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1.
Plant Cell ; 36(1): 112-135, 2023 Dec 21.
Artículo en Inglés | MEDLINE | ID: mdl-37770034

RESUMEN

Reactive oxygen species (ROS) play an essential role in plant growth and responses to environmental stresses. Plant cells sense and transduce ROS signaling directly via hydrogen peroxide (H2O2)-mediated posttranslational modifications (PTMs) on protein cysteine residues. Here, we show that the H2O2-mediated cysteine oxidation of NAC WITH TRANS-MEMBRANE MOTIF1-LIKE 1 (GmNTL1) in soybean (Glycine max) during salt stress promotes its release from the endoplasmic reticulum (ER) membrane and translocation to the nucleus. We further show that an oxidative posttranslational modification on GmNTL1 residue Cys-247 steers downstream amplification of ROS production by binding to and activating the promoters of RESPIRATORY BURST OXIDASE HOMOLOG B (GmRbohB) genes, thereby creating a feed-forward loop to fine-tune GmNTL1 activity. In addition, oxidation of GmNTL1 Cys-247 directly promotes the expression of CATION H+ EXCHANGER 1 (GmCHX1)/SALT TOLERANCE-ASSOCIATED GENE ON CHROMOSOME 3 (GmSALT3) and Na+/H+ Antiporter 1 (GmNHX1). Accordingly, transgenic overexpression of GmNTL1 in soybean increases the H2O2 levels and K+/Na+ ratio in the cell, promotes salt tolerance, and increases yield under salt stress, while an RNA interference-mediated knockdown of GmNTL1 elicits the opposite effects. Our results reveal that the salt-induced oxidation of GmNTL1 promotes its relocation and transcriptional activity through an H2O2-mediated posttranslational modification on cysteine that improves resilience of soybean against salt stress.


Asunto(s)
Glycine max , Tolerancia a la Sal , Glycine max/genética , Tolerancia a la Sal/genética , Peróxido de Hidrógeno/metabolismo , Factores de Transcripción/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Cisteína/metabolismo , Estrés Fisiológico/genética , Plantas Modificadas Genéticamente/metabolismo , Regulación de la Expresión Génica de las Plantas
2.
J Biol Chem ; 299(12): 105481, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-38041932

RESUMEN

Singlet oxygen (1O2) has a very short half-life of 10-5 s; however, it is a strong oxidant that causes growth arrest and necrotic lesions on plants. Its signaling pathway remains largely unknown. The Arabidopsis flu (fluorescent) mutant accumulates a high level of 1O2 and shows drastic changes in nuclear gene expression. Only two plastid proteins, EX1 (executer 1) and EX2 (executer 2), have been identified in the singlet oxygen signaling. Here, we found that the transcription factor abscisic acid insensitive 4 (ABI4) binds the promoters of genes responsive to 1O2-signals. Inactivation of the ABI4 protein in the flu/abi4 double mutant was sufficient to compromise the changes of almost all 1O2-responsive-genes and rescued the lethal phenotype of flu grown under light/dark cycles, similar to the flu/ex1/ex2 triple mutant. In addition to cell death, we reported for the first time that 1O2 also induces cell wall thickening and stomatal development defect. Contrastingly, no apparent growth arrest was observed for the flu mutant under normal light/dim light cycles, but the cell wall thickening (doubled) and stomatal density reduction (by two-thirds) still occurred. These results offer a new idea for breeding stress tolerant plants.


Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Pared Celular/metabolismo , Regulación de la Expresión Génica de las Plantas , Luz , Oxígeno Singlete/metabolismo , Transcriptoma , Estomas de Plantas/metabolismo
3.
Biotechnol Bioeng ; 117(11): 3499-3507, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-32691870

RESUMEN

Squalene is a lipophilic and non-volatile triterpene with many industrial applications for food, pharmaceuticals, and cosmetics. Metabolic engineering focused on optimization of the production pathway suffer from little success in improving titers because of a limited space of the cell membrane accommodating the lipophilic product. Extension of cell membrane would be a promising approach to overcome the storage limitation for successful production of squalene. In this study, Escherichia coli was engineered for squalene production by overexpression of some membrane proteins. The highest production of 612 mg/L was observed in the engineered E. coli with overexpression of Tsr, a serine chemoreceptor protein, which induced invagination of inner membrane to form multilayered structure. It was also observed an increase in unsaturated fatty acid in membrane lipids composition, suggesting cellular response to maintain membrane fluidity against squalene accumulation in the engineered strain. This study potentiates the capability of E. coli for squalene production and provides an effective strategy for the enhanced production of such compounds.


Asunto(s)
Membrana Celular , Escherichia coli , Ingeniería Metabólica/métodos , Escualeno/metabolismo , Membrana Celular/química , Membrana Celular/metabolismo , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Quimiotácticas Aceptoras de Metilo/genética , Proteínas Quimiotácticas Aceptoras de Metilo/metabolismo , Ácido Mevalónico/metabolismo
4.
Int J Mol Sci ; 21(19)2020 Oct 08.
Artículo en Inglés | MEDLINE | ID: mdl-33050099

RESUMEN

Nitrogen (N), phosphorus (P), and potassium (K) are three essential macro-elements for plant growth and development. Used to improve yield in agricultural production, the excessive use of chemical fertilizers often leads to increased production costs and ecological environmental pollution. Vitamins C and E are antioxidants that play an important role in alleviating abiotic stress. However, there are few studies on alleviating oxidative stress caused by macro-element deficiency. Here, we used Arabidopsis vitamin E synthesis-deficient mutant vte4 and vitamin C synthesis-deficient mutant vtc1 on which exogenous vitamin E and vitamin C, respectively, were applied at the bolting stage. In the deficiency of macro-elements, the Arabidopsis chlorophyll content decreased, malondialdehyde (MDA) content and relative electric conductivity increased, and reactive oxygen species (ROS) accumulated. The mutants vtc1 and vte4 are more severely stressed than the wild-type plants. Adding exogenous vitamin E was found to better alleviate stress than adding vitamin C. Vitamin C barely affected and vitamin E significantly inhibited the synthesis of ethylene (ETH) and jasmonic acid (JA) genes, thereby reducing the accumulation of ETH and JA that alleviated the senescence caused by macro-element deficiency at the later stage of bolting in Arabidopsis. A deficiency of macro-elements also reduced the yield and germination rate of the seeds, which were more apparent in vtc1 and vte4, and adding exogenous vitamin C and vitamin E, respectively, could restore them. This study reported, for the first time, that vitamin E is better than vitamin C in delaying seedling senescence caused by macro-element deficiency in Arabidopsis.


Asunto(s)
Antioxidantes/farmacología , Arabidopsis/efectos de los fármacos , Arabidopsis/genética , Ácido Ascórbico/farmacología , Resistencia a la Enfermedad/efectos de los fármacos , Plantones/efectos de los fármacos , Vitamina E/farmacología , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Ciclopentanos/antagonistas & inhibidores , Ciclopentanos/metabolismo , Etilenos/antagonistas & inhibidores , Etilenos/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Malondialdehído/metabolismo , Estrés Oxidativo/efectos de los fármacos , Oxilipinas/antagonistas & inhibidores , Oxilipinas/metabolismo , Enfermedades de las Plantas/prevención & control , Hojas de la Planta/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Plantones/genética , Plantones/metabolismo , Semillas/metabolismo , Transducción de Señal/efectos de los fármacos , Factores de Tiempo
5.
Nitric Oxide ; 76: 6-15, 2018 06 01.
Artículo en Inglés | MEDLINE | ID: mdl-29510200

RESUMEN

Nitric oxide (NO) has a general inhibitory effects on chlorophyll biosynthesis, especially to the step of 5-aminolevulinic acid (ALA) biosynthesis and protochlorophyllide (Pchlide) to chlorophyllide (Chlide) conversion (responsible by the NADPH:Pchlide oxidoreductase POR). Previous study suggested that barley large POR aggregates may be generated by dithiol oxidation of cysteines of two POR monomers, which can be disconnected by some reducing agents. POR aggregate assembly may be correlated with seedling greening in barley, but not in Arabidopsis. Thus, NO may affect POR activity and seedling greening differently between Arabidopsis and barley. We proved this assumption by non-denaturing gel-analysis and reactive oxygen species (ROS) monitoring during the greening. NO treatments cause S-nitrosylation to POR cysteine residues and disassembly of POR aggregates. This modification reduces POR activity and induces Pchlide accumulation and singlet oxygen generation upon dark-to-high-light shift (and therefore inducing photobleaching lesions) in barley leaf apex, but not in Arabidopsis seedlings. ROS staining and ROS-related-gene expression detection confirmed that superoxide anion and singlet oxygen accumulated in barley etiolated seedlings after the NO treatments, when exposed to a fluctuating light. The data suggest that POR aggregate assembly may be correlated with barley chlorophyll biosynthesis and redox homeostasis during greening. Cysteine S-nitrosylation may be one of the key reasons for the NO-induced inhibition to chlorophyll biosynthetic enzymes.


Asunto(s)
Arabidopsis/metabolismo , Clorofilidas/biosíntesis , Hordeum/metabolismo , Óxido Nítrico/metabolismo , Oxígeno Singlete/metabolismo
6.
Transgenic Res ; 25(4): 507-16, 2016 08.
Artículo en Inglés | MEDLINE | ID: mdl-26951193

RESUMEN

Bombyxin (BBX) is an insulin-like peptide exists in the silkworm Bombyx mori. Our previous studies on the effects of inhibiting BBX-B8 expression found that BBX-B8 is important for the development of organ, reproduction and trehalose metabolism in the silkworms. In this paper, we investigated the expression profile of the BBX-B8 gene and effect of BBX-B8 overexpression on the development, body weight, silk protein synthesis and egg diapause of B. mori to further understand BBX-B8 functions. BBX-B8 gene expression could be detected in the brains, midguts, anterior silkglands, ovaries, testes, fat bodies, hemolymph, malpighian tubules and embryos by RT-PCR, however it was mainly expressed in the brain. Western blots showed that the change in BBX-B8 expression was not obvious in the brain of 1- to 4-day-old larvae of fifth instar silkworms, but expression increased substantially at 5- to 6-day-old larvae of fifth instar silkworms. Transgenic silkworms overexpressing BBX-B8 were obtained by introducing non-transposon transgenic vector pIZT-B8 containing a BBX-B8 gene driven by Orgyia pseudotsugata nucleopolyhedrovirus IE2 promoter into the genome. Development duration of the transgenic silkworms was delayed by 2.5-3.5 days. Cocoon shell weight of transgenic silkworms was reduced by 4.79 % in females and 7.44 % in males, pupal weight of transgenic silkworms was reduced 6.75 % in females and 13.83 % in males compared to non-transgenic silkworms, and 5.56-14.29 % of transgenic moths laid nondiapausing eggs. All results indicated that BBX-B8 plays an important role in the development, silk protein synthesis and egg diapause of silkworm.


Asunto(s)
Animales Modificados Genéticamente/genética , Bombyx/crecimiento & desarrollo , Diapausa/genética , Neuropéptidos/genética , Seda/biosíntesis , Animales , Peso Corporal/genética , Bombyx/genética , Bombyx/fisiología , Femenino , Regulación del Desarrollo de la Expresión Génica , Larva , Masculino , Regiones Promotoras Genéticas , Seda/genética
7.
Front Psychol ; 14: 1291711, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-38259527

RESUMEN

Objectives: Parents are one of the main social agents that shape young athletes' experiences and participation in sports, but they are also the least explored group in the literature. Therefore, the purpose of this study was to conduct a systematic review of research on the role of parents in the motivation of young athletes. Method: The systematic literature review consisted of four electronic databases from which 29 articles published in English and in full-text form in peer-reviewed journals between 1999 and 2023 were retrieved. Results: A total of 29 studies met the eligibility criteria. These studies collectively surveyed 9,185 young athlete participants and 2,191 parent participants. The sample comprised 26 quantitative studies and 3 qualitative studies. The findings underscore that parents play both unique and synergistic multidimensional roles in motivating young athletes. Parents' positive goals and values, autonomy-supportive parenting styles, moderate parental involvement, positive parent-child relationships, and a parent-initiated task climate are identified as optimal parenting strategies. Conclusion: While parents undeniably play a crucial role in motivating young athletes, the manner and extent of their involvement are key.

8.
Adv Mater ; 35(37): e2203684, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35735048

RESUMEN

Diverse microscopic ionic dynamics help mediate the ability of a biological neural network to handle complex tasks with low energy consumption. Thus, rich internal ionic dynamics in memristors based on transition metal oxide are expected to provide a unique and useful platform for implementing energy-efficient neuromorphic computing. To this end, a titanium oxide (TiOx )-based interface-type dynamic memristor and an niobium oxide (NbOx )-based Mott memristor are integrated as an artificial dendrite and spike-firing soma, respectively, to construct a dendritic neuron unit for realizing high-efficiency spatial-temporal information processing. Further, a dendritic neural network is hardware-implemented for spatial-temporal information processing to highlight the computational advantages achieved by incorporating dendritic functions in the network. Human motion recognition is demonstrated using the Nanyang Technological University-Red Green Blue (NTU-RGB) dataset as a benchmark spatial-temporal task; it shows a nearly 20% improvement in accuracy for the memristors-based hardware incorporating dendrites and a 1000× advantage in power efficiency compared to that of the graphics processing unit (GPU). The dendritic neuron developed in this study can be considered a critical building block for implementing more bio-plausible neural networks that can manage complex spatial-temporal tasks with high efficiency.

9.
Front Public Health ; 11: 1164744, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37124786

RESUMEN

Background: Despite years of public cardiopulmonary resuscitation (CPR) training efforts, the training rate and survival following out-of-hospital cardiac arrest (OHCA) have increased modestly in China. Access is imperative to increase the public CPR training rate, which is determined by both demand- (e.g., the lay public) and supply-side (e.g., CPR trainers) factors. We aimed to explore the demand and supply determinants of access to CPR training for the lay public in China. Methods: Qualitative semi-structured interviews were conducted with 77 laypeople (demand side) and eight key stakeholders from CPR training institutions (supply side) in Shanghai, China. The interview guide was informed by Levesque et al. healthcare access framework. Data were transcribed, quantified, described, and analyzed through thematic content analysis. Results: On the demand side, the laypeople's ability to perceive their need and willingness for CPR training was strong. However, they failed to access CPR training mainly due to the lack of information on where to get trained. Overestimation of skills, optimism bias, and misconceptions impeded laypeople from attending training. On the supply side, trainers were able to meet the needs of the trainees with existing resources, but they relied on participants who actively sought out and registered for training and lacked an understanding of the needs of the public for marketing and encouraging participation in the training. Conclusion: Insufficient information and lack of initiative on the demand side, lack of motivation, and understanding of public needs on the supply side all contributed to the persistently low CPR training rate in China. Suppliers should integrate resources, take the initiative to increase the CPR training rate, innovate training modes, expand correct publicity, and establish whole-process management of training programs.


Asunto(s)
Reanimación Cardiopulmonar , Paro Cardíaco Extrahospitalario , Humanos , Reanimación Cardiopulmonar/educación , China , Paro Cardíaco Extrahospitalario/terapia
10.
Nat Commun ; 14(1): 6262, 2023 10 07.
Artículo en Inglés | MEDLINE | ID: mdl-37805547

RESUMEN

Plant height is a key agronomic trait that affects yield and is controlled by both phytohormone gibberellin (GA) and ultraviolet-B (UV-B) irradiation. However, whether and how plant height is modulated by UV-B-mediated changes in GA metabolism are not well understood. It has not been reported that the E3 ubiquitin ligase Anaphase Promoting Complex/Cyclosome (APC/C) is involved in the regulation of plant growth in response to environmental factors. We perform a forward genetic screen in soybean and find that a mutation in Glycine max Increased Leaf Petiole Angle1 (GmILPA1), encoding a subunit of the APC/C, lead to dwarfism under UV-B irradiation. UV-B promotes the accumulation of GmILPA1, which ubiquitinate the GA catabolic enzyme GA2 OXIDASE-like (GmGA2ox-like), resulting in its degradation in a UV-B-dependent manner. Another E3 ligase, GmUBL1, also ubiquitinate GmGA2ox-like and enhance the GmILPA1-mediated degradation of GmGA2ox-like, which suggest that GmILPA1-GmGA2ox-like module counteract the UV-B-mediated reduction of bioactive GAs. We also determine that GmILPA1 is a target of selection during soybean domestication and breeding. The deletion (Indel-665) in the promoter might facilitate the adaptation of soybean to high UV-B irradiation. This study indicates that an evolutionary GmILPA1 variant has the capability to develop ideal plant architecture with soybean cultivars.


Asunto(s)
Glycine max , Ubiquitina-Proteína Ligasas , Ubiquitina-Proteína Ligasas/genética , Ubiquitina-Proteína Ligasas/metabolismo , Glycine max/genética , Glycine max/metabolismo , Giberelinas , Fitomejoramiento , Ciclosoma-Complejo Promotor de la Anafase , Plantas/metabolismo , Hojas de la Planta/metabolismo
11.
Transgenic Res ; 21(1): 101-11, 2012 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-21533901

RESUMEN

The silk gland of the silkworm is a highly specialized organ that has the wonderful ability to synthesize and secrete silk protein. To express human granucyto-macrophage colony-stimulating factor (hGM-CSF) in the posterior silk glands of gene-targeted silkworms, a targeting vector pSK-FibL-L-A3GFP-PH-GMCSF-LPA-FibL-R was constructed, harboring a 1.2 kb portion of the left homogenous arm (FibL-L), a 0.5 kb portion of the right homogenous arm (FibL-R), fibroin H-chain-promoter-driven hGM-CSF and silkworm actin 3-promoter-driven gfp. The targeting vector was then introduced into the eggs of silkworm, and the transgenic silkworms were verified by PCR and DNA hybridization after being screened for the gfp gene. Western blotting analysis using an antibody against hGM-CSF demonstrated a specific band with a molecular weight of 22 kD in the silk glands of the G3 generation transgenic silkworms. The level of expression of hGM-CSF in the posterior silk glands of the G3 generation transgenic silkworms was approximately 2.70 ng/g of freeze-dried powdered posterior silk gland. These results showed that the heterologous gene could be introduced into the silkworm genome and expressed successfully. Further more, the exogenous genes existing in the G5 transgenic silkworm identified by PCR confirmed its integration stability. In addition, the silk glands containing expressed hGM-CSF performed the function of significantly increasing leukocyte count of CY-treated mice in a time-and-dose-dependent manner.


Asunto(s)
Bombyx/genética , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Proteínas Recombinantes/genética , Animales , Animales Modificados Genéticamente , Glándulas Exocrinas/metabolismo , Exones , Regulación de la Expresión Génica , Vectores Genéticos , Factor Estimulante de Colonias de Granulocitos y Macrófagos/metabolismo , Factor Estimulante de Colonias de Granulocitos y Macrófagos/farmacología , Humanos , Recuento de Leucocitos , Ratones , Ratones Endogámicos ICR , Peso Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología
12.
Mol Biol Rep ; 39(5): 5967-76, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22207176

RESUMEN

VASA is considered to be one of the most reliable molecular marker of germ cells. In order to study the Bombyx mori vasa-like gene (Bmvlg), the cDNAs of Bmvlg were cloned and sequenced, and the results showed that the Bmvlg gene from the fifth instar larval testes had four alternative splicing isoforms. The open reading frame (ORF) of the longest isoform was composed of 1,806 nucleotides encoding 601 amino acid residues and contained some known conserved domains. The other three isoforms had complete ORF, suggesting that the Bmvlg gene had several alternative splicing forms, completely different from that of Drosophila melanogaster. The results of sequencing demonstrated that the Bmvlg gene promoter had several elements conserved in eukaryotic and gonadal tissue-specific promoters. To detect the specificity of the Bmvlg promoter, a transient expression vector pSK-vlg-DsRed-polyA with a red fluorescent protein gene (DsRed), controlled by the Bmvlg promoter and a vector pIZT/V5-His-vlg-DsRed containing a Bmvlg fused with DsRed driven by the Bmvlg promoter, was constructed, respectively. Red fluorescence could be observed in some transfected BmN cells derived from silkworm ovaries and in the eggs injected with the vector pSK-vlg-DsRed-polyA, but red fluorescence could not be detected in the tissues of silkworm larva, after the transient expression vector was injected into blood, suggesting the Bmvlg promoter had gonadal tissue specificity. The transcription levels of Bmvlg in gonads of the fourth and fifth instar larvae were determined by fluorescent quantitative PCR, and the results revealed that the expression level of the Bmvlg gene in testes was slightly higher than that in ovaries. The expression levels of Bmvlg were lower in the fourth instar larva than that in the fifth instar larvae. Moreover, subcellular localization experiments showed that Bmvlg mainly existed in cytoplasm. These results provided new clues for understanding the function of the Bmvlg gene.


Asunto(s)
Empalme Alternativo/genética , Bombyx/enzimología , Bombyx/genética , ARN Helicasas DEAD-box/genética , Genes de Insecto/genética , Regiones Promotoras Genéticas/genética , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bombyx/citología , Secuencia Conservada/genética , ARN Helicasas DEAD-box/química , ARN Helicasas DEAD-box/metabolismo , Vectores Genéticos/genética , Proteínas de Insectos/química , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Datos de Secuencia Molecular , Óvulo/citología , Óvulo/enzimología , Transporte de Proteínas , Homología de Secuencia de Aminoácido , Fracciones Subcelulares/enzimología , Transcripción Genética
13.
Can J Microbiol ; 58(7): 872-83, 2012 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-22712678

RESUMEN

A novel Bombyx mori cypovirus 1 isolated from infected silkworm larvae and tentatively assigned as Bombyx mori cypovirus 1 isolate Suzhou (BmCPV-SZ). The complete nucleotide sequences of genomic segments S1-S10 from BmCPV-SZ were determined. All segments possessed a single open reading frame; however, bioinformatic evidence suggested a short overlapping coding sequence in S1. Each BmCPV-SZ segment possessed the conserved terminal sequences AGUAA and GUUAGCC at the 5' and 3' ends, respectively. The conserved A/G at the -3 position in relation to the AUG codon could be found in the BmCPV-SZ genome, and it was postulated that this conserved A/G may be the most important nucleotide for efficient translation initiation in cypoviruses (CPVs). Examination of the putative amino acid sequences encoded by BmCPV-SZ revealed some characteristic motifs. Homology searches showed that viral structural proteins VP1, VP3, and VP4 had localized homologies with proteins of Rice ragged stunt virus , a member of the genus Oryzavirus within the family Reoviridae. A phylogenetic tree based on RNA-dependent RNA polymerase sequences demonstrated that CPV is more closely related to Rice ragged stunt virus and Aedes pseudoscutellaris reovirus than to other members of Reoviridae, suggesting that they may have originated from common ancestors.


Asunto(s)
Bombyx/virología , Genoma Viral/genética , Filogenia , Reoviridae/clasificación , Reoviridae/genética , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Animales , Biología Computacional , Larva/virología , Datos de Secuencia Molecular , Sistemas de Lectura Abierta/genética , Homología de Secuencia , Proteínas Virales/genética
14.
J Insect Sci ; 12: 4, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22934965

RESUMEN

In the past few decades, the mouse has been used as a mammalian model for hyperuricemia and gout, which has increased not only in prevalence, but also in clinical complexity, accentuated in part by a dearth of novel advances in treatments for hyperuricemia and gouty arthritis. However, the use of mice for the development of gouty therapeutic drugs creates a number of problems. Thus, identification and evaluation of the therapeutic effects of chemicals in an alternative animal model is desirable. In the present study, the effects of gouty therapeutic drugs on lowering the content of uric acid and inhibiting activity of xanthine oxidase were evaluated by using a silkworm model, Bombyx mori L. (Lepidoptera: Bombycidae). The results showed that the effectiveness of oral administration of various gouty therapeutic drugs to 5(th) instar silkworms is consistent with results for human. The activity of xanthine oxidase of silkworm treated with allopurinol was lower, and declined in a dose-dependent manner compared with control silkworms, while sodium bicarbonate failed at inhibiting the activity of xanthine oxidase. The concentration of uric acid in the both hemolymph and fat body declined by 90 and 95% at six days post-administration with 25 mg/mL of allopurinol, respectively (p < 0.01), while the concentration of uric acid in both the hemolymph and fat body also declined by 81 and 95% at six days post-administration with 25 mg/mL of sodium bicarbonate, respectively (p < 0.01). Moreover, the epidermis of silkworm treated with allopurinol or sodium bicarbonate became transparent compared with the negative control group. These results suggest that silkworm larva can be used as an animal model for screening and evaluation of gouty therapeutic drugs.


Asunto(s)
Bombyx/efectos de los fármacos , Evaluación Preclínica de Medicamentos/métodos , Supresores de la Gota/farmacología , Gota/tratamiento farmacológico , Modelos Animales , Animales , Bombyx/crecimiento & desarrollo , Bombyx/metabolismo , Relación Dosis-Respuesta a Droga , Cuerpo Adiposo/metabolismo , Hemolinfa/metabolismo , Proteínas de Insectos/metabolismo , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Larva/metabolismo , Ácido Úrico/metabolismo , Xantina Oxidasa/metabolismo
15.
Adv Mater ; 34(42): e2205926, 2022 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-36027579

RESUMEN

Efficient exciton diffusion and charge transport play a vital role in advancing the power conversion efficiency (PCE) of organic solar cells (OSCs). Here, a facile strategy is presented to simultaneously enhance exciton/charge transport of the widely studied PM6:Y6-based OSCs by employing highly emissive trans-bis(dimesitylboron)stilbene (BBS) as a solid additive. BBS transforms the emissive sites from a more H-type aggregate into a more J-type aggregate, which benefits the resonance energy transfer for PM6 exciton diffusion and energy transfer from PM6 to Y6. Transient gated photoluminescence spectroscopy measurements indicate that addition of BBS improves the exciton diffusion coefficient of PM6 and the dissociation of PM6 excitons in the PM6:Y6:BBS film. Transient absorption spectroscopy measurements confirm faster charge generation in PM6:Y6:BBS. Moreover, BBS helps improve Y6 crystallization, and current-sensing atomic force microscopy characterization reveals an improved charge-carrier diffusion length in PM6:Y6:BBS. Owing to the enhanced exciton diffusion, exciton dissociation, charge generation, and charge transport, as well as reduced charge recombination and energy loss, a higher PCE of 17.6% with simultaneously improved open-circuit voltage, short-circuit current density, and fill factor is achieved for the PM6:Y6:BBS devices compared to the devices without BBS (16.2%).

16.
Mol Biol Rep ; 38(5): 3173-80, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-20127514

RESUMEN

For mass production of urease B subunit (UreB) and heat shock protein A subunit (HspA) of Helicobacter pylori with Bombyx mori nuclear polyhedrosis virus (BmNPV) baculovirus expression system (BES) and to determine whether they could be used as an oral vaccine against H. pylori, besides, to determine the time course of expressed recombinant protein and the optimum acquisition time directly through green fluorescence, HspA and enhanced green fluorescence protein (EGFP) genes were cloned into vector pFastBacDual to form donor vector pFastBacDual-(EGFP) (HspA), UreB gene was cloned into vector pFastBacDual to form donor vector pFastBacDual-UreB,then they were transformed into E. coli BmDH10Bac to obtain the recombinant Bacmid-(EGFP) (HspA) and Bacmid-UreB respectively. They were used to transfect BmN cells and generated the recombinant baculovirus BmNPV-(EGFP) (HspA) and BmNPV-UreB. Using these recombinant baculovirus BmNPV-(EGFP) (HspA) and BmNPV-UreB inoculated the silkworm pupae, a recombinant HspA and UreB protein were expressed in silkworm pupae, which were around 13 and 62 kDa in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. After oral immunization of mice, serum specific IgG antibodies against HspA and UreB in vaccine group were much higher than that in mock and native silkworm powder control groups. The results indicated that the expressed recombinant HspA and UreB in silkworm pupae would possess good immunogenicity. In addition, when EGFP and HspA proteins were expressed, a direct correlation between the increase in intensity of fluorescence and HspA concentration.


Asunto(s)
Proteínas Bacterianas/inmunología , Bombyx/microbiología , Proteínas de Choque Térmico/inmunología , Helicobacter pylori/inmunología , Subunidades de Proteína/inmunología , Pupa/microbiología , Ureasa/inmunología , Animales , Proteínas Bacterianas/genética , Bombyx/crecimiento & desarrollo , Bombyx/inmunología , Femenino , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Proteínas de Choque Térmico/genética , Helicobacter pylori/química , Humanos , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Subunidades de Proteína/química , Subunidades de Proteína/genética , Pupa/inmunología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , Ureasa/química , Ureasa/genética
17.
J Agric Food Chem ; 69(44): 13135-13142, 2021 Nov 10.
Artículo en Inglés | MEDLINE | ID: mdl-34709805

RESUMEN

α-Santalene belongs to a class of natural compounds with many physiological functions and medical applications. Advances in metabolic engineering enable non-native hosts (e.g., Escherichia coli) to produce α-santalene, the precursor of sandalwood oil. However, imbalances in enzymatic activity often result in a metabolic burden on hosts and repress the synthetic capacity of the desired product. In this work, we manipulated ribosome binding sites (RBSs) to optimize an α-santalene synthetic operon in E. coli, and the best engineered E. coli NA-IS3D strain could produce α-santalene at a titer of 412 mg·L-1. Concerning the observation of the inverse correlation between indole synthesis and α-santalene production, this study speculated that indole-associated amino acid metabolism would be competitive to the synthesis of α-santalene rather than indole toxicity itself. The deletion of tnaA could lead to a 1.5-fold increase in α-santalene production to a titer of 599 mg·L-1 in E. coli tnaA- NA-IS3D. Our results suggested that the optimization of RBS sets of the synthetic module and attenuation of the competitive pathway are promising approaches for improving the production of terpenoids including α-santalene.


Asunto(s)
Escherichia coli , Ingeniería Metabólica , Escherichia coli/genética , Aceites de Plantas , Sesquiterpenos Policíclicos , Sesquiterpenos
18.
Front Plant Sci ; 12: 794020, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-35154184

RESUMEN

Soil stress, such as salinity, is a primary cause of global crop yield reduction. Existing crop phenotyping platforms cannot fully meet the specific needs of phenomics studies of plant response to soil stress in terms of throughput, environmental controllability, or root phenotypic acquisition. Here, we report the WinRoots, a low-cost and high-throughput plant soil cultivation and phenotyping system that can provide uniform, controlled soil stress conditions and accurately quantify the whole-plant phenome, including roots. Using soybean seedlings exposed to salt stress as an example, we demonstrate the uniformity and controllability of the soil environment in this system. A high-throughput multiple-phenotypic assay among 178 soybean cultivars reveals that the cotyledon character can serve as a non-destructive indicator of the whole-seedling salt tolerance. Our results demonstrate that WinRoots is an effective tool for high-throughput plant cultivation and soil stress phenomics studies.

19.
Mol Biol Rep ; 37(6): 2599-608, 2010 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19728146

RESUMEN

The complete nucleotide sequence of the mitogenome of Bombyx mandarina strain Qingzhou was determined. The circular genome is 15,717 bp long and has the typical gene organization and order of lepidopteran mitogenomes. All protein-coding sequences are initiated with a typical ATN codon, except the COI gene, which has a 4-bp TTAG putative initiator codon. Eleven of the 13 protein-coding gene have a complete termination codon (all TAA), but the remaining two genes terminate with incomplete codons. All transfer RNAs (tRNAs) have a clover-leaf structure typical of the mitochondrial tRNAs, and some of them have a mismatch in the four-stem-and-loop structure. The length of the A + T rich region of B. mandarina strain Qingzhou is 495 bp, shorter than that of B. mandarina strain Tsukuba (747 bp) but similar to that of Bombyx mori. Phylogenetic analysis based on the whole mitochondrial genome sequences of the available sequenced species (B. mori strains C-108, Aojuku, Backokjam, and Xiafang, B. mandarina strains Tsukuba, Ankang, and Qingzhou, and Antheraea pernyi) shows the origin of the domesticated silkmoth B. mori to be the Chinese B. mandarina. Nuclear mitochondrial pseudogene sequences were detected in the nuclear genome of B. mori with the MEGA BLAST search program. A phylogenetic analysis of these nuclear mitochondrial pseudogene sequences suggests that B. mori was domesticated independently in different areas and periods.


Asunto(s)
Bombyx/genética , Genoma Mitocondrial/genética , Filogenia , Secuencia Rica en At/genética , Animales , Secuencia de Bases , Núcleo Celular/genética , ADN Mitocondrial/genética , Evolución Molecular , Genes de Insecto/genética , Datos de Secuencia Molecular , Sistemas de Lectura Abierta/genética , Seudogenes/genética , ARN Ribosómico/genética , ARN de Transferencia/genética , Seda/biosíntesis , Factores de Tiempo
20.
Sci Total Environ ; 740: 139911, 2020 Oct 20.
Artículo en Inglés | MEDLINE | ID: mdl-32927563

RESUMEN

Dissolved humic substances (DHS) are ubiquitous in surface and subsurface aquatic environments and greatly affect the redox transformation of organic contaminants as reactants and/or electron transfer mediators. However, little is known about the quantitative relationship between the mediation efficiency of DHS and the physicochemical properties of DHS. Using sulfide-induced nitrobenzene reduction as a model system, we measured the reduction rate of nitrobenzene in the presence of 12 different DHS (20 mgC·L-1), including 4 commercial humic substances (Suwannee River humic and fulvic acids and Pahokee Peat humic and fulvic acids) and 8 soil humic substances collected as leachates from a wide variety of soils. In addition to the UV-vis absorption and fluorescence spectra, the electron donating/accepting capacities (EDC/EAC) of the tested DHS were measured using an electrochemical approach. A significant linear correlation (r = 0.99, P < .0001) was observed between the observed pseudo-first-order rate constant (kobs) of nitrobenzene reduction and the sum of EDC and EAC which is defined as electron transfer capacity (ETC) of DHS. A relatively good positive correlation (r = 0.69, P < .2) was shown between the kobs and the specific UV-absorbance at 254 nm (SUVA254), whereas no good correlation was shown between the kobs and the fluorescence of the C1-C4 components identified by the excitation emission matrices and parallel factor (EEM-PARAFAC) analysis. This study provides a new framework for accurate prediction of the capability of DHS in mediating the redox transformation of organic contaminants. CAPSULE: A significant linear correlation exists between the kinetics of nitrobenzene reduction by sulfide and electron transfer capacity of mediating dissolved humic substances.

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