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1.
Environ Sci Technol ; 58(14): 6296-6304, 2024 Apr 09.
Artigo em Inglês | MEDLINE | ID: mdl-38556999

RESUMO

Anaerobic digestion (AD) is an important biological resource recovery process, where microorganisms play key roles for material transformation. There has been some knowledge about the prokaryotic community and antibiotic resistance genes (ARGs) in AD, but there has been very limited knowledge of phages. In this study, samples from a full-scale AD plant were collected over 13 months, sequenced, and analyzed for viral and prokaryotic metagenomes. Totally, 3015 viral operational taxonomic units (vOTUs) were detected, mostly assigned to Caudoviricetes. The phage community had faster temporal variation than the prokaryotic community. Warm seasons harbored a higher abundance of both temperate phages and broad host-range phages. Seven ARGs of 6 subtypes were carried by 20 vOTUs, a representative ermT gene was synthesized and expressed, and the resistance activity in the host was examined, confirming the real activity of virus-carried ARGs in the AD process. Some of the ARGs were horizontally transferred between the phage and prokaryotic genomes. However, phage infection was not found to contribute to ARG transfer. This study provided an insight into the ecological patterns of the phage community, confirmed the antibiotic resistance activity of virus-carried ARGs, evaluated the contribution of phages on the ARG prevalence, and laid the foundation for the control strategies of the community and antibiotic resistance in the AD process.


Assuntos
Bacteriófagos , Esgotos , Bacteriófagos/genética , Antibacterianos/farmacologia , Anaerobiose , Prevalência , Resistência Microbiana a Medicamentos/genética , Genes Bacterianos
2.
Nano Lett ; 20(10): 6966-6973, 2020 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-32870015

RESUMO

We explore the ultrafast optical response of graphene subjected to intense (∼106 V/cm) local (∼10 nm) electric fields. Nanoscale gating of graphene is achieved using a voltage-biased, SrTiO3-based conductive nanowire junction "written" directly under the graphene and isolated from it by an insulating ultrathin (<2 nm) LaAlO3 barrier. Upon illumination with ultrafast visible-to-near-infrared (VIS-NIR) light pulses, the local field from the nanojunction creates a strong gate-tunable second-order nonlinearity in the graphene and produces a substantial difference-frequency (DFG) and sum-frequency generation (SFG) response detected by the nanojunction. Spectrally sharp, gate-tunable extinction features (>99.9%) are observed in the VIS-NIR and SFG spectral ranges, in parameter regimes that are positively correlated with the enhanced nonlinear response. The observed graphene-light interaction and nonlinear response are of fundamental interest and open the way for future exploitation in graphene-based optical devices such as phase shifters, modulators, and nanoscale THz sources.

3.
Virol J ; 17(1): 78, 2020 06 17.
Artigo em Inglês | MEDLINE | ID: mdl-32552884

RESUMO

BACKGROUND: The low pathogenic H9N2 AIV caused the serious impact on the poultry industry and public safety. Our purpose was to investigate the molecular evolutionary characteristics of the new isolated H9N2 virus and investigate the intracellular target protein of H9N2 AIV replication in sensitive cells. METHODS: AIV A/chicken/Shandong/LY1/2017 (H9N2) was isolated from the cloaca of the healthy chicken in Shandong, and the full-length eight gene segments of this isolated H9N2 AIV were amplified by RT-PCR and analyzed. MDCK cells were used as the target cell model, and VOPBA assay and LC-MS/MS were carried out to identify the virus-binding protein of H9N2 AIV. MDCK cells were pre-treated with the special antibody and siRNA, and treated with H9N2 AIV to detect the virus replication. Additionally, Vimentin-pcDNA3.0 was successfully constructed, and transinfected into MDCK cells, and then H9N2 AIV mRNA was detected with RT-PCR. RESULTS: Phylogenetic analysis revealed that HA, NA, PB2, PB1, PA, NP and M seven genes of the isolated H9N2 AIV were derived from A/Chicken/Shanghai/F/98, while NS gene was derived from A/Duck/Hong Kong/Y439/97. The cleavage site sequence of HA gene of the isolated H9N2 AIV was a PARSSR G pattern, and the left side sequence (224 ~ 229) of receptor binding site was NGQQGR pattern, which were similar to that of A/Chicken/Shanghai/F/98. Following VOPBA assay, we found one protein of about 50KDa binding to H9N2 AIV, and the results of LC-MS/MS analysis proved that vimentin was the vital protein binding to H9N2 AIV. The pre-incubation of the specific antibody and siRNA decreased the viral RNA level in MDCK cells treated with H9N2 AIV. Furthermore, we found that over-expressed vimentin increased H9N2 AIV replication in MDCK cells. CONCLUSIONS: These findings suggested that the isolated H9N2 AIV might be a recent clinical common H9N2 strain, and vimentin protein might be one vital factor for H9N2 AIV replication in MDCK cells, which might be a novel target for design and development of antiviral drug.


Assuntos
Evolução Molecular , Vírus da Influenza A Subtipo H9N2/genética , Filogenia , Vimentina/farmacologia , Proteínas Virais/genética , Replicação Viral/efeitos dos fármacos , Animais , Galinhas/virologia , China , Cães , Vírus da Influenza A Subtipo H9N2/fisiologia , Células Madin Darby de Rim Canino , Aves Domésticas/virologia , Doenças das Aves Domésticas/virologia
4.
J Asian Nat Prod Res ; 22(3): 217-224, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30740993

RESUMO

A phytochemical investigation on the 75% EtOH extract of the aerial parts of Lycopodiastrum casuarinoides resulted in the isolation of three new lycodine alkaloids, 16-hydroxy-9-oxo-lycocasuarinine D (1), 6α-hydroxy-16-dehydroxy-lycocasuarinine A (2), and 6α,16-dihydroxy-lycocasuarinine B (3). Structural elucidation of all the compounds was performed by spectral methods such as 1D- and 2D-NMR, infrared, ultraviolet, and HR-ESI-MS. The isolated alkaloids were tested in vitro for cytotoxic potential against six lung cancer cell lines. Consequently, alkaloid 1 exhibited cytotoxicity against all the tested tumor cell lines with IC50 values less than 20 µM.[Formula: see text].


Assuntos
Alcaloides , Inibidores da Colinesterase , Linhagem Celular Tumoral , Compostos Heterocíclicos de 4 ou mais Anéis , Estrutura Molecular , Componentes Aéreos da Planta
5.
Vet Res ; 50(1): 64, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533803

RESUMO

The bursa of Fabricius (BF) is the acknowledged central humoural immune organ unique to birds and plays a vital role in B lymphocyte development. In addition, the unique molecular immune features of bursal-derived biological peptides involved in B cell development are rarely reported. In this paper, a novel bursal heptapeptide (BP7) with the sequence GGCDGAA was isolated from the BF and was shown to enhance the monoclonal antibody production of a hybridoma. A mouse immunization experiment showed that mice immunized with an AIV antigen and BP7 produced strong antibody responses and cell-mediated immune responses. Additionally, BP7 stimulated increased mRNA levels of sIgM in immature mouse WEHI-231 B cells. Gene microarray results confirmed that BP7 regulated 2465 differentially expressed genes in BP7-treated WEHI-231 cells and induced 13 signalling pathways and various immune-related functional processes. Furthermore, we found that BP7 stimulated WEHI-231 cell autophagy and AMPK-ULK1 phosphorylation and regulated Bcl-2 protein expression. Finally, chicken immunization showed that BP7 enhanced the potential antibody and cytokine responses to the AIV antigen. These results suggested that BP7 might be an active biological factor that functions as a potential immunopotentiator, which provided some novel insights into the molecular mechanisms of the effects of bursal peptides on immune functions and B cell differentiation.


Assuntos
Proteínas Aviárias/genética , Linfócitos B/imunologia , Bolsa de Fabricius/imunologia , Galinhas/imunologia , Imunidade Celular , Ativação Linfocitária/imunologia , Animais , Formação de Anticorpos , Proteínas Aviárias/imunologia , Feminino , Regulação da Expressão Gênica/imunologia , Imunização , Imunoglobulina M/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Análise Serial de Proteínas , RNA Mensageiro/metabolismo
6.
Zhongguo Zhong Yao Za Zhi ; 44(4): 717-722, 2019 Feb.
Artigo em Zh | MEDLINE | ID: mdl-30989884

RESUMO

Eight C_(19)-diterpenoid alkaloids( 1-8) were isolated from the ethyl acetate soluble fraction of 95% ethanol extract of the ground roots of Aconitum austroyunnanense through various column chromatographies on silica gel,ODS,Sephadex LH-20 and MCI gel.Their structures were elucidated as 14α-benzoyloxy-13ß,15α-dihydroxy-1α,6α,8ß,16ß,18-pentamethoxy-19-oxoaconitan( 1),N-deethylaconitine( 2),spicatine B( 3),leucanthumsine A( 4),acofamine B( 5),macrorhynine B( 6),aconitilearine( 7),and ambiguine( 8) based on their chemical and physicochemical properties and spectroscopic data. Compound 1 was a new compound and alkaloids 2-8 were isolated from this plant for the first time. Some isolated alkaloids were tested in vitro for cytotoxic potential by employing the MTT method. As a result,alkaloid 1 exhibited weak cytotoxic activity against three tested tumor cell lines( A-549,He La,and Hep G2) with IC_(50) values less than 20 µmol·L~(-1).


Assuntos
Aconitum , Alcaloides , Diterpenos , Estrutura Molecular , Raízes de Plantas
7.
BMC Immunol ; 19(1): 29, 2018 10 24.
Artigo em Inglês | MEDLINE | ID: mdl-30355335

RESUMO

BACKGROUND: Excretory-secretory products released by Echinococcus granulosus protoscoleces (EgPSC-ESPs) are well-known to regulate T cell responses. However, their direct influence on the differentiation of B cell subsets remains largely elusive. This study investigated the effects of EgPSC-ESPs on the differentiation of IL-10-producing B cells (B10), and explored the possible role of Toll-like receptor 2 (TLR-2) signaling in this process. RESULTS: In comparison to phosphate buffered saline (PBS), B cells exposed to the excretory-secretory products (ESPs) generated higher percentages of B10 cells, with higher expression of IL-10 mRNA, and larger amount of IL-10 production, which were in a dose dependent way. The mRNA and protein expression of TLR-2 in the ESPs-stimulated B cells were significantly higher than those in PBS, which was consistent to the results in B cells isolated from EgPSC infected mice. Moreover, TLR-2-/- B cells in response to ESPs stimulation expressed lower levels of IL-10 mRNA and produced undetectable IL-10 in comparison to those in normal B cells. In addition, Phosphatase and tensin homolog deleted on chromosome ten/AKT/Phosphatidylinositol-3 kinase (PTEN/AKT/PI3K) pathway was activated in ESPs-treated B cells, which was also dependent on TLR-2 signaling. Pam3CSK4, the agonist of TLR-2, could mock the effects of ESPs on the expression of PTEN, AKT and PI3K. CONCLUSION: Overall, this study revealed that TLR-2 signaling was required for B10 induction mediated by EgPSC-ESPs, which might be an immunomodulatory target against the parasite infection.


Assuntos
Antígenos de Helmintos/imunologia , Subpopulações de Linfócitos B/imunologia , Equinococose/imunologia , Echinococcus granulosus/imunologia , Interleucina-10/metabolismo , Receptor 2 Toll-Like/metabolismo , Animais , Interleucina-10/genética , Camundongos Endogâmicos C57BL , PTEN Fosfo-Hidrolase/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptor 2 Toll-Like/genética
8.
Zhongguo Zhong Yao Za Zhi ; 43(10): 2086-2090, 2018 May.
Artigo em Zh | MEDLINE | ID: mdl-29933675

RESUMO

Three aporphine-type alkaloids (1-3), three lycorine-type alkaloids (4-6), two crinane type alkaloids (7, 8) and one phenanthridine-type alkaloid (9) were isolated from the chloroform soluble fraction of 70% ethanol extract of the bulbs of Lycoris radiata through various column chromatographies over silica gel, ODS, Sephadex LH-20 and MCI. Their structures were elucidated as (+)-N-methoxylcarbonyl-1,2-methylenedioxyl-isocorydione (1), isocorydione (2), 8-demethyl-dehydrocrebanine (3), (+)-3-hydroxy-anhydrolycorine N-oxide (4), vasconine (5), pancratinine D (6), yemenine A (7), 11-O-acetylhaemanthamine (8), and 5,6-dihydro-5-methyl-2-hydroxyphenanthridine (9) based on their chemical and physicochemical properlies and spectroscopic data. Compound 1 was a new compound and alkaloids 2-9 were isolated and identified from this plant for the first time.


Assuntos
Alcaloides de Amaryllidaceae/isolamento & purificação , Lycoris/química , Alcaloides de Amaryllidaceae/química , Compostos Fitoquímicos/química , Compostos Fitoquímicos/isolamento & purificação , Extratos Vegetais/química , Raízes de Plantas/química
10.
Planta ; 239(4): 753-63, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24366682

RESUMO

Δ8-sphingolipid desaturase and Δ6-fatty acid desaturase share high protein sequence identity. Thus, it has been hypothesized that Δ6-fatty acid desaturase is derived from Δ8-sphingolipid desaturase; however, there is no direct proof. The substrate recognition regions of Δ6-fatty acid desaturase and Δ8-sphingolipid desaturase, which aid in understanding the evolution of these two enzymes, have not been reported. A blackcurrant Δ6-fatty acid desaturase and a Δ8-sphingolipid desaturase gene, RnD6C and RnD8A, respectively, share more than 80 % identity in their coding protein sequences. In this study, a set of fusion genes of RnD6C and RnD8A were constructed and expressed in yeast. The Δ6- and Δ8-desaturase activities of the fusion proteins were characterized. Our results indicated that (1) the exchange of the C-terminal 172 amino acid residues can lead to a significant decrease in both desaturase activities; (2) amino acid residues 114-174, 206-257, and 258-276 played important roles in Δ6-substrate recognition, and the last two regions were crucial for Δ8-substrate recognition; and (3) amino acid residues 114-276 of Δ6-fatty acid desaturase contained the substrate recognition site(s) responsible for discrimination between ceramide (a substrate of Δ8-sphingolipid desaturase) and acyl-PC (a substrate of Δ6-fatty acid desaturase). Substituting the amino acid residues 114-276 of RnD8A with those of RnD6C resulted in a gain of Δ6-desaturase activity in the fusion protein but a loss in Δ8-sphingolipid desaturase activity. In conclusion, several regions important for the substrate recognition of Δ8-sphingolipid desaturase and Δ6-fatty acid desaturase were identified, which provide clues in understanding the relationship between the structure and function in desaturases.


Assuntos
Ácidos Graxos Dessaturases/metabolismo , Ácidos Graxos/metabolismo , Oxirredutases/metabolismo , Saccharomyces cerevisiae/enzimologia , Sequência de Aminoácidos , Ácidos Graxos Dessaturases/genética , Modelos Moleculares , Mutagênese , Oxirredutases/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteínas Recombinantes de Fusão , Saccharomyces cerevisiae/genética , Especificidade por Substrato
11.
Bioresour Technol ; 394: 130267, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38154733

RESUMO

The long-term occurrence, dynamics and risk of antibiotic resistance genes (ARGs) in anaerobic digestion (AD) of excess sludge (ES) are not fully understood. Therefore, 13-month metagenomic monitoring was carried out in a full-scale AD plant. The highest ARG abundance and risk scores were observed in spring. AD achieved a 35 % removal rate for the total ARG abundance, but the risk score of AD sludge was not always lower than ES samples, because of the higher proportion of Rank I ARGs in AD sludge. ARGs showed less obvious patterns under linear models compared with microbial community, implying their chaotic dynamics, which was further confirmed by nonlinearity tests. Empirical dynamic modeling performed better than the autoregressive integrated moving average model for ARG dynamics, especially for those with simple and nonlinear dynamics. This study highlighted spring for its higher ARG abundance and risk, and recommended nonlinear models for revealing the dynamics of ARGs.


Assuntos
Antibacterianos , Esgotos , Antibacterianos/farmacologia , Anaerobiose , Genes Bacterianos/genética , Resistência Microbiana a Medicamentos/genética
12.
EBioMedicine ; 106: 105239, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38996766

RESUMO

BACKGROUND: Induction of donor-specific tolerance is a promising approach to achieve long-term graft patency in transplantation with little to no maintenance immunosuppression. Changes to the recipient's T cell receptor (TCR) repertoire are understood to play a pivotal role in the establishment of a robust state of tolerance in chimerism-based transplantation protocols. METHODS: We investigated changes to the TCR repertoires of patients participating in an ongoing prospective, controlled, phase I/IIa trial designed to evaluate the safety and efficacy of combination cell therapy in living donor kidney transplantation. Using high-throughput sequencing, we characterized the repertoires of six kidney recipients who also received bone marrow from the same donor (CKBMT), together with an infusion of polyclonal autologous Treg cells instead of myelosuppression. FINDINGS: Patients undergoing combination cell therapy exhibited partial clonal deletion of donor-reactive CD4+ T cells at one, three, and six months post-transplant, compared to control patients receiving the same immunosuppression regimen but no cell therapy (p = 0.024). The clonality, R20 and turnover rates of the CD4+ and CD8+ TCR repertoires were comparable in both groups, showing our protocol caused no excessive repertoire shift or loss of diversity. Treg clonality was lower in the case group than in control (p = 0.033), suggesting combination cell therapy helps to preserve Treg diversity. INTERPRETATION: Overall, our data indicate that combining Treg cell therapy with CKBMT dampens the alloimmune response to transplanted kidneys in humans in the absence of myelosuppression. FUNDING: This study was funded by the Vienna Science and Technology Fund (WWTF).


Assuntos
Deleção Clonal , Transplante de Rim , Humanos , Transplante de Rim/efeitos adversos , Masculino , Feminino , Pessoa de Meia-Idade , Adulto , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Terapia Baseada em Transplante de Células e Tecidos/métodos , Transplantados , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/metabolismo , Doadores de Tecidos , Linfócitos T/imunologia , Linfócitos T/metabolismo
13.
Vasc Endovascular Surg ; 57(3): 257-263, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36239775

RESUMO

Spontaneous rupture of the iliac vein is a rare cause of retroperitoneal hematoma. A misdiagnosis may delay the treatment and consequently put the patient in a life-threatening emergency. We report the case of a 73-year-old woman who presented with hemorrhagic shock from bleeding caused by a large left retroperitoneal hematoma. She was successfully treated with an endovascular approach by using a double bare stent technique. An extensive review of the literature was conducted and a total 44 articles with 50 patients were identified. Among these patients, 88.2% were women, 94.1% presented with a left-sided rupture, and the mortality rate was 13.7%. Spontaneous iliac vein rupture was more likely to occur in the left side in female patients. Conservative treatment was an option in hemodynamically stable patients. Exploratory laparotomy and surgical iliac vein repair was necessitated in most patients. Endovascular treatment including placement of covered stent and coil embolization had been widely used to treat spontaneous vein rupture since 2003. Double bare stent technique was also an effective alternative if a suitable covered stent was unavailable.


Assuntos
Hematoma , Veia Ilíaca , Humanos , Feminino , Idoso , Masculino , Ruptura Espontânea/complicações , Veia Ilíaca/cirurgia , Resultado do Tratamento , Hematoma/etiologia , Stents/efeitos adversos , Hemorragia Gastrointestinal
14.
Zhonghua Wai Ke Za Zhi ; 50(8): 704-8, 2012 Aug.
Artigo em Zh | MEDLINE | ID: mdl-23157902

RESUMO

OBJECTIVE: Using two antithrombotic treatment (clopidogrel vs. clopidogrel combined warfarin) strategies after femoral-popliteal artery angioplasty prospectively, to evaluate which strategy is more effective for the restenosis prevention. METHODS: Totally 50 patients referred for endovascular treatment (including the percutaneous transluminal angioplasty (PTA) and stent implantation) of the superficial femoral artery and popliteal artery from January 2008 to May 2009 were randomly divided into clopidogrel group (group A, 25 cases, 30 limbs) and clopidogrel plus warfarin group (group B, 25 cases, 33 limbs) before operation. Clinical outcomes and restenosis rate of the target lesions were evaluated at 3, 6 and 12 months after operation. RESULTS: Totally 88 patients were screened for participation in the study, 56 patients were included after the follow-up of 12 months. At 3 months, the rates of restenosis were 16.7% in group A and 18.2% in group B (χ² = 0.025, P = 0.874). At 6 months, the accumulated restenosis rates were 36.7% in group A and 36.4% in group B (χ² = 0.001, P = 0.98). At 12 months, the accumulated restenosis rates were 53.3% in group A and 42.4% in group B (χ² = 0.75, P = 0.387). Analysis for the critical limb ischemia sub-group showed that follow-up of 12 months, the accumulated restenosis rate was 8/10 in group A and 6/12 in group B (χ² = 1.023, P = 0.312). CONCLUSION: The clopidogrel alone treatment for PTA or PTA plus stent implantation of femoral popliteal artery has no statistically significant difference in comparison with the clopidogrel combined warfarin treatment in terms of the cumulative vascular restenosis rate at 3, 6, 12 months postoperatively.


Assuntos
Arteriopatias Oclusivas/prevenção & controle , Artéria Femoral , Artéria Poplítea , Ticlopidina/análogos & derivados , Varfarina/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Angioplastia com Balão , Arteriopatias Oclusivas/etiologia , Clopidogrel , Feminino , Artéria Femoral/cirurgia , Humanos , Masculino , Pessoa de Meia-Idade , Artéria Poplítea/cirurgia , Complicações Pós-Operatórias/prevenção & controle , Estudos Prospectivos , Ticlopidina/uso terapêutico
15.
Nat Commun ; 13(1): 3605, 2022 06 23.
Artigo em Inglês | MEDLINE | ID: mdl-35739125

RESUMO

The cell-type-specific recording and manipulation is instrumental to disentangle causal neural mechanisms in physiology and behavior and increasingly requires intersectional control; however, current approaches are largely limited by the number of intersectional features, incompatibility of common effectors and insufficient gene expression. Here, we utilized the protein-splicing technique mediated by intervening sequences (intein) and devised an intein-based intersectional synthesis of transactivator (IBIST) to selectively control gene expression of common effectors in multiple-feature defined cell types in mice. We validated the specificity and sufficiency of IBIST to control fluorophores, optogenetic opsins and Ca2+ indicators in various intersectional conditions. The IBIST-based Ca2+ imaging showed that the IBIST can intersect five features and that hippocampal neurons tune differently to distinct emotional stimuli depending on the pattern of projection targets. Collectively, the IBIST multiplexes the capability to intersect cell-type features and controls common effectors to effectively regulate gene expression, monitor and manipulate neural activities.


Assuntos
Inteínas , Optogenética , Animais , Inteínas/genética , Camundongos , Neurônios , Processamento de Proteína , Transativadores/genética
16.
J Exp Bot ; 61(6): 1827-38, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20231328

RESUMO

Gamma-linolenic acid (gamma-linolenic acid, GLA; C18:3 Delta(6, 9, 12)) belongs to the omega-6 family and exists primarily in several plant oils, such as evening primrose oil, blackcurrant oil, and borage oil. Delta(6)-desaturase is a key enzyme involved in the synthesis of GLA. There have been no previous reports on the genes encoding Delta(6)-desaturase in blackcurrant (Ribes nigrum L.). In this research, five nearly identical copies of Delta(6)-desaturase gene-like sequences, named RnD8A, RnD8B, RnD6C, RnD6D, and RnD6E, were isolated from blackcurrant. Heterologous expression in Saccharomyces cerevisiae and/or Arabidopsis thaliana confirmed that RnD6C/D/E were Delta(6)-desaturases that could use both alpha-linolenic acids (ALA; C18:3 Delta(9,12,15)) and linoleic acid (LA; C18:2 Delta(9,12)) precursors in vivo, whereas RnD8A/B were Delta(8)-sphingolipid desaturases. Expression of GFP tagged with RnD6C/D/E showed that blackcurrant Delta(6)-desaturases were located in the mitochondrion (MIT) in yeast and the endoplasmic reticulum (ER) in tobacco. GC-MS results showed that blackcurrant accumulated GLA and octadecatetraenoic acids (OTA; C18:4 Delta(6,9,12,15)) mainly in seeds and a little in other organs and tissues. RT-PCR results showed that RnD6C and RnD6E were expressed in all the tissues at a low level, whereas RnD6D was expressed at a high level only in seeds, leading to the accumulation of GLA and OTA in seeds. This research provides new insights to our understanding of GLA synthesis and accumulation in plants and the evolutionary relationship of this class of desaturases, and new clues as to the amino acid determinants which define precise enzyme activity.


Assuntos
Ácidos Graxos Dessaturases/metabolismo , Plantas Geneticamente Modificadas/enzimologia , Ribes/enzimologia , Sequência de Aminoácidos , Ácidos Graxos Dessaturases/química , Ácidos Graxos Dessaturases/genética , Cromatografia Gasosa-Espectrometria de Massas , Dados de Sequência Molecular , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Ribes/genética , Ribes/metabolismo , Homologia de Sequência de Aminoácidos , Nicotiana/enzimologia , Nicotiana/genética , Nicotiana/metabolismo
17.
Immunol Res ; 68(1): 28-38, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32248343

RESUMO

This study aimed to investigate whether the classic hepatoprotective drug polyene phosphatidylcholine (PPC) regulates macrophage polarization and explores the potential role of TLR-2 in this process. In RAW264.7 macrophages and murine bone marrow-derived macrophages (BMDMs) stimulated by lipopolysaccharide (LPS), PPC significantly inhibited the production of IL-6, TNF-α, and the mRNA expression of M1-type macrophage markers. Consistently, PPC reduced the mRNA expression of several key enzymes in the pathways of glycolysis and lipid synthesis while increasing the expression of key enzymes associated with lipid oxidation. Moreover, blocking the glycolytic pathway using 2-deoxy-D-glucose (2-DG) significantly enhanced the anti-inflammatory effect of PPC. However, inhibition of lipid oxidation using GW9662 (an inhibitor of PPAR-γ) and GW6471 (an inhibitor of PPAR-α) abolished the anti-inflammatory effect of PPC. Interestingly, TLR-2 expression in macrophages was significantly downregulated after exposure to PPC. Moreover, pre-activation of TLR-2 hampered the anti-inflammatory effect of PPC. In addition, PPC did not inhibit the secretion of IL-6 and TNF-α in TLR-2-/- BMDMs that were activated by LPS. This was consistent with the increased expression of M1 markers and glycolytic and lipid synthesis enzymes but decreased lipid oxidation-related enzymes. These results showed that PPC inhibits the differentiation of M1-type macrophages, which was most likely related to TLR-2-mediated metabolic reprogramming.


Assuntos
Anti-Inflamatórios/farmacologia , Macrófagos/fisiologia , Fosfatidilcolinas/farmacologia , Receptor 2 Toll-Like/metabolismo , Animais , Diferenciação Celular , Reprogramação Celular , Feminino , Interleucina-6/metabolismo , Peroxidação de Lipídeos , Lipopolissacarídeos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células RAW 264.7 , Transdução de Sinais , Células Th1/imunologia , Receptor 2 Toll-Like/genética , Fator de Necrose Tumoral alfa/metabolismo
18.
Ying Yong Sheng Tai Xue Bao ; 31(4): 1349-1356, 2020 Apr.
Artigo em Zh | MEDLINE | ID: mdl-32530211

RESUMO

Green land is important carrier in cities. We investigated the effects of soil permeability improvement and purification of pollutants under different matrix composition amendments in Xixian New Area, Shaanxi Province. Six materials, including biochar, vermiculite, perlite, compost, polyacrylamide (PAM) and coir, were mixed with basic materials that were composed of soil and sand (volume ratio of 4:4), and then incubated for 30 days. We analyzed bulk density, saturated hydraulic conductivity, saturated moisture, and total nitrogen (TN), total phosphorus (TP), chemical oxygen demand (COD), copper (Cu), zinc (Zn), cadmium (Cd) concentrations in pollutants. For biochar and compost with volume ratio of 0.5:1.5 (1) and 1:1 (2), 1%PAM application had the best effect on soil permeability (including bulk density, saturated hydraulic conductivity, and saturated moisture). The effect purification of TN and COD was better under the condition of biochar and compost (1) with 0.5% PAM amendment than that with 1% PAM amendment. Under biochar and compost (1) with 1%PAM treatment, the adsorption rate of Zn, Cd, Cu was 99.9%, 99.7%, and 97.2%, respectively. In general, biochar and compost (volume ratio of 0.5:1.5) with 1% PAM could be recommended as the media of the sponge city green land in Xixian New Area.


Assuntos
Compostagem , Poluentes Ambientais , Poluentes do Solo , Carvão Vegetal , Cidades , Permeabilidade , Solo
19.
Protein Pept Lett ; 26(5): 348-356, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30816077

RESUMO

BACKGROUND: The Bursa of Fabricius is an acknowledged central humoral immune organ unique to birds, which provides an ideal research model on the immature B cell development. OBJECTIVE: In this article, our motivation is to study the role on sIgM and establish the molecular basis and functional processes of Bursal Hexapeptide (BHP) in avian immature B cells DT40 cell lines. METHODS: In this article, we detected the expressions of sIgM mRNA with qPCR in DT40 cells with BHP treatment, and investigated the gene expression profiles of BHP-treated DT40 cells, employing microarray analyses. Also, to validate the differentially expressed genes, we performed KEGG pathway and Gene Ontology analysis in the BHP-treated DT40 cells. Finally, we comparatively analyzed the similar regulated genes and their involved immune functional processes between DT40 cell and mouse immature B cell line WEHI231 cell with BHP treatment. RESULTS: Following the proposed framework, we proved that the BHP enhanced the mRNA expression levels of IgM in DT40 cells, and induced 460 upregulated genes and 460 downregulated genes in BHP-treated DT40 cells. The pathway analysis showed that the differentially regulated genes in DT40 cell line with BHP treatment were involved in 12 enrichment pathways, in which Toll-like receptor signaling pathway was the vital pathways, and cytokine-cytokine receptor interaction and Jak-STAT signaling pathway were another two important pathways in BHP-treated DT40 cells. Moreover, BHP induced the immune related biological processes in BHP-treated DT40 cells, including T cell related, cytokine related, lymphocyte related, and innate immune response GO terms. Finally, the comparatively analysis showed that there were two downregulated genes GATA3 and IFNG to be found co-existed among the differentially expressed genes in BHP-treated DT40 cell and WEHI231 cells, which shared some same immune related functional processes in both cell lines. CONCLUSION: After the applying the framework, we proved the inducing roles and the gene expression profiles of BHP on avian immature B cells, and verified some molecular basis from the KEGG and GO analysis. These results provided the insight for mechanism on immature B cell differentiation, and offer the essential direction for the vaccine improvement.


Assuntos
Oligopeptídeos/metabolismo , Células Precursoras de Linfócitos B/metabolismo , Animais , Bolsa de Fabricius , Linhagem Celular , Galinhas , Imunidade Inata , Imunoglobulina M/genética , Imunoglobulina M/metabolismo , Camundongos , Oligopeptídeos/farmacologia , Células Precursoras de Linfócitos B/efeitos dos fármacos , Células Precursoras de Linfócitos B/imunologia , RNA Mensageiro/metabolismo , Transdução de Sinais , Especificidade da Espécie , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T/metabolismo
20.
Huan Jing Ke Xue ; 40(11): 5164-5172, 2019 Nov 08.
Artigo em Zh | MEDLINE | ID: mdl-31854586

RESUMO

To evaluate the emission characteristics of carbon dioxide (CO2) and nitrous oxide (N2O) in the Loess Plateau, a field in situ study was conducted from July 2017 to July 2018 under two land-use types (15 year old apple orchard and wheat field) using static chamber-gas chromatographic techniques. Four treatments were conducted in this experiment as follows:apple orchard with fertilization (AF), apple orchard without fertilization (ACK), wheat field with fertilization (WF), and wheat field without fertilization (WCK). The results showed that CO2 and N2O emissions varied significantly with the season, and the emission peaks appeared after rainfall and fertilization. The cumulative amount of CO2 and N2O emissions from the AF treatment were 7.14% and 461.4% higher than that of the WF treatment, respectively. However, the cumulative amount of CO2 emissions under the ACK treatment was 10.41% lower than that of the WCK treatment, whereas the cumulative amount of N2O emissions was 109.5% higher than that of the WCK treatment. The N2O emission flux from the orchard was significantly positively correlated with soil temperature and moisture (P<0.01). The CO2 emission fluxes from the orchard and wheat field were significantly positively correlated with topsoil temperature (P<0.05) but were not correlated with topsoil moisture. Thus, the combination of field management and environmental factors affected soil CO2 and N2O emissions. The fertilizer regime and soil hydrothermal conditions were the main factors influencing the characteristics of CO2 and N2O emissions under different land-use types.

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