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1.
Plant Physiol ; 195(3): 2354-2371, 2024 Jun 28.
Artículo en Inglés | MEDLINE | ID: mdl-38501602

RESUMEN

Teosinte branched 1/Cycloidea/Proliferating cell factor (TCP) transcription factors function in abiotic stress responses. However, how TCPs confer salt tolerance is unclear. Here, we characterized a TCP transcription factor, BpTCP20, that responds to salt stress in birch (Betula platyphylla Suk). Plants overexpressing BpTCP20 displayed increased salt tolerance, and Bptcp20 knockout mutants displayed reduced salt tolerance relative to the wild-type (WT) birch. BpTCP20 conferred salt tolerance by mediating stomatal closure and reducing reactive oxygen species (ROS) accumulation. Chromatin immunoprecipitation sequencing showed that BpTCP20 binds to NeuroD1, T-box, and two unknown elements (termed TBS1 and TBS2) to regulate target genes. In birch, salt stress led to acetylation of BpTCP20 acetylation at lysine 259. A mutated BpTCP20 variant (abolished for acetylation, termed BpTCP20259) was overexpressed in birch, which led to decreased salt tolerance compared with plants overexpressing BpTCP20. However, BpTCP20259-overexpressing plants still displayed increased salt tolerance relative to untransformed WT plants. BpTCP20259 showed reduced binding to the promoters of target genes and decreased target gene activation, leading to decreased salt tolerance. In addition, we identified dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex (BpPDCE23), an acetyltransferase that interacts with and acetylates BpTCP20 to enhance its binding to DNA motifs. Together, these results suggest that BpTCP20 is a transcriptional regulator of salt tolerance, whose activity is modulated by BpPDCE23-mediated acetylation.


Asunto(s)
Betula , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas , Tolerancia a la Sal , Factores de Transcripción , Tolerancia a la Sal/genética , Acetilación , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Betula/genética , Betula/metabolismo , Betula/fisiología , Acetiltransferasas/metabolismo , Acetiltransferasas/genética , Plantas Modificadas Genéticamente , Especies Reactivas de Oxígeno/metabolismo
2.
J Fluoresc ; 34(1): 425-436, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37284963

RESUMEN

A novel reagent named 4-(N-methyl-1,3-dioxo-benzoisoquinolin-6-yl-oxy)benzene sulfonyl chloride (MBIOBS-Cl) for the determination of estrogens in food samples by high-performance liquid chromatography (HPLC) with fluorescence detection has been developed. Estrogens could be easily labeled by MBIOBS-Cl in Na2CO3-NaHCO3 buffer solution at pH 10.0. The complete labeling reaction for estrogens could be accomplished within five minutes, the corresponding derivatives exhibited strong fluorescence with the maximum excitation and emission wavelengths at 249 nm and 443 nm, respectively. The derivatization conditions, such as the molar ratio of reagent to estrogens, derivatization time, pH, temperature, and buffers were optimized. Derivatives were sufficiently stable to be efficiently analyzed by HPLC with a reversed-phase Agilent ZORBAX 300SB-C18 column with a good baseline resolution. Excellent linear correlations were obtained for all estrogen derivatives with correlation coefficients greater than 0.9998. Ultrasonic-Assisted extraction was used to optimize the extraction of estrogens from meat samples with a recovery higher than 82%. The detection limits (LOD, S/N = 3) of the method ranged from 0.95 to 3.3 µg· kg-1. The established method, which is fast, simple, inexpensive, and environment friendly, can be successfully applied for the detection of four steroidal estrogens from meat samples with little matrix interference.


Asunto(s)
Estrógenos , Carne , Estrógenos/análisis , Cromatografía Líquida de Alta Presión/métodos , Carne/análisis
3.
Appl Microbiol Biotechnol ; 108(1): 340, 2024 May 23.
Artículo en Inglés | MEDLINE | ID: mdl-38777914

RESUMEN

Horizontal gene transfer occurs frequently in bacteria, but the mechanism driving activation and optimization of the expression of horizontally transferred genes (HTGs) in new recipient strains is not clear. Our previous study found that spontaneous tandem DNA duplication resulted in rapid activation of HTGs. Here, we took advantage of this finding to develop a novel technique for tandem gene duplication, named tandem gene duplication selected by activation of horizontally transferred gene in bacteria (TDAH), in which tandem duplication was selected by the activation of horizontally transferred selectable marker gene. TDAH construction does not contain any reported functional elements based on homologous or site-specific recombination and DNA amplification. TDAH only contains an essential selectable marker for copy number selection and 9-bp-microhomology border sequences for precise illegitimate recombination. One transformation and 3 days were enough to produce a high-copy strain, so its procedure is simple and fast. Without subsequent knockout of the endogenous recombination system, TDAH could also generate the relatively stable high-copy tandem duplication for plasmid-carried and genome-integrated DNA. TDAH also showed an excellent capacity for increase gene expression and worked well in different industrial bacteria. We also applied TDAH to select the optimal high copy number of ribA for vitamin B2 production in E. coli; the yield was improved by 3.5 times and remained stable even after 12 subcultures. TDAH is a useful tool for recombinant protein production and expression optimization of biosynthetic pathways. KEY POINTS: • We develop a novel and efficient technique (TDAH) for tandem gene duplication in bacterium. TDAH is based on the mechanism of HTG rapid activation. TDAH does not contain any reported functional elements based on homologous recombination and DNA amplification. TDAH only contains an essential selectable marker for copy number selection, so its construction and procedure are very simple and fast. • TDAH is the first reported selected and stable tandem-gene-duplication technique in which the selected high-copy plasmid-carried and genome-integrated DNA could remain stable without the subsequent knockout of recombination system. • TDAH showed an excellent capacity for regulating gene expression and worked well in different industrial bacteria, indicating it is a useful tool for recombinant protein production and expression optimization of biosynthetic pathways. • TDAH was applied to select the optimal high copy number of ribA for vitamin B2 production in E. coli; the yield was improved by 3.5-fold and remained stable even after 12 subcultures.


Asunto(s)
Escherichia coli , Duplicación de Gen , Transferencia de Gen Horizontal , Plásmidos , Escherichia coli/genética , Escherichia coli/metabolismo , Plásmidos/genética , Bacterias/genética , Bacterias/metabolismo , Dosificación de Gen , Recombinación Genética
4.
Pestic Biochem Physiol ; 201: 105865, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38685241

RESUMEN

Fluvalinate is widely used in the control of Varroa destructor, but its residues in colonies threaten honeybees. The effect of fluvalinate-induced dysbiosis on honeybee-related gene expression and the gut microenvironment of honeybees has not yet been fully elucidated. In this study, two-day-old larvae to seven-day-old adult worker bees were continuously fed different amounts of fluvalinate-sucrose solutions (0, 0.5, 5, and 50 mg/kg), after which the expression levels of two immune-related genes (Hymenoptaecin and Defensin1) and three detoxication-related genes (GSTS3, CAT, and CYP450) in worker bees (1, 7, and 20 days old) were measured. The effect of fluvalinate on the gut microbes of worker bees at seven days old also was explored using 16S rRNA Illumina deep sequencing. The results showed that exposure of honeybees to the insecticide fluvalinate affected their gene expression and gut microbial composition. As the age of honeybees increased, the effect of fluvalinate on the expression of Hymenoptaecin, CYP450, and CAT decreased, and the abundance of honeybee gut bacteria was affected by increasing the fluvalinate concentration. These findings provide insights into the synergistic defense of honeybee hosts against exogenous stresses in conjunction with honeybee gut microbes.


Asunto(s)
Péptidos Catiónicos Antimicrobianos , Microbioma Gastrointestinal , Insecticidas , Nitrilos , Piretrinas , Animales , Abejas/efectos de los fármacos , Abejas/microbiología , Microbioma Gastrointestinal/efectos de los fármacos , Piretrinas/farmacología , Piretrinas/toxicidad , Insecticidas/farmacología , Insecticidas/toxicidad , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Sistema Enzimático del Citocromo P-450/metabolismo , ARN Ribosómico 16S/genética
5.
Ecotoxicol Environ Saf ; 254: 114716, 2023 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-36870311

RESUMEN

Acetamiprid is a neonicotinoid insecticide used in crop protection worldwide. Such widespread application can pose risks to pollinator insects, particularly to honeybees (Apis mellifera); therefore, the evaluation of the harmful effects of acetamiprid is necessary. Recent studies report behavior and gene expression dysfunction in honeybees, related to acetamiprid contamination. However, most studies do not consider potential metabolism disorders. To examine the effects of sublethal acetamiprid doses on the hemolymph metabolism of honeybees, worker bee larvae(2 days old) were fed with sucrose water containing different concentrations of acetamiprid (0, 5, and 25 mg/L) until capped (6 days old). The hemolymph (200 µL) of freshly capped larvae was collected for liquid chromatography-mass spectrometry (LC-MS). Overall, increasing acetamiprid exposure induced greater metabolic variations in worker bee larvae(treated groups compared to untreated). In the positive ion mode, 36 common differential metabolites in the acetamiprid-treated groups were screened from the identified differential metabolites. Of these, 19 metabolites were upregulated, and 17 were downregulated. 10 common differential metabolites were screened in the negative ion mode. 3 metabolites were upregulated, and 7 metabolites were downregulated. These common metabolites included traumatic acid, indole etc. These commonly differentiated metabolites were classified as compounds with biological roles, lipids, and phytochemical compounds, and others. The metabolic pathways of common differentiated metabolites with significant differences (P < 0.05) included the metabolism of tryptophan, purines, phenylalanine, etc. As the concentration of acetamiprid increased, the content of traumatic acid increased, the content of tryptophan metabolite l-kynurenine and indole decreased, and the content of lipids also decreased. Our results revealed that the damage to honeybee larvae increased when the acetamiprid solution formulations residue in their food had a concentration higher than 5 mg/L, causing metabolic disorders in various substances in larvae. Analysis of these metabolic processes can provide a theoretical basis for further research on the metabolism of acetamiprid-treated honeybees and elucidate the detoxification mechanisms.


Asunto(s)
Insecticidas , Triptófano , Abejas , Animales , Larva , Neonicotinoides/toxicidad , Insecticidas/toxicidad , Lípidos
6.
Mol Cell Biochem ; 477(9): 2161-2171, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-33417162

RESUMEN

Circular RNAs (circRNAs) have emerged as a multifunctional class of RNAs, while there is limited knowledge on their functions in the development of cancers. Herein, we performed the current study to probe into the regulatory mechanism of circ_0044516 in malignant behaviors of gastric cancer (GC) cells with the involvement of microRNA (miR)-149-5p/human antigen R (HuR) axis. Firstly, the expression levels of circ_0044516 in GC cell lines and normal gastric mucosal epithelial cells were determined by qRT-PCR, and GC cell lines with the highest expression of circ_0044516 were screened for further cell experiments. Subsequently, the biological functions of silenced circ_0044516 in GC were identified by CCK-8, colony formation, and transwell assays. Xenograft mouse models were established for in vivo verification. Furthermore, luciferase reporter, RIP, RNA pull-down assay and rescue experiments were performed to explore the sponge regulatory mechanism of circ_0044516. circ_0044516 was suggested to be highly expressed in GC cell lines, and circ_0044516 could promote GC cell proliferation, migration and invasion, as well as in vivo tumor growth. In addition, silenced circ-0044516 reversed the promotive roles in cell viability caused by overexpressed HuR. Furthermore, circ_0044516 mainly localized in the cytoplasm, which may act as a miR-149-5p sponge to modulate HuR expression, thereby playing an essential role in GC development. This study suggests that circ_0044516 may promote HuR expression through sponging miR-149-5p, thereby playing a part in GC progression.


Asunto(s)
MicroARNs , Neoplasias Gástricas , Animales , Línea Celular Tumoral , Proliferación Celular/genética , Modelos Animales de Enfermedad , Regulación Neoplásica de la Expresión Génica , Humanos , Ratones , MicroARNs/genética , MicroARNs/metabolismo , Neoplasias Gástricas/metabolismo
7.
Small ; 15(48): e1901507, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-31168960

RESUMEN

As novel fluorescent nanomaterials, carbon dots (CDs) exhibit excellent photostability, good biocompatibility, and high quantum yield (QY). Their superior properties make them promising candidates for biomedical assays and therapy. Among them, the red-emission (>600 nm) CDs have attracted increasing attention in the past years due to their little damage to the biological matrix, deep tissue penetration, and minimum autofluorescence background of biosamples. This Review, summarizes the recent progress of far-red to near-infrared (NIR) CDs from the preparation and their biological applications. The challenges in designing far-red and NIR CDs and their further applications in biomedical fields are also discussed.


Asunto(s)
Biotecnología , Carbono/química , Rayos Infrarrojos , Puntos Cuánticos/química , Imagen Óptica , Solventes/química
8.
Plant Cell Physiol ; 59(5): 946-965, 2018 May 01.
Artículo en Inglés | MEDLINE | ID: mdl-29420810

RESUMEN

Trihelix transcription factors are characterized by containing a conserved trihelix (helix-loop-helix-loop-helix) domain that binds to GT elements required for light response, and they play roles in light stress and in abiotic stress responses. However, only a few of them have been functionally characterized. In the present study, we characterized the function of AST1 (Arabidopsis SIP1 clade Trihelix1) in response to salt and osmotic stress. AST1 shows transcriptional activation activity, and its expression is induced by osmotic and salt stress. A conserved sequence highly present in the promoters of genes regulated by AST1 was identified, which was bound by AST1, and termed the AGAG-box with the sequence [A/G][G/A][A/T]GAGAG. Additionally, AST1 also binds to some GT motifs including the sequence of GGTAATT, TACAGT, GGTAAAT and GGTAAA, but failed in binding to the sequence of GTTAC and GGTTAA. Chromatin immunoprecipitation combined with quantitative real-time reverse transcription-PCR analysis suggested that AST1 binds to the AGAG-box and/or some GT motifs to regulate the expression of stress tolerance genes, resulting in reduced reactive oxygen species, Na+ accumulation, stomatal apertures, lipid peroxidation, cell death and water loss rate, and increased proline content and reactive oxygen species scavenging capability. These physiological changes affected by AST1 finally improve salt and osmotic tolerance.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiología , Motivos de Nucleótidos/genética , Ósmosis , Tolerancia a la Sal , Factores de Transcripción/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Secuencia de Bases , Muerte Celular , Desecación , Perfilación de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Técnicas de Inactivación de Genes , Genes de Plantas , Genotipo , Malondialdehído/metabolismo , Estomas de Plantas/fisiología , Plantas Modificadas Genéticamente , Potasio/metabolismo , Prolina/metabolismo , Unión Proteica , Transporte de Proteínas , Especies Reactivas de Oxígeno/metabolismo , Reproducibilidad de los Resultados , Tolerancia a la Sal/genética , Análisis de Secuencia de ARN , Sodio/metabolismo , Fracciones Subcelulares/metabolismo , Factores de Tiempo , Factores de Transcripción/genética
9.
J Environ Sci (China) ; 65: 327-334, 2018 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-29548404

RESUMEN

As an efficient method for ammonium (NH4+) removal, contact catalytic oxidation technology has drawn much attention recently, due to its good low temperature resistance and short start-up period. Two identical filters were employed to compare the process for ammonium removal during the start-up period for ammonium removal in groundwater (Filter-N) and surface water (Filter-S) treatment. Two types of source water (groundwater and surface water) were used as the feed waters for the filtration trials. Although the same initiating method was used, Filter-N exhibited much better ammonium removal performance than Filter-S. The differences in catalytic activity among these two filters were probed using X-ray diffraction (XRD), scanning electron microscopy (SEM), X-ray photoelectron spectroscopy (XPS), and compositional analysis. XRD results indicated that different manganese oxide species were formed in Filter-N and Filter-S. Furthermore, the Mn3p XPS spectra taken on the surface of the filter films revealed that the average manganese valence of the inactive manganese oxide film collected from Filter-S (FS-MnOx) was higher than in the film collected from Filter-N (FN-MnOx). Mn(IV) was identified as the predominant oxidation state in FS-MnOx and Mn(III) was identified as the predominant oxidation state in FN-MnOx. The results of compositional analyses suggested that polyaluminum ferric chloride (PAFC) used during the surface water treatment was an important factor in the mineralogy and reactivity of MnOx. This study provides the theoretical basis for promoting the wide application of the technology and has great practical significance.


Asunto(s)
Compuestos de Amonio/química , Restauración y Remediación Ambiental/métodos , Filtración , Agua Dulce/química , Compuestos de Manganeso/química , Óxidos/química , Contaminantes Químicos del Agua/química , Compuestos de Amonio/análisis , Catálisis , Agua Subterránea/química , Microscopía Electrónica de Rastreo , Oxidación-Reducción , Espectroscopía de Fotoelectrones , Contaminantes Químicos del Agua/análisis
10.
Plant Mol Biol ; 93(4-5): 369-387, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-27975189

RESUMEN

KEY MESSAGE: ANAC069 binds to the DNA sequence of C[A/G]CG[T/G] to regulate the expression of genes, resulting in decreased ROS scavenging capability and proline biosynthesis, which contribute to increased sensitivity to salt and osmotic stress. NAM-ATAF1/2 and CUC2 (NAC) proteins are plant-specific transcription factors that play important roles in abiotic stress responses. In the present study, we characterized the physiological and regulatory roles of Arabidopsis thaliana ANAC069 in response to abiotic stresses. Arabidopsis plants overexpressing ANAC069 displayed increased sensitivity to abscisic acid, salt, and osmotic stress. Conversely, ANAC069 knockdown plants showed enhanced tolerance to salt and osmotic stress, but no change in ABA sensitivity. Further studies showed that ANAC069 inhibits the expression of SOD, POD, GST, and P5CS genes. Consequently, the transcript level of ANAC069 correlated negatively with the reactive oxygen species (ROS) scavenging ability and the proline level. The genes regulated by ANAC069 were further studied using a gene chip on a genome-wide scale, and 339 and 226 genes up- and downregulated by ANAC069 were identified. Analysis of the promoters of the genes affected by ANAC069 suggested that ANAC069 regulates the expression of genes mainly through interacting with the DNA sequence C[A/G]CG[T/G] in response to abiotic stresses. Collectively, our data suggest that ANAC069 could recognize C[A/G]CG[T/G] sequences to regulate the expression of genes that negatively regulates salt and osmotic stress tolerance by decreasing ROS scavenging capability and proline biosynthesis.


Asunto(s)
Adaptación Fisiológica/genética , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Proteínas de la Membrana/genética , Tolerancia a la Sal/genética , Ácido Abscísico/farmacología , Adaptación Fisiológica/efectos de los fármacos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Secuencia de Bases , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Proteínas de la Membrana/metabolismo , Mutación , Presión Osmótica , Reguladores del Crecimiento de las Plantas/farmacología , Plantas Modificadas Genéticamente , Prolina/biosíntesis , Regiones Promotoras Genéticas/genética , Unión Proteica , Especies Reactivas de Oxígeno/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tolerancia a la Sal/efectos de los fármacos , Cloruro de Sodio/farmacología , Estrés Fisiológico , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismo , Agua/metabolismo
11.
J Environ Sci (China) ; 57: 402-410, 2017 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-28647261

RESUMEN

Na-rich birnessite (NRB) was synthesized by a simple synthesis method and used as a high-efficiency adsorbent for the removal of ammonium ion (NH4+) from aqueous solution. In order to demonstrate the adsorption performance of the synthesized material, the effects of contact time, pH, initial ammonium ion concentration, and temperature were investigated. Adsorption kinetics showed that the adsorption behavior followed the pseudo second-order kinetic model. The equilibrium adsorption data were fitted to Langmuir and Freundlich adsorption models and the model parameters were evaluated. The monolayer adsorption capacity of the adsorbent, as obtained from the Langmuir isotherm, was 22.61mg NH4+-N/g at 283K. Thermodynamic analyses showed that the adsorption was spontaneous and that it was also a physisorption process. Our data revealed that the higher NH4+ adsorption capacity could be primarily attributed to the water absorption process and electrostatic interaction. Particularly, the high surface hydroxyl-content of NRB enables strong interactions with ammonium ion. The results obtained in this study illustrate that the NRB is expected to be an effective and economically viable adsorbent for ammonium ion removal from aqueous system.


Asunto(s)
Compuestos de Amonio/química , Óxidos/química , Sodio/química , Contaminantes Químicos del Agua/química , Adsorción , Compuestos de Amonio/análisis , Concentración de Iones de Hidrógeno , Cinética , Termodinámica , Contaminantes Químicos del Agua/análisis , Purificación del Agua/métodos
12.
Guang Pu Xue Yu Guang Pu Fen Xi ; 36(5): 1389-95, 2016 May.
Artículo en Zh | MEDLINE | ID: mdl-30001012

RESUMEN

The UV-B radiation on the surface of our planet has been enhanced due to gradual thinning of ozone layer. The change of solar spectrum UV-B radiation will cause damage to all kinds of terrestrial plants at certain degree. In this paper, taking breeding sorghum (Sorghum bicolor (L.Moench))variety Longza No.5 as sample, 40 µW·cm-2 UV-B radiation treatment was conducted on sorghum seedlings at two-leaf and one-heart stage and different time courses; then after a 2 d recovering, photosynthetic parameters were measured with a photosynthetic apparatus; the activities of antioxidant enzymes were detected as well. Our results revealed that, as the dosages of UV-B increasing, leaf browning injury was aggravated, plants dwarfing and significantly were reduced fresh weight and dry weight were observed; anthocyanin content was significantly increased; chlorophyll and carotenoid content significantly were reduced and net photosynthetic rate and chlorophyll fluorescence parameters were decreased. Meanwhile, with the increase in UV-B dosages, stomatal conductance, intercellular CO2 concentration and transpiration rate showed "down - up - down" trend; the activities of SOD and GR presented "down - up" changes; activities of POD and CAT demonstrated "down - up - down", and APX, GPX showed an "up - down - up" pattern. It is worth to note that, under the four-dose treatment, a sharp decline in net photosynthesis in sorghum seedlings was observed at 6 h UV-B treatment (equals to 2.4 J·m-2), and an obvious turning point was also found for other photosynthetic parameters and activities of antioxidant enzymes at the same time point. In summary, the results indicated that the enhanced UV-B radiation directly accounted for the damages in photosynthesis system including photosynthetic pigment content, net photosynthetic rate and chlorophyll fluorescence parameters of sorghum; the antioxidant system showed different responses to UV-B radiation below or above 6 h treatment: ASA-GSH cycle was more sensitive to low-dose UV-B radiation, while high-dose UV-B radiation not only undermined the photosynthesis system, but also triggered plant enzymatic and non-enzymatic antioxidant systems, resulting in leaf browning and necrosis,biomass accumulation reduction, plant dwarfing and even death.


Asunto(s)
Sorghum , Antioxidantes , Biomasa , Clorofila , Fotosíntesis , Hojas de la Planta , Plantones , Rayos Ultravioleta
13.
BMC Bioinformatics ; 16: 332, 2015 Oct 16.
Artículo en Inglés | MEDLINE | ID: mdl-26475308

RESUMEN

BACKGROUND: The high-throughput sequencing technology, RNA-Seq, has been widely used to quantify gene and isoform expression in the study of transcriptome in recent years. Accurate expression measurement from the millions or billions of short generated reads is obstructed by difficulties. One is ambiguous mapping of reads to reference transcriptome caused by alternative splicing. This increases the uncertainty in estimating isoform expression. The other is non-uniformity of read distribution along the reference transcriptome due to positional, sequencing, mappability and other undiscovered sources of biases. This violates the uniform assumption of read distribution for many expression calculation approaches, such as the direct RPKM calculation and Poisson-based models. Many methods have been proposed to address these difficulties. Some approaches employ latent variable models to discover the underlying pattern of read sequencing. However, most of these methods make bias correction based on surrounding sequence contents and share the bias models by all genes. They therefore cannot estimate gene- and isoform-specific biases as revealed by recent studies. RESULTS: We propose a latent variable model, NLDMseq, to estimate gene and isoform expression. Our method adopts latent variables to model the unknown isoforms, from which reads originate, and the underlying percentage of multiple spliced variants. The isoform- and exon-specific read sequencing biases are modeled to account for the non-uniformity of read distribution, and are identified by utilizing the replicate information of multiple lanes of a single library run. We employ simulation and real data to verify the performance of our method in terms of accuracy in the calculation of gene and isoform expression. Results show that NLDMseq obtains competitive gene and isoform expression compared to popular alternatives. Finally, the proposed method is applied to the detection of differential expression (DE) to show its usefulness in the downstream analysis. CONCLUSIONS: The proposed NLDMseq method provides an approach to accurately estimate gene and isoform expression from RNA-Seq data by modeling the isoform- and exon-specific read sequencing biases. It makes use of a latent variable model to discover the hidden pattern of read sequencing. We have shown that it works well in both simulations and real datasets, and has competitive performance compared to popular methods. The method has been implemented as a freely available software which can be found at https://github.com/PUGEA/NLDMseq.


Asunto(s)
ARN/metabolismo , Empalme Alternativo , Exones , Secuenciación de Nucleótidos de Alto Rendimiento , Humanos , Internet , Modelos Genéticos , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , ARN/química , ARN/genética , Análisis de Secuencia de ARN , Transcriptoma , Interfaz Usuario-Computador
14.
Insects ; 15(5)2024 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-38786900

RESUMEN

In the global apiculture industry, reward feeding and supplementary feeding are essential for maintaining bee colonies. Beekeepers provide artificial supplements to their colonies, typically in the form of either a honey-water solution or sugar syrup. Owing to cost considerations associated with beekeeping, most beekeepers opt for sugar syrup. However, the effects of different types of artificial sugar supplements on bee colonies and their subsequent impact on honey composition remain unclear. To address this gap, this study compared the chemical composition, antioxidant capacity, and nutritional potency of three types of honey: honey derived from colonies fed sugar syrup (sugar-based product, SP) or a honey-water solution (honey-sourced honey, HH) and naturally sourced honey (flower-sourced honey, FH), which served as the control. The results revealed that FH outperformed HH and SP in terms of total acidity, sugar content, total protein content, and antioxidant capacity, and HH outperformed SP. Regarding nutritional efficacy, including the lifespan and learning and memory capabilities of worker bees, FH exhibited the best outcomes, with no significant differences observed between HH and SP. This study underscores the importance of sugar source selection in influencing honey quality and emphasizes the potential consequences of substituting honey with sugar syrup in traditional apiculture practices.

15.
Front Immunol ; 15: 1353111, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38440726

RESUMEN

Background: Myocardial infarction (MI) caused by severe coronary artery disease has high incidence and mortality rates, making its prevention and treatment a central and challenging aspect of clinical work for cardiovascular practitioners. Recently, researchers have turned their attention to a novel mechanism of cell death caused by Cu2+, cuproptosis. Methods: This study integrated data from three MI-related bulk datasets downloaded from the Gene Expression Omnibus (GEO) database, and identified 16 differentially expressed genes (DEGs) related to cuproptosis by taking intersection of the 6378 DEGs obtained by differential analysis with 49 cuproptosis-related genes. Four hub genes, Dbt, Dlat, Ube2d1 and Ube2d3, were screened out through random forest analysis and Lasso analysis. In the disease group, Dbt, Dlat, and Ube2d1 showed low expression, while Ube2d3 exhibited high expression. Results: Focusing on Ube2d3 for subsequent functional studies, we confirmed its high expression in the MI group through qRT-PCR and Western Blot detection after successful construction of a MI mouse model by left anterior descending (LAD) coronary artery ligation, and further clarified the correlation of cuproptosis with MI development by detecting the levels of cuproptosis-related proteins. Moreover, through in vitro experiments, Ube2d3 was confirmed to be highly expressed in oxygen-glucose deprivation (OGD)-treated cardiomyocytes AC16. In order to further clarify the role of Ube2d3, we knocked down Ube2d3 expression in OGD-treated AC16 cells, and confirmed Ube2d3's promoting role in the hypoxia damage of AC16 cells by inducing cuproptosis, as evidenced by the detection of MTT, TUNEL, LDH release and cuproptosis-related proteins. Conclusion: In summary, our findings indicate that Ube2d3 regulates cuproptosis to affect the progression of MI.


Asunto(s)
Enfermedad de la Arteria Coronaria , Infarto del Miocardio , Animales , Ratones , Western Blotting , Muerte Celular , Biología Computacional , Glucosa , Infarto del Miocardio/genética
16.
ACS Appl Mater Interfaces ; 16(6): 6849-6858, 2024 Feb 14.
Artículo en Inglés | MEDLINE | ID: mdl-38293917

RESUMEN

Rationally designing highly catalytic and stable nanozymes for metabolite monitoring is of great importance because of their huge potential in early disease diagnosis. Herein, a novel nanozyme based on hierarchically structured CuS/ZnS with a highly efficient peroxidase (POD)-mimic capability was developed and synthesized for multiple metabolite determination and recognition via the plasmon-stimulated biosensor array strategy. The designed nanozyme can simultaneously harvest plasmon triggered hot electron-hole pairs and generate photothermal properties, leading to a sharply boosted POD-mimic capability under 808 nm laser irradiation. Interestingly, because of the interaction diversity of the metabolite with POD-like nanomaterials, the unique inhibitory effect of metabolites on the POD-mimic activity could be the signal response as the differentiation. Thus, utilizing TMB as a typical chromogenic substrate in the addition of H2O2, the designed colorimetric biosensor array can produce diverse fingerprints for the three vital metabolisms (cysteine (Cys), ascorbic acid (AA), and glutathione (GSH)), which can be precisely identified by principal component analysis (PCA). Notably, a distinct fingerprint of a single metabolite with different levels and metabolite mixtures is also achieved with a detection limit of 1 µM. Most importantly, cell lysis could be effectively discriminated by the biosensor assay, implying its great potential in clinical diagnosis.


Asunto(s)
Técnicas Biosensibles , Colorimetría , Peróxido de Hidrógeno/química , Peroxidasa/química , Peroxidasas/metabolismo , Colorantes/química
17.
Sci Total Environ ; : 174332, 2024 Jun 29.
Artículo en Inglés | MEDLINE | ID: mdl-38950630

RESUMEN

Cathodic electroactive bacteria (C-EAB) which are capable of accepting electrons from solid electrode provide fresh avenues for pollutant removal, biosensor design and electrosynthesis. This review systematically summarized the burgeoning applications of the C-EAB over the past decade, including 1) removal of nitrate, aromatic derivatives and metal ions; 2) biosensing based on biocathode; 3) electrosynthesis of CH4, H2, organic carbon, NH3 and protein. In addition, the mechanisms of electron transfer by the C-EAB are also classified and summarized. Extracellular electron transfer and interspecies electron transfer have been introduced, and the electron transport mechanism of typical C-EAB, such as Shewanella oneidensis MR-1, has been combed in detail. By bringing to light this cutting-edge area of the C-EAB, this review aims to stimulate more interest and research on not only exploring great potential applications of these electron-accepting bacteria, but also developing steady and scalable processes harnessing biocathodes.

18.
Math Biosci Eng ; 20(7): 11875-11894, 2023 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-37501424

RESUMEN

The density peak clustering algorithm (DPC) requires manual determination of cluster centers, and poor performance on complex datasets with varying densities or non-convexity. Hence, a novel density peak clustering algorithm is proposed for the automatic selection of clustering centers based on K-nearest neighbors (AKDPC). First, the AKDPC classifies samples according to their mutual K-nearest neighbor values into core and non-core points. Second, the AKDPC uses the average distance of K nearest neighbors of a sample as its density. The smaller the average distance is, the higher the density. Subsequently, it selects the highest density sample among all unclassified core points as a center of the new cluster, and the core points that satisfy the merging condition are added to the cluster until no core points satisfy the condition. Afterwards, the above steps are repeated to complete the clustering of all core points. Lastly, the AKDPC labels the unclassified non-core points similar to the nearest points that have been classified. In addition, to prove the validity of AKDPC, experiments on manual and real datasets are conducted. By comparing the AKDPC with classical clustering algorithms and excellent DPC-variants, this paper demonstrates that AKDPC presents higher accuracy.

19.
Front Cell Dev Biol ; 11: 1156923, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37181751

RESUMEN

Maternal effects are an evolutionary strategy used to improve offspring quality. In an example of maternal effects in honey bees (Apis mellifera), mother queens produce larger eggs in queen cells than in worker cells in order to breed better daughter queens. In our current study, morphological indexes, reproductive tissues, and the egg-laying ability of newly reared queens reared with eggs laid in queen cells (QE), eggs laid in worker cells (WE), and 2-day-old larvae in worker cells (2L) were evaluated. In addition, morphological indexes of offspring queens and working performance of offspring workers were examined. The thorax weight, number of ovarioles, egg length, and number of laid eggs and capped broods of QE were significantly higher than those of WE and 2L, indicating that the reproductive capacity of QE group was better than that of other groups. Furthermore, offspring queens from QE had larger thorax weights and sizes than those from the other two groups. Offspring worker bees from QE also had larger body sizes and greater pollen-collecting and royal jelly-producing abilities than those of other two groups. These results demonstrate that honey bees display profound maternal effects on queen quality that can be transmitted across generations. These findings provide a basis for improving queen quality, with implications in apicultural and agricultural production.

20.
Math Biosci Eng ; 20(9): 15830-15858, 2023 Jul 31.
Artículo en Inglés | MEDLINE | ID: mdl-37919991

RESUMEN

Minimum spanning tree (MST)-based clustering algorithms are widely used to detect clusters with diverse densities and irregular shapes. However, most algorithms require the entire dataset to construct an MST, which leads to significant computational overhead. To alleviate this issue, our proposed algorithm R-MST utilizes representative points instead of all sample points for constructing MST. Additionally, based on the density and nearest neighbor distance, we improved the representative point selection strategy to enhance the uniform distribution of representative points in sparse areas, enabling the algorithm to perform well on datasets with varying densities. Furthermore, traditional methods for eliminating inconsistent edges generally require prior knowledge about the number of clusters, which is not always readily available in practical applications. Therefore, we propose an adaptive method that employs mutual neighbors to identify inconsistent edges and determine the optimal number of clusters automatically. The experimental results indicate that the R-MST algorithm not only improves the efficiency of clustering but also enhances its accuracy.

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