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BACKGROUND: Boron (B) is an essential micronutrient for plants. Inappropriate B supply detrimentally affects the productivity of numerous crops. Understanding of the molecular responses of plants to different B supply levels would be of significance in crop improvement and cultivation practices to deal with the problem. RESULTS: We conducted a comprehensive analysis of the transcriptome and proteome of tobacco seedlings to investigate the expression changes of genes/proteins in response to different B supply levels, with a particular focus on B deficiency. The global gene and protein expression profiles revealed the potential mechanisms involved in the responses of tobacco to B deficiency, including up-regulation of the NIP5;1-BORs module, complex regulation of genes/proteins related to cell wall metabolism, and up-regulation of the antioxidant machinery. CONCLUSION: Our results demonstrated that B deficiency caused severe morphological and physiological disorders in tobacco seedlings, and revealed dynamic expression changes of tobacco genes/proteins in response to different B supply levels, especially to B deficiency, thus offering valuable insights into the molecular responses of tobacco to B deficiency.
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Boro , Nicotiana , Proteoma , Transcriptoma , Boro/deficiência , Boro/metabolismo , Nicotiana/genética , Nicotiana/metabolismo , Proteoma/metabolismo , Regulação da Expressão Gênica de Plantas , Plântula/genética , Plântula/metabolismo , Plântula/crescimento & desenvolvimento , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Perfilação da Expressão GênicaRESUMO
The extraction of ammonia (NH3) through electrocatalytic nitrate reduction reaction (NO3 -RR) represents a sustainable avenue in NH3 generation and utilization. However, the catalytic efficiency of the NO3 -RR is hindered by the sluggish kinetics. This study first theoretically found that phosphide-based heterostructure can alter the adsorption structure of intermediates in the nitrate-to-ammonia process, thereby achieving precise regulation of the energy barrier in the rate-determining step. Based on theoretical design, a novel Co-doped Fe2P@NiP2 heterojunction catalyst is successfully synthesized, which deliver a notable NH3 yield rate of 0.395 mmol h-1 cm-2 at -0.7 V versus RHE, as well as a remarkable ammonia Faraday efficiency of 97.2% at -0.6 V versus RHE. Experimental and theoretical results further confirm that redistributing electrons and shifting the center of the d-band upwards through interfacial doping modulate intermediates adsorption strength and inhibition of hydrogen evolution, leading to excellent performance in NO3 --to-NH3. Further integrating the Co-Fe2P@NiP2 catalyst into a Zn-nitrate battery exhibits a substantial voltage output of 1.49 V and a commendable power density of 13.2 mW cm-2. The heteroatom-doped heterojunction strategy provides a versatile route for developing advanced catalysts, thereby broadening the horizons of electrocatalytic methodologies for nitrate reduction and ammonia synthesis.
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The ALMT1 transporter aids malate secretion, chelating Al3+ ions to form nontoxic Al-malate complexes, believed to exclude Al from the roots. However, the extent to which malate secreted by ALMT1 is solely used for the exclusion of Al3+ or can be reutilized by plant roots for internal Al tolerance remains uncertain. In our investigation, we explored the impact of malate secretion on both external and internal Al resistance in Arabidopsis thaliana. Additionally, we delved into the mechanism by which the tonoplast-localized bacterial-type ATP-binding cassette (ABC) transporter complex STAR1/ALS3 promotes the degradation of the Al resistance transcription factor STOP1 to regulate ALMT1 expression. Our study demonstrates that the level of secreted malate influences whether the Al-malate complex is excluded from the roots or transported into root cells. The nodulin 26-like intrinsic protein (NIP) subfamily members NIP1;1 and NIP1;2, located in the plasma membrane, coordinate with STAR1/ALS3 to facilitate Al-malate transport from root apoplasm to the symplasm and eventually to the vacuoles for the internal Al detoxification. ALS3-dependent STAR1 interacts with and promotes the degradation of STOP1, regulating malate exudation. Our findings demonstrate the dual roles of malate exudation in external Al exclusion and Al absorption for internal Al detoxification.
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Transcriptional regulation is a crucial component of plant adaptation to numerous different stresses; however, its role in how plants adapt to low-boron (B) stress remains unclear. In this study, we show that the C2H2-type transcription factor SENSITIVE TO PROTON RHIZOTOXICITY1 (STOP1) in Arabidopsis is essential for improving plant growth under low-B conditions. STOP1 and the boric acid-channel protein NOD26-LIKE MAJOR INTRINSIC PROTEIN5;1 (NIP5;1) were found to co-localize in root epidermal cells, and STOP1 binds to the 5´-untranslated region of NIP5;1 to activate its expression and enhance B uptake by the roots. Overexpression of STOP1 increased tolerance to low-B stress by up-regulating NIP5;1 transcript levels. Further genetic analyses revealed that STOP1 and NIP5;1 function together in the same pathway to confer low-B tolerance. These results highlight the importance of the STOP1-NIP5;1 module in improving plant growth under low-B conditions.
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Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Transporte Biológico , Boro , Regulação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Raízes de Plantas/genética , Fatores de Transcrição/genéticaRESUMO
BACKGROUND: In Korea, there are no surveillance programs for vaccines that are not included in the national immunization program (NIP), and vaccine safety monitoring in the adult population is inadequate. This study aimed to establish a safety monitoring system for non-NIP vaccines in adults. METHODS: Frequently administered non-NIP vaccines were selected. Individuals were included if they received at least one of the selected vaccines at a participating institution and provided informed consent. Solicited and unsolicited adverse events were monitored using questionnaires sent through text messages on days 1, 3, 7, 28, and 90 post-vaccination. Selected adverse events of special interest (AESIs) were monitored monthly by retrospective review of electronic medical records. Causality was assessed according to the Korea Disease Control and Prevention Agency guidelines. RESULTS: Four vaccines (tetanus-diphtheria-pertussis [Tdap], pneumococcal conjugate 13-valent [PCV13], live zoster vaccine [ZVL], and recombinant zoster vaccine [RZV]) were selected, and their safety profiles were monitored at four tertiary hospitals and 10 primary care clinics. The response rates of the questionnaires on post-vaccination days 1, 7, 28, and 90 were 99.2%, 93.6%, 81.0%, and 48.7%, respectively. Of 555 AESI identified over 10 months, 10 cases received one of the selected non-NIP vaccines within 90 days of the event. CONCLUSION: We are establishing the first safety monitoring system for selected non-NIP vaccines in Korea since September 2022 and report its progress as of July 2023. However, continuous government support is essential for its maintenance and improvement.
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Vacina contra Herpes Zoster , Tétano , Adulto , Humanos , Vacinas Pneumocócicas , Vacinação/efeitos adversos , Vacinas Sintéticas , Programas de Imunização , República da CoreiaRESUMO
The complexity of the underwater environment enables significant energy consumption of sensor nodes for communication with base stations in underwater wireless sensor networks (UWSNs), and the energy consumption of nodes in different water depths is unbalanced. How to improve the energy efficiency of sensor nodes and meanwhile balance the energy consumption of nodes in different water depths in UWSNs are thus urgent concerns. Therefore, in this paper, we first propose a novel hierarchical underwater wireless sensor transmission (HUWST) framework. We then propose a game-based, energy-efficient underwater communication mechanism in the presented HUWST. It improves the energy efficiency of the underwater sensors personalized according to the various water depth layers of sensor locations. In particular, we integrate the economic game theory in our mechanism to trade off variations in communication energy consumption due to sensors in different water depth layers. Mathematically, the optimal mechanism is formulated as a complex nonlinear integer programming (NIP) problem. A new energy-efficient distributed data transmission mode decision algorithm (E-DDTMD) based on the alternating direction method of multipliers (ADMM) is thus further proposed to tackle this sophisticated NIP problem. The systematic simulation results demonstrate the effectiveness of our mechanism in improving the energy efficiency of UWSNs. Moreover, our presented E-DDTMD algorithm achieves significantly superior performance to the baseline schemes.
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Redes de Comunicação de Computadores , Tecnologia sem Fio , Simulação por Computador , Fenômenos Físicos , ÁguaRESUMO
A Ni-P amorphous alloy was deposited on a low carbon steel substrate via electroless plating. Further, the prepared samples were crystallized under the high temperature with a range from 200 °C to 500 °C in air for 1 h. The crystallization process was studied via XRD, AFM, and XPS, and anodic electrochemical behavior was investigated by potentiostatic methods in a 3.5 wt% NaCl solution. The experimental results indicate that the diffusion, dissolution, and enrichment of the component elements in the Ni-P alloy are essential during crystallization because the various corrosion behaviors corresponding to Ni and P are directly affected. More importantly, under the 400 °C treatment, H2PO2- was enriched in the alloy, which effectively hinders the anodic dissolution of nickel and forms a complete adsorption layer on the surface of the alloy. Our results demonstrate that P can effectively block the anodic dissolution of Ni during the corrosion process, and the crystallization process can effectively promote the surface enrichment of P to improve the corrosion resistance of the coating.
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Despite increasing research on psychotherapy preferences, the preferences of psychotherapy trainees are largely unknown. Moreover, differences in preferences between trainees and their patients could (a) hinder symptom improvement and therapy success for patients and (b) represent significant obstacles in the early career and development of future therapists.We compared the preferences of n = 466 psychotherapy trainees to those of n = 969 laypersons using the Cooper-Norcross Inventory of Preferences. Moreover, we compared preferences between trainees in cognitive-behavioural therapy (CBT) and psychodynamic trainees.We found significant differences between both samples in 13 of 18 items, and three of four subscales. Psychotherapy trainees preferred less therapist directiveness (d = 0.58), more emotional intensity (d = 0.74), as well as more focused challenge (d = 0.35) than laypeople. CBT trainees preferred more therapist directiveness (d = 2.00), less emotional intensity (d = 0.51), more present orientation (d = 0.76) and more focused challenge (d = 0.33) than trainees in psychodynamic/psychoanalytic therapy.Overall, the results underline the importance of implementing preference assessment and discussion during psychotherapy training. Moreover, therapists of different orientations seem to cover a large range of preferences for patients, in order to choose the right fit.
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Terapia Cognitivo-Comportamental , Terapia Psicanalítica , Humanos , Psicoterapia/métodos , Terapia Cognitivo-Comportamental/educação , EmoçõesRESUMO
OBJECTIVE: The Cooper-Norcross Inventory of Preferences (C-NIP) is a brief, multidimensional measure of clients' therapy preferences. This study aimed to examine the factor structure and measurement invariance of the C-NIP. METHOD: Fifteen datasets (N = 10,088 observations) representing the C-NIP in nine language versions were obtained from authors of psychometric studies. Confirmatory factor analysis and exploratory structural equation modeling were used to analyze the data. RESULTS: None of the proposed models adequately fit the data. Therefore, a new model was developed that sufficiently fit most of the C-NIP version 1.1 datasets. The new model was invariant up to the strict and means levels across genders, ages, and psychotherapy experience but only up to the metric level across translations. CONCLUSIONS: The C-NIP can be used to compare men and women, people of diverse ages, and people with some vs. no experience with psychotherapy. Lower reliabilities of the C-NIP scales are a limitation.
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NGATHA-Like 1 (NGAL1) transcription factor has been identified as a gene regulated through AUG-stop-mediated boron (B)-dependent translation stall; however, its function in B response remains unknown. Here, we show that NGAL1 plays an important role in the maintenance of B transport under both low- and high-B conditions in Arabidopsis thaliana. NGAL1 mRNA is accumulated predominantly in shoots in response to B stress. Independent ngal1 mutants carrying transferred DNA (T-DNA) and Ds-transposon insertions exhibit reduced B concentrations in aerial tissues and produce shortened and reduced number of siliques when B supply is limited. Furthermore, the expression of B transporter genes including nodulin 26-like intrinsic protein 6; 1 (NIP6;1), NIP5;1, NIP7;1 and borate exporter 1 (BOR1) is significantly decreased in ngal1 mutants under low-B condition, suggesting that NGAL1 is required for the transcript accumulation of B transporter genes to facilitate B transport and distribution under B limitation. Under high-B condition, ngal1 mutants exhibit reduced growth and increased B concentration in their shoots. The accumulation of BOR4 mRNA, a B transporter required for B efflux to soil, is significantly reduced in roots of ngal1 plants under high-B condition, suggesting that NGAL1 is involved in the upregulation of BOR4 in response to excess B. Together, our results indicate that NGAL1 is involved in the transcriptional regulation of B transporter genes to facilitate B transport and distribution under both low- and high-B conditions.
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Aquaporinas , Proteínas de Arabidopsis , Arabidopsis , Antiporters/metabolismo , Aquaporinas/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Boro/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras/metabolismo , Raízes de Plantas/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de Transcrição/metabolismoRESUMO
Perovskite solar cells (PSCs) offer advantages over widely deployed silicon solar cells in terms of ease of fabrication; however, the device is still under rigorous materials optimization for cell performance, stability, and cost. In this work, we explore a version of a PSC by replacing the polymeric hole transport layer (HTL) such as Spiro-OMeTAD, P3HT, and PEDOT: PSS with a more air-stable metal oxide, viz., nitrogen-doped titanium dioxide (TiO2:N). Numerical simulations on formamidinium (FA)-based PSCs in the FTO/TiO2/FAPbI3/Ag configuration have been carried out to depict the behaviour of the HTL as well as the effect of absorber layer thickness (∆t) on photovoltaic parameters. The results show that the cell output increases when the HTL bandgap increases from 2.5 to 3.0 eV. By optimizing the absorber layer thickness and the gradient in defect density (Nt), the device structure considered here can deliver a maximum power conversion efficiency of ~21.38% for a lower HTL bandgap (~2.5 eV) and ~26.99% for a higher HTL bandgap of ~3.0 eV. The results are validated by reproducing the performance of PSCs employing commonly used polymeric HTLs, viz. Spiro-OMeTAD, P3HT, and PEDOT: PSS as well as high power conversion efficiency in the highly crystalline perovskite layer. Therefore, the present study provides high-performing, cost-effective PSCs using TiO2:N.
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Triarylamine-substituted bithiophene (BT-4D), terthiophene (TT-4D), and quarterthiophene (QT-4D) small molecules are synthesized and used as low-cost hole-transporting materials (HTMs) for perovskite solar cells (PSCs). The optoelectronic, electrochemical, and thermal properties of the compounds are investigated systematically. The BT-4D, TT-4D, and QT-4D compounds exhibit thermal decomposition temperature over 400 °C. The n-i-p configured perovskite solar cells (PSCs) fabricated with BT-4D as HTM show the maximum power conversion efficiency (PCE) of 19.34% owing to its better hole-extracting properties and film formation compared to TT-4D and QT-4D, which exhibit PCE of 17% and 16%, respectively. Importantly, PSCs using BT-4D demonstrate exceptional stability by retaining 98% of its initial PCE after 1186 h of continuous 1 sun illumination. The remarkable long-term stability and facile synthetic procedure of BT-4D show a great promise for efficient, stable, and low-cost HTMs for PSCs for commercial applications.
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Boron (B) is an essential micronutrient for plant growth and development. Jasmonic acid (JA) plays pivotal roles in plant growth, but the underlying molecular mechanism of JA involvement in B-deficiency-induced root growth inhibition is yet to be explored. In this study, we investigated the response of JA to B deficiency and the mechanism of JAR1-dependent JA signaling in root growth inhibition under B deficiency in Arabidopsis. B deficiency enhanced JA signaling in roots, and root growth inhibition was partially restored by JA biosynthesis inhibition. The jar1-1 (jasmonate-resistant 1, JAR1) mutant, and mutants of coronatine-insensitive 1 (coi1-2) and myc2 defective in JA signaling showed insensitivity to B deficiency. The ethylene-overproduction mutant eto1 and ethylene-insensitive mutant etr1 showed sensitivity and insensitivity to B deficiency, respectively, suggesting that ethylene is involved in the inhibition of primary root growth under B deficiency. Furthermore, after a decline in levels of EIN3, which may contribute to root growth, ethylene signaling was weakened in the jar1-1 mutant root under B deficiency. Under B deficiency, B concentrations were increased in the roots and shoots of the jar1-1 mutant, owing to the large root system and its activity. Therefore, our findings revealed that JA, which is involved in the inhibition of root growth under B deficiency, is regulated by JAR1-activated JA and ethylene signaling pathways.
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Proteínas de Arabidopsis , Arabidopsis , Boro/deficiência , Raízes de Plantas/crescimento & desenvolvimento , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Ciclopentanos , Regulação da Expressão Gênica de Plantas , Nucleotidiltransferases , OxilipinasRESUMO
Theory is proposed for nanohole siliconpin/nipphotodetector (PD) physics, promising devices in the future data communications and lidar applications. Photons and carriers have wavelengths of 1µm and 5 nm, respectively. We propose vertical nanoholes having 2D periodicity with a feature size of 1µm will produce photons slower than those in bulk silicon, but carriers are unchanged. Close comparison to experiments validates this view. First, we study steady state nanohole PD current as a function of illumination power, and results are attributed to the voltage drop partitions in the PD and electrodes. Nanohole PD voltage drop depends on illumination, but series resistance voltage drop does not, and this explains experiments well. Next, we study transient characteristics for the sudden termination of light illumination. Nanohole PDs are much faster than flat PDs, and this is because the former produces much less slow diffusion minority carriers. In fact, most photons have already been absorbed in thei-layer in nanohole PDs, resulting in much less diffusion minority carriers at the bottom highly doped layer. Why diffusion in PDs is slow and that in bipolar junction transistors is quick is discussed in appendix.
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BACKGROUND: In Arabidopsis, the aluminum (Al) exclusion mechanism is mainly facilitated by ALMT1-mediated malate exudation and MATE-mediated citrate releases from the root. Recently, we have demonstrated that coordinated functioning between an ALMT1-mediated Al exclusion mechanism, via exudation of malate from the root tip, and a NIP1;2-facilitated internal detoxification mechanism, via removal of Al from the root cell wall and subsequent root-to-shoot Al translocation, plays critical roles in achieving overall Al resistance. However, the genetic relationship between ALMT1 and NIP1;2 in these processes remained unclear. RESULTS: Through genetic and physiological analyses, we demonstrate that unlike ALMT1 and MATE, which function independently and additively, ALMT1 and NIP1;2 show an epistatic relationship in Al resistance. These results indicate that ALMT1 and NIP1;2 function in the same biochemical pathway, whereas ALMT1 and MATE in different ones. CONCLUSION: The establishment of the epistatic relationship and the coordinated functioning between the ALMT1 and NIP1;2-mediated exclusion and internal detoxification mechanisms are pivotal for achieving overall Al resistance in the non-accumulating Arabidopsis plant. We discuss and emphasize the indispensable roles of the root cell wall for the implementation of the Al exclusion mechanism and for the establishment of an epistatic relationship between the ALMT1-mediated exclusion mechanism and the NIP1;2-facilitated internal detoxification mechanism.
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Alumínio/metabolismo , Aquaporinas/genética , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Epistasia Genética , Transportadores de Ânions Orgânicos/genética , Aquaporinas/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Inativação Metabólica , Transportadores de Ânions Orgânicos/metabolismoRESUMO
Under high boron (B) conditions, nodulin 26-like intrinsic protein 5;1 (NIP5;1) mRNA, a boric acid channel, is destabilized to avoid excess B entry into roots of Arabidopsis thaliana. In this regulation, the minimum upstream open reading frame (uORF), AUGUAA, in its 5'-untranslated region (5'-UTR) is essential, and high B enhances ribosome stalling at AUGUAA and leads to suppression of translation and mRNA degradation. This B-dependent AUGUAA-mediated regulation occurs also in animal transient expression and reticulocyte lysate translation systems. Thus, uncovering the ubiquitousness of B-dependent unique regulation is important to reveal the evolution of translational regulation. In the present study, we examined the regulation in Saccharomyces cerevisiae. Reporter assay showed that in yeast, carrying ATGTAA in 5'-UTR of NIP5;1 upstream of the reporter gene, the relative reporter activities were reduced significantly under high B conditions compared with control, whereas deletion of ATGTAA abolished such responses. This suggests that AUGUAA mediates B-dependent regulation of translation in Saccharomyces cerevisiae. Moreover, the deletion of ATGTAA resulted in up to 10-fold increase in general reporter activities indicating the suppression effect of AUGUAA on translation of the main ORF. Interestingly, mRNA level of the reporter gene was not affected by B in both yeast cells with and without AUGUAA. This finding reveals that in yeast, unlike the case in plants, mRNA degradation is not associated with AUGUAA regulation. Together, results suggest that B-dependent AUGUAA-mediated translational regulation is common among eukaryotes.
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Aminoácidos/genética , Boro/metabolismo , Boro/farmacologia , Regulação Fúngica da Expressão Gênica , Saccharomyces cerevisiae/efeitos dos fármacos , Saccharomyces cerevisiae/genética , Regiões 5' não Traduzidas , Fases de Leitura Aberta , Biossíntese de Proteínas , RNA Mensageiro/metabolismo , Saccharomyces cerevisiae/metabolismoRESUMO
Deficiency of the essential nutrient boron (B) in the soil is one of the most widespread micronutrient deficiencies worldwide, leading to developmental defects in root and shoot tissues of plants, and severe yield reductions in many crops. Despite this agricultural importance, the underlying mechanisms of how B shapes plant developmental and morphological processes are still not unequivocally understood in detail. This review evaluates experimental approaches that address our current understanding of how B influences plant morphological processes by focusing on developmental defects observed under B deficiency. We assess what is known about mechanisms that control B homeostasis and specifically highlight: (i) limitations in the methodology that is used to induce B deficiency; (ii) differences between mutant phenotypes and normal plants grown under B deficiency; and (iii) recent research on analyzing interactions between B and phytohormones. Our analysis highlights the need for standardized methodology to evaluate the roles of B in the cell wall versus other parts of the cell.
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Boro/deficiência , Desenvolvimento Vegetal , Plantas/metabolismo , Ácidos Bóricos/metabolismo , Reguladores de Crescimento de Plantas/metabolismoRESUMO
Osteoclasts derived from the monocyte/macrophage hematopoietic lineage regulate bone resorption, a process balanced by bone formation in the continual renewal of the skeletal system. As dysfunctions of these cells result in bone metabolic diseases such as osteoporosis and osteopetrosis, the exploration of the mechanisms regulating their differentiation is a priority. A potential mechanism may involve long noncoding RNAs (lncRNAs), which are known to regulate various cell biology activities, including proliferation, differentiation, and apoptosis. The expression of the lncRNA AK077216 (Lnc-AK077216) is significantly upregulated during osteoclastogenesis identified by microarray and verified by qPCR. Up- and downregulation of Lnc-AK077216, respectively promotes and inhibits osteoclast differentiation, bone resorption, and the expression of related genes on the basis of tartrate-resistant acid phosphatase staining, qPCR, and western blot results. In addition, Lnc-AK077216 suppresses NIP45 expression and promotes the expression of NFATc1, an essential transcription factor during osteoclastogenesis. Besides, it was found that the expression of Lnc-AK077216 and Nfatc1 is upregulated, whereas Nip45 expression is downregulated in bone marrow and spleen tissues of ovariectomized mice. The results suggest that Lnc-AK077216 regulates NFATc1 expression and promotes osteoclast formation and function, providing a novel mechanism of osteoclastogenesis and a potential biomarker or a new drug target for osteoporosis.
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Reabsorção Óssea , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Macrófagos/efeitos dos fármacos , Fatores de Transcrição NFATC/metabolismo , Osteoclastos/efeitos dos fármacos , Osteogênese/efeitos dos fármacos , Osteoporose Pós-Menopausa/enzimologia , Ligante RANK/farmacologia , RNA Longo não Codificante/metabolismo , Animais , Apoptose/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/genética , Macrófagos/enzimologia , Macrófagos/patologia , Camundongos , Camundongos Endogâmicos C57BL , Fatores de Transcrição NFATC/genética , Osteoclastos/enzimologia , Osteoclastos/patologia , Osteoporose Pós-Menopausa/genética , Osteoporose Pós-Menopausa/patologia , Ovariectomia , Células RAW 264.7 , RNA Longo não Codificante/genética , Transdução de SinaisRESUMO
Macrolide antibiotics have immunomodulatory activities, including suppression of cytokine production, cell adhesion molecule expression, and mucin production. These immunomodulatory activities improve the symptoms of respiratory diseases associated with chronic inflammation. However, the underlying molecular mechanism(s) is not well understood yet. To address this, we prepared clarithromycin (CAM)-conjugated Sepharose and examined bound cellular proteins by proteome analysis. We identified mitochondrial proteins 4-nitrophenylphosphatase domain and non-neuronal synaptosomal associated protein 25-like protein homolog (NIP-SNAP)-1 and -2 and very long-chain acyl-CoA dehydrogenase (VLCAD) as CAM-binding proteins. Production of proinflammatory cytokines (IL-8 and IL-6) induced by lipopolysaccharides (LPSs) and Pam3-CSK4 in human epithelial cell lines BEAS-2B and T24 were suppressed by knockdown of NIP-SNAP-1 or -2, and partly by knockdown of VLCAD. Also, knockdown of NIP-SNAP-1 or -2 in various cell lines suppressed LPS-induced expression of IL-8 and IL-6 mRNA and NF-κB activity. Thus, CAM suppresses NF-κB-mediated proinflammatory cytokine production by interacting with mitochondrial proteins, NIP-SNAP-1 and -2.
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Claritromicina/farmacologia , Citocinas/biossíntese , Fatores Imunológicos/farmacologia , Proteínas de Membrana/metabolismo , Proteínas Mitocondriais/metabolismo , Fosfoproteínas/metabolismo , Proteínas/metabolismo , Acil-CoA Desidrogenase de Cadeia Longa/antagonistas & inibidores , Acil-CoA Desidrogenase de Cadeia Longa/genética , Acil-CoA Desidrogenase de Cadeia Longa/metabolismo , Citocinas/genética , Técnicas de Silenciamento de Genes , Células HEK293 , Humanos , Peptídeos e Proteínas de Sinalização Intercelular , Peptídeos e Proteínas de Sinalização Intracelular , Lipopolissacarídeos/farmacologia , Proteínas de Membrana/antagonistas & inibidores , Proteínas de Membrana/genética , NF-kappa B/metabolismo , Fosfoproteínas/antagonistas & inibidores , Fosfoproteínas/genética , Ligação Proteica , Proteínas/antagonistas & inibidores , Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteína 25 Associada a Sinaptossoma/metabolismo , Receptores Toll-Like/agonistasRESUMO
NIP-SNAP-1 and -2 are ubiquitous proteins thought to be associated with maintenance of mitochondrial function, neuronal transmission, and autophagy. However, their physiological functions remain largely unknown. To elucidate their functional importance, we screened for proteins that interact with NIP-SNAP-1 and -2, resulting in identification of HSP60 and P62/SQSTM1 as binding proteins. NIP-SNAP-1 and -2 localized in the mitochondrial inner membrane space, whereas HSP60 localized in the matrix. Native gel electrophoresis and filter trap assays revealed that human HSP60 prevented aggregation of newly synthesized NIP-SNAP-2 in an in vitro translation system. Moreover, expression levels of NIP-SNAP-1 and -2 in cells were decreased by knockdown of HSP60, but not HSP10. These findings indicate that HSP60 promotes folding and maintains the stability of NIP-SNAP-1 and -2.