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BACKGROUND: Glioblastoma (GBM) characterized by immune escape is the most malignant primary brain tumors, which has strong immunosuppressive effect. Programmed death ligand-1 (PD-L1) is a recognized immunosuppressive member on the surface of tumor cells, and plays a crucial role in immune evasion of tumors. Actually, little is known about the regulation of PD-L1 expression in GBM. Insulin-like growth factor binding protein 3 (IGFBP3) is upregulated in GBM and is related to poor patient prognosis. However, it remains unclear whether IGFBP3 plays a role in the regulation of PD-L1 expression in GBM. METHODS: The role of IGFBP3 in the glioma immune microenvironment was investigated using the CIBERSORT algorithm. The correlation between IGFBP3 and PD-L1 expression was analyzed using TCGA and CGGA databases. QRT-PCR, immunoblotting and RNA-seq were used to examine the regulatory effect of IGFBP3 on PD-L1 expression. Co-culture assay, cell counting kit (CCK-8), qRT-PCR, ELISA and flow cytometry were performed to explore the function of IGFBP3 in inducing immunosuppression. The biological role of IGFBP3 was verified using immunohistochemical, immunofluorescence and mice orthotopic tumor model. RESULTS: In this study, we analyzed immune cells infiltration in gliomas and found that IGFBP3 may be associated with an immunosuppressive microenvironment. Then, by analyzing TCGA and CGGA databases, our results showed that IGFBP3 and PD-L1 expression were positively correlated in GBM patients, but not in LGG patients. In vitro experiments conducted on different GBM cell lines revealed that the overexpression of IGFBP3 led to an increase in PD-L1 expression, which was reversible upon knockdown IGFBP3. Mechanistically, IGFBP3 activated the JAK2/STAT3 signaling pathway, leading to an increase in PD-L1 expression. Additionally, co-culture experiments results showed IGFBP3 overexpression induced upregulation of PD-L1 expression promoted apoptosis in Jurkat cells, and this effect was blocked by IGFBP3 antibody and PDL-1 inhibitors. Importantly, in vivo experiments targeting IGFBP3 suppressed tumor growth and significantly prolonged the survival of mice. CONCLUSIONS: This research demonstrated IGFBP3 is a novel regulator for PD-L1 expression in GBM, and identified a new mechanism by which IGFBP3 regulates immune evasion through PD-L1, suggesting that IGFBP3 may be a potential novel target for GBM therapy.
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BACKGROUND: Diabetes can lead to extensive damage to the enteric nervous system (ENS), causing gastrointestinal motility disorders. However, there is currently a lack of effective treatments for diabetes-induced ENS damage. Enteric neural precursor cells (ENPCs) closely regulate the structural and functional integrity of the ENS. L-Fucose, is a dietary sugar that has been showed to effectively ameliorate central nervous system injuries, but its potential for ameliorating ENS damage and the involvement of ENPCs in this process remains uncertain. METHODS: Genetically engineered mice were generated for lineage tracing of ENPCs in vivo. Using diabetic mice in vivo and high glucose-treated primary ENPCs in vitro, the effects of L-Fucose on the injured ENS and ENPCs was evaluated by assessing gastrointestinal motility, ENS structure, and the differentiation of ENPCs. The key signaling pathways in regulating neurogenesis and neural precursor cells properties, transforming growth factor-ß (TGF-ß) and its downstream signaling pathways were further examined to clarify the potential mechanism of L-Fucose on the injured ENS and ENPCs. RESULTS: L-Fucose improved gastrointestinal motility in diabetic mice, including increased defecation frequency (p < 0.05), reduced total gastrointestinal transmission time (p < 0.001) and bead expulsion time (p < 0.05), as well as enhanced spontaneous contractility and electric field stimulation-induced contraction response in isolated colonic muscle strips (p < 0.001). The decrease in the number of neurons and glial cells in the ENS of diabetic mice were reversed by L-Fucose treatment. More importantly, L-Fucose treatment significantly promoted the proportion of ENPCs differentiated into neurons and glial cells both in vitro and in vivo, accompanied by inhibiting SMAD2 phosphorylation. CONCLUSIONS: L-Fucose could promote neurogenesis and gliogenesis derived from ENPCs by inhibiting the SMAD2 signaling, thus facilitating ENS regeneration and gastrointestinal motility recovery in type 1 diabetic mice. Video Abstract.
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Diabetes Mellitus Experimental , Diabetes Mellitus Tipo 1 , Sistema Nervioso Entérico , Células-Madre Neurales , Ratones , Animales , Fucosa/farmacología , Fucosa/metabolismo , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Diabetes Mellitus Tipo 1/metabolismo , Neuronas/metabolismo , Sistema Nervioso Entérico/metabolismo , Transducción de SeñalRESUMEN
Although nanopores can be used for single-molecule sequencing of nucleic acids using low-cost portable devices, the characterization of proteins and their modifications has yet to be established. Here, we show that hydrophilic or glycosylated peptides translocate too quickly across FraC nanopores to be recognized. However, high ionic strengths (i.e., 3 M LiCl) and low pH (i.e., pH 3) together with using a nanopore with a phenylalanine at its constriction allows the recognition of hydrophilic peptides, and to distinguish between mono- and diglycosylated peptides. Using these conditions, we devise a nanopore method to detect, characterize, and quantify post-translational modifications in generic proteins, which is one of the pressing challenges in proteomic analysis.
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Nanoporos , Glicosilación , Nanotecnología , Péptidos/química , Proteínas , ProteómicaRESUMEN
Coactivator-associated arginine methyltransferase 1 (CARM1) is a member of the family of protein arginine methyltransferases. CARM1 catalyzes methyl group transfer from the cofactor S-adenosyl-l-methionine (AdoMet) to both histone and nonhistone protein substrates. CARM1 is involved in a range of cellular processes, mainly involving RNA transcription and gene regulation. As the aberrant expression of CARM1 has been linked to tumorigenesis, the enzyme is a potential therapeutic target, leading to the development of inhibitors and tool compounds engaging with CARM1. To evaluate the effects of these compounds on the activity of CARM1, sensitive and specific analytical methods are needed. While different methods are currently available to assess the activity of methyltransferases, these assays mainly focus on either the measurement of the cofactor product S-adenosyl-l-homocysteine (AdoHcy) or employ radioactive or expensive reagents, each with their own advantages and limitations. To complement the tools currently available for the analysis of CARM1 activity, we here describe the development of a convenient assay employing peptide substrates derived from poly(A)-binding protein 1 (PABP1). This operationally straightforward liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based approach allows for the direct detection of substrate methylation with minimal workup. The method was validated, and its value in characterizing CARM1 activity and inhibition was demonstrated through a comparative analysis involving a set of established small molecules and peptide-based CARM1 inhibitors.
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Reactive astrocytes play a complex role in multiple sclerosis, and the astrocytes reactivity is an important factor in the pathogenesis of pain. It is of great significance to explore the genesis and development mechanism of pain in the early stage of multiple sclerosis (MS) for early intervention of the disease. This study aims to explore astrocyte reactivity at different stages of the experimental autoimmune encephalomyelitis (EAE) model, a mouse model of MS, and the role of astrocytes in the pain in the early stage of the EAE. In this study, we demonstrated that spinal dorsal horn astrocytes were activated in the pre-clinical stage of EAE mice, and the inhibition of spinal cord astrocyte reactivity effectively alleviates pain symptoms in EAE mice. On the other hand, spinal cord microglia were not directly participated in the early EAE pain. Moreover, the ion channel LRRC8A mediated the reactivity of spinal dorsal horn astrocytes by regulating the STAT3 pathway, therefore playing a role in the early pain of EAE.
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Encefalomielitis Autoinmune Experimental , Esclerosis Múltiple , Neuralgia , Ratones , Animales , Encefalomielitis Autoinmune Experimental/patología , Astrocitos/metabolismo , Asta Dorsal de la Médula Espinal/metabolismo , Asta Dorsal de la Médula Espinal/patología , Neuralgia/metabolismo , Médula Espinal/patología , Esclerosis Múltiple/patología , Ratones Endogámicos C57BL , Proteínas de la Membrana/metabolismoRESUMEN
BACKGROUND: The nucleotide oligomerization domain (NOD)-like receptor family pyrin domain containing 3 (NLRP3) in dorsal root ganglion (DRG) contributes to pain hypersensitivity in multiple neuropathic pain models, but the function of the NLRP3 in diabetic neuropathic pain (DNP) and the regulation mechanism are still largely unknown. Epigenetic regulation plays a vital role in the controlling of gene expression. Ten-eleven translocation methylcytosine dioxygenase 2 (TET2) is a DNA demethylase that contributes to transcriptional activation. TET2 is also involved in high glucose (HG)-induced pathology. METHODS: DNP was induced in mice via the intraperitoneal injection of streptozotocin (STZ) for five consecutive days and the mechanical threshold was evaluated in STZ-diabetic mice by using von Frey hairs. The expression level of the NLRP3 pathway and TET2 in DRG were determined through molecular biology experiments. The regulation of the NLRP3 pathway by TET2 was examined in in vitro and in vivo conditions. RESULTS: In the present research, we first established the DNP model and found that NLRP3 pathway was activated in DRG. The treatment of NLRP3 inhibitor MCC950 alleviated the mechanical allodynia of DNP mice. Then we revealed that in STZ-diabetic mice DRG, the genomic DNA was demethylated, and the expression of DNA demethylase TET2 was increased evidently. Using RNA-sequencing analysis, we found that the expression of Txnip, a gene that encodes a thioredoxin-interacting protein (TXNIP) which mediates NLRP3 activation, was elevated in the DRG after STZ treatment. In addition, knocking down of TET2 expression in DRG using TET2-siRNA suppressed the mRNA expression of Txnip and subsequently inhibited the expression/activation of NLRP3 inflammasome in vitro and in vivo as well as relieved the pain sensitivity of DNP animals. CONCLUSION: The results suggested that the upregulation of the TXNIP/NLRP3 pathway by TET2 in DRG was involved in the pain hypersensitivity of the DNP model.
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Diabetes Mellitus Experimental , Neuropatías Diabéticas , Dioxigenasas , Neuralgia , Ratones , Animales , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Ganglios Espinales/metabolismo , Diabetes Mellitus Experimental/complicaciones , Diabetes Mellitus Experimental/metabolismo , Regulación hacia Arriba , Activación Transcripcional , Dioxigenasas/genética , Dioxigenasas/metabolismo , Epigénesis Genética , Estreptozocina , Neuralgia/metabolismo , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismoRESUMEN
Neuropathic pain is one of the most common conditions requiring treatment worldwide. Salidroside (SAL), a phenylpropanoid glucoside extracted from Rhodiola, has been suggested to produce an analgesic effect in chronic pain. However, whether SAL could alleviate pain hypersensitivity after peripheral nerve injury and its mode of action remains unclear. Several studies suggest that activation of the spinal NOD-like receptor protein 3 (NLRP3) inflammasome and its related proteins contribute to neuropathic pain's pathogenesis. This study investigates the time course of activation of spinal NLRP3 inflammasome axis in the development of neuropathic pain and also whether SAL could be an effective treatment for this type of pain by modulating NLRP3 inflammasome. In the chronic constriction injury (CCI) mice model, spinal NLRP3 inflammasome-related proteins and TXNIP, the mediator of NLRP3, were upregulated from the 14th to the 28th day after injury. The TXNIP and NLRP3 inflammasome-related proteins were mainly present in neurons and microglial cells in the spinal dorsal horn after CCI. Intraperitoneal injection of SAL at 200 mg/kg for 14 consecutive days starting from the 7th day of CCI injury could ameliorate mechanical and thermal hypersensitivity in the CCI model. Moreover, SAL inhibited the activation of the TXNIP/NLRP3 inflammasome axis and mitigated the neuronal loss of spinal dorsal horn induced by nerve injury. These results indicate that SAL could produce analgesic and neuroprotective effects in the CCI model of neuropathic pain.
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Proteína con Dominio Pirina 3 de la Familia NLR , Neuralgia , Animales , Proteínas Portadoras , Proteínas de Ciclo Celular/metabolismo , Constricción , Glucósidos/farmacología , Glucósidos/uso terapéutico , Inflamasomas/metabolismo , Ratones , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteínas NLR/metabolismo , Neuralgia/tratamiento farmacológico , Neuralgia/etiología , Neuralgia/metabolismo , Fenoles , Ratas , Ratas Sprague-Dawley , Tiorredoxinas/metabolismoRESUMEN
BACKGROUND: The role of Helicobacter pylori (H. pylori) infection in the development of non-alcoholic fatty liver disease (NAFLD) remains controversial. The exact relationship requires further investigation. This study aimed to determine the association between them in China. METHODS: A retrospective study was conducted on 71,633 participants who underwent physical examinations. 13C urea breath test (13C-UBT) was conducted to detect H. pylori infection, and ultrasonography was used to detect NAFLD. RESULTS: Body mass index (BMI), blood pressure (BP), and triglyceride (TG) levels were higher in participants with H. pylori infection than in those without H. pylori infection. While the levels of high-density lipoprotein cholesterol (HDL-C) for participants with H. pylori infection was lower than without H. pylori infection (P < 0.001). After adjusting for confounding factors (age, sex, BMI, BP, Scr, BUN, LDL-C, HDL-C, triglycerides, FBG and HbA1c), multivariate logistic regression analysis indicated that there was no independent relationship between them (P = 0.574). Subgroup analysis (stratified by sex, age, BMI, hypertension, diabetes and dyslipidemia) showed that H. pylori infection was not included as an independent risk factor for NAFLD. Moreover, the different grades of NAFLD were not related to H. pylori infection. CONCLUSIONS: These results indicate that H. pylori infection is not an independent risk factor for NAFLD in China.
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Infecciones por Helicobacter , Helicobacter pylori , Enfermedad del Hígado Graso no Alcohólico , China/epidemiología , Infecciones por Helicobacter/complicaciones , Infecciones por Helicobacter/epidemiología , Humanos , Estudios Retrospectivos , Factores de RiesgoRESUMEN
The phenomenon that the anaerobic system is inhibited by acid has always been a bottleneck hindering the application of anaerobic digestion (AD) technology. We tried to introduce electrolysis into AD to improve the acidification resistance, and eventually the productivity of the energy. In a batch fermentation device, the ability of electrochemical anaerobic digestion (EAD) to resist acidification was evaluated in current intensity, electrode potential, AC impedance, microbial community, pH value, and volatile fatty acids (VFAs). The results showed that the average concentration of VFAs in EAD was 32.9% lower than that in AD, the energy efficiency of EAD is 53.25% higher than AD, indicating that EAD has stronger anti-acidification ability and energy conversion efficiency than AD. When the EAD reaches a steady state, the current intensity fluctuates in the range of 7-12 mA, the electrode potential difference is maintained at 600 ± 5 mV, and the internal resistance decreases from 3333.3 ± 16Ω at startup to 68.9 ± 1.4Ω at the steady state, indicating that the EAD has stronger resistance to acidification may be due to the degradation of some VFAs on the electrode surface. Furthermore, the 16S rRNA sequencing analysis showed that the dominant electricity-producing bacteria on EAD anode surface were Clostridium, Hydrogenophaga and Trichloromonas, with a relative abundance of 40.32%, while the relative abundance of electrogenic bacteria in AD bulk solution and EAD bulk solution were about 1/2 and 1/4 that of EAD anode film, suggesting that the electricity-producing bacteria on the electrode surface play an important role in the degradation of VFAs.
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Reactores Biológicos , Ácidos Grasos Volátiles , Anaerobiosis , Reactores Biológicos/microbiología , Electrólisis , Concentración de Iones de Hidrógeno , ARN Ribosómico 16S/genética , ARN Ribosómico 16S/metabolismo , Aguas del AlcantarilladoRESUMEN
17α-ethinylestradiol (17α-EE2) is frequently detected in water bodies due to its use being widespread in the treatment of prostate and breast cancer and in the control of alopecia, posing a threat to humans and aquatic organisms. However, studies on its toxicity to Chlorella pyrenoidosa have been limited to date. This study investigated the effects of 17α-EE2 on the growth, photosynthetic activity, and antioxidant system of C. pyrenoidosa and revealed related molecular changes using transcriptomic analysis. The cell density of algae was inhibited in the presence of 17α-EE2, and cell morphology was also altered. Photosynthetics were damaged, while reactive oxygen species (ROS), superoxide dismutase (SOD), and malondialdehyde (MDA) content increased. Further transcriptomic analysis revealed that the pathways of photosynthesis and DNA replication were affected at three concentrations of 17α-EE2, but several specific pathways exhibited various behaviors at different concentrations. Significant changes in differentially expressed genes and their enrichment pathways showed that the low-concentration group was predominantly impaired in photosynthesis, while the higher-concentration groups were biased towards oxidative and DNA damage. This study provides a better understanding of the cellular and molecular variations of microalgae under 17α-EE2 exposure, contributing to the environmental risk assessment of such hazardous pollutants on aquatic organisms.
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Chlorella , Microalgas , Contaminantes Químicos del Agua , Chlorella/genética , Chlorella/metabolismo , Etinilestradiol/toxicidad , Humanos , Masculino , Microalgas/metabolismo , Transcriptoma , Contaminantes Químicos del Agua/análisisRESUMEN
The dynamic interplay of post-translational modifications (PTMs) in chromatin provides a communication system for the regulation of gene expression. An increasing number of studies have highlighted the role that such crosstalk between PTMs plays in chromatin recognition. In this study, (bio)chemical and structural approaches were applied to specifically probe the impact of acetylation of Lys18 in the histone H3 tail peptide on peptide recognition by the protein methyltransferase coactivator-associated arginine methyltransferase 1 (CARM1). Peptidomimetics that recapitulate the transition state of protein arginine N-methyltransferases, were designed based on the H3 peptide wherein the target Arg17 was flanked by either a free or an acetylated lysine. Structural studies with these peptidomimetics and the catalytic domain of CARM1 provide new insights into the binding of the H3 peptide within the enzyme active site. While the co-crystal structures reveal that lysine acetylation results in minor conformational differences for both CARM1 and the H3 peptide, acetylation of Lys18 does lead to additional interactions (Van der Waals and hydrogen bonding) and likely reduces the cost of desolvation upon binding, resulting in increased affinity. Informed by these findings a series of smaller peptidomimetics were also prepared and found to maintain potent and selective CARM1 inhibition. These findings provide new insights both into the mechanism of crosstalk between arginine methylation and lysine acetylation as well as towards the development of peptidomimetic CARM1 inhibitors.
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Diseño de Fármacos , Inhibidores Enzimáticos/farmacología , Lisina/antagonistas & inhibidores , Peptidomiméticos/farmacología , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , Acetilación , Animales , Cristalografía por Rayos X , Inhibidores Enzimáticos/síntesis química , Inhibidores Enzimáticos/química , Lisina/metabolismo , Ratones , Modelos Moleculares , Peptidomiméticos/síntesis química , Peptidomiméticos/química , Conformación Proteica , Proteína-Arginina N-Metiltransferasas/metabolismo , Especificidad por SustratoRESUMEN
Protein arginine N-methyltransferases (PRMTs) methylate arginine residues in target proteins using the ubiquitous methyl donor S-adenosyl-l-methionine (AdoMet) as a cofactor. PRMTs play important roles in both healthy and disease states and as such inhibition of PRMTs has gained increasing interest. A primary challenge in the development of PRMT inhibitors is achieving specificity for the PRMT of interest as the active sites are highly conserved for all nine members of the PRMT family. Notably, PRMTs show very little redundancy in vivo due to their specific sets of protein substrates. However, relatively little is known about the interactions of PRMTs with their protein substrates that drive this substrate specificity. We here describe the extended application of a methodology recently developed in our group for the production of peptide-based transition state mimicking PRMT inhibitors. Using this approach, an adenosine moiety, mimicking that of the AdoMet cofactor, is covalently linked to the guanidine side chain of a target arginine residue contained in a peptidic fragment derived from a PRMT substrate protein. Using this approach, histone H4 tail peptide-based transition state mimics were synthesized wherein the adenosine group was linked to the Arg3 residue. H4R3 is a substrate for multiple PRMTs, including PRMT1 and PRMT6. The inhibition results obtained with these new H4-based transition state mimics show low micromolar IC50 values against PRMT1 and PRMT6, indicating that the methodology is applicable to the broader family of PRMTs.
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Arginina/química , Inhibidores Enzimáticos/química , Proteína-Arginina N-Metiltransferasas/antagonistas & inhibidores , S-Adenosilmetionina/química , Dominio Catalítico , Histonas/química , Concentración 50 Inhibidora , Metilación , Péptidos/síntesis química , Péptidos/química , Proteína-Arginina N-Metiltransferasas/química , Especificidad por SustratoRESUMEN
Microalgae have been used for the removal of heavy metals or synthetic organics; however, the simultaneous removal of both types of compounds is always technically difficult. In this study, a green algae, Chlamydomonas reinhardtii, was first used to simultaneously remove hexavalent chromium [Cr(VI)] and o-nitrophenol (ONP), and the balance among biomass, oxidative damage and removal rate was also investigated. The results showed that treatment with Cr(VI) or ONP decreased the photosynthetic and superoxide dismutase activities and increased the production of reactive oxygen species (ROS) and malondialdehyde content. However, combined treatment with Cr(VI) (≤4 mg/L) and ONP (≤15 mg/L) significantly decreased ROS generation and alleviated cell damage in C. reinhardtii. In addition, the removal rates of Cr(VI) and ONP by C. reinhardtii cells significantly increased from 37.4% to 54.9% and from 35.8% to 45.9%, respectively, and the cells could be reused at least four times. Moreover, the increased acidity in the medium and Cr(VI) reductase content in C. reinhardtii caused Cr(VI) to be reduced to Cr(III). The addition of an exogenous antioxidant decreased the removal rates of Cr(VI) and ONP. These results indicated that the presence of Cr(VI) could induce ROS generation in C. reinhardtii and enhance ONP degradation, which consumed ROS, alleviated cell damage, and thus benefited Cr(VI) reduction. As a result, C. reinhardtii could be used as a theoretical candidate for the simultaneous removal of combined Cr(VI) and ONP contamination.
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Biodegradación Ambiental , Chlamydomonas reinhardtii/metabolismo , Cromo/metabolismo , Contaminantes Ambientales/metabolismo , Nitrofenoles/metabolismo , Biomasa , Metales Pesados/metabolismoRESUMEN
The increased anthropogenic activities in the Tibetan Plateau may threaten the river environmental safety. However, limited information is available on the Lhasa River in the Tibetan Plateau, which is known as the remaining pure land on Earth. Here, we firstly investigated the distribution patterns of bacterial and archaeal communities in sediments in response to dam construction and sewage discharge along the reaches of the Lhasa River. The total organic carbon, total Nitrogen (N), nitrate and ammonium contents and the relative abundance of bacteria and archaea significantly increased in reservoir sites in comparison with sites below dam, and they also gradually increased from upstream to downstream in sewage discharge sites. By contrast, the diversity of sediment bacteria and archaea in reservoir sites were significantly less than that in sites below dam and sewage discharge sites at Operational Taxonomic Units (OTUs) level. The dominant species were water-bloom cyanobacteria in the reservoir area of Zhikong Dam and Proteobacteria in the sewage discharge sites, which were significantly correlated with the nutrient concentration. The abundance of nitrogen functional genes significantly also increased in reservoir sites and the downstream of sewage discharge areas. These results suggested that dam construction and sewage discharge caused the increase of sediment bacterial communities and nutrient levels and potentially induced eutrophication in the Lhasa River.
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Archaea/aislamiento & purificación , Sedimentos Geológicos/microbiología , Proteobacteria/aislamiento & purificación , Ríos/química , Ríos/microbiología , Aguas del Alcantarillado/microbiología , Compuestos de Amonio/análisis , Archaea/genética , Eutrofización , Nitrógeno/análisis , Proteobacteria/genética , ARN Ribosómico 16S/genética , Tibet , Contaminantes Químicos del Agua/análisisRESUMEN
The radiation-induced damage to mitochondrial oxidative respiratory chain could lead to generating of superoxide anions (O2-) and secondary reactive oxygen species (ROS), which are the major resources of continuous ROS production after radiation. Scavenging radiation-induced ROS effectively can help mitochondria to maintain their physiological function and relief cells from oxidative stress. Dihydropyridines (DHPs) are biomimetic hydrogen sources that could protect cells against radiation damage. In this study, we designed and synthetized three novel mitochondrial-targeted dihydropyridines (Mito-DHPs) that utilize the mitochondrial membrane potential to enter the organelle and scavenge ROS. MitoTracker confirmed Mito-DHPs accumulation in mitochondria, and the DCFH-DA assay demonstrated effective ROS scavenging activity. In addition, the γ-H2AX and comet assay demonstrated the ability of Mito-DHPs to protect against both radiation and ROS-induced DNA strand breaks. Furthermore, Mito-DHP1 proved to be non-toxic and displayed significant radioprotection activity (p < 0.05) in vitro. Mito-DHPs are therefore promising antioxidants that could penetrate the membrane of mitochondria, scavenge excessive ROS, and protect cells against radiation-induced oxidative damage.
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Dihidropiridinas/farmacología , Depuradores de Radicales Libres/farmacología , Mitocondrias/metabolismo , Protectores contra Radiación/farmacología , Especies Reactivas de Oxígeno/metabolismo , Animales , Células CHO , Cricetulus/metabolismo , Dihidropiridinas/química , Femenino , Depuradores de Radicales Libres/química , Células HeLa , Humanos , Especies Reactivas de Oxígeno/químicaRESUMEN
Threonine is a nutritionally essential amino acid (EAA) for the growth and development of humans and other nonruminant animals and must be provided in diets to sustain life. The aim of this study was to synthesize threonine in mammalian cells through transgenic techniques. To achieve this goal, we combined the genes involved in bacterial threonine biosynthesis pathways into a single open reading frame separated by self-cleaving peptides (2A) and then linked it into a transposon system (piggyBac). The plasmids pEF1a-IRES-GFP-E2F-his and pEF1a-IRES-GFP-M2F-his expressed Escherichia coli homoserine kinase and threonine synthase efficiently in mouse cells and enabled cells to synthesize threonine from homoserine. This biosynthetic pathway occurred with a low level of efficiency in transgenic mice. Three transgenic mice were identified by Southern blot from 72 newborn mice, raising the possibility that a high level of expression of these genes in mouse embryos might be lethal. The results indicated that it is feasible to synthesize threonine in animal cells using genetic engineering technology. Further work is required to improve the efficiency of this method for introducing genes into mammals. We propose that the transgenic technology provides a promising means to enhance the synthesis of nutritionally EAAs in farm animals and to eliminate or reduce supplementation of these nutrients in diets for livestock, poultry and fish.
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Aminoácidos Esenciales , Regulación Enzimológica de la Expresión Génica , Aminoácidos Esenciales/biosíntesis , Aminoácidos Esenciales/genética , Animales , Escherichia coli/enzimología , Escherichia coli/genética , Ratones , Ratones Transgénicos , Células 3T3 NIHRESUMEN
BACKGROUND: Viral infection elicits the type I interferon (IFN-I) response in host cells and subsequently inhibits viral infection through inducing hundreds of IFN-stimulated genes (ISGs) that counteract many steps in the virus life cycle. However, most of ISGs have unclear functions and mechanisms in viral infection. Thus, more work is required to elucidate the role and mechanisms of individual ISGs against different types of viruses. RESULTS: Herein, we demonstrate that poliovirus receptor-like protein4 (PVRL4) is an ISG strongly induced by IFN-I stimulation and various viral infections. Overexpression of PVRL4 protein broadly restricts growth of enveloped RNA and DNA viruses, including vesicular stomatitis virus (VSV), herpes simplex virus 1 (HSV-1), influenza A virus (IAV) and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) whereas deletion of PVRL4 in host cells increases viral infections. Mechanistically, it suppresses viral entry by blocking viral-cellular membrane fusion through inhibiting endosomal acidification. The vivo studies demonstrate that Pvrl4-deficient mice were more susceptible to the infection of VSV and IAV. CONCLUSION: Overall, our studies not only identify PVRL4 as an intrinsic broad-spectrum antiviral ISG, but also provide a candidate host-directed target for antiviral therapy against various viruses including SARS-CoV-2 and its variants in the future.
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Herein, the preparation process, morphology, structure, and magnetic properties of La1.85Sr0.15CuO4 (LSCO) cobweb-like nanofibers are reported. LSCO nanofibers with a regular grain size distribution are successfully prepared via electrospinning, followed by calcination. We conducted morphology analysis and elemental distribution using electron microscopy and energy-dispersive X-ray spectroscopy (EDS), respectively. Additionally, magnetic property testing was performed using a vibrating sample magnetometer (VSM) to confirm the superconducting properties of the samples. Interestingly, our samples exhibited a superconducting transition temperature, Tc, of 25.21 K, which showed some disparity compared to similar works. Furthermore, we observed a ferromagnetic response at low temperatures in the superconducting nanofibers. We attribute these phenomena to the effects generated by surface states of nanoscale superconducting materials.
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To analyze the effect of various drying treatments (microwave drying (MD), hot air drying (HAD), vacuum drying (VD), and vacuum freeze drying (VFD)) on taste compounds in Penaeus vannamei, relevant indicators such as free amino acids, 5'-nucleotides, and organic acids were performed. Multidimensional infrared spectroscopy (MM-IR) results found that there were notable variations in taste properties of P. vannamei. There were 18 autocorrelation peaks in 3400-900 cm-1 were screened using second-derivative infrared spectroscopy (SD-IR) and two-dimensional correlation infrared spectroscopy (2DCOS-IR). Variations in functional groups were the major contributors to taste profiles. The TAV of glutamic acid (Glu), guanine (GMP), and inosinemonphosphate (IMP) were greater than one and had notable impacts on taste profiles. VD had the highest equivalent umami value, followed by VFD, HAD, and MD. This study may provide a theoretical guide for the production of dried P. vannamei products on an industrial scale.
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BACKGROUND: The study of changes in the microbiome in chronic pancreatitis (CP) and pancreatic ductal adenocarcinoma (PDAC) holds significant potential for developing noninvasive diagnostic tools as well as innovative interventions to alter the progression of diseases. This systematic review and meta-analysis aimed to analyze in detail the taxonomic and functional characteristics of the gut microbiome in patients with CP and PDAC. METHODS: Two researchers conducted a systematic search across public databases to gather all published research up to June 2023. Diversity and gut microbiota composition are the main outcomes we focus on. RESULTS: This meta-analysis included 14 studies, involving a total of 1511 individuals in the PDAC (n=285), CP (n=342), and control (n=649) groups. Our results show a significant difference in the composition of gut microbiota between PDAC/CP patients compared to healthy controls (HC), as evidenced by a slight decrease in α-diversity, including Shannon (SMD=-0.33; P=0.002 and SMD=-0.59; P<0.001, respectively) and a statistically significant ß-diversity (P<0.05). The pooled results showed that at the phylum level, the proportion of Firmicutes was lower in PDAC and CP patients than in HC patients. At the genus level, more than two studies demonstrated that 4 genera were significantly increased in PDAC patients compared to HC (e.g., Escherichia-Shigella and Veillonella). CP patients had an increase in 4 genera (e.g., Escherichia-Shigella and Klebsiella) and a decrease in 8 genera (e.g., Coprococcus and Bifidobacterium) compared to HC. Functional/metabolomics results from various studies also showed differences between PDAC/CP patients and HC. In addition, this study found no significant differences in gut microbiota between PDAC and CP patients. CONCLUSIONS: Current evidence suggests changes in gut microbiota is associated with PDAC/CP, commonly reflected by a reduction in beneficial species and an increase in the pathogenic species. Further studies are needed to confirm these findings and explore therapeutic possibilities.