However, by the orogastric route, immunization with the chimera elicited a superior functional immune response compared to an equivalent admixture of dscCfaE and CTB, supporting further investigation of the chimera as an ETEC vaccine candidate.


Assuntos
Toxina da Cólera , Escherichia coli Enterotoxigênica/imunologia , Vacinas contra Escherichia coli/imunologia , Proteínas de Fímbrias , Proteínas Recombinantes de Fusão , Adesinas Bacterianas/imunologia , Adesinas Bacterianas/metabolismo , Adjuvantes Imunológicos/administração & dosagem , Animais , Quimera/genética , Toxina da Cólera/genética , Toxina da Cólera/imunologia , Toxina da Cólera/metabolismo , Feminino , Proteínas de Fímbrias/genética , Proteínas de Fímbrias/imunologia , Proteínas de Fímbrias/metabolismo , Imunização , Imunogenicidade da Vacina , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/metabolismo
10.
Chemosphere ; 249: 126540, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32208221

RESUMO

Hybridization is common between invasive and native species and may be accompanied by invasive evolution. The hybrid of Sphagneticola trilobata (alien invasive species) and Sphagneticola calendulacea (indigenous congener) was found in South China. According to previous studies, the hybrid performed weak environmental adaptability in comparison with parents. However, based on the results from this study, the hybridization significantly improved the tolerance of the hybrid to cadmium (Cd) stress (200 µmol L-1). Under Cd stress, the hybrid lines showed lowest level of oxidative damage and the highest level of photosynthetic efficiency. Compared with the parents, the hybrid utilized more active detoxification strategies, such as the cell walls of the leaves and roots adsorbed 88% and 95% Cd, respectively, reducing the amount of Cd entering cells; moreover, most of the Cd that entered cells was transformed into less toxic chemical forms through the reduction of the highly toxic chemical forms; furthermore, it accumulated a large number of phytochelatins to bind Cd2+ and reduced the damage of organelles by Cd2+. The results demonstrate that hybridization between S. trilobata and S. calendulacea improved the adaptability of the new hybrid species to Cd stress and may pose a greater threat to the survival of the native parent species in the presence of serious water and soil pollution.


Assuntos
Adaptação Fisiológica/fisiologia , Asteraceae/fisiologia , Cádmio/toxicidade , Poluentes do Solo/toxicidade , Adsorção , Asteraceae/metabolismo , Quimera , China , Tolerância a Medicamentos , Espécies Introduzidas , Hibridização de Ácido Nucleico , Oxirredução , Fotossíntese , Fitoquelatinas/metabolismo , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo
11.
PLoS One ; 15(2): e0229503, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32092127

RESUMO

BACKGROUND: Cattleyak are the hybrid offspring between cattle and yak and combine yak hardiness with cattle productivity. Much attempt has been made to examine the mechanisms of male sterility caused by spermatogenic arrest, but yet there is no research systematically and precisely elucidated testis gene expression profiling between cattleyak and yak. METHODS: To explore the higher resolution comparative transcriptome map between the testes of yak and cattleyak, and further analyze the mRNA expression dynamics of spermatogenic arrest in cattleyak. We characterized the comparative transcriptome profile from the testes of yak and cattleyak using high-throughput sequencing. Then we used quantitative analysis to validate several differentially expressed genes (DEGs) in testicular tissue and spermatogenic cells. RESULTS: Testis transcriptome profiling identified 6477 DEGs (2919 upregulated and 3558 downregulated) between cattleyak and yak. Further analysis revealed that the marker genes and apoptosis regulatory genes for undifferentiated spermatogonia were upregulated, while the genes for differentiation maintenance were downregulated in cattleyak. A majority of DEGs associated with mitotic checkpoint, and cell cycle progression were downregulated in cattleyak during spermatogonial mitosis. Furthermore, almost all DEGs related to synaptonemal complex assembly, and meiotic progression presented no sign of expression in cattleyak. Even worse, dozens of genes involved in acrosome formation, and flagellar development were dominantly downregulated in cattleyak. CONCLUSION: DEGs indicated that spermatogenic arrest of cattleyak may originate from the differentiation stage of spermatogonial stem cells and be aggravated during spermatogonial mitosis and spermatocyte meiosis, which contributes to the scarcely presented sperms in cattleyak.


Assuntos
Azoospermia/congênito , Quimera/genética , Infertilidade Masculina/genética , Animais , Azoospermia/genética , Bovinos/genética , Perfilação da Expressão Gênica/métodos , Perfilação da Expressão Gênica/veterinária , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Masculino , Meiose/genética , Espermatócitos/metabolismo , Espermatogênese/genética , Espermatogônias/metabolismo , Espermatozoides/metabolismo , Testículo/metabolismo , Transcriptoma/genética
12.
Hastings Cent Rep ; 50(1): 46, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-32068274
13.
Hastings Cent Rep ; 50(1): 46, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-32068278

RESUMO

The writer responds to the article "Human-Animal Chimeras: The Moral Insignificance of Uniquely Human Capacities," by Julian J. Koplin, in the September-October 2019 issue of the Hastings Center Report.


Assuntos
Quimera , Princípios Morais , Animais , Humanos
14.
Proc Natl Acad Sci U S A ; 117(10): 5329-5338, 2020 03 10.
Artigo em Inglês | MEDLINE | ID: mdl-32094185

RESUMO

Accumulating evidence suggests participation of RNA-binding proteins with intrinsically disordered domains (IDPs) in the DNA damage response (DDR). These IDPs form liquid compartments at DNA damage sites in a poly(ADP ribose) (PAR)-dependent manner. However, it is greatly unknown how the IDPs are involved in DDR. We have shown previously that one of the IDPs RBM14 is required for the canonical nonhomologous end joining (cNHEJ). Here we show that RBM14 is recruited to DNA damage sites in a PARP- and RNA polymerase II (RNAPII)-dependent manner. Both KU and RBM14 are required for RNAPII-dependent generation of RNA:DNA hybrids at DNA damage sites. In fact, RBM14 binds to RNA:DNA hybrids. Furthermore, RNA:DNA hybrids and RNAPII are detected at gene-coding as well as at intergenic areas when double-strand breaks (DSBs) are induced. We propose that the cNHEJ pathway utilizes damage-induced transcription and intrinsically disordered protein RBM14 for efficient repair of DSBs.


Assuntos
Quebras de DNA de Cadeia Dupla , Reparo do DNA por Junção de Extremidades , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Proteínas Intrinsicamente Desordenadas/metabolismo , Proteínas de Ligação a RNA/metabolismo , RNA/metabolismo , Quimera , Células HEK293 , Humanos , Autoantígeno Ku/metabolismo , Hibridização de Ácido Nucleico , Domínios Proteicos , RNA/genética , RNA Polimerase II/metabolismo , Proteínas de Ligação a RNA/química , Proteínas de Ligação a RNA/genética
15.
Proc Biol Sci ; 287(1918): 20192563, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31937228

RESUMO

The mammalian placenta is both the physical interface between mother and fetus, and the source of endocrine signals that target the maternal hypothalamus, priming females for parturition, lactation and motherhood. Despite the importance of this connection, the effects of altered placental signalling on the maternal brain are insufficiently studied. Here, we show that placental dysfunction alters gene expression in the maternal brain, with the potential to affect maternal behaviour. Using a cross between the house mouse and the Algerian mouse, in which hybrid placental development is abnormal, we sequenced late-gestation placental and maternal medial preoptic area transcriptomes and quantified differential expression and placenta-maternal brain co-expression between normal and hybrid pregnancies. The expression of Fmn1 and Drd3 was significantly altered in the brains of females exposed to hybrid placentas. Most strikingly, expression patterns of placenta-specific gene families and Drd3 in the brains of house mouse females carrying hybrid litters matched those of female Algerian mice, the paternal species in the cross. Our results indicate that the paternally derived placental genome can influence the expression of maternal-fetal communication genes, including placental hormones, suggesting an effect of the offspring's father on the mother's brain.


Assuntos
Encéfalo/fisiologia , Expressão Gênica/fisiologia , Comportamento Materno , Placenta/fisiologia , Animais , Quimera , Feminino , Camundongos , Gravidez
16.
Am J Pathol ; 190(3): 577-585, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31953037

RESUMO

Psoriasis is an autoinflammatory/autoimmune skin disease and the epitome of an exaggerated primary inflammatory response in the surface barrier tissue. Despite the efficacy of dimethyl fumarate, an electrophilic drug for psoriasis management, there is a paucity of mechanistic evidence in vivo. In response to electrophiles, the Kelch-like erythroid cell-derived protein with cap-n-collar homology-associated protein 1/nuclear factor erythroid 2-related factor 2 (NRF2) system mediates a myriad of cytoprotective mechanisms, including the regulation of excessive inflammatory response and epidermal differentiation. Because the psoriasiform tissue reaction comprises neutrophil infiltration and parakeratotic scaling, it is hypothesized that Nrf2 not only regulates inflammatory responses but also maintains epidermal differentiation, a hallmark of epidermal homeostasis. By using the imiquimod-induced cutaneous inflammation model, an exaggerated inflammatory response and impaired epidermal differentiation in Nrf2-/- mice was detected. Dimethyl fumarate treatment in Nrf2+/+ mice attenuated a psoriasiform tissue reaction and rescued epidermal differentiation, which was not observed in Nrf2-/- mice. In accordance with the fact that psoriasis plaques form well-demarcated parakeratotic lesions in association with the psoriasiform tissue reaction, the lesional skin showed reduced expression levels of NRF2 and its downstream target genes compared with nonlesional skin. In conclusion, Nrf2 attenuates the psoriasiform tissue reaction and underscores the mechanistic legitimacy of the electrophile-based approach for the management of psoriasis.


Assuntos
Imiquimode/efeitos adversos , Inflamação/patologia , Fator 2 Relacionado a NF-E2/metabolismo , Paraceratose/patologia , Psoríase/patologia , Animais , Diferenciação Celular , Células Cultivadas , Quimera , Epiderme/efeitos dos fármacos , Epiderme/patologia , Feminino , Homeostase/efeitos dos fármacos , Humanos , Imuno-Histoquímica , Inflamação/induzido quimicamente , Queratinócitos/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fator 2 Relacionado a NF-E2/genética , Paraceratose/induzido quimicamente , Psoríase/induzido quimicamente , Pele/efeitos dos fármacos , Pele/patologia
17.
Nucleic Acids Res ; 48(4): 1764-1778, 2020 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-31965184

RESUMO

Chimeric RNAs and their encoded proteins have been traditionally viewed as unique features of neoplasia, and have been used as biomarkers and therapeutic targets for multiple cancers. Recent studies have demonstrated that chimeric RNAs also exist in non-cancerous cells and tissues, although large-scale, genome-wide studies of chimeric RNAs in non-diseased tissues have been scarce. Here, we explored the landscape of chimeric RNAs in 9495 non-diseased human tissue samples of 53 different tissues from the GTEx project. Further, we established means for classifying chimeric RNAs, and observed enrichment for particular classifications as more stringent filters are applied. We experimentally validated a subset of chimeric RNAs from each classification and demonstrated functional relevance of two chimeric RNAs in non-cancerous cells. Importantly, our list of chimeric RNAs in non-diseased tissues overlaps with some entries in several cancer fusion databases, raising concerns for some annotations. The data from this study provides a large repository of chimeric RNAs present in non-diseased tissues, which can be used as a control dataset to facilitate the identification of true cancer-specific chimeras.


Assuntos
Biomarcadores , Quimera/genética , RNA/genética , Quimera/classificação , Humanos , Neoplasias/genética , RNA/química , RNA/classificação
18.
BMC Plant Biol ; 20(1): 43, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31996141

RESUMO

BACKGROUND: Grapevine is a crop of major economic importance, yet little is known about the regulation of shoot development in grapevine or other perennial fruits crops. Here we combine genetic and genomic tools to identify candidate genes regulating shoot development in Vitis spp. RESULTS: An F2 population from an interspecific cross between V. vinifera and V. riparia was phenotyped for shoot development traits, and three Quantitative Trait Loci (QTLs) were identified on linkage groups (LGs) 7, 14 and 18. Around 17% of the individuals exhibited a dwarfed phenotype. A transcriptomic study identified four candidate genes that were not expressed in dwarfed individuals and located within the confidence interval of the QTL on LG7. A deletion of 84,482 bp was identified in the genome of dwarfed plants, which included these four not expressed genes. One of these genes was VviCURLY LEAF (VviCLF), an orthologue of CLF, a regulator of shoot development in Arabidopsis thaliana. CONCLUSIONS: The phenotype of the dwarfed grapevine plants was similar to that of clf mutants of A. thaliana and orthologues of the known targets of CLF in A. thaliana were differentially expressed in the dwarfed plants. This suggests that CLF, a major developmental regulator in A. thaliana, also controls shoot development in grapevine.


Assuntos
Proteínas de Arabidopsis/genética , Proteínas de Homeodomínio/genética , Brotos de Planta/crescimento & desenvolvimento , Vitis , Quimera , Mapeamento Cromossômico , Genes de Plantas , Genoma de Planta , Fenótipo , Locos de Características Quantitativas , Transcriptoma/genética , Vitis/genética
19.
BMC Med Genet ; 21(1): 15, 2020 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-31964351

RESUMO

BACKGROUND: Proteus syndrome (PS) is an extremely rare disease characterized by excessive chimeric growth of cells, and progressive and irregular asymmetrical hyperplasia. CASE PRESENTATION: Herein, a PS case with atypical clinical features and syndromes was reported, to improve the understanding of the diagnosis and treatment of the disease. The case was a 3-year-and-11-month-old male child. He was admitted due to a primary diagnosis of McCune-Albright syndrome. After admission, the lesion samples from the milk coffee spots, and nodular thickening skin at hands and feet were subjected to genetic screening. Genetic testing results confirmed the diagnosis of PS. CONCLUSIONS: Based on the clinical manifestations, laboratory tests, imaging data, and literature reviewing, the etiology, diagnosis, treatment and prognosis of PS have been analyzed and discussed.


Assuntos
Diagnóstico Diferencial , Displasia Fibrosa Poliostótica/genética , Síndrome de Proteu/diagnóstico , Doenças Raras/diagnóstico , Proliferação de Células , Criança , Pré-Escolar , Quimera/genética , Displasia Fibrosa Poliostótica/fisiopatologia , Humanos , Lactente , Masculino , Síndrome de Proteu/fisiopatologia , Doenças Raras/fisiopatologia
20.
Proc Natl Acad Sci U S A ; 117(6): 2886-2893, 2020 02 11.
Artigo em Inglês | MEDLINE | ID: mdl-31988135

RESUMO

Transcriptome profiling by RNA sequencing (RNA-seq) has been widely used to characterize cellular status, but it relies on second-strand complementary DNA (cDNA) synthesis to generate initial material for library preparation. Here we use bacterial transposase Tn5, which has been increasingly used in various high-throughput DNA analyses, to construct RNA-seq libraries without second-strand synthesis. We show that Tn5 transposome can randomly bind RNA/DNA heteroduplexes and add sequencing adapters onto RNA directly after reverse transcription. This method, Sequencing HEteRo RNA-DNA-hYbrid (SHERRY), is versatile and scalable. SHERRY accepts a wide range of starting materials, from bulk RNA to single cells. SHERRY offers a greatly simplified protocol and produces results with higher reproducibility and GC uniformity compared with prevailing RNA-seq methods.


Assuntos
DNA/genética , RNA/genética , Análise de Sequência de RNA/métodos , Quimera/genética , DNA Complementar/genética , Biblioteca Gênica , Células HEK293 , Células HeLa , Humanos , Análise de Célula Única , Transposases/metabolismo
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA