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1.
J Biol Chem ; 300(2): 105638, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38199570

RESUMEN

The inflammasome is a large multiprotein complex that assembles in the cell cytoplasm in response to stress or pathogenic infection. Its primary function is to defend the cell and promote the secretion of pro-inflammatory cytokines, including IL-1ß and IL-18. Previous research has shown that in immortalized bone marrow-derived macrophages (iBMDMs) inflammasome assembly is dependent on the deacetylase HDAC6 and the aggresome processing pathway (APP), a cellular pathway involved in the disposal of misfolded proteins. Here we used primary BMDMs from mice in which HDAC6 is ablated or impaired and found that inflammasome activation was largely normal. We also used human peripheral blood mononuclear cells and monocyte cell lines expressing a synthetic protein blocking the HDAC6-ubiquitin interaction and impairing the APP and found that inflammasome activation was moderately affected. Finally, we used a novel HDAC6 degrader and showed that inflammasome activation was partially impaired in human macrophage cell lines with depleted HDAC6. Our results therefore show that HDAC6 importance in inflammasome activation is context-dependent.


Asunto(s)
Inflamasomas , Leucocitos Mononucleares , Animales , Humanos , Ratones , Línea Celular , Histona Desacetilasa 6/genética , Histona Desacetilasa 6/metabolismo , Inflamasomas/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Leucocitos Mononucleares/metabolismo , Macrófagos/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/genética , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Transporte de Proteínas/fisiología
2.
Opt Express ; 32(3): 4650-4667, 2024 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-38297661

RESUMEN

Aerosol intensive optical properties, including lidar ratio and particle depolarization ratio, are of vital importance for aerosol typing. However, aerosol intensive optical properties at near-infrared wavelength are less exploited by atmospheric lidar measurements, because of the comparably small backscatter cross section of Raman-scattering and a low efficiency of signal detection compared to what is commonly available at 355 nm and 532 nm. To obtain accurate optical properties of aerosols at near-infrared wavelength, we considered three factors: Raman-spectra selection, detector selection, and interference-filter optimization. Rotational Raman scattering has been chosen for Raman signal detection, because of the higher cross-section compared to vibrational Raman scattering. The optimization of the properties of the interference filter are based on a comprehensive consideration of both signal-to-noise ratio and temperature dependence of the simulated lidar signals. The interference filter that has eventually been chosen uses the central wavelength at 1056 nm and a filter bandwidth (full-width-at-half-maximum) of 6 nm. We built a 3-channel 1064-nm rotational Raman lidar. In this paper two methods are proposed to test the temperature dependence of the signal-detection unit and to evaluate the quality of the Raman signals. We performed two measurements to test the quality of the detection channel: cirrus clouds in the free troposphere and aerosols in the planetary boundary layer. Our analysis of the measured Raman signals shows a negligible temperature dependence of the Raman signals in our system. For cirrus measurements, the Raman signal profile did not show crosstalk even for the case of strong elastic backscatter from clouds, which was about 100 times larger than Rayleigh scattering in the case considered here. The cirrus-mean extinction-to-backscatter ratio (lidar ratio) was 27.8 ± 10.0 sr (1064 nm) at a height of 10.5-11.5 km above ground. For the aerosols in the planetary boundary layer, we found the mean lidar ratio of 38.9 ± 7.0 sr at a height of 1.0-3.0 km above ground.

3.
Opt Express ; 32(9): 14963-14977, 2024 Apr 22.
Artículo en Inglés | MEDLINE | ID: mdl-38859159

RESUMEN

The vertical profiles of aerosol or mixed-phase cloud optical properties (e.g. extinction coefficient) at 1064 nm are difficult to obtain from lidar observations. Based on the techniques of rotational Raman signal at 1058 nm described by Haarig et al. [Atmos. Meas. Tech.9, 4269 (2016)10.5194/amt-9-4269-2016], we have developed a novel rotational Raman polarization lidar at 1064 nm at Wuhan University. In this design, we optimized the central wavelength of the rotational Raman channel to 1056 nm with a bandwidth of 6 nm to increase the signal-to-noise ratio and minimize the temperature dependence of the extracted rotational Raman spectrum. And then separated elastic polarization channels (1064 nm Parallel, P and 1064 nm Cross, S) into near range (low 1064 nm P and 1064 nm S) and far range detection channels (high 1064 nm P and 1064 nm S) to extend the dynamic range of lidar observation. Silicon single photon avalanche diodes (SPAD) working at photon counting mode were applied to improve the quantum efficiency and reduce the electronic noise, which resulted in quantum efficiency of 2.5%. With a power of 3 W diode pumped pulsed Nd:YAG laser and aperture of 250 mm Cassegrain telescope, the detectable range can cover the atmosphere from 0.3 km to the top troposphere (about 12-15 km). To the best of our knowledge, the design of this novel lidar system is described and the mixed-phase cloud and aerosol optical properties observations of backscatter coefficients, extinction coefficients, lidar ratio and depolarization ratio at 1064 nm were performed as demonstrations of the system capabilities.

4.
J Exp Bot ; 75(5): 1547-1564, 2024 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-37976184

RESUMEN

Legume nodules express multiple leghemoglobins (Lbs) and non-symbiotic hemoglobins (Glbs), but how they are regulated is unclear. Here, we study the regulation of all Lbs and Glbs of Lotus japonicus in different physiologically relevant conditions and mutant backgrounds. We quantified hemoglobin expression, localized reactive oxygen species (ROS) and nitric oxide (NO) in nodules, and deployed mutants deficient in Lbs and in the transcription factors NLP4 (associated with nitrate sensitivity) and NAC094 (associated with senescence). Expression of Lbs and class 2 Glbs was suppressed by nitrate, whereas expression of class 1 and 3 Glbs was positively correlated with external nitrate concentrations. Nitrate-responsive elements were found in the promoters of several hemoglobin genes. Mutant nodules without Lbs showed accumulation of ROS and NO and alterations of antioxidants and senescence markers. NO accumulation occurred by a nitrate-independent pathway, probably due to the virtual disappearance of Glb1-1 and the deficiency of Lbs. We conclude that hemoglobins are regulated in a gene-specific manner during nodule development and in response to nitrate and dark stress. Mutant analyses reveal that nodules lacking Lbs experience nitro-oxidative stress and that there is compensation of expression between Lb1 and Lb2. They also show modulation of hemoglobin expression by NLP4 and NAC094.


Asunto(s)
Lotus , Nitratos , Nitratos/metabolismo , Lotus/fisiología , Especies Reactivas de Oxígeno/metabolismo , Hemoglobinas/genética , Hemoglobinas/metabolismo , Leghemoglobina/metabolismo , Óxido Nítrico/metabolismo , Simbiosis , Nódulos de las Raíces de las Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas
5.
New Phytol ; 239(5): 1989-2006, 2023 09.
Artículo en Inglés | MEDLINE | ID: mdl-37329247

RESUMEN

Legume nodules produce large quantities of heme required for the synthesis of leghemoglobin (Lb) and other hemoproteins. Despite the crucial function of Lb in nitrogen fixation and the toxicity of free heme, the mechanisms of heme homeostasis remain elusive. Biochemical, cellular, and genetic approaches were used to study the role of heme oxygenases (HOs) in heme degradation in the model legume Lotus japonicus. Heme and biliverdin were quantified and localized, HOs were characterized, and knockout LORE1 and CRISPR/Cas9 mutants for LjHO1 were generated and phenotyped. We show that LjHO1, but not the LjHO2 isoform, is responsible for heme catabolism in nodules and identify biliverdin as the in vivo product of the enzyme in senescing green nodules. Spatiotemporal expression analysis revealed that LjHO1 expression and biliverdin production are restricted to the plastids of uninfected interstitial cells. The nodules of ho1 mutants showed decreased nitrogen fixation, and the development of brown, rather than green, nodules during senescence. Increased superoxide production was observed in ho1 nodules, underscoring the importance of LjHO1 in antioxidant defense. We conclude that LjHO1 plays an essential role in degradation of Lb heme, uncovering a novel function of nodule plastids and uninfected interstitial cells in nitrogen fixation.


Asunto(s)
Lotus , Fijación del Nitrógeno , Fijación del Nitrógeno/genética , Lotus/metabolismo , Hemo Oxigenasa (Desciclizante)/genética , Hemo Oxigenasa (Desciclizante)/metabolismo , Biliverdina/metabolismo , Leghemoglobina/genética , Simbiosis/genética , Nódulos de las Raíces de las Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas
6.
New Phytol ; 238(5): 2113-2129, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36945893

RESUMEN

Legumes establish symbioses with rhizobia by forming nitrogen-fixing nodules. Nitrate is a major environmental factor that affects symbiotic functioning. However, the molecular mechanism of nitrate-induced nodule senescence is poorly understood. Comparative transcriptomic analysis reveals an NAC-type transcription factor in Lotus japonicus, LjNAC094, that acts as a positive regulator in nitrate-induced nodule senescence. Stable overexpression and mutant lines of NAC094 were constructed and used for phenotypic characterization. DNA-affinity purification sequencing was performed to identify NAC094 targeting genes and results were confirmed by electrophoretic mobility shift and transactivation assays. Overexpression of NAC094 induces premature nodule senescence. Knocking out NAC094 partially relieves nitrate-induced degradation of leghemoglobins and abolishes nodule expression of senescence-associated genes (SAGs) that contain a conserved binding motif for NAC094. Nitrate-triggered metabolic changes in wild-type nodules are largely affected in nac094 mutant nodules. Induction of NAC094 and its targeting SAGs was almost blocked in the nitrate-insensitive nlp1, nlp4, and nlp1 nlp4 mutants. We conclude that NAC094 functions downstream of NLP1 and NLP4 by regulating nitrate-induced expression of SAGs. Our study fills in a key gap between nitrate and the execution of nodule senescence, and provides a potential strategy to improve nitrogen fixation and stress tolerance of legumes.


Asunto(s)
Lotus , Nódulos de las Raíces de las Plantas , Nódulos de las Raíces de las Plantas/metabolismo , Nitratos/farmacología , Nitratos/metabolismo , Factores de Transcripción/metabolismo , Fijación del Nitrógeno/genética , Lotus/metabolismo , Simbiosis/fisiología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas
7.
Opt Express ; 31(19): 30040-30065, 2023 Sep 11.
Artículo en Inglés | MEDLINE | ID: mdl-37710556

RESUMEN

Multi-wavelength Raman lidar has been widely used in profiling aerosol optical properties. The accuracy of measured aerosol optical properties largely depends on sophisticated lidar data retrieval algorithms. Commonly to retrieve aerosol optical properties of Raman lidar, the extinction-related Ångström exponent (EAE) is assumed (to be 1). This value usually generally differs from the true value (called EAE deviation) and adds uncertainty to the retrieved aerosol optical properties. Lidar-signal noise and EAE-deviation are two important error sources for retrieving aerosol optical properties. As the measurement accuracy of Raman lidar has been greatly improved in recent years, the influence of signal noise on retrieval results becomes relatively small, and the uncertainty of retrieved aerosol optical properties caused by an EAE-deviation becomes nonnegligible, especially in scenes that EAE deviation is large. In this study, an iteration retrieval algorithm is proposed to obtain more reliable EAE based on multi-wavelength Raman lidar. Results from this iteration are more precise values of aerosol optical properties. Three atmospheric scenarios where aerosol distribution and the values of EAE vary widely were simulated with a Monte Carlo method to analyze the characteristics and robustness of the iterative algorithm. The results show that the proposed iterative algorithm can eliminate the systematic errors of aerosol optical properties retrieved by traditional retrieval method. The EAEs after iteration does converge to the true value, and the accuracy of aerosol optical properties can be greatly improved, especially for the particle backscatter coefficient and lidar ratio, which has been improved by more than 10% in most cases, and even more than 30%. In addition, field observations data of a three-wavelength Raman lidar are analyzed to illustrate the necessity and reliability of the proposed iterative retrieval algorithm.

8.
PLoS Biol ; 18(11): e3000872, 2020 11.
Artículo en Inglés | MEDLINE | ID: mdl-33186350

RESUMEN

Metabolic reprogramming to fulfill the biosynthetic and bioenergetic demands of cancer cells has aroused great interest in recent years. However, metabolic reprogramming for cancer metastasis has not been well elucidated. Here, we screened a subpopulation of breast cancer cells with highly metastatic capacity to the lung in mice and investigated the metabolic alternations by analyzing the metabolome and the transcriptome, which were confirmed in breast cancer cells, mouse models, and patients' tissues. The effects and the mechanisms of nucleotide de novo synthesis in cancer metastasis were further evaluated in vitro and in vivo. In our study, we report an increased nucleotide de novo synthesis as a key metabolic hallmark in metastatic breast cancer cells and revealed that enforced nucleotide de novo synthesis was enough to drive the metastasis of breast cancer cells. An increased key metabolite of de novo synthesis, guanosine-5'-triphosphate (GTP), is able to generate more cyclic guanosine monophosphate (cGMP) to activate cGMP-dependent protein kinases PKG and downstream MAPK pathway, resulting in the increased tumor cell stemness and metastasis. Blocking de novo synthesis by silencing phosphoribosylpyrophosphate synthetase 2 (PRPS2) can effectively decrease the stemness of breast cancer cells and reduce the lung metastasis. More interestingly, in breast cancer patients, the level of plasma uric acid (UA), a downstream metabolite of purine, is tightly correlated with patient's survival. Our study uncovered that increased de novo synthesis is a metabolic hallmark of metastatic breast cancer cells and its metabolites can regulate the signaling pathway to promote the stemness and metastasis of breast cancer.


Asunto(s)
Neoplasias de la Mama/metabolismo , Células Madre Neoplásicas/metabolismo , Nucleótidos/metabolismo , Adulto , Animales , Neoplasias de la Mama/genética , Línea Celular Tumoral , China , GMP Cíclico/metabolismo , Proteínas Quinasas Dependientes de GMP Cíclico/metabolismo , Femenino , Perfilación de la Expresión Génica/métodos , Humanos , Sistema de Señalización de MAP Quinasas/fisiología , Metabolómica/métodos , Ratones , Ratones Endogámicos BALB C , Nucleótidos/biosíntesis , Purinas , Ribosa-Fosfato Pirofosfoquinasa/metabolismo , Transducción de Señal
9.
J Sleep Res ; 32(3): e13762, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36325765

RESUMEN

Sleep-disordered breathing (SDB) is highly prevalent, but its rates of diagnosis and treatment are low. As a heterogeneous condition, it is associated with delayed diagnosis and/or suboptimal treatment. We aimed to determine distinct SDB clusters and examine their association with medical care-seeking behaviour and sleep habits in a community-dwelling population in South China. Participants were enrolled in the 'Guangdong Sleep Health Study'. Five distinct community sites were investigated. Participant information was collected, and overnight sleep monitoring was performed. Latent class analysis was performed to classify patients with SDB based on patterns of SDB-related symptoms and signs. Overall, 1,524 patients with SDB were evaluated. Four distinct clusters were identified: 'minimally symptomatic' (Cluster 2), which was the most dominant subtype (41.6%), followed by 'pure insomnia/fewer daytime symptoms' (Cluster 4; 24.7%), 'insomnia/multiple daytime symptoms' (Cluster 3; 17.8%), and 'upper airway symptoms with sleepiness' (Cluster 1; 15.9%). The overall medical care-seeking rate attributable to sleep and breathing disorders was only 3.3%: 10.3% in Cluster 3, 2.5% in Cluster 1, 2.1% in Cluster 4, and 1.3% in Cluster 2, in which Cluster 3 was the highest and Cluster 2 was the lowest (adjusted p < 0.05). Regarding the sleep habit of going to bed and waking up at a consistent time, Cluster 3 exhibited the worst performance and Cluster 2 the best. In conclusion, distinct phenotypic subtypes were identified in community-dwelling patients with SDB. Tailored strategies to encourage medical care-seeking, early identification, and optimisation of treatment are necessary considering the different subtypes.


Asunto(s)
Síndromes de la Apnea del Sueño , Trastornos del Inicio y del Mantenimiento del Sueño , Humanos , Polisomnografía , Trastornos del Inicio y del Mantenimiento del Sueño/complicaciones , Síndromes de la Apnea del Sueño/diagnóstico , Síndromes de la Apnea del Sueño/epidemiología , Síndromes de la Apnea del Sueño/terapia , Sueño , China/epidemiología
10.
Phytother Res ; 37(9): 3820-3838, 2023 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-37421347

RESUMEN

We conducted a meta-analysis to evaluate the clinical efficacy of berberine (BBR) in treating acute ischemic stroke (AIS), explore its anti-inflammatory effects, and assess its potential applications for AIS patients. We comprehensively searched nine databases from inception until July 1, 2022, to identify clinical trials investigating the use of BBR in treating AIS. We performed statistical analyses using RevMan5.4 software and focused on primary outcomes such as inflammatory markers as well as secondary outcomes including immune system indicators, relevant biomarkers, carotid artery atherosclerosis, and adverse reactions. Our analysis included data from 17 clinical trials involving 1670 patients with AIS. Our results revealed that BBR in combination with conventional treatment significantly reduced levels of high-sensitivity C-reactive protein (hs-CRP), macrophage migration inhibitory factor (MIF), interleukin-6 (IL-6), complement C3, hypoxic inducible factor-1 α (HIF-1α), cysteine protease-3 (Caspase-3), the national institutes of health stroke scale (NIHSS), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), carotid intima-media thickness (IMT), the number of unstable plaques, and carotid crouse score on ultrasound when compared with conventional treatment alone. Furthermore, combining BBR with conventional treatment may improve the overall effective rate. Therefore, our findings suggest that BBR can be used as an adjuvant therapy for AIS due to its ability to reduce inflammatory cytokine levels, providing a novel therapeutic option for AIS. However, larger randomized controlled trials are necessary to confirm these results.


Asunto(s)
Aterosclerosis , Berberina , Accidente Cerebrovascular Isquémico , Accidente Cerebrovascular , Humanos , Accidente Cerebrovascular Isquémico/tratamiento farmacológico , Berberina/uso terapéutico , Grosor Intima-Media Carotídeo , Triglicéridos , Aterosclerosis/tratamiento farmacológico , Accidente Cerebrovascular/tratamiento farmacológico
11.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 40(6): 1142-1151, 2023 Dec 25.
Artículo en Zh | MEDLINE | ID: mdl-38151937

RESUMEN

Alzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive impairment, with the predominant clinical diagnosis of spatial working memory (SWM) deficiency, which seriously affects the physical and mental health of patients. However, the current pharmacological therapies have unsatisfactory cure rates and other problems, so non-pharmacological physical therapies have gradually received widespread attention. Recently, a novel treatment using 40 Hz light flicker stimulation (40 Hz-LFS) to rescue the cognitive function of model animals with AD has made initial progress, but the neurophysiological mechanism remains unclear. Therefore, this paper will explore the potential neural mechanisms underlying the modulation of SWM by 40 Hz-LFS based on cross-frequency coupling (CFC). Ten adult Wistar rats were first subjected to acute LFS at frequencies of 20, 40, and 60 Hz. The entrainment effect of LFS with different frequency on neural oscillations in the hippocampus (HPC) and medial prefrontal cortex (mPFC) was analyzed. The results showed that acute 40 Hz-LFS was able to develop strong entrainment and significantly modulate the oscillation power of the low-frequency gamma (lγ) rhythms. The rats were then randomly divided into experimental and control groups of 5 rats each for a long-term 40 Hz-LFS (7 d). Their SWM function was assessed by a T-maze task, and the CFC changes in the HPC-mPFC circuit were analyzed by phase-amplitude coupling (PAC). The results showed that the behavioral performance of the experimental group was improved and the PAC of θ-lγ rhythm was enhanced, and the difference was statistically significant. The results of this paper suggested that the long-term 40 Hz-LFS effectively improved SWM function in rats, which may be attributed to its enhanced communication of different rhythmic oscillations in the relevant neural circuits. It is expected that the study in this paper will build a foundation for further research on the mechanism of 40 Hz-LFS to improve cognitive function and promote its clinical application in the future.


Asunto(s)
Memoria a Corto Plazo , Enfermedades Neurodegenerativas , Humanos , Adulto , Ratas , Animales , Memoria a Corto Plazo/fisiología , Ratas Wistar , Hipocampo , Corteza Prefrontal
12.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi ; 40(6): 1135-1141, 2023 Dec 25.
Artículo en Zh | MEDLINE | ID: mdl-38151936

RESUMEN

With the widespread use of electrical equipment, cognitive functions such as working memory (WM) could be severely affected when people are exposed to 50 Hz electromagnetic fields (EMF) for long term. However, the effects of EMF exposure on WM and its neural mechanism remain unclear. In the present paper, 15 rats were randomly assigned to three groups, and exposed to an EMF environment at 50 Hz and 2 mT for a different duration: 0 days (control group), 24 days (experimental group I), and 48 days (experimental group II). Then, their WM function was assessed by the T-maze task. Besides, their local field potential (LFP) in the media prefrontal cortex (mPFC) was recorded by the in vivo multichannel electrophysiological recording system to study the power spectral density (PSD) of θ and γ oscillations and the phase-amplitude coupling (PAC) intensity of θ-γ oscillations during the T-maze task. The results showed that the PSD of θ and γ oscillations decreased in experimental groups I and II, and the PAC intensity between θ and high-frequency γ (hγ) decreased significantly compared to the control group. The number of days needed to meet the task criterion was more in experimental groups I and II than that of control group. The results indicate that long-term exposure to EMF could impair WM function. The possible reason may be the impaired communication between different rhythmic oscillations caused by a decrease in θ-hγ PAC intensity. This paper demonstrates the negative effects of EMF on WM and reveals the potential neural mechanisms from the changes of PAC intensity, which provides important support for further investigation of the biological effects of EMF and its mechanisms.


Asunto(s)
Campos Electromagnéticos , Memoria a Corto Plazo , Humanos , Ratas , Animales , Memoria a Corto Plazo/fisiología , Campos Electromagnéticos/efectos adversos , Corteza Prefrontal , Cognición
13.
J Clin Immunol ; 42(2): 375-393, 2022 02.
Artículo en Inglés | MEDLINE | ID: mdl-34825286

RESUMEN

Both DNA damage response and methylation play a crucial role in antigen receptor recombination by creating a diverse repertoire in developing lymphocytes, but how their defects relate to T cell repertoire and phenotypic heterogeneity of immunodeficiency remains obscure. We studied the TCR repertoire in patients with the mutation in different genes (ATM, DNMT3B, ZBTB24, RAG1, DCLRE1C, and JAK3) and uncovered distinct characteristics of repertoire diversity. We propose that early aberrancies in thymus T cell development predispose to the heterogeneous phenotypes of the immunodeficiency spectrum. Shorter CDR3 lengths in ATM-deficient patients, resulting from a decreased number of nucleotide insertions during VDJ recombination in the pre-selected TCR repertoire, as well as the increment of CDR3 tyrosine residues, lead to the enrichment of pathology-associated TCRs, which may contribute to the phenotypes of ATM deficiency. Furthermore, patients with DNMT3B and ZBTB24 mutations who exhibit discrepant phenotypes present longer CDR3 lengths and reduced number of known pathology-associated TCRs.


Asunto(s)
Síndromes de Inmunodeficiencia , Linfocitos T , Regiones Determinantes de Complementariedad/química , Regiones Determinantes de Complementariedad/genética , Reparación del ADN/genética , Humanos , Síndromes de Inmunodeficiencia/diagnóstico , Síndromes de Inmunodeficiencia/genética , Metilación , Proteínas Represoras/genética
14.
Biochem Biophys Res Commun ; 589: 107-115, 2022 01 22.
Artículo en Inglés | MEDLINE | ID: mdl-34902746

RESUMEN

Breast cancer is prone to relapse and metastasize to many vital organs, contributing to most of the breast cancer-related death and accentuating the importance of systematic identification of key factors regulating the metastasis of breast cancer. In this study, we performed a genome-wide CRISPR/Cas9 knock out screen in an orthotopic murine model of breast cancer for essential genes monitoring the progression and metastasis of breast cancer. We found one member of the zinc finger protein (ZNF) family, i.e., ZNF319, was among the top candidate genes. We further confirmed the lower expression of ZNF319 in the tumor tissue of breast cancer patients by analyzing tissue sections with IHC staining and TCGA database. Consistently, higher expression of ZNF319 correlates with better clinical outcome in almost all subtypes of breast cancer. Moreover, knocking down or overexpressing ZNF319 in breast cancer cells dramatically affects the breast cancer growth and metastasis capacity both in vitro and in vivo, suggesting ZNF319 functions as a strong suppressor of breast cancer progression. Lastly, the transcriptome analysis on ZNF319-silenced breast cancer cells shows that ZNF319 is involved in multiple crucial signaling pathways and biological processes, especially in cell cycle and proliferation. GO and KEGG analyses of our RNA-seq results reveal the up-regulation of E2F and G2/M related genes in ZNF319-silenced cells, suggesting that ZNF319 monitors the cell cycle during the breast cancer progression through the regulation of the E2F target genes and G2/M checkpoint. In summary, our study identifies ZNF319 as a novel metastasis suppressor gene arresting tumor cell cycle in breast cancer and thus presents a novel potential therapeutic target for breast cancer treatment.


Asunto(s)
Neoplasias de la Mama , Sistemas CRISPR-Cas , Técnicas de Inactivación de Genes , Genes Supresores de Tumor , Genoma Humano , Animales , Femenino , Humanos , Ratones , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Línea Celular Tumoral , Proliferación Celular/genética , Sistemas CRISPR-Cas/genética , Fase G2 , Mitosis , Metástasis de la Neoplasia
15.
Biol Reprod ; 107(4): 1026-1034, 2022 10 11.
Artículo en Inglés | MEDLINE | ID: mdl-35774023

RESUMEN

G protein-coupled estrogen receptor (GPER), a seven-transmembrane G protein-coupled receptor, mediates the rapid pre-genomic signaling actions of estrogen and derivatives thereof. The expression of GPER is extensive in mammal male reproductive system. However, the functional role of GPER in mouse sperm has not yet been well recognized. This study revealed that GPER was expressed at the acrosome and the mid-flagellum of the mouse sperm. The endogenous GPER ligand 17ß-estradiol and the selective GPER agonist G1 increased intracellular Ca2+ concentration ([Ca2+]i) in mouse sperm, which could be abolished by G15, an antagonist of GPER. In addition, the G1-stimulated Ca2+ response was attenuated by interference with the phospholipase C (PLC) signaling pathways or by blocking the cation channel of sperm (CatSper). Chlortetracycline staining assay showed that the activation of GPER increased the incidence of acrosome-reacted sperm. Conclusively, GPER was located at the acrosome and mid-flagellum of the mouse sperm. Activation of GPER triggered the elevation of [Ca2+]i through PLC-dependent Ca2+ mobilization and CatSper-mediated Ca2+ influx, which promoted the acrosome reaction of mouse sperm.


Asunto(s)
Reacción Acrosómica , Clortetraciclina , Animales , Calcio/metabolismo , Clortetraciclina/metabolismo , Estradiol/metabolismo , Estrógenos/metabolismo , Proteínas de Unión al GTP/metabolismo , Ligandos , Masculino , Mamíferos/metabolismo , Ratones , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Semen/metabolismo , Espermatozoides/metabolismo , Fosfolipasas de Tipo C/metabolismo
16.
Blood Cells Mol Dis ; 96: 102666, 2022 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-35567997

RESUMEN

The link between hyperuricemia (HUA) and the risk of venous thromboembolism (VTE) has been well established. However, the mechanisms of thrombus generation and the effect of HUA on procoagulant activity (PCA) of erythrocytes remain unclear no matter in uremia or hyperuricemia. Here, phosphatidylserine (PS) exposure, microparticles (MPs) release, cytosolic Ca2+, TMEM16F expression, reactive oxygen species (ROS) and lipid peroxidation of erythrocyte were detected by flow cytometer. PCA was assessed by coagulation time, purified coagulation complex and fibrin production assays. The fibrin formation was observed by scanning electron microscopy (SEM). We found that PS exposure, MPs generation, TMEM16F expression and consequent PCA of erythrocyte in HUA patients significantly increased compared to those in healthy volunteers. Furthermore, high UA induced PS exposure, and MPs release of erythrocyte in concentration and time-dependent manners in vitro, which enhanced the PCA of erythrocyte and was inhibited by lactadherin, a PS inhibitor. Additionally, using SEM, we also observed compact fibrin clots with highly-branched networks and thin fibers supported by red blood cells (RBCs) and RBC-derived MPs (RMPs). Importantly, we demonstrated UA enhanced the production of ROS and lipid peroxidation and reduced the generation of glutathione (GSH) of erythrocyte, which enhanced TMEM16F activity and followed PS externalization and RMPs formation. Collectively, these results suggest that Ca2+-dependent TMEM16F activation may be responsible for UA-induced PS exposure and MPs release of RBC, which thereby contribute to the prothrombotic risk in HUA.

17.
Opt Express ; 30(20): 36235-36253, 2022 Sep 26.
Artículo en Inglés | MEDLINE | ID: mdl-36258557

RESUMEN

Vortex beam has the potential to significantly improve the performance of lidar (light detection and ranging) and optical communication applications in which low signal-to-noise ratio (SNR) limits the detection/transmission range. The vortex beam method allows for spatially separating the coherent light (laser signal) from the incoherent light (the background radiation and multiple-scattered light) of the received signal. This paper presents results of a simulation model in which the optical vortex acts as an optical filter. We present instrument parameters that describe the filtering effect, e.g., the form of the vortex phase modulation function, the topological charge of the vortex and the focal length of a virtual Fresnel lens that is used for optical filtering. Preliminary experimental results show that the background radiation within the spectral filter bandwidth can be suppressed by as much as 95%. At the same time, we retain 97% of the coherent laser signal. Our simulation model will be used in future design of lidar instruments and optical communication systems in which the optical vortex method is used for optical filtering of the detected signals.

18.
Vet Res ; 53(1): 82, 2022 Oct 12.
Artículo en Inglés | MEDLINE | ID: mdl-36224663

RESUMEN

Innate immunity plays an essential role in preventing the invasion of pathogenic microorganisms. However, innate immunity is a double-edged sword, whose excessive activation is detrimental to immune homeostasis and even leads to a "cytokine storm" of the infected host. The host develops a series of negative regulatory mechanisms to balance the immune response. Here, we report a negative regulatory mechanism of chicken innate immunity mediated by miRNA. In the GEO database, we found that miR-126-5p was markedly up-regulated in chickens infected by RNA viruses. Upregulation of miR-126-5p by RNA virus was then further shown via both a cell model and in vivo tests. Overexpression of miR-126-5p significantly inhibited the expression of interferon and inflammatory cytokine-related genes induced by RNA viruses. The opposite result was achieved after the knockdown of miR-126-5p expression. Bioinformatics analysis identified TRAF3 as candidate target gene of miR-126-5p. Experimentally, miR-126-5p can target TRAF3, as shown by the effects of miR-126-5p on the endogenous expression of TRAF3, and by the TRAF3 3'UTR driven luciferase reporter assay. Furthermore, we demonstrated that miR-126-5p negatively regulated innate immunity by blocking the MAVS-TRAF3-TBK1 axis, with a co-expression assay. Overall, our results suggest that miR-126-5p is involved in the negative regulation of chicken innate immunity, which might contribute to maintaining immune balance.


Asunto(s)
MicroARNs , Factor 3 Asociado a Receptor de TNF , Regiones no Traducidas 3' , Animales , Antivirales , Pollos/genética , Pollos/metabolismo , Citocinas/metabolismo , Inmunidad Innata/genética , Interferones/metabolismo , Luciferasas/genética , Luciferasas/metabolismo , MicroARNs/genética , MicroARNs/metabolismo , Factor 3 Asociado a Receptor de TNF/genética , Factor 3 Asociado a Receptor de TNF/metabolismo
19.
Environ Sci Technol ; 56(22): 16240-16248, 2022 Nov 15.
Artículo en Inglés | MEDLINE | ID: mdl-36322385

RESUMEN

Fabricating two-dimensional transition-metal dichalcogenide (TMD)-based unique composites is an effective way to boost the overall physical and chemical properties, which will be helpful for the efficient and fast capture of elemental mercury (Hg0) over a wide temperature range. Herein, we constructed a defect-rich Cu2WS4 nano-homojunction decorated on covalent organic frameworks (COFs) with abundant S vacancies. Highly well-dispersed and uniform Cu2WS4 nanoparticles were immobilized on COFs strongly via an ion pre-anchored strategy, consequently exhibiting enhanced Hg0 removal performance. The saturation adsorption capacity of Cu2WS4@COF composites (21.60 mg·g-1) was 9 times larger than that of Cu2WS4 crystals, which may be ascribed to more active S sites exposed in hybrid interfaces formed in the Cu2WS4 nano-homojunction and between Cu2WS4 nanoparticles and COFs. More importantly, such hybrid materials reduced adsorption deactivation at high temperatures, having a wide operating temperature range (from 40 to 200 °C) owing to the thermostability of active S species immobilized by both physical confined and chemical interactions in COFs. Accordingly, this work not only provides an effective method to construct uniform TMD-based sorbents for mercury capture but also opens a new realm of advanced COF hybrid materials with designed functionalities.

20.
J Environ Sci (China) ; 115: 1-9, 2022 May.
Artículo en Inglés | MEDLINE | ID: mdl-34969439

RESUMEN

Fe3O4-based materials are widely used for magnetic separation from wastewater. However, they often suffer from Fe-leaching behavior under acidic conditions, decreasing their activity and limiting sustainable practical applications. In this study, covalent organic frameworks (COFs) were used as the shell to protect the Fe3O4 core, and the Fe3O4@COF core-shell composites were synthesized for As(III) removal from acid wastewater. The imine-linked COFs can in situ grow on the surface of the Fe3O4 core layer by layer with [COFs/Fe3O4]mol ratio of up to 2:1. The Fe-leaching behavior was weakened over a wide pH range of 1-13. Moreover, such composites keep their magnetic characteristic, making them favorable for nanomaterial separation. As(III) batch adsorption experiments results indicated that, when COFs are used as the shell for the Fe3O4 core, a balance between As(III) removal efficiencies and the thickness of the COF shell exists. Higher As(III) removal efficiencies are obtained when the [COFs/Fe3O4]mol ratios were < 1.5:1, but thicker COF shells were not beneficial for As(III) removal. Such composites also exhibited better As(III) removal performances in the pH range of 1-7. Over a wide pH range, the zeta potential of Fe3O4@COF core-shell composites becomes more positive, which benefits the capture of negative arsenic ions. In addition, thinner surface COFs were favorable for mass transfer and facilitating the reaction of Fe and As elements. Our study highlights the promise of using COFs in nanomaterial surface protection and achieving As(III) depth removal under acidic conditions.


Asunto(s)
Estructuras Metalorgánicas , Fenómenos Magnéticos , Magnetismo , Extracción en Fase Sólida , Aguas Residuales
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