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1.
FASEB J ; 38(3): e23472, 2024 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-38329323

RESUMEN

Allergic asthma development and pathogenesis are influenced by airway epithelial cells in response to allergens. Heme oxygenase-1 (HO-1), an inducible enzyme responsible for the breakdown of heme, has been considered an appealing target for the treatment of chronic inflammatory diseases. Herein, we report that alleviation of allergic airway inflammation by HO-1-mediated suppression of pyroptosis in airway epithelial cells (AECs). Using house dust mite (HDM)-induced asthma models of mice, we found increased gasdermin D (GSDMD) in the airway epithelium. In vivo administration of disulfiram, a specific inhibitor of pore formation by GSDMD, decreased thymic stromal lymphopoietin (TSLP) release, T helper type 2 immune response, alleviated airway inflammation, and reduced airway hyperresponsiveness (AHR). HO-1 induction by hemin administration reversed these phenotypes. In vitro studies revealed that HO-1 restrained GSDMD-mediated pyroptosis and cytokine TSLP release in AECs by binding Nuclear Factor-Kappa B (NF-κB) p65 RHD domain and thus controlling NF-κB-dependent pyroptosis. These data provide new therapeutic indications for purposing HO-1 to counteract inflammation, which contributes to allergic inflammation control.


Asunto(s)
Asma , Hemo-Oxigenasa 1 , FN-kappa B , Animales , Ratones , Citocinas/metabolismo , Células Epiteliales/metabolismo , Hemo-Oxigenasa 1/metabolismo , Inflamación/metabolismo , FN-kappa B/metabolismo , Piroptosis , Linfopoyetina del Estroma Tímico
2.
Epilepsy Behav ; 152: 109641, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38286099

RESUMEN

OBJECTIVE: To evaluate the therapeutic efficacy and safety of agomelatine for treating the sleep and mood disorders in epilepsy patients. METHODS: Retrospective data were derived from 113 epilepsy patients for at least 8 weeks. All the subjects were divided into two groups, one was treated with agomelatine, the other was treated with escitalopram. Their depression and anxiety states were assessed by Hamilton Depression (HAMD) and Hamilton Anxiety (HAMA) Scales. Sleep quality was assessed by the Pittsburgh Sleep Quality Index (PSQI). RESULTS: The HAMA, HAMD and PSQI scores in both groups significantly declined after the treatments with agomelatine and escitalopram. However, the agomelatine group exhibited greater improvement in terms of HAMA and PSQI scores compared to the escitalopram group. No severe adverse events were observed in agomelatine group. SIGNIFICANCE: Agomelatine performed better in HAMA and PSQI scores compared to escitalopram, where no significant increase in seizure frequency or side effects were observed. Possibly, agomelatine presents a promising therapeutic option for treating the sleep or mood disorders in epilepsy patients.


Asunto(s)
Trastorno Depresivo Mayor , Epilepsia , Humanos , Estudios Retrospectivos , Escitalopram , Resultado del Tratamiento , Sueño , Trastornos del Humor/etiología , Trastornos del Humor/inducido químicamente , Acetamidas/efectos adversos , Epilepsia/complicaciones , Epilepsia/tratamiento farmacológico , Epilepsia/inducido químicamente
3.
Inflamm Res ; 72(1): 89-105, 2023 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-36331571

RESUMEN

AIM: To explore the role and mechanism of human adipose-derived mesenchymal stem cells (hAD-MSCs) in the treatment of osteoarthritis (OA). METHODS: OA hulth model of Sprague Dawley (SD) rats and 20 ng/ml TNF-α treated chondrocytes were used as models of OA in vivo and in vitro, respectively. hAD-MSCs were administrated in the articular cavity by injection in vivo and co-cultured with chondrocytes using transwell in vitro. Haematoxylin and eosin staining and Safranin-O/Fast green staining were performed to detect tissue destruction and histopathology. Scanning electron microscopy and transmission electron microscopy were used to observe the ultrastructure of chondrocytes. The pyroptosis signaling pathway-related proteins were detected by immunohistochemistry, immunofluorescence, qRT-PCR and Western blot. And small interference technology was used to study the mechanism in depth. RESULTS: hAD-MSCs could delay the development of rat OA, improve the pathological changes of joints, inhibit the expression of NLRP3, Caspase1, GSDMD and TNFR1. In vitro, the expression of pyroptosis signal proteins in chondrocytes was significantly elevated when stimulated with TNF-α, the level of inflammatory factors such as IL-1ß, IL-18 was increased, and the cell morphology was significantly destroyed. While co-cultured with hAD-MSCs, these syndromes were reversed. Knockout of TNFR1 also returned the upregulation of pyroptosis signals which caused by TNF-α. CONCLUSION: These results demonstrated that hAD-MSCs could inhibit pyroptosis signaling pathway of chondrocytes induced by TNF-α, which have raised our understanding of the role of hAD-MSCs as promising therapy for the management of OA.


Asunto(s)
Células Madre Mesenquimatosas , Osteoartritis , Humanos , Ratas , Animales , Condrocitos/metabolismo , Receptores Tipo I de Factores de Necrosis Tumoral , Piroptosis , Factor de Necrosis Tumoral alfa/metabolismo , Células Cultivadas , Ratas Sprague-Dawley , Osteoartritis/metabolismo , Células Madre Mesenquimatosas/metabolismo
4.
Arch Virol ; 168(4): 118, 2023 Mar 23.
Artículo en Inglés | MEDLINE | ID: mdl-36952055

RESUMEN

A novel negative-sense single-stranded RNA virus, tentatively named "rose-associated cytorhabdovirus" (RaCV), was identified by high-throughput sequencing. RaCV is 16,067 nucleotides in length and contains eight open reading frames (ORFs 1-8) encoding a nucleocapsid protein (N), a putative phosphoprotein (P), a putative P3 protein (P3), a putative P4 protein (P4), a putative matrix protein (M), a glycoprotein (G), a putative P7 protein (P7), and an RNA-dependent RNA polymerase (L), respectively. The coding genes are flanked by a 3' leader sequence (228 nt) and a 5' trailer sequence (251 nt) and are separated by conserved intergenic junctions (3'-AUUCUUUUUG(N)nCUN-5'). Phylogenetic analysis showed that RaCV clustered with yerba mate virus A (YmVA) within the cytorhabdovirus clade, and it exhibited low a degree of nt sequence similarity (<40% identity) to other rhabdoviruses. Amino acid sequence comparisons between the putative proteins of RaCV and the corresponding proteins of other cytorhabdoviruses showed that the sequence identity levels were far below the species demarcation cutoff of 80% for cytorhabdoviruses. These results suggest that RaCV should be classified as a new member of the genus Cytorhabdovirus.


Asunto(s)
Rhabdoviridae , Rosa , Filogenia , Genoma Viral , Rhabdoviridae/genética , Proteínas Virales/genética , Sistemas de Lectura Abierta , ARN Viral/genética , Enfermedades de las Plantas
5.
Plant Cell ; 31(6): 1367-1384, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-30962395

RESUMEN

Saline-alkali soil is a major environmental constraint impairing plant growth and crop productivity. In this study, we identified a Ca2+ sensor/kinase/plasma membrane (PM) H+-ATPase module as a central component conferring alkali tolerance in Arabidopsis (Arabidopsis thaliana). We report that the SCaBP3 (SOS3-LIKE CALCIUM BINDING PROTEIN3)/CBL7 (CALCINEURIN B-LIKE7) loss-of-function plants exhibit enhanced stress tolerance associated with increased PM H+-ATPase activity and provide fundamental mechanistic insights into the regulation of PM H+-ATPase activity. Consistent with the genetic evidence, interaction analyses, in vivo reconstitution experiments, and determination of H+-ATPase activity indicate that interaction of the Ca2+ sensor SCaBP3 with the C-terminal Region I domain of the PM H+-ATPase AHA2 (Arabidopsis thaliana PLASMA MEMBRANE PROTON ATPASE2) facilitates the intramolecular interaction of the AHA2 C terminus with the Central loop region of the PM H+-ATPase to promote autoinhibition of H+-ATPase activity. Concurrently, direct interaction of SCaPB3 with the kinase PKS5 (PROTEIN KINASE SOS2-LIKE5) stabilizes the kinase-ATPase interaction and thereby fosters the inhibitory phosphorylation of AHA2 by PKS5. Consistently, yeast reconstitution experiments and genetic analysis indicate that SCaBP3 provides a bifurcated pathway for coordinating intramolecular and intermolecular inhibition of PM H+-ATPase. We propose that alkaline stress-triggered Ca2+ signals induce SCaBP3 dissociation from AHA2 to enhance PM H+-ATPase activity. This work illustrates a versatile signaling module that enables the stress-responsive adjustment of plasma membrane proton fluxes.


Asunto(s)
Arabidopsis/metabolismo , Membrana Celular/metabolismo , Raíces de Plantas/metabolismo , ATPasas de Translocación de Protón/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Raíces de Plantas/genética , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , ATPasas de Translocación de Protón/genética , Transducción de Señal/genética , Transducción de Señal/fisiología
6.
Sensors (Basel) ; 22(20)2022 Oct 21.
Artículo en Inglés | MEDLINE | ID: mdl-36298433

RESUMEN

The key to evaluating the health status of cable-stayed bridges lies in the accuracy of cable force measurement. When measuring the cable force using the conventional frequency method, the clearance between the bracing cable and the protective tube is typically disregarded. Moreover, due to their large size, existing vibration sensors are difficult to install into protective tubes for steel strand-type bracing cables to measure the cable force. To address the above difficulties, a type of thin rod vibration sensor only 5 mm in diameter was designed based on the high sensitivity of Fiber Bragg grating (FBG), and high-throughput data processing software for engineering calculation (EC) was self-developed. Then, the recognition principle of the thin rod vibration sensor was theoretically analyzed and a step-by-step tension test was carried out. The results demonstrated that the relative error of the cable force measured by the thin rod vibration sensor within 12.865 Hz was less than 5% and the sensitivity reached 28.7 pm/Hz, indicating its high measurement precision. Upon subsequent application of the thin rod vibration sensor to a monitoring test in the field, the relative error of the fundamental frequency between artificial and natural excitations was less than 4%. In addition, the error relative to both the theoretical frequency and the third-party sampling frequency was less than 5%, further verifying the accuracy and applicability for monitoring the cable force of bridges under natural excitation. Compared with the traditional cantilever FBG sensor, the improved sensor with supporting data processing software has the advantages of small cross-section, high reliability, and good sensitivity. The research results can provide a reference for the subsequent accurate measurement of cable force and the development of a supporting sensor data processing system.


Asunto(s)
Fibras Ópticas , Vibración , Reproducibilidad de los Resultados , Acero
7.
J Allergy Clin Immunol ; 148(6): 1545-1558, 2021 12.
Artículo en Inglés | MEDLINE | ID: mdl-33957164

RESUMEN

BACKGROUND: Exosomes have emerged as a vital player in cell-cell communication; however, whether airway epithelial cell (AEC)-generated exosomes participate in asthma development remains unknown. OBJECTIVE: Our aims were to characterize the AEC-secreted exosomes and the potentially functional protein(s) that may contribute to the proinflammatory effects of AEC exosomes in the dendritic cell (DC)-dominant airway allergic models and to confirm their clinical significance in patients with asthma. METHODS: Mice were treated with exosomes derived from house dust mite (HDM)-stimulated AECs (HDM-AEC-EXOs) or monocyte-derived DCs primed by HDM and/or contactin-1 (CNTN1). The numbers of DCs in the lung were determined by flow cytometry. Proteomic analysis of purified HDM-AEC-EXOs was performed. CNTN1 small interfering RNA was designed to probe its role in airway allergy, and γ-secretase inhibitor was used to determine involvement of the Notch pathway. RESULTS: HDM-AEC-EXOs facilitate the recruitment, proliferation, migration, and activation of monocyte-derived DCs in cell culture and in mice. CNTN1 in exosomes is a critical player in asthma pathology. RNA interference-mediated silencing and pharmaceutical inhibitors characterize Notch2 receptor as necessary for relaying the CNTN1 signal to activate TH2 cell/TH17 cell immune response. Studies of patients with asthma also support existence of the CNTN1-Notch2 axis that has been observed in cell and mouse models. CONCLUSION: This study's findings reveal a novel role for CNTN1 in asthma pathogenesis mediated through exosome secretion, indicating a potential strategy for the treatment of allergic airway inflammation.


Asunto(s)
Asma/inmunología , Contactina 1/metabolismo , Células Dendríticas/inmunología , Exosomas/metabolismo , Hipersensibilidad/inmunología , Mucosa Respiratoria/metabolismo , Células Th2/inmunología , Animales , Antígenos Dermatofagoides/inmunología , Movimiento Celular , Proliferación Celular , Células Cultivadas , Contactina 1/genética , Humanos , Ratones , Ratones Endogámicos C57BL , Monocitos/citología , ARN Interferente Pequeño/genética , Receptor Notch2/genética , Receptor Notch2/metabolismo
8.
Int J Mol Sci ; 23(19)2022 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-36232879

RESUMEN

Enterocytozoon hepatopenaei (EHP) is the pathogen of hepatopancreatic microsporidiosis (HPM) in shrimp. The diseased shrimp Litopenaeus vannamei exhibits a slow growth syndrome, which causes severe economic losses. Herein, 4D label-free quantitative proteomics was employed to analyze the hepatopancreas of L. vannamei with a light (EHPptp2 < 103 copies/50 ng hpDNA, L group) and heavy (EHPptp2 > 104 copies/50 ng hpDNA, H group) load of EHP to better understand the pathogenesis of HPM. Exactly 786 (L group) and 1056 (H group) differentially expressed proteins (DEPs) versus the EHP-free (C group) control were mainly clustered to lipid metabolism, amino acid metabolism, and energy production processing. Compared with the L group, the H group exhibited down-regulation significantly in lipid metabolism, especially in the elongation and degradation of fatty acid, biosynthesis of unsaturated fatty acid, metabolism of α-linolenic acid, sphingolipid, and glycerolipid, as well as juvenile hormone (JH) degradation. Expression pattern analysis showed that the degree of infection was positively correlated with metabolic change. About 479 EHP proteins were detected in infected shrimps, including 95 predicted transporters. These findings suggest that EHP infection induced the consumption of storage lipids and the entire down-regulation of lipid metabolism and the coupling energy production, in addition to the hormone metabolism disorder. These were ultimately responsible for the stunted growth.


Asunto(s)
Hepatopáncreas , Penaeidae , Aminoácidos , Animales , Regulación hacia Abajo , Enterocytozoon , Hormonas , Hormonas Juveniles , Metabolismo de los Lípidos , Proteómica , Esfingolípidos , Ácido alfa-Linolénico
9.
J Invertebr Pathol ; 183: 107600, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33961882

RESUMEN

The single-celled pathogen Nosema bombycis, that can infect silkworm Bombyx mori and other lepidoptera including Spodoptera, is the first identified Microsporidia which has diplokaryotic nuclei throughout the life cycle. Septin proteins can form highly ordered filaments, bundles or ring structures related to the cytokinesis in fungi. Here, three septin proteins (NbSeptin1, NbSeptin2 and NbSeptin3) from Nosema bombycis CQ I are described. These proteins, appear to be conserved within the phylum Microsporidia. NbSeptins transcripts were detected throughout the pathogen developmental cycle and were significantly enhanced from second days of infection, which lead to our hypothesis that NbSeptins play a role in merogony. Immunofluorescence assay (IFA) revealed a broad distribution of NbSeptins in meronts and partly co-localization of NbSeptins. Interestingly, in some of meronts, NbSeptin2 and NbSeptin3 showed localization between the nuclei of the diplokaryon. Yeast two-hybrid and co-immunoprecipitation analysis verified that NbSeptins can interact with each other. Our findings suggest that NbSeptins can cooperate in the proliferation stage of Nosema bombycis and contribute towards the understanding of the rols of septins in microsporidia development.


Asunto(s)
Nosema/fisiología , Septinas/genética , Esporas Fúngicas/fisiología , Secuencia de Aminoácidos , Animales , Bombyx/crecimiento & desarrollo , Bombyx/microbiología , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Larva/crecimiento & desarrollo , Larva/microbiología , Nosema/genética , Nosema/crecimiento & desarrollo , Filogenia , Septinas/química , Septinas/metabolismo , Alineación de Secuencia
10.
J Invertebr Pathol ; 169: 107310, 2020 01.
Artículo en Inglés | MEDLINE | ID: mdl-31862268

RESUMEN

Nosema bombycis is an obligate intracellular pathogen that can be transmitted vertically from infected females to eggs, resulting in congenital infections in embryos. Here we investigated the proliferation characteristics of N. bombycis in silkworm embryos using a histopathological approach and deep RNA sequencing. We found that N. bombycis proliferated mainly around yolk granules at the early stage of the embryonic development, 1-2 days post oviposition (dpo). At 4-6 dpo, a portion of N. bombycis in different stages adjacent to the embryo were packaged into the newly formed intestinal lumen, while the remaining parasites continued to proliferate around yolk granules. In the newly hatched larvae (9 dpo), the newly formed spores accumulated in the gut lumen and immediately were released into the environment via the faeces. Transcriptional profiling of N. bombycis further confirmed multiplication of N. bombycis throughout every stage of embryonic development. Additionally, the increased transcriptional level of spore wall proteins and polar tube proteins from 4 dpo indicated an active formation of mature spores. Taken together, our results have provided a characterization of the proliferation of this intracellular microsporidian pathogen in congenitally infected embryos leading to vertical transmission.


Asunto(s)
Bombyx/microbiología , Interacciones Huésped-Patógeno , Nosema/fisiología , Animales , Bombyx/embriología , Bombyx/crecimiento & desarrollo , Embrión no Mamífero/microbiología , Larva/crecimiento & desarrollo , Larva/microbiología , RNA-Seq
11.
Proc Natl Acad Sci U S A ; 114(17): 4388-4393, 2017 04 25.
Artículo en Inglés | MEDLINE | ID: mdl-28396416

RESUMEN

The mitochondrial calcium uniporter is a Ca2+-activated Ca2+ channel complex mediating mitochondrial Ca2+ uptake, a process crucial for Ca2+ signaling, bioenergetics, and cell death. The uniporter is composed of the pore-forming MCU protein, the gatekeeping MICU1 and MICU2 subunits, and EMRE, a single-pass membrane protein that links MCU and MICU1 together. As a bridging subunit required for channel function, EMRE could paradoxically inhibit uniporter complex formation if expressed in excess. Here, we show that mitochondrial mAAA proteases AFG3L2 and SPG7 rapidly degrade unassembled EMRE using the energy of ATP hydrolysis. Once EMRE is incorporated into the complex, its turnover is inhibited >15-fold. Protease-resistant EMRE mutants produce uniporter subcomplexes that induce constitutive Ca2+ leakage into mitochondria, a condition linked to debilitating neuromuscular disorders in humans. The results highlight the dynamic nature of uniporter subunit assembly, which must be tightly regulated to ensure proper mitochondrial responses to intracellular Ca2+ signals.


Asunto(s)
Canales de Calcio/metabolismo , Señalización del Calcio/fisiología , Calcio/metabolismo , Regulación de la Expresión Génica/fisiología , Canales de Calcio/genética , Eliminación de Gen , Células HEK293 , Células HeLa , Humanos , Péptido Hidrolasas/metabolismo , Subunidades de Proteína
12.
J Invertebr Pathol ; 164: 59-65, 2019 06.
Artículo en Inglés | MEDLINE | ID: mdl-31039370

RESUMEN

Nosema bombycis, the pathogen of silkworm pébrine, causes enormous economic losses to sericulture. As such, quarantine of commercial silkworm eggs represents an important safeguard to the silkworm industry. Here, we established a user-friendly detection system based on a nucleic acid lateral flow strip (NAFLS) that combines polymerase chain reaction (PCR) and a colloidal gold strip. PCR primers were designed based on the sequence of LSU rDNA of N. bombycis and has favourable specificity for common microsporidian isolates in silkworms. The forward and reverse primers were labeled on the 5' end with biotin and carboxyfluorescein (FAM), respectively. Genomic DNA was extracted from egg samples and was used as a template for PCR, followed by subsequent detection by NALFS. The detection limit of purified N. bombycis genomic DNA was 1 pg, 100× more sensitive than that of agarose gel electrophoresis (AGE). Furthermore, the sensitivity of detection of simulated "infected" silkworm eggs was 10-100× higher than that of AGE. NALFS detected infection in 27 of 29 samples of silkworm eggs oviposited by female moths infected in lab; ≥2% infected eggs per batch are detected as positive, while ≥40% infected eggs per batch are required for detection by AGE. Collectively, NALFS is easy to use and has great potential for widespread use in the detection of N. bombycis in silkworm egg production.


Asunto(s)
Bombyx/microbiología , Microsporidiosis/diagnóstico , Nosema/aislamiento & purificación , Animales , Óvulo/microbiología , Patología Molecular/métodos
13.
Sensors (Basel) ; 19(2)2019 Jan 17.
Artículo en Inglés | MEDLINE | ID: mdl-30658412

RESUMEN

This paper presents a smart "e-nose" device to monitor indoor hazardous air. Indoor hazardous odor is a threat for seniors, infants, children, pregnant women, disabled residents, and patients. To overcome the limitations of using existing non-intelligent, slow-responding, deficient gas sensors, we propose a novel artificial-intelligent-based multiple hazard gas detector (MHGD) system that is mounted on a motor vehicle-based robot which can be remotely controlled. First, we optimized the sensor array for the classification of three hazardous gases, including cigarette smoke, inflammable ethanol, and off-flavor from spoiled food, using an e-nose with a mixing chamber. The mixing chamber can prevent the impact of environmental changes. We compared the classification results of all combinations of sensors, and selected the one with the highest accuracy (98.88%) as the optimal sensor array for the MHGD. The optimal sensor array was then mounted on the MHGD to detect and classify the target gases without a mixing chamber but in a controlled environment. Finally, we tested the MHGD under these conditions, and achieved an acceptable accuracy (70.00%).

14.
Plant Cell ; 27(7): 2032-41, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-26198069

RESUMEN

Stomata are natural openings through which many pathogenic bacteria enter plants. Successful bacterial pathogens have evolved various virulence factors to promote stomatal opening. Here, we show that the Pseudomonas syringae type III effector protein AvrB induces stomatal opening and enhances bacterial virulence in a manner dependent on RPM1-INTERACTING4 (RIN4), which promotes stomatal opening by positively regulating the Arabidopsis plasma membrane H(+)-ATPase (AHA1), which is presumed to directly regulate guard cell turgor pressure. In support of a role of AHA1 in AvrB-induced stomatal opening, AvrB enhances ATPase activity in plants. Unexpectedly, AHA1 promotes the interaction between the jasmonate (JA) receptor CORONATINE INSENSITIVE1 (COI1) and JASMONATE ZIM-DOMAIN (JAZ) proteins and enhances JA signaling. JA signaling is required for optimum stomatal infection in AHA1-active plants. Similarly, AvrB also induces the COI1-JAZ9 interaction and the degradation of multiple JAZ proteins. AvrB-induced stomatal opening and virulence require the canonical JA signaling pathway, which involves the COI1 and NAC transcription factors. The findings thus point to a previously unknown pathway exploited by P. syringae that acts upstream of COI1 to regulate JA signaling and stomatal opening.


Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimología , Proteínas Bacterianas/metabolismo , Ciclopentanos/metabolismo , Oxilipinas/metabolismo , Estomas de Plantas/microbiología , ATPasas de Translocación de Protón/metabolismo , Pseudomonas syringae/patogenicidad , Transducción de Señal , Aminoácidos/farmacología , Arabidopsis/efectos de los fármacos , Arabidopsis/microbiología , Membrana Celular/efectos de los fármacos , Membrana Celular/enzimología , Prueba de Complementación Genética , Indenos/farmacología , Mutación/genética , Enfermedades de las Plantas/microbiología , Estomas de Plantas/efectos de los fármacos , Unión Proteica/efectos de los fármacos , Proteolisis/efectos de los fármacos , Pseudomonas syringae/efectos de los fármacos , Transducción de Señal/efectos de los fármacos , Factores de Transcripción/metabolismo , Virulencia/efectos de los fármacos
15.
Infect Immun ; 85(3)2017 03.
Artículo en Inglés | MEDLINE | ID: mdl-28031263

RESUMEN

All microsporidia possess a unique, highly specialized invasion mechanism that involves the polar tube and spore wall. The interaction between spore wall proteins (SWPs) and polar tube proteins (PTPs) in the formation, arrangement, orderly orientation, and function of the polar tube and spore wall remains to be determined. This study was undertaken to examine the protein interactions of Nosema bombycis SWP7 (NbSWP7), NbSWP9, and PTPs. Coimmunoprecipitation, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and yeast two-hybrid data demonstrated that NbSWP9, but not NbSWP7, interacts with NbPTP1 and NbPTP2. Furthermore, immunoelectron microscopy (IEM) showed that NbSWP9 was localized mainly in the developing polar tube of sporoblasts, while NbSWP7 was found randomly in the cytoplasm. However, both NbSWP9 and NbSWP7 were located in the polar tube and spore wall of N. bombycis mature spores. The reason why NbSWP7 was localized to the polar tube may be due to the interaction between NbSWP9 and NbSWP7. Interestingly, the majority of NbSWP9, but not NbSWP7, accumulated in the beginning part of the extruded polar tube and the ruptured spore wall called the anchoring disk (AD) when the mature spores germinated under weak-alkaline environmental stimulation. Additionally, anti-NbSWP9 antibody reduced spore germination in a dose-dependent manner. In conclusion, our study further confirmed that NbSWP9 is a scaffolding protein that not only anchors and holds the polar tube but also tethers the polar tube to the spore wall.


Asunto(s)
Proteínas Fúngicas/metabolismo , Nosema/fisiología , Esporas Fúngicas , Pared Celular/metabolismo , Nosema/ultraestructura , Unión Proteica
16.
J Exp Bot ; 68(11): 2951-2962, 2017 05 17.
Artículo en Inglés | MEDLINE | ID: mdl-28582540

RESUMEN

Plasma membrane (PM) H+-ATPase is essential for plant growth and development. Various environmental stimuli regulate its activity, a process that involves many protein cofactors. However, whether endogenous small molecules play a role in this regulation remains unknown. Here, we describe a bio-guided isolation method to identify endogenous small molecules that regulate PM H+-ATPase activity. We obtained crude extracts from Arabidopsis seedlings with or without salt treatment and then purified them into fractions based on polarity and molecular mass by repeated column chromatography. By evaluating the effect of each fraction on PM H+-ATPase activity, we found that fractions containing the endogenous, free unsaturated fatty acids oleic acid (C18:1), linoleic acid (C18:2), and linolenic acid (C18:3) extracted from salt-treated seedlings stimulate PM H+-ATPase activity. These results were further confirmed by the addition of exogenous C18:1, C18:2, or C18:3 in the activity assay. The ssi2 mutant, with reduced levels of C18:1, C18:2, and C18:3, displayed reduced PM H+-ATPase activity. Furthermore, C18:1, C18:2, and C18:3 directly bound to the C-terminus of the PM H+-ATPase AHA2. Collectively, our results demonstrate that the binding of free unsaturated fatty acids to the C-terminus of PM H+-ATPase is required for its activation under salt stress. The bio-guided isolation model described in this study could enable the identification of new endogenous small molecules that modulate essential protein functions, as well as signal transduction, in plants.


Asunto(s)
Arabidopsis/enzimología , ATPasas de Translocación de Protón/metabolismo , Bioensayo , Fraccionamiento Celular , Membrana Celular/metabolismo , Conductividad Eléctrica , Activación Enzimática , Ácido Linoleico/metabolismo , Ácido Oléico/metabolismo , Ácido alfa-Linolénico/metabolismo
17.
Anal Biochem ; 462: 13-8, 2014 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-24953010

RESUMEN

In this paper, a new capillary electrophoresis (CE) separation and detection method was developed for the chiral separation of the four major Cinchona alkaloids (quinine/quinidine and cinchonine/cinchonidine) using hydroxypropyl-ß-cyclodextrin (HP-ß-CD) and chiral ionic liquid ([TBA][L-ASP]) as selectors. Separation parameters such as buffer concentrations, pH, HP-ß-CD and chiral ionic liquid concentrations, capillary temperature, and separation voltage were investigated. After optimization of separation conditions, baseline separation of the three analytes (cinchonidine, quinine, cinchonine) was achieved in fewer than 7 min in ammonium acetate background electrolyte (pH 5.0) with the addition of HP-ß-CD in a concentration of 40 mM and [TBA][L-ASP] of 14 mM, while the baseline separation of cinchonine and quinidine was not obtained. Therefore, the first-order derivative electropherogram was applied for resolving overlapping peaks. Regression equations revealed a good linear relationship between peak areas in first-order derivative electropherograms and concentrations of the two diastereomer pairs. The results not only indicated that the first-order derivative electropherogram was effective in determination of a low content component and of those not fully separated from adjacent ones, but also showed that the ionic liquid appeared to be a very promising chiral selector in CE.


Asunto(s)
Ácido Aspártico/química , Alcaloides de Cinchona/aislamiento & purificación , Electroforesis Capilar/métodos , Líquidos Iónicos/química , Compuestos de Amonio Cuaternario/química , beta-Ciclodextrinas/química , 2-Hidroxipropil-beta-Ciclodextrina , Adsorción , Tampones (Química) , Alcaloides de Cinchona/química , Electricidad , Electroósmosis , Concentración de Iones de Hidrógeno , Estereoisomerismo , Temperatura , Factores de Tiempo
18.
Nanoscale Adv ; 6(8): 2209-2216, 2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38633048

RESUMEN

Flexible electrode films with good film-forming properties, large deformation ability, high conductivity, and strong charge and discharge capabilities are crucial for ionic electroactive polymer soft actuators. However, there are still challenges in preparing high-quality electrode films that can combine well with the intermediate polyelectrolyte to form high-performance soft actuators. Herein, we propose an advanced sandwich ionic electroactive actuator utilizing self-standing bacterial cellulose (BC) reinforced poly(3,4-ethylenedioxythiophene)/poly(4-styrenesulfonate) (PP) doped with graphene oxide (GO) conductive composite electrodes and a Nafion ion-exchange membrane via a hot-pressing method. The prepared BC-PP-GO electrodes have good film-forming properties with a Young's modulus of 1360 MPa and a high conductivity of 150 S cm-1. The hot-pressed BC-PP-GO/Nafion ionic actuator exhibited a large bending displacement of 6.2 mm (1 V, 0.1 Hz) with a long-term actuation stability up to 95% over 360 cycles without degradation. Furthermore, we introduced the actuator's potential applications including bionic grippers, flies, and fish, providing more opportunities for the development of next-generation micromanipulators and biomimetic microrobots in cm-scale space.

19.
Food Chem Toxicol ; 188: 114656, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38615797

RESUMEN

In recent years, with the acceleration of industrialization, the decline of male fertility caused by heavy metal pollution has attracted much attention. However, whether the inhibition of testicular function after cadmium exposure is reversible remains to be studied. In this study, we constructed rat models of cadmium exposure and dis-exposure, and collected relative samples to observe the changes of related indicators. The results showed that cadmium exposure could reduce the fertility, inhibit the hypothalamic-pituitary-testis axis and activate hypothalamic-pituitary-adrenal axis function, the testicular GR/PI3K-AKT/AMPK signal was abnormal, cell proliferation was inhibited and apoptosis was enhanced. Four weeks after the exposure was stopped, the fertility was still decreased, testicular testosterone synthesis and spermatogenesis were inhibited, cell proliferation was inhibited and apoptosis was enhanced, but all of them were reversed. After eight weeks of cadmium exposure, the above indicators were observed to return to normal. At the same time, by giving different concentrations of corticosterone to spermatogonium, we confirmed that corticosterone may regulate the proliferation and apoptosis of spermatogonium through GR/PI3K-AKT/AMPK signal. In this study, the reproductive toxicity of cadmium, a metal environmental pollutant, was analyzed in depth to provide a new theoretical and experimental basis for ensuring male reproductive health.


Asunto(s)
Apoptosis , Cadmio , Ratas Sprague-Dawley , Testículo , Masculino , Animales , Cadmio/toxicidad , Testículo/efectos de los fármacos , Testículo/metabolismo , Ratas , Apoptosis/efectos de los fármacos , Testosterona/metabolismo , Espermatogénesis/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-akt/genética , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Sistema Hipotálamo-Hipofisario/metabolismo , Fosfatidilinositol 3-Quinasas/metabolismo , Fosfatidilinositol 3-Quinasas/genética , Corticosterona , Transducción de Señal/efectos de los fármacos
20.
Mol Plant Pathol ; 25(1): e13408, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-38041680

RESUMEN

Citrus exocortis viroid (CEVd) and citrus bark cracking viroid (CBCVd) are two important viroids that infect citrus plants and frequently occur as mixed infections in orchards. However, the mechanism of antagonism between the two viroids in mixed infections remains unclear. The CEVd/CBCVd-citron system and small RNA sequencing (sRNA-seq) were used to study the antagonism. When CBCVd was inoculated before CEVd, the CEVd titre was significantly reduced and the symptoms were attenuated. Viroid-derived sRNAs (vd-sRNAs) from CEVd and CBCVd were predominantly 21-nucleotide (nt) and 22-nt in length and had similar 5' base biases. Homologous sequences of the two viroids in the terminal right (TR) region are rich in vd-sRNAs, and the high frequency vd-sRNAs selected from the CBCVd TR region can be used to degrade the transcripts of CEVd in vivo directly. These results suggest that RNA silencing may play an important role in the antagonism of the two viroids, thus deepening our understanding of the molecular interaction of long noncoding RNAs in woody plants.


Asunto(s)
Citrus , Coinfección , Viroides , Viroides/genética , Interferencia de ARN , Coinfección/genética , Corteza de la Planta , Enfermedades de las Plantas/genética , Nucleótidos
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