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1.
Artículo en Inglés | MEDLINE | ID: mdl-32058017

RESUMEN

The vitellogenin receptor (VgR) plays a critical role in egg development by mediating endocytosis of the major yolk protein precursor vitellogenin (Vg). Therefore, identifying the VgR of beneficial insects and its characterization could lead to the development of novel egg production strategies to enhance their commercial values. Here, we present the cloning, expression, and functional characterization of the VgR from an economically important eri silkworm, Samia ricini. The complete mRNA sequence was 6002 bp with an ORF of 5484 bp, encoding a protein of 1827 amino acids. Sequence analyses revealed that the SrVgR contained all of the conservative structural motifs characteristics of LDLR family members. The SrVgR was specifically expressed in the ovary, and the mRNA level increased steadily in pupal stages, reached its peak on day 9, and then declined to a bare minimum in adults. RNA interference (RNAi) clearly reduced the VgR transcript levels, disrupted the ovarian development resulting in malformed ovarioles and abnormal development of eggs. Taken together, these data provide conclusive evidence for the essential roles of VgR in insect reproduction.


Asunto(s)
Bombyx/metabolismo , Proteínas del Huevo/metabolismo , Proteínas de Insectos/metabolismo , Receptores de Superficie Celular/metabolismo , Secuencia de Aminoácidos , Animales , Bombyx/genética , Clonación Molecular , Proteínas del Huevo/genética , Femenino , Proteínas de Insectos/genética , Ovario/crecimiento & desarrollo , Ovario/metabolismo , ARN Mensajero/metabolismo , Receptores de Superficie Celular/genética , Homología de Secuencia de Aminoácido
2.
Life Sci ; 248: 117451, 2020 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-32088213

RESUMEN

AIMS: Electro-acupuncture pretreatment (EAP) plays a protective role in myocardial ischemia (MI) injury. However, the underlying mechanism remains unclear. A growing body of evidence suggests postinfarction inflammatory response directly affects the remodeling of ventricular function. The purpose of this study was to investigate whether EAP alleviates MI through NLRP3 inflammasome inhibition. MATERIALS AND METHODS: We constructed an AMI model by ligating the left anterior descending (LAD) coronary artery after 3 days of EAP with C57BL/6 mice. Echocardiography and TTC staining were employed to evaluate cardiac function and infarct size after 24 h of ischemia. HE staining and immunohistochemistry were employed to determine inflammatory level. Then, inflammasome activation was detected by western blotting, and macrophage polarization and neutrophil infiltration were observed by flow cytometry. KEY FINDINGS: Our preliminary findings showed that EAP reduced the infarct area and increased fractional shortening (FS) and ejection fraction (EF) and decreased the degree of inflammation after AMI injury. Meanwhile, EAP inhibited the expression of NLRP3, cleaved caspase-1 and IL-1ß in ischemia myocardial tissue, companied by inhibiting the expression of F4/80+, CD11b+, CD206low macrophages and activated M2 macrophage, and decreasing Ly-6G+CD11b+ neutrophils in ischemia myocardial and spleen tissue. SIGNIFICANCE: EAP inhibits the activation of NLRP3 inflammasome, promotes M2 polarization of macrophages and reduces the recruitment of neutrophils in damaged myocardium, thereby decreases the infarct size and improves the cardiac function.


Asunto(s)
Electroacupuntura/métodos , Inflamasomas/inmunología , Precondicionamiento Isquémico Miocárdico , Isquemia Miocárdica/genética , Isquemia Miocárdica/terapia , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Animales , Antígenos Ly/genética , Antígenos Ly/inmunología , Antígeno CD11b/genética , Antígeno CD11b/inmunología , Proteínas de Unión al Calcio/genética , Proteínas de Unión al Calcio/inmunología , Caspasa 1/genética , Caspasa 1/inmunología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Inflamasomas/genética , Inflamación , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Lectinas Tipo C/genética , Lectinas Tipo C/inmunología , Macrófagos/inmunología , Macrófagos/patología , Masculino , Lectinas de Unión a Manosa/genética , Lectinas de Unión a Manosa/inmunología , Ratones , Ratones Endogámicos C57BL , Isquemia Miocárdica/inmunología , Isquemia Miocárdica/patología , Miocardio/inmunología , Miocardio/patología , Proteína con Dominio Pirina 3 de la Familia NLR/inmunología , Infiltración Neutrófila , Neutrófilos/inmunología , Neutrófilos/patología , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/inmunología , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/inmunología , Transducción de Señal
3.
Cancer Sci ; 111(1): 47-58, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31710162

RESUMEN

Breast cancer is the most prevalent malignancy among women. Although endocrine therapy is effective, the development of endocrine resistance is a major clinical challenge. The tumor microenvironment (TME) promotes tumor malignancy, and tumor-associated macrophages (TAM) within the TME play a crucial role in endocrine resistance. Herein, we aimed to elucidate the relationship between TAM and the endocrine-resistant phenotype of breast cancer. Macrophages were cultured with conditioned medium (CM) from tamoxifen-sensitive (MCF7-S) or -resistant (MCF7-R) MCF7 breast cancer cells. M2 polarization was detected by CD163 immunofluorescence. To determine the effect on endocrine resistance, MCF7 cells were cultured in the supernatant of different TAM, and then treated with tamoxifen. CC-chemokine ligand 2 (CCL2) immunohistochemistry was carried out on pathological sections from 100 patients with invasive estrogen receptor-positive breast cancer. We found that macrophages cultured in the CM of MCF7-S and MCF7-R cells were induced into TAM, with a more obvious M2 polarization in the latter. Tamoxifen resistance was increased by culture in TAM medium. TAM secreted CCL2, which increased endocrine resistance in breast cancer cells through activation of the PI3K/Akt/mTOR signaling pathway. High expression of CCL2 was correlated with infiltration of CD163+macrophages (r = 0.548, P < .001), and patients with high CCL2 expression presented shorter progression-free survival than those with low CCL2 expression (P < .05). We conclude that CCL2 secreted by TAM activates PI3K/Akt/mTOR signaling and promotes an endocrine resistance feedback loop in the TME, suggesting that CCL2 and TAM may be novel therapeutic targets for patients with endocrine-resistant breast cancer.


Asunto(s)
Neoplasias de la Mama/genética , Quimiocina CCL2/genética , Resistencia a Antineoplásicos/genética , Macrófagos/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Proteínas Proto-Oncogénicas c-akt/genética , Antígenos CD/genética , Antígenos de Diferenciación Mielomonocítica/genética , Antineoplásicos Hormonales/farmacología , Neoplasias de la Mama/tratamiento farmacológico , Neoplasias de la Mama/metabolismo , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Femenino , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Células MCF-7 , Supervivencia sin Progresión , Receptores de Superficie Celular/genética , Transducción de Señal/genética , Serina-Treonina Quinasas TOR/genética , Tamoxifeno/farmacología , Microambiente Tumoral/efectos de los fármacos , Microambiente Tumoral/genética
4.
J Basic Microbiol ; 60(1): 72-81, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31737922

RESUMEN

Salmonella Enteritidis is an important foodborne pathogen that can infect a wide range of animal species including human beings, resulting in great losses to commercial husbandry and human health. CirA is an outer membrane receptor involved in iron uptake and colicin1A/B-mediated competitive killing. Although iron uptake is crucial to bacterial virulence, limited literature is available about the role of CirA in infection. In the present work, we aimed to evaluate the role of CirA during S. Enteritidis infection. For this purpose, we generated a CirA-deficient mutant of the S. Enteritidis strain C50336 and examined its biological characteristics. The results showed that cirA gene inactivation caused sharply decreased biofilm formation and apparently impaired antibiotic resistance. Furthermore, the cirA gene deletion mutant showed markedly reduced adhesion and invasion to human epithelial cell line Caco-2 cells and decreased proliferation in mouse macrophage cell line RAW264.7 cells. Moreover, attenuated virulence was determined by a mouse model, with an LD50 increase of approximately 1,000-fold. These data indicated that CirA plays critical roles in the S. Enteritidis infection process.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/metabolismo , Receptores de Superficie Celular/metabolismo , Salmonella enteritidis/fisiología , Salmonella enteritidis/patogenicidad , Virulencia/genética , Animales , Antibacterianos/farmacología , Adhesión Bacteriana , Proteínas de la Membrana Bacteriana Externa/genética , Biopelículas/crecimiento & desarrollo , Células CACO-2 , Colicinas , Farmacorresistencia Bacteriana , Humanos , Macrófagos/microbiología , Ratones , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana , Células RAW 264.7 , Receptores de Superficie Celular/deficiencia , Receptores de Superficie Celular/genética , Salmonelosis Animal/microbiología , Salmonella enteritidis/efectos de los fármacos
5.
Insect Biochem Mol Biol ; 117: 103284, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31760135

RESUMEN

Carbon dioxide has been used in traps for more than six decades to monitor mosquito populations and help make informed vector management decisions. CO2 is sensed by gustatory receptors (GRs) housed in neurons in the maxillary palps. CO2-sensitive GRs have been identified from the vinegar fly and mosquitoes, but it remains to be resolved whether these receptors respond to CO2 or bicarbonate. As opposed to the vinegar fly, mosquitoes have three GR subunits, but it is assumed that subunits GR1 and GR3 form functional receptors. In our attempt to identify the chemical species that bind these receptors, we discovered that GR2 and GR3 are essential for receptor function and that GR1 appears to function as a modulator. While Xenopus oocytes coexpressing Culex quinquefasciatus subunits CquiGR1/3 and CquiGR1/2 were not activated, CquiGR2/3 gave robust responses to sodium bicarbonate. Interestingly, CquiGR1/2/3-coexpressing oocytes gave significantly lower responses. That the ternary combination is markedly less sensitive than the GR2/GR3 combination was also observed with orthologs from the yellow fever and the malaria mosquito. By comparing responses of CquiGR2/CquiGR3-coexpressing oocytes to sodium bicarbonate samples (with or without acidification) and measuring the concentration of aqueous CO2, we showed that there is a direct correlation between dissolved CO2 and receptor response. We then concluded that subunits GR2 and GR3 are essential for these carbon dioxide-sensitive receptors and that they are activated by CO2 per se, not bicarbonate.


Asunto(s)
Dióxido de Carbono/metabolismo , Culex/genética , Regulación de la Expresión Génica , Proteínas de Insectos/genética , Receptores de Superficie Celular/genética , Animales , Animales Modificados Genéticamente/genética , Culex/metabolismo , Femenino , Proteínas de Insectos/metabolismo , Oocitos , Receptores de Superficie Celular/metabolismo , Xenopus laevis/genética
6.
Cell Mol Life Sci ; 77(3): 497-509, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31254045

RESUMEN

Cell-free DNA (cfDNA) is described to mirror intratumoral heterogeneity and gives insight about clonal evolution for improved therapeutic decisions. We sequenced cfDNA of a hormone receptor-positive, HER2-negative metastatic breast cancer (MBC) cohort with a high coverage to examine the prevalence and relevance of any detected variant. cfDNA of 44 MBC patients was isolated, followed by library construction using a customized targeted DNA panel with integrated unique molecular indices analyzing AKT1, AR, BRCA1, BRCA2, EGFR, ERCC4, ERBB2, ERBB3, ESR1, FGFR1, KRAS, MUC16, PIK3CA, PIK3R1, PTEN, PTGFR, and TGFB1. CfDNA was sequenced on the NextSeq® 550 platform (Illumina) and variants were analyzed with Ingenuity Variant Analysis (QIAGEN). We evaluated cfDNA variants in 40 of the 44 hormone receptor-positive and HER2-negative patients with a high mean coverage of 22,000×, resulting in MUC16, BRCA2, ERBB3, and AR variant calling in > 90% of the patients. 47% of all AR variants were pathogenic and at least one pathogenic or likely pathogenic variant was detected in each patient. A specific BRCA1 variant and > 3.5 pathogenic variants significantly associated with a reduced survival after diagnosis of metastasis. Longitudinal monitoring revealed an increase of pathogenic and likely pathogenic PIK3CA and ESR1 variant allele frequency under everolimus and exemestane, 8 months before proof of therapy failure by visual staging in one exemplary case. The identification of new variants with high prevalence, prognostic value, and dynamics under treatment by deep sequencing of cfDNA might empower sensitive monitoring and personalized therapeutic decisions.


Asunto(s)
Neoplasias de la Mama/genética , Ácidos Nucleicos Libres de Células/genética , Variación Genética/genética , Receptores de Superficie Celular/genética , Alelos , Proteína BRCA1/genética , Proteína BRCA2/genética , Biomarcadores de Tumor/genética , Fosfatidilinositol 3-Quinasa Clase I/genética , Receptor alfa de Estrógeno/genética , Femenino , Humanos
7.
Insect Mol Biol ; 29(1): 92-103, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31456272

RESUMEN

Ninety-four putative G protein-coupled receptors (GPCRs) were identified in the Musca domestica genome. They were annotated and compared with their homologues in Drosophila melanogaster. Phylogenetic analyses of the GPCRs from both species revealed that several family members shared a closer relationship based on the domain architecture. The expression profiles of these genes were examined by quantitative real-time PCR amongst three strains of the house fly, a near-isogenic line strain with imidacloprid resistance (N-IRS), the corresponding susceptible strain (CSS) and another strain derived from field populations with imidacloprid resistance (IRS). We found that five GPCR genes were upregulated in the N-IRS and eight GPCR genes were upregulated in the IRS strains compared to the CSS strain. The transgenic lines of D. melanogaster with the GPCR genes (LOC101899380 in the N-IRS strain and LOC101895664 in the IRS strain) exhibited significantly increased tolerance to imidacloprid, and higher expression of cytochrome P450 genes. Bioinformatic analysis of LOC101899380 was carried out based on its full-length nucleic acid sequence and putative amino acid sequence, and it was named Methuselah-like10 (Mthl10) owing to its homology with D. melanogaster Mthl10. A cell-base cell counting kit-8 toxicity assay demonstrated that the expression of the GPCR gene LOC101899380 in Spodoptera frugiperda (Sf9) cells using a baculovirus-mediated expression system can elevate the cell tolerance to imidacloprid, indirectly supporting the hypothesis that the GPCR gene LOC101899380 plays some role in imidacloprid resistance. These results should be useful for furthering understanding of the regulatory pathway by which house flies develop resistance.


Asunto(s)
Moscas Domésticas/genética , Resistencia a los Insecticidas/genética , Insecticidas/farmacología , Neonicotinoides/farmacología , Nitrocompuestos/farmacología , Receptores de Superficie Celular/genética , Animales , Animales Modificados Genéticamente , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , Drosophila melanogaster/genética , Moscas Domésticas/efectos de los fármacos , Receptores de Superficie Celular/metabolismo , Células Sf9
8.
Plant Mol Biol ; 102(3): 271-285, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31838617

RESUMEN

KEY MESSAGE: H2 prolonged the vase life and improved the vase quality of cut roses through repressing endogenous ethylene production and alleviating ethylene signal transduction during the entire senescing period. Recently, the application of hydrogen gas (H2) was shown to improve postharvest quality and longevity in perishable horticultural products, but the specific regulation mechanism remains obscure. Here, endogenous ethylene production and the expression of genes in ethylene biosynthesis and signalling pathway were investigated to explore the crosstalk between H2 and ethylene during the senescence of cut roses. Our results revealed that addition of exogenous ethylene by ethephon accelerated the senescence of cut roses, in which 100 mg L-1 ethephon displayed the most obvious senescent phenotype. While the applied different concentrations (1%, 10%, 50% and 100%) of hydrogen-rich water (HRW) conducted different affects in alleviating the senescence of cut roses, and 1% HRW displayed the best ornamental quality and the longest vase life by reducing ethylene production, supported by the decrease of 1-aminocyclopropene-1-carboxylate (ACC) accumulation, ACC synthase (ACS) and ACC oxidase (ACO) activities, and Rh-ACS3 and Rh-ACO1 expressions in ethylene biosynthesis. In addition, HRW increased the transcripts of ethylene receptor genes Rh-ETR1 at blooming period from day 4 to day 6 and suppressed Rh-ETR3 at senescence phase at day 8 after harvest. Furthermore, the relevant affection of HRW on Rh-ETR1 and Rh-ETR3 expressions still existed when the ethylene production was compromised by adequate addition of exogenous ethylene in HRW-treated cut rose petals, and HRW directly repressed the protein level of Rh-ETR3 in a transient expression assay. Overall, the results suggested that H2 is involved in neutralizing ethylene-mediated postharvest in cut flowers.


Asunto(s)
Etilenos/antagonistas & inhibidores , Etilenos/biosíntesis , Flores/efectos de los fármacos , Hidrógeno/farmacología , Rosa/efectos de los fármacos , Rosa/metabolismo , Aminoácidos Cíclicos/metabolismo , Flores/enzimología , Flores/genética , Flores/crecimiento & desarrollo , Regulación Enzimológica de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Hidrógeno/metabolismo , Liasas/genética , Liasas/metabolismo , Compuestos Organofosforados , Fenotipo , Reguladores del Crecimiento de las Plantas , Proteínas de Plantas/genética , Receptores de Superficie Celular/genética , Rosa/enzimología , Rosa/genética , Transducción de Señal
9.
Immunity ; 52(1): 123-135.e6, 2020 01 14.
Artículo en Inglés | MEDLINE | ID: mdl-31859049

RESUMEN

The immune system monitors the health of cells and is stimulated by necrosis. Here we examined the receptors and ligands driving this response. In a targeted screen of C-type lectin receptors, a Clec2d reporter responded to lysates from necrotic cells. Biochemical purification identified histones, both free and bound to nucleosomes or neutrophil extracellular traps, as Clec2d ligands. Clec2d recognized poly-basic sequences in histone tails and this recognition was sensitive to post-translational modifications of these sequences. As compared with WT mice, Clec2d-/- mice exhibited reduced proinflammatory responses to injected histones, and less tissue damage and improved survival in a hepatotoxic injury model. In macrophages, Clec2d localized to the plasma membrane and endosomes. Histone binding to Clec2d did not stimulate kinase activation or cytokine production. Rather, histone-bound DNA stimulated endosomal Tlr9-dependent responses in a Clec2d-dependent manner. Thus, Clec2d binds to histones released upon necrotic cell death, with functional consequences to inflammation and tissue damage.


Asunto(s)
Histonas/metabolismo , Lectinas Tipo C/inmunología , Lectinas Tipo C/metabolismo , Hígado/lesiones , Necrosis/patología , Receptores de Superficie Celular/inmunología , Receptores de Superficie Celular/metabolismo , Animales , Apoptosis/inmunología , Endosomas/metabolismo , Células HEK293 , Humanos , Células Jurkat , Lectinas Tipo C/genética , Macrófagos/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neutrófilos/inmunología , Receptores de Superficie Celular/genética , Receptor Toll-Like 9/inmunología
10.
Protein Pept Lett ; 27(1): 60-66, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31362652

RESUMEN

BACKGROUND: Cathelicidins are a family of Host Defense Peptides (HDPs), that play an important role in the innate immune response. They exert both broad-spectrum antimicrobial activity against pathogens, and strong immunomodulatory functions that affect the response of innate and adaptive immune cells. OBJECTIVE: The aim of this study was to investigate immunomodulation by the chicken cathelicidin CATH-2 and compare its activities to those of the human cathelicidin LL-37. METHODS: Chicken macrophages and chicken monocytes were incubated with cathelicidins. Activation of immune cells was determined by measuring surface markers Mannose Receptor Ctype 1 (MRC1) and MHC-II. Cytokine production was measured by qPCR and nitric oxide production was determined using the Griess assay. Finally, the effect of cathelicidins on phagocytosis was measured using carboxylate-modified polystyrene latex beads. RESULTS: CATH-2 and its all-D enantiomer D-CATH-2 increased MRC1 and MHC-II expression, markers for antigen presentation, on primary chicken monocytes, whereas LL-37 did not. D-CATH- 2 also increased the MRC1 and MHC-II expression if a chicken macrophage cell line (HD11 cells) was used. In addition, LPS-induced NO production by HD11 cells was inhibited by CATH-2 and D-CATH-2. CONCLUSION: These results are a clear indication that CATH-2 (and D-CATH-2) affect the activation state of monocytes and macrophages, which leads to optimization of the innate immune response and enhancement of the adaptive immune response.


Asunto(s)
Biomarcadores/metabolismo , Catelicidinas/metabolismo , Macrófagos/metabolismo , Monocitos/metabolismo , Secuencia de Aminoácidos , Animales , Presentación de Antígeno/efectos de los fármacos , Péptidos Catiónicos Antimicrobianos/metabolismo , Línea Celular , Pollos , Citocinas/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Antígenos de Histocompatibilidad Clase II/genética , Antígenos de Histocompatibilidad Clase II/metabolismo , Humanos , Inmunidad Innata/efectos de los fármacos , Inmunomodulación/efectos de los fármacos , Lectinas Tipo C/genética , Lectinas Tipo C/metabolismo , Lectinas de Unión a Manosa/genética , Lectinas de Unión a Manosa/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo
11.
Arch Insect Biochem Physiol ; 103(1): e21636, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31612557

RESUMEN

As a member of the low-density lipoprotein receptor (LDLR) superfamily, vitellogenin (Vg) receptor (VgR) is responsible for the uptake of Vg into developing oocytes and is a potential target for pest control. Here, a full-length VgR complementary DNA (named as CsVgR) was isolated and characterized in the rice stem borer, Chilo suppressalis. The composite CsVgR gene contained an open reading frame of 5,484 bp encoding a protein of 1,827 amino acid residues. Structural analysis revealed that CsVgR contained two ligand-binding domains (LBDs) with four Class A (LDLRA ) repeats in LBD1 and seven in LBD2, which was structurally different from most non-Lepidopteran insect VgRs having five repeats in LBD1 and eight in LBD2. The developmental expression analysis showed that CsVgR messenger RNA expression was first detectable in 3-day-old pupae, sharply increased in newly emerged female adults, and reached a peak in 2-day-old female adults. Consistent with most other insects VgRs, CsVgR was exclusively expressed in the ovary. Notably, injection of dsCsVgR into late pupae resulted in fewer follicles in the ovarioles as well as reduced fecundity, suggesting a critical role of CsVgR in female reproduction. These results may contribute to the development of RNA interference-mediated disruption of reproduction as a control strategy of C. suppressalis.


Asunto(s)
Proteínas del Huevo/genética , Mariposas Nocturnas/genética , Receptores de Superficie Celular/genética , Animales , Proteínas del Huevo/química , Proteínas del Huevo/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Proteínas de Insectos/metabolismo , Mariposas Nocturnas/crecimiento & desarrollo , Mariposas Nocturnas/metabolismo , Ovario/crecimiento & desarrollo , Ovario/metabolismo , Filogenia , Interferencia de ARN , ARN Mensajero/metabolismo , Receptores de Superficie Celular/química , Receptores de Superficie Celular/metabolismo , Análisis de Secuencia de Proteína
12.
DNA Cell Biol ; 39(1): 63-68, 2020 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-31855460

RESUMEN

To investigate the contribution of de novo variants to Tourette disorder (TD) probands in China. Whole-exome sequencing (WES) conducted on 15 child-parent trios (45 samples) detected 25 coding de novo variants, including 2 de novo Likely Gene Disrupting (LGD) variants and 6 Missense3 variants. The de novo LGD variants were consistently associated with TD risk (Fisher's exact test OR 2.69; p = 0.1952), although statistical significance was not achieved due to the small sample size. We then assessed the relationship between the genetic events and phenotypic data by comparing Yale Global Tic Severity Scale (YGTSS) scores. The TD probands with damaging variants (defined as LGD variants and Mis3 variants) had significantly higher YGTSS scores, suggesting more severe tic symptoms (p = 0.019). We also observed a hit for a damaging compound heterozygous (CH) mutation in CELSR3, a high-confidence TD risk gene, in one of the TD probands. To our knowledge, this is the first study to investigate de novo variants in TD in a Chinese population. Our results showed that de novo LGD variants contributed to TD risk in our cohort and that TD probands with de novo damaging variants have more severe symptoms. Furthermore, our observation of damaging CH mutations in CELSR3 in an individual affected with TD further strengthened the confidence in a role for this gene in TD etiology.


Asunto(s)
Predisposición Genética a la Enfermedad/genética , Mutación , Síndrome de Tourette/genética , Secuenciación del Exoma Completo/métodos , Adulto , Grupo de Ascendencia Continental Asiática/genética , Cadherinas/genética , China , Estudios de Cohortes , Femenino , Predisposición Genética a la Enfermedad/etnología , Heterocigoto , Humanos , Masculino , Receptores de Superficie Celular/genética , Síndrome de Tourette/etnología , Adulto Joven
13.
J Insect Sci ; 19(6)2019 Nov 01.
Artículo en Inglés | MEDLINE | ID: mdl-31812980

RESUMEN

Vitellogenin receptor (VgR) mediates the intake of vitellin via oocytes, thus exerting an important role in vitellogenesis. In this study, reverse transcription-polymerase chain reaction (RT-PCR) and rapid-amplification of cDNA ends techniques were adopted to clone the CiVgR gene, namely the VgR gene of Calliptamus italicus, i.e., Orthopteran. The full length of CiVgR was 5,589 bp, and the open reading frame was estimated to be 5,265 bp, which encoded 1,754 amino acids (aa). Sequence alignment analysis showed that CiVgR belonged to the superfamily of low-density lipoprotein receptor genes, which contained several conserved domains, including ligand-binding domains, epidermal growth factor precursor homology domains, transmembrane domains, and cytoplasmic domains. However, no O-linked sugar domain was identified. Phylogenetic analysis showed that CiVgR had the closest genetic relationship to Blattarias. RT-PCR showed that CiVgR was only specifically expressed in the ovarian tissue of females. quantitative real time polymerase chain reaction showed that the transcription of CiVgR already appeared in the fourth-instar nymph of C. italicus, which gradually increased after adult emergence, peaked at the previtellogenesis stage, and then started to decrease. The expression pattern of CiVgR was closely associated with vitellogenesis. The findings of this study further our understanding of the molecular mechanisms involved in the reproduction of C. italicus, and provide new ideas to control this insect.


Asunto(s)
Proteínas del Huevo/metabolismo , Saltamontes/metabolismo , Receptores de Superficie Celular/metabolismo , Secuencia de Aminoácidos , Animales , Proteínas del Huevo/genética , Femenino , Masculino , Filogenia , Receptores de Superficie Celular/genética , Reproducción , Homología de Secuencia de Aminoácido
14.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi ; 36(12): 1153-1157, 2019 Dec 10.
Artículo en Chino | MEDLINE | ID: mdl-31813136

RESUMEN

OBJECTIVE: The phenotype and genetics of three patients with autosomal recessive polycystic kidney disease (ARPKD) at childhood, teenage and advanced age were analyzed. METHODS: Next generation sequencing (NGS) was applied to all the probands. PCR and Sanger sequencing were used to verify the suspicious gene variants screened by NGS in the probands and their family members, and one of the family got prenatal diagnosis. RESULTS: Through NGS, PCR and Sanger sequencing, the 5-yr proband in pedigree 1 was shown to carry compound heterozygous variants of c.5935G>A(p.G1979R) and c.5428G>T(p.E1810X) of PKHD1, originated from his parents; In pedigree 2, the 17-ys proband was detected with c.5512T>C(p.Y1838H) and c.5935G>A(p.G1979R) variants of PKHD1 orginated from her parents, and her mother also got prenatal diagnosis during the second trimester; In pedigree 3, the 70-ys female proband was found with variants c.11314C>T (p.R3772X) and c.3860T>G (p.V1287G) of PKHD1. CONCLUSION: The three pedigrees were diagnosed as ARPKD caused by PKHD1 variants. Five types of variants were detected, c.5935G>A and c.11314C>T were the known pathogenic variants, while c.5512T>C, c.5428G>T and c.3860T>G were not reported previously. Considering the complexity of the genetics and phenotypes of the cystic renal diseases, genetic diagnosis is crucial to give accurate etiological diagnosis, which may benefit the clinic management.


Asunto(s)
Riñón Poliquístico Autosómico Recesivo/genética , Receptores de Superficie Celular/genética , Adolescente , Anciano , Preescolar , Femenino , Humanos , Masculino , Mutación , Fenotipo , Embarazo
15.
BMC Neurol ; 19(1): 253, 2019 Oct 27.
Artículo en Inglés | MEDLINE | ID: mdl-31656175

RESUMEN

BACKGROUND: We encountered two unrelated individuals suffering from neurological disorders, including epilepsy and scoliosis. CASE PRESENTATION: Whole-exome sequencing identified the same recurrent, de novo, pathogenic variant in NUS1 [NM_138459.4:c.691 + 1C > A] in both individuals. This variant is located in the conserved cis-prenyltransferase domain of the nuclear undecaprenyl pyrophosphate synthase 1 gene (NUS1), which encodes the Nogo-B receptor, an essential catalyst for protein glycosylation. This variant was confirmed to create a new splice donor site, resulting in aberrant RNA splicing resulting in a 91-bp deletion in exon 3 in both individuals. The mutant mRNA was partially degraded by nonsense mediated mRNA decay. To date, only four de novo variants and one homozygous variant have been reported in NUS1, which cause developmental and epileptic encephalopathy, early onset Parkinson's disease, and a congenital disorder of glycosylation. Seven patients, including our two patients, have presented with epileptic seizures and intellectual disabilities. CONCLUSIONS: Our study strongly supports the finding that this recurrent, de novo, variant in NUS1 causes developmental and epileptic encephalopathy with involuntary movement, ataxia and scoliosis.


Asunto(s)
Ataxia/genética , Epilepsia/genética , Mioclonía/genética , Receptores de Superficie Celular/genética , Escoliosis/genética , Femenino , Humanos , Discapacidad Intelectual/genética , Masculino , Mutación , Linaje , Sitios de Empalme de ARN
16.
PLoS Biol ; 17(10): e3000466, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-31658245

RESUMEN

The pre- and postsynaptic membranes comprising the synaptic junction differ in protein composition. The membrane trafficking mechanisms by which neurons control surface polarization of synaptic receptors remain poorly understood. The sorting receptor Sortilin-related CNS expressed 1 (SorCS1) is a critical regulator of trafficking of neuronal receptors, including the presynaptic adhesion molecule neurexin (Nrxn), an essential synaptic organizer. Here, we show that SorCS1 maintains a balance between axonal and dendritic Nrxn surface levels in the same neuron. Newly synthesized Nrxn1α traffics to the dendritic surface, where it is endocytosed. Endosomal SorCS1 interacts with the Rab11 GTPase effector Rab11 family-interacting protein 5 (Rab11FIP5)/Rab11 interacting protein (Rip11) to facilitate the transition of internalized Nrxn1α from early to recycling endosomes and bias Nrxn1α surface polarization towards the axon. In the absence of SorCS1, Nrxn1α accumulates in early endosomes and mispolarizes to the dendritic surface, impairing presynaptic differentiation and function. Thus, SorCS1-mediated sorting in dendritic endosomes controls Nrxn axonal surface polarization required for proper synapse development and function.


Asunto(s)
Proteínas de Unión al Calcio/genética , Corteza Cerebral/metabolismo , Moléculas de Adhesión de Célula Nerviosa/genética , Neuronas/metabolismo , Receptores de Superficie Celular/genética , Membranas Sinápticas/metabolismo , Transmisión Sináptica/genética , Animales , Proteínas de Unión al Calcio/metabolismo , Polaridad Celular , Corteza Cerebral/citología , Embrión de Mamíferos , Endocitosis , Endosomas/metabolismo , Regulación de la Expresión Génica , Células HEK293 , Humanos , Ratones , Ratones Endogámicos C57BL , Proteínas Mitocondriales/genética , Proteínas Mitocondriales/metabolismo , Moléculas de Adhesión de Célula Nerviosa/metabolismo , Neuronas/ultraestructura , Cultivo Primario de Células , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Transporte de Proteínas , Ratas , Ratas Wistar , Receptores de Superficie Celular/metabolismo , Membranas Sinápticas/ultraestructura , Proteínas de Unión al GTP rab/genética , Proteínas de Unión al GTP rab/metabolismo
17.
Anticancer Res ; 39(10): 5653-5662, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31570463

RESUMEN

BACKGROUND/AIM: Factors influencing fulvestrant efficacy may be useful in selecting the optimal treatment regimen for postmenopausal Japanese women with metastatic/recurrent HR-positive, HER2-negative breast cancer. PATIENTS AND METHODS: We retrospectively evaluated progression-free and overall survival (PFS and OS) in 100 fulvestrant-treated patients according to metastatic site. RESULTS: Median PFS was significantly better in patients with non-visceral (bone and regional metastases; 22.8 months) vs. visceral metastasis (lung, liver, and other organs; 8.2 months; p=0.024), although median OS did not differ (p=0.922). Median PFS in patients with lung metastasis (20.8 months) and non-visceral metastasis (22.8 months) were comparable; patients with liver metastasis (6.1 months) and other organ metastases (3.7 months) had worse prognoses. CONCLUSION: Patients with non-visceral metastases had a better prognosis than those with visceral metastases. Fulvestrant induced a longer PFS in patients with non-visceral metastasis, and also in those with lung metastasis without liver or other organ involvement.


Asunto(s)
Neoplasias de la Mama/tratamiento farmacológico , Fulvestrant/uso terapéutico , Metástasis de la Neoplasia/prevención & control , Recurrencia Local de Neoplasia/tratamiento farmacológico , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Femenino , Humanos , Persona de Mediana Edad , Metástasis de la Neoplasia/genética , Metástasis de la Neoplasia/patología , Recurrencia Local de Neoplasia/genética , Recurrencia Local de Neoplasia/patología , Pronóstico , Supervivencia sin Progresión , Receptor ErbB-2/genética , Receptores de Superficie Celular/genética , Estudios Retrospectivos
18.
Nat Commun ; 10(1): 4456, 2019 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-31575859

RESUMEN

Fertilization is essential for species survival. Although Izumo1 and Juno are critical for initial interaction between gametes, additional molecules necessary for sperm:egg fusion on both the sperm and the oocyte remain to be defined. Here, we show that phosphatidylserine (PtdSer) is exposed on the head region of viable and motile sperm, with PtdSer exposure progressively increasing during sperm transit through the epididymis. Functionally, masking phosphatidylserine on sperm via three different approaches inhibits fertilization. On the oocyte, phosphatidylserine recognition receptors BAI1, CD36, Tim-4, and Mer-TK contribute to fertilization. Further, oocytes lacking the cytoplasmic ELMO1, or functional disruption of RAC1 (both of which signal downstream of BAI1/BAI3), also affect sperm entry into oocytes. Intriguingly, mammalian sperm could fuse with skeletal myoblasts, requiring PtdSer on sperm and BAI1/3, ELMO2, RAC1 in myoblasts. Collectively, these data identify phosphatidylserine on viable sperm and PtdSer recognition receptors on oocytes as key players in sperm:egg fusion.


Asunto(s)
Oocitos/metabolismo , Fagocitos/metabolismo , Fosfatidilserinas/metabolismo , Interacciones Espermatozoide-Óvulo/fisiología , Espermatozoides/metabolismo , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas Angiogénicas/metabolismo , Animales , Antígenos CD36/metabolismo , Proteínas del Citoesqueleto/metabolismo , Epidídimo , Femenino , Humanos , Masculino , Proteínas de la Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Modelos Animales , Mioblastos Esqueléticos , Proteínas del Tejido Nervioso/metabolismo , Neuropéptidos/metabolismo , Fosfatidilserinas/genética , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Tirosina Quinasa c-Mer/metabolismo , Proteína de Unión al GTP rac1/metabolismo
19.
Enzyme Microb Technol ; 131: 109418, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31615663

RESUMEN

A novel carbohydrate binding module (CBM) was identified in a ß-1,3-xylanase from Flammeovirga pacifica, which showed only 25.0% sequence identity with the reported CBMs with the coverage of 36.4%. To verify its function, a truncated ß-1,3-xylanase (Xy13088-T) and a carbohydrate binding module (CBM3088) were expressed and purified. The thermostability and catalytic efficiency of the Xy13088-T declined significantly when compared with the full-length one, with the decreasing of the half-life and catalytic efficiency (Kcat/Km) by 90%. Interestingly, the CBM3088 showed the binding ability to ß-1,3-xylan only when Ca2+ existed, which was different from the reported CBMs of ß-1,3-xylanases. The maximum amount of CBM3088 binding to ß-1,3-xylan was 9.65 µmol/g of ß-1,3-xylan. The residues probably involved in the binding to the ß-1,3-xylan and Ca2+ were addressed by bioinformatics analysis.


Asunto(s)
Bacteroidetes/enzimología , Calcio/metabolismo , Receptores de Superficie Celular/genética , Receptores de Superficie Celular/metabolismo , Xilosidasas/genética , Xilosidasas/metabolismo , Cationes Bivalentes/metabolismo , Estabilidad de Enzimas , Cinética , Unión Proteica , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Eliminación de Secuencia , Temperatura Ambiental , Xilanos/metabolismo
20.
Plant Sci ; 289: 110269, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31623779

RESUMEN

Ethylene is a key element of plant physiology, thus ethylene research is important for both, fundamental research and agriculture. Previous work on ethylene receptors focused on expression level and protein interaction, but knowledge on regulation of gene transcription is scarce. Promoters of mango ethylene receptor genes (pMiERS1a, pMiERS1b) were analysed particularly regarding responsiveness to hormones. The promoter sequences reveal some variation and they were characterized by identifying functional regulatory candidate modules via truncated-promoter approach. Based on ectopic gene expression studies in transgenic Arabidopsis and Nicotiana it is demonstrated that both promoters are positively responsive to ethylene. For pMiERS1a the AHBP/DOFF1 module is linked to ethylene responsiveness, while for pMiERS1b it is the module MYBL/OPAQ1. A negative gene regulation in response to abscisic acid (ABA) is linked to MYBL/DOFF2. A positive response to indole-3-acetic acid (IAA) was found for GTBX/MYCL1, containing the motifs IBOX/IDDF/TEFB, which are present in this combination only in pMiERS1b, but not in pMiERS1a. Conclusively, the general response of the ethylene receptor genes is conserved, but similar regulation can be linked to different modules. Further, a minor variation in a transcription factor binding site (TFBS) motif within an overall conserved module type can lead to a different expression.


Asunto(s)
Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Mangifera/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Proteínas de Plantas/genética , Receptores de Superficie Celular/genética , Tabaco/genética , Arabidopsis/metabolismo , Etilenos/metabolismo , Mangifera/metabolismo , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Plantas Modificadas Genéticamente/metabolismo , Receptores de Superficie Celular/metabolismo , Tabaco/metabolismo
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