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1.
FASEB J ; 37(4): e22831, 2023 04.
Artículo en Inglés | MEDLINE | ID: mdl-36856728

RESUMEN

The metabolic benefits of intermittent fasting (IF) have been well recognized. However, limited studies have examined the relationship between long-term maternal IF before pregnancy and offspring health. In this study, a C57BL/6J mouse model of long-term IF before pregnancy was established: 4-week-old female mice were subjected to alternate-day fasting for 12 weeks and resumed normal diet after mating. Female mice in the control group were fed ad libitum. Offspring mice were weaned at 6 weeks of age and fed a normal chow diet or a 60% high-fat diet. The effects of long-term pre-pregnancy IF on offspring metabolism and its underlying mechanism were examined. We found that neonatal IF offspring weighted significantly less relevant to control mice. This difference gradually disappeared as a result of catch-up growth. In the IF offspring, adipose tissue mass was significantly increased. This alteration was associated with a considerable deterioration in glucose tolerance. No significant difference in food intake was observed. Further, lipid deposition as well as triglyceride contents in the liver were greatly increased. Maternal IF significantly decreased levels of DNA methyltransferase in the liver of offspring. DNA methylation modifications of molecules associated with the mTORC1 signaling pathway were significantly altered, leading to the significant inhibition of mTORC1 signaling. Overexpression of S6K1 activated hepatic mTORC1 signaling and reversed the metabolic dysfunction in IF offspring. In conclusion, long-term pre-pregnancy IF increases hepatic steatosis and adiposity, as well as impairs glucose metabolism in adult offspring. This occurs through DNA methylation-dependent suppression of hepatic mTORC1 signaling activity.


Asunto(s)
Hígado Graso , Ayuno Intermitente , Femenino , Embarazo , Animales , Ratones , Ratones Endogámicos C57BL , Transducción de Señal
2.
Biochem Biophys Res Commun ; 645: 30-39, 2023 02 19.
Artículo en Inglés | MEDLINE | ID: mdl-36680934

RESUMEN

Pseudomonas aeruginosa is a Gram-negative bacterium capable of widespread niches, which is also one of the main bacteria that cause patient infection. The metabolic diversity of Pseudomonas aeruginosa is an essential factor in adapting to a variety of environments. Based on the previous studies, adaptive genetic variation in the glycerol kinase GlpK, the glycerol 3-phosphotransferase, contributes to the fitness of bacteria in human bodies, such as Mycobacterium tuberculosis and Escherichia coli. Thus, this study aimed to explore the molecular evolution and function of glpK in P. aeruginosa. Using extensive population genomic data, we have identified the prevalence of two glpK copies in P. aeruginosa that clustered into distinct branches, which were later known as Clade 1 and 2. The evolution analysis revealed that glpK in Clade 1 derived from an ancestral P. aeruginosa species and the other from an ancient horizontal gene transfer event. In addition, we confirmed that the GlpK in Clade 2 still retained glycerol kinase activity but was much weaker than that of GlpK in Clade 1. We demonstrated the importance of the critical amino acid Q70 in GlpK glycerol kinase activity by point mutation. Furthermore, Co-expression network analysis implied that the two glpK copies of P. aeruginosa regulate separate networks and may be a strategy to improve fitness in P. aeruginosa.


Asunto(s)
Glicerol Quinasa , Pseudomonas aeruginosa , Humanos , Glicerol/metabolismo , Glicerol Quinasa/genética , Glicerol Quinasa/metabolismo , Fosforilación , Pseudomonas aeruginosa/enzimología , Pseudomonas aeruginosa/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo
3.
Opt Express ; 31(4): 6426-6452, 2023 Feb 13.
Artículo en Inglés | MEDLINE | ID: mdl-36823899

RESUMEN

For a long time, due to the difficulty of obtaining accurate propagation trajectories, the research on creeping waves is limited to canonical geometries or simple targets, which leads to the situation that it is relatively mature in theoretical research on creeping waves, while the practical application scope of creeping waves for complex targets is narrow. In this paper, a thorough electromagnetic computation method for creeping waves on complex planar mesh model is systematically proposed. This approach broadens the field of creeping waves applications due to the generality of planar mesh models in electromagnetic engineering. The contents consist of the tracing of creeping waves, the calculation of the diffraction field, and the coupling effect with other scattering mechanisms. Aiming at the trajectory of creeping waves, we propose a set of tracing algorithms that enable rapid, real-time tracing based on analytical geometry and related computer graphics algorithms. Utilizing information such as vertices, triangles, and topological relations in the mesh model, one can recover the mathematical properties of the surfaces of the model and then, the corresponding parameters can be obtained. Therefore, the uniform geometrical theory of diffraction (UTD) can be used to accurately calculate the diffraction field. Moreover, for complex targets, the multiple coupling effect caused by creeping waves is the main source of radar echoes in many cases, which is not unimportant. Hence based on the electromagnetic accurate modeling, the coupling mechanism of creeping waves and various scattering mechanisms are studied. The research content is expected to have high application values in target recognition and characteristics.

4.
Sensors (Basel) ; 23(17)2023 Aug 30.
Artículo en Inglés | MEDLINE | ID: mdl-37688000

RESUMEN

In this paper, we propose to extract the motions of different human limbs by using interferometric radar based on the micro-Doppler-Range signature (mDRS). As we know, accurate extraction of human limbs in motion has great potential for improving the radar performance on human motion detection. Because the motions of human limbs usually overlap in the time-Doppler plane, it is extremely hard to separate human limbs without other information such as the range or the angle. In addition, it is also difficult to identify which part of the body each signal component belongs to. In this work, the overlaps of multiple components can be solved, and the motions from different limbs can be extracted and classified as well based on the extracted micro-Doppler-Range trajectories (MDRTs) along with a proposed three-dimensional constant false alarm (3D-CFAR) detection. Three experiments are conducted with three different people on typical human motions using a 77 GHz radar board of 4 GHz bandwidth, and the results are validated by the measurements of a Kinect sensor. All three experiments were repeatedly conducted for three different people of different heights to test the repeatability and robust of the proposed approach, and the results met our expectations very well.


Asunto(s)
Extremidades , Radar , Humanos , Interferometría , Movimiento (Física) , Ultrasonografía Doppler
5.
J Environ Sci (China) ; 126: 794-805, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36503804

RESUMEN

In this study, the effects of a diesel oxidation catalyst (DOC) coupled with a catalyzed diesel particulate filter (CDPF) with different catalyst loadings on the power, fuel consumption, gaseous and particulate emissions from a non-road diesel engine were investigated. Results showed that the after-treatment had a negligible effect on the power and fuel consumption. The reduction effect of the DOC on the CO and hydrocarbon (HC) increased with the engine load. Further reductions occurred coupling with the CDPF. Increasing the catalyst loading resulted in a more significant reduction in the HC emissions than CO emissions. The DOC could increase the NO2 proportion to 37.9%, and more NO2 was produced when coupled with the CDPF below 250°C; above 250°C, more NO2 was consumed. The after-treatment could reduce more than 99% of the particle number (PN) and 98% of the particle mass (PM). Further reductions in the PN and PM occurred with a higher CDPF catalyst loading. The DOC had a better reduction effect on the nucleation particles than the accumulation ones, but the trend reversed with the CDPF. The DOC shifted the particle size distribution (PSD) to larger particles with an accumulation particle proportion increasing from 13% to 20%, and the geometric mean diameter (GMD) increased from 18.2 to 26.0 nm. The trend reversed with the CDPF and the accumulation particle proportion declined to less than 10%. A lower catalyst loading on the CDPF led to a higher proportion of nucleation particles and a smaller GMD.


Asunto(s)
Polvo , Gases
6.
Microb Ecol ; 83(4): 1049-1058, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-34302509

RESUMEN

Understanding the composition of microorganismal communities hosted by insect pests is an important prerequisite for revealing their functions and developing new pest control strategies. Although studies of the structure of the microbiome of Nilaparvata lugens have been published, little is known about the dynamic changes in this microbiome across different developmental stages, and an understanding of the core microbiota is still lacking. In this study, we investigated the dynamic changes in bacteria and fungi in different developmental stages of N. lugens using high-throughput sequencing technology. We observed that the microbial diversity in eggs and mated adults was higher than that in nymphs and unmated adults. We also observed a notable strong correlation between fungal and bacterial α-diversity, which suggests that fungi and bacteria are closely linked and may perform functions collaboratively during the whole developmental period. Arsenophonus and Hirsutella were the predominant bacterial and fungal taxa, respectively. Bacteria were more conserved than fungi during the transmission of the microbiota between developmental stages. Compared with that in the nymph and unmated adult stages of N. lugens, the correlation between bacterial and fungal communities in the mated adult and egg stages was stronger. Moreover, the core microbiota across all developmental stages in N. lugens was identified, and there were more bacterial genera than fungal genera; notably, the core microbiota of eggs, nymphs, and mated and unmated adults showed distinctive functional enrichment. These findings highlight the potential value of further exploring microbial functions during different developmental stages and developing new pest management strategies.


Asunto(s)
Hemípteros , Microbiota , Animales , Bacterias/genética , Hemípteros/microbiología , Secuenciación de Nucleótidos de Alto Rendimiento , Ninfa/microbiología
7.
Appl Opt ; 61(14): 4013-4021, 2022 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-36256074

RESUMEN

Biophotons in the nervous system are a potential carrier of neural signals. Previous experiments and studies indicated that biophotons are closely related to the neuronal activity and can propagate along myelinated axons. We establish a multilayer electromagnetic simulation model and demonstrate that the myelinated axon waveguide has low attenuation and low dispersion and operates in a narrow bandwidth on the order of 10 nm. We also find that the operating wavelength of the waveguide is almost linearly related to the axon diameter and the number of myelin layers. Each additional layer of the myelin sheath causes the operating wavelength of the myelinated axon waveguide to shift 52.3 nm to the long-wave direction, while an increase in the axon diameter of 1.0 µm causes the operating wavelength to shift 94.5 nm to the short-wave direction. These findings well explain the tendency of the spectral redshift among different species and the spectral blueshift during the aging process of mice. Via the analysis method in this paper, we can predict the wavelength of the propagating biophotons based on the neural structure.


Asunto(s)
Axones , Vaina de Mielina , Animales , Ratones , Axones/fisiología , Neuronas , Simulación por Computador , Fenómenos Electromagnéticos
8.
World J Microbiol Biotechnol ; 38(7): 126, 2022 Jun 06.
Artículo en Inglés | MEDLINE | ID: mdl-35666348

RESUMEN

Nitrogen metabolism is the most basic process of material and energy metabolism in living organisms, and processes involving the uptake and use of different nitrogen sources are usually tightly regulated at the transcriptional and post-transcriptional levels. Bacterial regulatory noncoding RNAs are novel post-transcriptional regulators that repress or activate the expression of target genes through complementarily pairing with target mRNAs; therefore, these noncoding RNAs play an important regulatory role in many physiological processes, such as bacterial substance metabolism and stress response. In recent years, a study found that noncoding RNAs play a vital role in the post-transcriptional regulation of nitrogen metabolism, which is currently a hot topic in the study of bacterial nitrogen metabolism regulation. In this review, we present an overview of recent advances that increase our understanding on the regulatory roles of bacterial noncoding RNAs and describe in detail how noncoding RNAs regulate biological nitrogen fixation and nitrogen metabolic engineering. Furthermore, our goal is to lay a theoretical foundation for better understanding the molecular mechanisms in bacteria that are involved in environmental adaptations and metabolically-engineered genetic modifications.


Asunto(s)
ARN Pequeño no Traducido , Bacterias/genética , Bacterias/metabolismo , Regulación Bacteriana de la Expresión Génica , Nitrógeno/metabolismo , ARN Bacteriano/genética , ARN Bacteriano/metabolismo , ARN Pequeño no Traducido/genética , ARN Pequeño no Traducido/metabolismo
9.
J Sci Food Agric ; 101(1): 334-340, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-32627840

RESUMEN

BACKGROUND: Perennial legumes cultivated under irrigation in the Mountain West USA have non-fibrous carbohydrate (NFC) concentrations exceeding 400 g kg-1 , a level commonly found in concentrate-based ruminant diets. Our objective was to determine the influence of NFC concentration and plant secondary compounds on in vitro rumen digestion of grass, legume and forb forages compared with digestion of their isolated neutral detergent fiber (NDF) fraction. Forages were composited from ungrazed paddocks of rotationally stocked, irrigated monoculture pastures between May and August 2016, frozen in the field, freeze-dried, and ground. RESULTS: The maximum rate (RMax ) of gas production was greater for the legumes alfalfa (ALF; Medicago sativa L.) and birdsfoot trefoil (BFT; Lotus corniculatus L.) than for the legume cicer milkvetch (CMV; Astragalus cicer L.) the grass meadow brome (MBG; Bromus riparius Rehm.) and the non-legume forb small burnet (SMB; Sanguisorba minor Scop.), and intermediate for the legume sainfoin (SNF; Onobrychis viciifolia Scop.). The RMax of isolated NDF was greatest for BFT and CMV, intermediate for ALF, SNF and SMB and least for MBG. CONCLUSIONS: More than 900 g of organic matter kg-1 dry matter of legumes was digested after 96 h. Across forages, the extent of whole-plant digestion increased with NFC and crude protein concentrations, decreased with NDF concentrations, and was modulated by secondary compounds. The extent of digestion of isolated NDF decreased with concentration of lignin and residual tannins. © 2020 Society of Chemical Industry.


Asunto(s)
Carbohidratos/química , Bovinos/metabolismo , Medicago sativa/metabolismo , Rumen/metabolismo , Alimentación Animal/análisis , Animales , Metabolismo de los Hidratos de Carbono , Digestión , Lotus/química , Lotus/metabolismo , Medicago sativa/química , Poaceae/química , Poaceae/metabolismo
10.
World J Microbiol Biotechnol ; 37(10): 177, 2021 Sep 15.
Artículo en Inglés | MEDLINE | ID: mdl-34524580

RESUMEN

Pseudomonas stutzeri A1501 is a model strain used to study associative nitrogen fixation, and it possesses the nitrogen regulatory NtrC protein in the core genome. Nitrogen sources represent one of the important factors affecting the efficiency of biological nitrogen fixation in the natural environment. However, the regulation of NtrC during nitrogen metabolism in P. stutzeri A1501 has not been clarified. In this work, a phenotypic analysis of the ntrC mutant characterized the roles of NtrC in nitrogen metabolism and the oxidative stress response of P. stutzeri A1501. To systematically identify NtrC-controlled gene expression, RNA-seq was performed to further analyse the gene expression differences between the wild-type strain and the ∆ntrC mutant under nitrogen fixation conditions. A total of 1431 genes were found to be significantly altered by ntrC deletion, among which 147 associative genes had NtrC-binding sites, and the pathways for nitrogen fixation regulation, nitrogenous compound acquisition and catabolism and nitrate assimilation were discussed. Furthermore, the oxidative stress-related gene (katB), which was upregulated by ntrC deletion, was suggested to be a potential target gene of NtrC, thus highlighting the importance of NtrC in nitrogenase protection against oxygen damage. Based on these findings, we propose that NtrC is a high-ranking element in the regulatory network of P. stutzeri A1501 that controls a variety of nitrogen metabolic and oxidative stress responsive traits required for adaptation to complex rhizosphere environments.


Asunto(s)
Nitrógeno/metabolismo , Pseudomonas stutzeri , Factores de Transcripción/genética , Proteínas Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Mutación , Fijación del Nitrógeno , Pseudomonas stutzeri/genética , Pseudomonas stutzeri/metabolismo , RNA-Seq , Rizosfera
11.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao ; 43(6): 975-979, 2021 Dec 30.
Artículo en Zh | MEDLINE | ID: mdl-34980340

RESUMEN

Tuberculous peritonitis(TBP)is currently one of the common manifestations of extrapulmonary tuberculosis.Due to the atypical clinical features,diverse types of diseases to be distinguished,and limited detection methods,TBP is difficult to be diagnosed and the fatality caused by delayed diagnosis increases significantly.We studied the current research status of TBP and found that T cells spot test,abdominal CT,and laparoscopic biopsy were of high diagnostic value for TBP.However,the application of ascites Xpert-MTB/RIF-ultra assay,ascites ADA,and whole-body positron emission tomography/computed tomography remained to be studied.Serum CA125 helps to judge the efficacy of anti-tuberculosis treatment.


Asunto(s)
Mycobacterium tuberculosis , Peritonitis Tuberculosa , Tuberculosis , Ascitis , Biopsia , Humanos , Peritonitis Tuberculosa/diagnóstico , Sensibilidad y Especificidad , Tuberculosis/diagnóstico
12.
J Environ Sci (China) ; 84: 166-173, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31284908

RESUMEN

The primary purpose of this study was to investigate the effect of a catalyzed continuously regenerating trap (CCRT) system composed of a diesel oxidation catalyst (DOC) and a catalyzed diesel particulate filter (CDPF) on the main gaseous and particulate emissions from an urban diesel bus, as well as the durability performance of the CCRT system. Experiments were conducted based on a heavy chassis dynamometer, and a laboratory activity test as well as X-ray photoelectron spectroscopy (XPS) test were applied to evaluate the changes of the aged CCRT catalyst. Results showed that the CCRT could reduce the CO by 71.5% and the total hydrocarbons (THC) by 88.9%, and meanwhile promote the oxidation of NO. However, the conversion rates for CO and THC dropped to 25.1% and 55.1%, respectively, after the CCRT was used for one year (~60,000 km), and the NO oxidation was also weakened. For particulate emissions, the CCRT could reduce 97.4% of the particle mass (PM) and almost 100% of the particle number (PN). The aging of the CCRT resulted in a reduced PM trapping efficiency but had no observable effect on the PN; however, it increased the proportion of nucleation mode particles. The activity test results indicated that the deterioration of the CCRT was directly relevant to the increase in the light-off temperatures of the catalyst for CO, C3H8 and NO2. In addition, the decreased concentrations of the active components Pt2+ and Pt4+ in the catalyst are also important factors in the CCRT deterioration.


Asunto(s)
Filtración/instrumentación , Emisiones de Vehículos/prevención & control , Monóxido de Carbono/análisis , Catálisis , Hidrocarburos/análisis , Óxidos de Nitrógeno/análisis , Material Particulado/análisis , Emisiones de Vehículos/análisis
13.
J Cell Biochem ; 119(4): 3394-3403, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29130544

RESUMEN

Adult neural stem cells (NSCs) are a group of multi-potent, self-renewing progenitor cells that contribute to the generation of new neurons and oligodendrocytes. Three subtypes of NSCs can be isolated based on the stages of the NSC lineage, including quiescent neural stem cells (qNSCs), activated neural stem cells (aNSCs) and neural progenitor cells (NPCs). Although it is widely accepted that these three groups of NSCs play different roles in the development of the nervous system, their molecular signatures are poorly understood. In this study, we applied the Monte-Carlo Feature Selection (MCFS) method to identify the gene expression signatures, which can yield a Matthews correlation coefficient (MCC) value of 0.918 with a support vector machine evaluated by ten-fold cross-validation. In addition, some classification rules yielded by the MCFS program for distinguishing above three subtypes were reported. Our results not only demonstrate a high classification capacity and subtype-specific gene expression patterns but also quantitatively reflect the pattern of the gene expression levels across the NSC lineage, providing insight into deciphering the molecular basis of NSC differentiation.


Asunto(s)
Astrocitos/citología , Perfilación de la Expresión Génica/métodos , Redes Reguladoras de Genes , Células-Madre Neurales/clasificación , Algoritmos , Linaje de la Célula , Células Cultivadas , Humanos , Método de Montecarlo , Máquina de Vectores de Soporte
14.
World J Microbiol Biotechnol ; 34(8): 109, 2018 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-29971547

RESUMEN

Glutathione-S-transferase (GST) genes exist widely in plants and play major role in metabolic detoxification of exogenous chemical substances and oxidative stress. In this study, 14 sunflower GST genes (HaGSTs) were identified based on the sunflower transcriptome database that we had constructed. Full-length cDNA of 14 HaGTSs were isolated from total RNA by reverse transcription PCR (RT-PCR). Sunflower was received biotic stress (Sclerotinia sclerotiorum) and abiotic stress (NaCl, low-temperature, drought and wound). GST activity was measured by using the universal substrate. The results showed that most of the HaGSTs were up-regulated after NaCl and PEG6000-induced stresses, while a few HaGSTs were up-regulated after S. sclerotiorum, hypothermia and wound-induced stressed, and there was correlation between the changes of GST activity and the expression of HaGSTs, indicating that HaGSTs may play regulatory role in the biotic and abiotic stress responses. 14 HaGSTs from sunflower were identified, and the expression of HaGSTs were tissue-specific and played regulatory roles in both stress and abiotic stress.


Asunto(s)
Glutatión Transferasa/genética , Glutatión Transferasa/aislamiento & purificación , Glutatión Transferasa/fisiología , Helianthus/genética , Helianthus/fisiología , Estrés Fisiológico , Clonación Molecular , Frío , ADN Complementario/aislamiento & purificación , Sequías , Regulación de la Expresión Génica de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/fisiología , Genes de Plantas , Glutatión Transferasa/clasificación , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/fisiología , Análisis de Secuencia , Cloruro de Sodio , Transcriptoma , Regulación hacia Arriba
15.
Asian-Australas J Anim Sci ; 31(1): 63-70, 2018 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-28728360

RESUMEN

OBJECTIVE: The aim of the study was to isolate gossypol-degrading bacteria and to assess its potential for gossypol degradation. METHODS: Rumen liquid was collected from fistulated cows grazing the experimental pasture. Approximately 1 mL of the rumen liquid was spread onto basal medium plates containing 2 g/L gossypol as the only source of carbon and was then cultured at 39°C to isolate gossypol-degrading bacteria. The isolated colonies were cultured for 6 h and then their size and shape observed by microscope and scanning electron microscope. The 16S rRNA gene of isolated colonies was sequenced and aligned using National Center for Biotechnology Information-Basic Local Alignment Search Tool. The various fermentation conditions, initial pH, incubation temperature, inoculum level and fermentationperiod were analyzed in cottonseed meal (CSM). The crude protein (CP), total gossypol (TG), and free gossypol (FG) were determined in CSM after fermentation with isolated strain at 39°C for 72 h. RESULTS: Screening results showed that a single bacterial isolate, named Rumen Bacillus Subtilis (RBS), could use gossypol as a carbon source. The bacterium was identified by 16S rDNA sequencing as being 98% homologous to the sequence of Bacillus subtilis strain GH38. The optimum fermentation conditions were found to be 72 h, 39°C, pH 6.5, moisture 50%, inoculum level 107 cell/g. In the optimum fermentation conditions, the FG and TG content in fermented CSM decreased 78.86% and 49% relative to the control. The content of CP and the essential amino acids of the fermented CSM increased respectively, compared with the control. CONCLUSION: The isolation of a gossypol-degrading bacterium from the cow rumen is of great importance for gossypol biodegradation and may be a valuable potential source for gossypol-degradation of CSM.

16.
Biotechnol Lett ; 39(11): 1649-1655, 2017 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-28852892

RESUMEN

OBJECTIVE: To study the effects of recombinant neuritin expressed by Pichia pastoris GS115 on the senescence, apoptosis, proliferation, and migration associated with rat bone marrow-derived mesenchymal stem cells (BMSCs). RESULTS: Recombinant neuritin was purified by Ni-affinity chromatography and identified by western blot and MALDI-TOF spectrometry. The effects of recombinant neuritin on senescence, apoptosis, proliferation, and migration of rat BMSCs WERE investigated. ß-Galactosidase staining indicated that recombinant neuritin administration significantly inhibited BMSCs senescence at 1 µg neuritin/ml. Additionally, recombinant neuritin reduced the number of apoptotic cells at the early stage according to Annexin V/propidium iodide staining and inhibited cell proliferation according to MTT assay results. Moreover wound healing assay results showed that recombinant neuritin promoted BMSCs migration in the neuritin-treatment group. CONCLUSION: Recombinant neuritin affects the senescence, apoptosis, proliferation, migration of rat BMSCs. Our findings offer insight into neuritin function outside of the nervous system.


Asunto(s)
Células Madre Mesenquimatosas/citología , Neuropéptidos/genética , Proteínas Recombinantes/farmacología , Animales , Apoptosis , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Células Cultivadas , Senescencia Celular/efectos de los fármacos , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/farmacología , Células Madre Mesenquimatosas/efectos de los fármacos , Neuropéptidos/farmacología , Ratas
17.
Biochem Biophys Res Commun ; 457(4): 627-34, 2015 Feb 20.
Artículo en Inglés | MEDLINE | ID: mdl-25613865

RESUMEN

In this study, a necrosis-inducing protein was purified from the culture filtrate of the necrotrophic fungus Botrytis cinerea BC-98 strain. Secreted proteins were collected and fractionated by liquid chromatography. The fraction with the highest necrosis-inducing activity was further purified. A glycoprotein named BcGs1 was identified by 2D electrophoresis and mass spectrometry. The BcGs1 protein consisted of 672 amino acids with a theoretical molecular weight of 70.487 kDa. Functional domain analysis indicated that BcGs1 was a glucan 1,4-alpha-glucosidase, a cell wall-degrading enzyme, with a Glyco_hydro_15 domain and a CBM20_glucoamylase domain. The BcGs1 protein caused necrotic lesions that mimicked a typical hypersensitive response and H2O2 production in tomato and tobacco leaves. BcGs1-treated plants exhibited resistance to B. cinerea, Pseudomonas syringae pv. tomato DC3000 and tobacco mosaic virus in systemic leaves. In addition, BcGs1 triggered elevation of the transcript levels of the defence-related genes PR-1a, TPK1b and Prosystemin. This is the first report of a Botrytis glucan 1,4-alpha-glucosidase triggering host plant immunity as an elicitor. These results lay a foundation for further study of the comprehensive interaction between plants and necrotrophic fungi.


Asunto(s)
Botrytis/fisiología , Proteínas Fúngicas/metabolismo , Glicoproteínas/metabolismo , Interacciones Huésped-Patógeno , Nicotiana/microbiología , Enfermedades de las Plantas/microbiología , Solanum lycopersicum/microbiología , Botrytis/química , Proteínas Fúngicas/química , Proteínas Fúngicas/aislamiento & purificación , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Glucano 1,4-alfa-Glucosidasa/química , Glucano 1,4-alfa-Glucosidasa/aislamiento & purificación , Glucano 1,4-alfa-Glucosidasa/metabolismo , Glicoproteínas/química , Glicoproteínas/aislamiento & purificación , Solanum lycopersicum/genética , Enfermedades de las Plantas/genética , Hojas de la Planta/genética , Hojas de la Planta/microbiología , Nicotiana/genética
18.
Appl Microbiol Biotechnol ; 99(19): 8035-43, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26048470

RESUMEN

Neuritin (also known as candidate plasticity gene 15 (cpg15)) is a neurotrophic factor that was recently discovered in a screen aimed at identifying genes involved in activity-dependent synaptic plasticity. Neuritin plays multiple roles in both neural development (Chen et al. Zhonghua Yan Ke Za Zhi 46:978-983 2010; Corriveau et al. J Neurosci 19:7999-8008 1999; Lee and Nedivi J Neurosci 22:1807-1815 2002) and synaptic plasticity (Fujino et al. Gene Dev 25:2674-2685 2011; Leslie and Nedivi Prog 14 Neurobiol 94:223-237 2011; Loebrich and Nedivi Physiol Rev 89:1079 2009). In this study, to produce bioactive, soluble recombinant human neuritin protein, a portion of NRN1 was cloned into the Pichia pastoris expression vector pPIC9K. The recombinant vector was then transformed into the methylotrophic yeast strain P. pastoris GS115, and a shaking flask method and His-tag purification strategy were utilized to express and purify neuritin protein. The resulting protein had a molecular mass of approximately 11 kDa, and subsequent functional analyses indicated that the purified neuritin promoted neurite outgrowth from embryonic chicken dorsal root ganglions, while also prolonging the survival of these ganglions, and from PC12 cells. These findings suggest that neuritin was expressed effectively in vitro and that this protein may play a role in stimulating neurite outgrowth of both dorsal root ganglions and PC12 cells. This study provides a novel strategy for the large-scale production of bioactive neuritin, which will enable further study of the biological function of this protein.


Asunto(s)
Factores de Crecimiento Nervioso/genética , Factores de Crecimiento Nervioso/aislamiento & purificación , Neuropéptidos/genética , Neuropéptidos/aislamiento & purificación , Pichia/genética , Animales , Células Cultivadas , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/aislamiento & purificación , Proteínas Ligadas a GPI/metabolismo , Proteínas Ligadas a GPI/farmacología , Expresión Génica , Humanos , Factores de Crecimiento Nervioso/metabolismo , Factores de Crecimiento Nervioso/farmacología , Neuritas/efectos de los fármacos , Neuropéptidos/metabolismo , Neuropéptidos/farmacología , Pichia/metabolismo , Ratas , Proteínas Recombinantes/genética , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología
19.
Cell Mol Biol Lett ; 20(5): 965-73, 2015 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-26751893

RESUMEN

Neuritin (Nrn1) is a neurotrophic factor that plays various roles in neural development and synaptic plasticity. In this study, the NRN1 gene was cloned and expressed in Escherichia coli and then recombinant neuritin protein was purified so that its neurobiological activity could be evaluated. The protein, which was obtained at a concentration of 0.45 mg/ml and > 90% purity, had the predicted molecular weight of 30 kDa, as determined via sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Western blot analysis confirmed that an anti-neuritin antibody could recognize the fusion protein. Subsequent functional analyses revealed that recombinant neuritin promoted neurite outgrowth in embryonic chicken dorsal root ganglia and PC12 cells. These results suggest that recombinant neuritin protein could be a valuable tool for inducing neurite regeneration, for instance in cases of spinal cord injury or neurological diseases.


Asunto(s)
Escherichia coli/metabolismo , Neuritas/fisiología , Neuropéptidos/metabolismo , Animales , Embrión de Pollo , Clonación Molecular , Electroforesis en Gel de Poliacrilamida , Proteínas Ligadas a GPI/química , Proteínas Ligadas a GPI/genética , Proteínas Ligadas a GPI/metabolismo , Ganglios Espinales/citología , Ganglios Espinales/efectos de los fármacos , Ganglios Espinales/metabolismo , Humanos , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Neuropéptidos/química , Neuropéptidos/genética , Células PC12 , Plásmidos/metabolismo , Ratas , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/aislamiento & purificación , Proteínas Recombinantes/farmacología
20.
Biotechnol Lett ; 36(5): 1069-78, 2014 May.
Artículo en Inglés | MEDLINE | ID: mdl-24563295

RESUMEN

We previously identified a novel protein elicitor, PebC1, from Botrytis cinerea and described its enhancement of plant growth, drought tolerance and disease resistance in tomato. Here, we have investigated the defense-associated molecular responses in Arabidopsis thaliana after treatment with recombinant PebC1. PebC1 was expressed in Escherichia coli. Recombinant protein treatments improved plant resistance to Botrytis infection and maintained plant defenses for more than 21 days. The purified protein at 10 µg ml(-1) activated extracellular medium alkalization (pH) and reactive oxygen species and nitric oxide generation and also induced defense gene expression. Arabidopsis mutants that are insensitive to salicylic acid had increased resistance to Botrytis infection after PebC1 treatment but PebC1 did not affect the resistance of mutants with jasmonic acid and ethylene transduction pathways. The results suggest that PebC1 can function as an activator of plant disease resistance and can promote disease resistance to Botrytis in A. thaliana through the ethylene signal transduction pathway.


Asunto(s)
Arabidopsis/efectos de los fármacos , Botrytis/genética , Resistencia a la Enfermedad/efectos de los fármacos , Proteínas Fúngicas/farmacología , Proteínas Recombinantes/farmacología , Adaptación Biológica/efectos de los fármacos , Arabidopsis/fisiología , Sequías , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Transducción de Señal/efectos de los fármacos , Estrés Fisiológico/efectos de los fármacos
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