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1.
BMC Microbiol ; 23(1): 331, 2023 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-37936065

RESUMO

The biological mechanisms underlying meat quality remain unclear. Currently, many studies report that the gastrointestinal microbiota is essential for animal growth and performance. However, it is uncertain which bacterial species are specifically associated with the meat quality traits. In this study, 16S rDNA and metagenomic sequencing were performed to explore the composition and function of microbes in various gastrointestinal segments of Tan sheep and Dorper sheep, as well as the relationship between microbiota and meat quality (specifically, the fatty acid content of the muscle). In the ruminal, duodenal, and colonic microbiome, several bacteria were uniquely identified in respective breeds, including Agrobacterium tumefaciens, Bacteroidales bacterium CF, and several members of the family Oscillospiraceae. The annotation of GO, KEGG, and CAZYme revealed that these different bacterial species were linked to the metabolism of glucose, lipids, and amino acids. Additionally, our findings suggested that 16 microbial species may be essential to the content of fatty acids in the muscle, especially C12:0 (lauric acid). 4 bacterial species, including Achromobacter xylosoxidans, Mageeibacillus indolicus, and Mycobacterium dioxanotrophicus, were positively correlated with C12:0, while 13 bacteria, including Methanobrevibacter millerae, Bacteroidales bacterium CF, and Bacteroides coprosuis were negatively correlated with C12:0. In a word, this study provides a basic data for better understanding the interaction between ruminant gastrointestinal microorganisms and the meat quality traits of hosts.


Assuntos
Microbioma Gastrointestinal , Microbiota , Ovinos , Animais , Microbioma Gastrointestinal/genética , Bactérias , Músculos , Ácidos Graxos/metabolismo , Bacteroidetes , Ácidos Láuricos/metabolismo
2.
Insect Mol Biol ; 32(3): 229-239, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36533988

RESUMO

Farnesyl/geranylgeranyl diphosphate synthases (FPPS/GGPPS) as the short-chain prenyltransferases catalyse the formation of the acyclic precursors (E)-FPP and (E)-GGPP for isoprenoid biosynthesis. Here, we first cloned the cDNAs encoding FPPS and GGPPS in the vetch aphid Megoura viciae (designated as MvFPPS and MvGGPPS). They had an open reading frame of 1185 and 930 bp in length, encoding 395 and 309 amino acids, with a theoretical isoelectric point of 6.52 and 6.21, respectively. Sequence alignment and phylogenetic analysis showed that MvFPPS and MvGGPPS shared the conserved aspartate-rich motifs characterized by all prenyltransferases identified to date and were clustered with their homologues in two large clades. RNA interference (RNAi) combined with gas chromatography/mass spectrometry (GC-MS) analysis showed that both MvFPPS and MvGGPPS were involved in the biosynthesis of alarm pheromone. Spatiotemporal expression profiling showed that the expression of MvFPPS and MvGGPPS was significantly higher in embryos than in other tissues. RNAi and GC-MS performed specifically in embryos corroborated the function of MvFPPS and MvGGPPS. In vitro, enzymatic activity assay and product analysis demonstrated that MvFPPS could catalysed the formation of (E)-FPP using DMAPP or (E)-GPP as the allylic cosubstrates in the presence of IPP, while MvGGPPS could only use (E)-GPP or (E)-FPP as cosubstrates. Functional interaction analysis using RNAi revealed that MvGGPPS exerts unidirectional functional compensation for MvFPPS. Moreover, it can regulate the biosynthesis of alarm pheromone by imposing a negative feedback regulation on MvFPPS. Our study helps to understand the molecular regulatory mechanism of terpenoid biosynthesis in the aphid.


Assuntos
Afídeos , Geraniltranstransferase , Animais , Geraniltranstransferase/genética , Geraniltranstransferase/química , Geraniltranstransferase/metabolismo , Afídeos/metabolismo , Feromônios , Filogenia
3.
J Immunol ; 207(8): 2179-2191, 2021 10 15.
Artigo em Inglês | MEDLINE | ID: mdl-34497150

RESUMO

Gut microbes play an important role in the development of host B cells. It has been controversial whether GALT is the development site of B cells in pigs. By investigating the relationship between gut microbes and the development of B cells in the GALT of piglets, we found, to our knowledge for the first time, that early B cells exist in the gut lamina propria (LP) in pigs at different ages. We further used Lactobacillus rhamnosus GG (LGG) to treat piglets. The results showed that LGG promotes the development of the early B lineage, affects the composition of the Ig CDR3 repertoires of B cells, and promotes the production of IgA in the intestinal LP. Additionally, we found that the p40 protein derived from LGG can activate the EGFR/AKT and NF-κB signaling pathways, inducing porcine intestinal epithelial cells (IPEC-J2) to secrete a proliferation-inducing ligand (APRIL), which promotes IgA production in B cells. Finally, we identified ARF4 and DIF3 as candidates for p40 receptors on IPEC-J2 by GST pull-down, liquid chromatography-mass spectrometry/mass spectrometry analysis, and coimmunoprecipitation. In conclusion, LGG could promote early B cell differentiation and development in the intestinal LP in piglets and might contribute to promoting IgA production via secretion of p40, which interacts with the membrane receptors on IPEC-J2 and induces them to secrete APRIL. Our study will provide insight to aid in better utilization of probiotics to increase human health.


Assuntos
Linfócitos B/imunologia , Proteínas de Bactérias/metabolismo , Microbioma Gastrointestinal/imunologia , Imunoglobulina A/metabolismo , Mucosa Intestinal/patologia , Lacticaseibacillus rhamnosus/imunologia , Mucosa/imunologia , Animais , Formação de Anticorpos , Diferenciação Celular , Linhagem Celular , Linhagem da Célula , Proteínas de Fluorescência Verde/metabolismo , NF-kappa B/metabolismo , Proteína Oncogênica v-akt/metabolismo , Transdução de Sinais , Suínos , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo
4.
J Synchrotron Radiat ; 29(Pt 5): 1241-1250, 2022 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-36073883

RESUMO

In synchrotron radiation X-ray imaging, the imaging field of view and spatial resolution are mutually restricted, which makes it impossible to have both a large field of view and high resolution when carrying out experiments. Constructing an oversampled image through the micro-scanning method and using the deconvolution algorithm to eliminate the point spread function introduced by pixel overlap can increase the resolution under a fixed imaging field of view, thereby improving the ratio of the field of view to the spatial resolution. In this paper, numerical simulation and synchrotron radiation experiments are carried out with a different number of micro-scanning steps. In numerical simulation experiments only affected by the image pixel size, as the number of micro-scanning steps increases, the ability of the oversampled image with deconvolution to improve the resolution is stronger. The achievable resolution of the oversampled image with deconvolution is basically the same as that of the sample image. In the synchrotron radiation experiments, the resolution of the oversampled image with deconvolution in the 2 × 2 mode is significantly improved. However, as the number of micro-scanning steps increases, the resolution improvement is limited, or even no longer improved. Finally, by analyzing the results of numerical simulation and synchrotron radiation experiments, three factors (four other factors affecting the resolution besides the camera resolution, translational accuracy of micro-scanning, and the signal-to-noise ratio of projections) affecting the micro-scanning method are proposed and verified by experiments.

5.
BMC Urol ; 22(1): 200, 2022 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-36496361

RESUMO

OBJECTIVE: The aim of the present study was to explore the effect of cytoplasmic transduction peptide (CTP)-phosphatase and tensin homolog (PTEN) on the proliferation, cell cycle, apoptosis, migration and invasion of bladder cancer cells and the underlying molecular mechanism. METHODS: A eukaryotic expression vector, pTT5-CTP-PTEN, was constructed. The constructed vector was transfected into HEK 293-6E cells to express a fusion protein, CTP-PTEN. The fusion protein was purified. 5637 bladder cancer cells were cocultured with purified CTP-PTEN fusion protein. Target gene expression, protein expression, cell proliferation, cell cycle, apoptosis, cell invasion and cell migration were examined by reverse transcription polymerase chain reaction (RT-PCR), western blot, MTT assay, flow cytometry, Transwell assay, and cell scratch assay, respectively. RESULTS: Both PTEN and CTP-PTEN fusion protein inhibited the proliferation, cell cycle, invasion and migration of bladder cancer cells and promoted the apoptosis of bladder cancer cells. The effect of CTP-PTEN was more significant. CONCLUSIONS: The fused expression of CTP and PTEN significantly increased the penetrability of the tumor suppressor gene PTEN into cancer cells. The CTP-PTEN fusion protein exhibited a significant carcinostatic effect on 5637 bladder cancer cells.


Assuntos
MicroRNAs , Neoplasias da Bexiga Urinária , Humanos , Células HEK293 , PTEN Fosfo-Hidrolase/genética , PTEN Fosfo-Hidrolase/metabolismo , Proliferação de Células , Movimento Celular , Apoptose , Neoplasias da Bexiga Urinária/genética , Linhagem Celular Tumoral , Peptídeos/genética , Peptídeos/metabolismo , MicroRNAs/genética , Regulação Neoplásica da Expressão Gênica
6.
J Acoust Soc Am ; 149(5): 2997, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-34241089

RESUMO

Detecting small defects in curved parts through classical monostatic pulse-echo ultrasonic imaging is known to be a challenge. Hence, a robot-assisted ultrasonic testing system with the track-scan imaging method is studied to improve the detecting coverage and contrast of ultrasonic images. To further improve the image resolution, we propose a visual geometry group-UNet (VGG-UNet) deep learning network to optimize the ultrasonic images reconstructed by the track-scan imaging method. The VGG-UNet uses VGG to extract advanced information from ultrasonic images and takes advantage of UNet for small dataset segmentation. A comparison of the reconstructed images on the simulation dataset with ground truth reveals that the peak signal-to-noise ratio (PSNR) and structural similarity index measure (SSIM) can reach 39 dB and 0.99, respectively. Meanwhile, the trained network is also robust against the noise and environmental factors according to experimental results. The experiments indicate that the PSNR and SSIM can reach 32 dB and 0.99, respectively. The resolution of ultrasonic images reconstructed by track-scan imaging method is increased approximately 10 times. All the results verify that the proposed method can improve the resolution of reconstructed ultrasonic images with high computation efficiency.

7.
Int J Hosp Manag ; 90: 102616, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32834353

RESUMO

The 7 Ps model is a very useful tool in helping service firms solve managerial issues in marketing. Guided by the 7 Ps marketing mix framework, a big-data, supervised machine learning analysis was performed with 1,148,062 English reviews of 37,092 Airbnb listings in San Francisco and New York City. The results disclose similar patterns in both markets, where travelers shared their experience about Service Product and Physical Evidence most often; Price and Promotion were the least mentioned elements. Furthermore, through a series of comparisons of Airbnb's 7 Ps marketing mix among the listings managed by different types of hosts, multi-unit and single-unit hosts seem to offer similar services with a small observable difference; whereas superhosts and the ordinary hosts deliver different services. This study makes valuable methodological contributions and provides practical marketing insights for hoteliers and the hosts and webmasters on home-sharing websites. Policymakers should pay special attention to multi-unit hosts.

8.
Opt Lett ; 44(6): 1411-1414, 2019 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-30874663

RESUMO

We propose and experimentally demonstrate a microwave photonic (MWP) notch filter based on a silica microsphere cavity. By using a high-Q-factor (∼1e7) cavity with a diameter of 132 um, the filter bandwidth can be easily decreased to 15 MHz in terms of simple fabrication and flexible coupling. Then we use the advanced modulation technique based on a dual parallel Mach-Zehnder modulator to further improve peak rejection (PR). The experimental results show that the MWP notch filter with its PR beyond 55 dB and frequency tunability range over 8 GHz has been achieved in combination with double-sideband modulation. To the best of our knowledge, this is a record for PR and bandwidth considered simultaneously for an MWP filter based on silica microcavities. Thus, the proposed MWP filter will be useful in the fields of microwave photonic signal processing, radar systems, etc.

9.
Mol Cell Probes ; 47: 101437, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31425739

RESUMO

Emetic Bacillus cereus is one of the causative agents of foodborne diseases which can cause vomiting-type food poisoning after ingestion of contaminated food. To minimize B. cereus food poisoning, propidium monoazide (PMA) combined with quantitative polymerase chain reaction (qPCR) called PMA-qPCR was applied for detecting viable emetic B. cereus in milk. The cereulide synthetase gene of emetic B. cereus (cesB) was chosen for the primer, and PMA treatment was optimized at 3 µg/mL to inhibit the PCR amplification of DNA from dead cells. Under optimized assay parameters, the limit of detection (LOD) using this method were 102 CFU/mL in both pure culture and in spiked milk matrix. The cycle threshold (Ct) values obtained for this assay was not significantly affected by the presence of non-target bacteria such as E. coli O157:H7 which indicated the high selectivity of the assay for emetic B. cereus. The PMA-qPCR assay used in this study has the potential for sensitive detection of viable emetic B. cereus in milk.


Assuntos
Bacillus cereus/isolamento & purificação , Proteínas de Bactérias/genética , Leite/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Azidas/química , Bacillus cereus/enzimologia , Bacillus cereus/genética , Microbiologia de Alimentos , Limite de Detecção , Propídio/análogos & derivados , Propídio/química , Especificidade da Espécie
10.
J Dairy Sci ; 102(6): 4945-4953, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30904294

RESUMO

A fluorescence assay combined with PCR, catalytic hairpin assembly (CHA), and graphene oxide (GO) was established to detect emetic Bacillus cereus in milk samples. The processes of the assay are not new, but components of the processes make the assay useful. Two partially complementary hairpin probes (H1 and FAM-H2) were designed according to the target single-strand DNA (ssDNA). The CHA reaction could be initiated only by the target ssDNA, which was generated by the denaturation of PCR amplicons. In the absence of the target ssDNA, CHA reaction could not be triggered, which caused the H1 and FAM-H2 adsorbing on the surface of GO and exhibiting a low fluorescence intensity. Addition of the target ssDNA resulted in opening of the hairpin H1 that subsequently hybridized with H2. Then, target ssDNA would be replaced from the H1 and recycled to promote another CHA reaction. Through the CHA reaction, multiple H1-H2 duplexes were generated that could not adsorb on the surface of GO. Thus, a strong fluorescence signal would be obtained. The assay showed a limit of detection for emetic B. cereus of 6.2 × 101 cfu/mL in pure culture and 5.9 × 102 cfu/mL in spiked milk without enrichment. By changing the PCR primer, the assay developed in this study had potential to detect other bacteria.


Assuntos
Bacillus cereus/isolamento & purificação , Grafite , Leite/microbiologia , Animais , Bacillus cereus/genética , Bacillus cereus/patogenicidade , Técnicas Biossensoriais , Catálise , DNA Bacteriano/química , DNA Bacteriano/isolamento & purificação , Fluorescência , Sequências Repetidas Invertidas , Reação em Cadeia da Polimerase
11.
Biochem Biophys Res Commun ; 495(2): 1864-1870, 2018 01 08.
Artigo em Inglês | MEDLINE | ID: mdl-29225169

RESUMO

Orai1-dependent Ca2+ entry plays an essential role in inflammatory response through regulating T cell and macrophage activation and neutrophil infiltration. However, whether Orai1 Ca2+ entry contributes to endothelial activation, one of the early steps of vascular inflammation, remains elusive. In the present study, we observed that knockdown of Orai1 reduced, whereas overexpression of Orai1 potentiated, TNFα-induced expression of adhesion molecules such as ICAM-1 and VCAM-1 in HUVECs, and subsequently blocked adhesion of monocyte to HUVECs. In vivo, Orai1 downregulation attenuated TNFα-induced ICAM-1 and VCAM-1 expression in mouse aorta and the levels of pro-inflammatory cytokines in the serum. In addition, Orai1 knockdown also dramatically decreased the expression of pro-inflammatory cytokines and neutrophil infiltration in the lung after TNFα treatment, and thus protected lung tissue injury. Notably, among all isoforms of nuclear factor of activated T cells (NFATs), TNFα only triggered NFATc4 nuclear accumulation in HUVECs. Knockdown of Orai1 or inhibition of calcineurin prevented TNFα-induced NFATc4 nuclear translocation and reduced ICAM-1 and VCAM-1 expression in HUVECs. Overexpression of NFATc4 further enhanced ICAM-1 and VCAM-1 expression induced by TNFα. Our study demonstrates that Orai1-Ca2+-calcineurin-NFATc4 signaling is an essential inflammatory pathway required for TNFα-induced endothelial cell activation and vascular inflammation. Therefore, Orai1 may be a potential therapeutic target for treatment of inflammatory diseases.


Assuntos
Aortite/imunologia , Calcineurina/imunologia , Cálcio/imunologia , Moléculas de Adesão Celular/imunologia , Endotélio Vascular/imunologia , Fatores de Transcrição NFATC/imunologia , Proteína ORAI1/imunologia , Animais , Aortite/patologia , Células Cultivadas , Regulação para Baixo/imunologia , Humanos , Mediadores da Inflamação/imunologia , Redes e Vias Metabólicas/imunologia , Camundongos , Camundongos Endogâmicos C57BL
12.
Appl Microbiol Biotechnol ; 102(19): 8403-8417, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30022263

RESUMO

Transmissible gastroenteritis coronavirus (TGEV) is one of the most severe threats to the swine industry. In this study, we constructed a suite of recombinant Lactobacillus plantarum with surface displaying the spike (S) protein coming from TGEV and fused with DC cells targeting peptides (DCpep) to develop an effective, safe, and convenient vaccine against transmissible gastroenteritis. Our research results found that the recombinant Lactobacillus plantarum (NC8-pSIP409-pgsA-S-DCpep) group expressing S fused with DCpep could not only significantly increase the percentages of MHC-II+CD80+ B cells and CD3+CD4+ T cells but also the number of IgA+ B cells and CD3+CD4+ T cells of ileum lamina propria, which elevated the specific secretory immunoglobulin A (SIgA) titers in feces and IgG titers in serum. Taken together, these results suggest that NC8-pSIP409-pgsA-S-DCpep expressing the S of TGEV fused with DCpep could effectively induce immune responses and provide a feasible original strategy and approach for the design of TGEV vaccines.


Assuntos
Proteínas de Bactérias/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , Lactobacillus plantarum/imunologia , Vírus da Gastroenterite Transmissível/imunologia , Animais , Anticorpos Antivirais/imunologia , Linfócitos B/imunologia , Gastroenterite Suína Transmissível/imunologia , Imunoglobulina A Secretora/imunologia , Imunoglobulina G/imunologia , Suínos , Linfócitos T/imunologia , Vacinas Virais/imunologia
13.
Appl Microbiol Biotechnol ; 102(12): 5077-5088, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29675804

RESUMO

Avian influenza virus (AIV) can infect poultry, mammals, and other hosts and causes enormous economic losses to the global poultry industry. In this study, to develop a novel and potent oral vaccine based on Lactobacillus plantarum (L. plantarum) for controlling the spread of AIV in the poultry industry, we constructed a recombinant L. plantarum strain displaying the 3M2e-HA2 protein of the influenza virus and determined the effect of N/pgsA'-3M2e-HA2 against AIV in chicks. We first confirmed that the 3M2e-HA2 fusion protein was expressed on the surface of L. plantarum via flow cytometry and immunofluorescence experiments. Our experimental results demonstrated that chicks immunized with N/pgsA'-3M2e-HA2 could induce specific humoral, mucosal, and T cell-mediated immune responses, eliciting the host body to protect itself against AIV. Additionally, compared to oral administration, the intranasal immunization of chicks with N/pgsA'-3M2e-HA2 provided a stronger immune response, resulting in a potent protective effect that hindered the loss of body weight, decreasing pulmonary virus titers and reducing lung and throat pathological damages. Thus, our results indicate that our novel approach is an effective method of vaccine design to promote mucosal immunity.


Assuntos
Antígenos Virais/imunologia , Vacinas contra Influenza/imunologia , Influenza Aviária/prevenção & controle , Lactobacillus plantarum/imunologia , Proteínas Recombinantes/imunologia , Imunidade Adaptativa/imunologia , Animais , Galinhas , Vírus da Influenza A/imunologia , Lactobacillus plantarum/genética , Proteínas Recombinantes/genética
14.
J Cell Mol Med ; 21(5): 904-915, 2017 05.
Artigo em Inglês | MEDLINE | ID: mdl-27878958

RESUMO

Increasing evidence supports that activation of store-operated Ca2+ entry (SOCE) is implicated in the chemoresistance of cancer cells subjected to chemotherapy. However, the molecular mechanisms underlying chemoresistance are not well understood. In this study, we aim to investigate whether 5-FU induces hepatocarcinoma cell death through regulating Ca2+ -dependent autophagy. [Ca2+ ]i was measured using fura2/AM dye. Protein expression was determined by Western blotting and immunohistochemistry. We found that 5-fluorouracil (5-FU) induced autophagic cell death in HepG2 hepatocarcinoma cells by inhibiting PI3K/AKT/mTOR pathway. Orai1 expression was obviously elevated in hepatocarcinoma tissues. 5-FU treatment decreased SOCE and Orai1 expressions, but had no effects on Stim1 and TRPC1 expressions. Knockdown of Orai1 or pharmacological inhibition of SOCE enhanced 5-FU-induced inhibition of PI3K/AKT/mTOR pathway and potentiated 5-FU-activated autophagic cell death. On the contrary, ectopic overexpression of Orai1 antagonizes 5-FU-induced autophagy and cell death. Our findings provide convincing evidence to show that Orai1 expression is increased in hepatocarcinoma tissues. 5-FU can induce autophagic cell death in HepG2 hepatocarcinoma cells through inhibition of SOCE via decreasing Orai1 expression. These findings suggest that Orai1 expression is a predictor of 5-FU sensitivity for hepatocarcinoma treatment and blockade of Orai1-mediated Ca2+ entry may be a promising strategy to sensitize hepatocarcinoma cells to 5-FU treatment.


Assuntos
Cálcio/metabolismo , Carcinoma Hepatocelular/metabolismo , Fluoruracila/farmacologia , Neoplasias Hepáticas/metabolismo , Proteína ORAI1/metabolismo , Autofagossomos/efeitos dos fármacos , Autofagossomos/metabolismo , Autofagia/efeitos dos fármacos , Carcinoma Hepatocelular/patologia , Regulação para Baixo/efeitos dos fármacos , Células Hep G2 , Humanos , Neoplasias Hepáticas/patologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais/efeitos dos fármacos , Serina-Treonina Quinases TOR/metabolismo
15.
Arterioscler Thromb Vasc Biol ; 36(4): 618-28, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26916730

RESUMO

OBJECTIVE: To determine the role of orai1 store-operated Ca(2+) entry in foam cell formation and atherogenesis. APPROACH AND RESULTS: Acute administration of oxidized low-density lipoprotein (oxLDL) activates an orai1-dependent Ca(2+) entry in macrophages. Chelation of intracellular Ca(2+), inhibition of orai1 store-operated Ca(2+) entry, or knockdown of orai1 dramatically inhibited oxLDL-induced upregulation of scavenger receptor A, uptake of modified LDL, and foam cell formation. Orai1-dependent Ca(2+) entry induces scavenger receptor A expression and foam cell formation through activation of calcineurin but not calmodulin kinase II. Activation of nuclear factor of activated T cells is not involved in calcineurin signaling to foam cell formation. However, oxLDL dephosohorylates and activates apoptosis signal-regulating kinase 1 in macrophages. Orai1 knockdown prevents oxLDL-induced apoptosis signal-regulating kinase 1 activation. Knockdown of apoptosis signal-regulating kinase 1, or inhibition of its downstream effectors, JNK and p38 mitogen-activated protein kinase, reduces scavenger receptor A expression and foam cell formation. Notably, orai1 expression is increased in atherosclerotic plaques of apolipoprotein E(-/-) mice fed with high-cholesterol diet. Knockdown of orai1 with adenovirus harboring orai1 siRNA or inhibition of orai1 Ca(2+) entry with SKF96365 for 4 weeks dramatically inhibits atherosclerotic plaque development in high-cholesterol diet feeding apolipoprotein E(-/-) mice. In addition, inhibition of orai1 Ca(2+) entry prevents macrophage apoptosis in atherosclerotic plaque. Moreover, the expression of inflammatory genes in atherosclerotic lesions and the infiltration of myeloid cells into the aortic sinus plaques are decreased after blocking orai1 signaling. CONCLUSIONS: Orai1-dependent Ca(2+) entry promotes atherogenesis possibly by promoting foam cell formation and vascular inflammation, rendering orai1 Ca(2+) channel a potential therapeutic target against atherosclerosis.


Assuntos
Anticolesterolemiantes/farmacologia , Aorta/efeitos dos fármacos , Doenças da Aorta/prevenção & controle , Aterosclerose/prevenção & controle , Bloqueadores dos Canais de Cálcio/farmacologia , Canais de Cálcio/efeitos dos fármacos , Cálcio/metabolismo , Colesterol/metabolismo , Células Espumosas/efeitos dos fármacos , Macrófagos Peritoneais/efeitos dos fármacos , Animais , Aorta/metabolismo , Aorta/patologia , Doenças da Aorta/genética , Doenças da Aorta/metabolismo , Doenças da Aorta/patologia , Apolipoproteínas E/deficiência , Apolipoproteínas E/genética , Apoptose/efeitos dos fármacos , Aterosclerose/genética , Aterosclerose/metabolismo , Aterosclerose/patologia , Calcineurina/metabolismo , Canais de Cálcio/genética , Canais de Cálcio/metabolismo , Quelantes de Cálcio/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Linhagem Celular Tumoral , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Células Espumosas/metabolismo , Células Espumosas/patologia , Humanos , Mediadores da Inflamação/metabolismo , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Lipoproteínas LDL/farmacologia , MAP Quinase Quinase Quinase 5/metabolismo , Macrófagos Peritoneais/metabolismo , Macrófagos Peritoneais/patologia , Camundongos Knockout , Proteína ORAI1 , Placa Aterosclerótica , Interferência de RNA , Receptores Depuradores Classe A/metabolismo , Fatores de Tempo , Transfecção , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
16.
Appl Microbiol Biotechnol ; 101(11): 4593-4603, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28353000

RESUMO

Avian influenza virus (AIV) is spreading worldwide and is a serious threat to the health of poultry and humans. In many countries, low pathogenic AIVs, such as H9N2, have become an enormous economic burden on the commercial poultry industry because they cause mild respiratory disease and decrease egg production. A recombinant Lactobacillus plantarum NC8 strain expressing NP-M1-DCpep from H9N2 AIV has been studied in a mouse model. However, it remains unknown whether this L. plantarum strain can induce an immune response and provide protection against H9N2 AIV in chickens. In this study, chickens that were orally vaccinated with NC8-pSIP409-NP-M1-DCpep exhibited significantly increased T cell-mediated immune responses and mucosal sIgA and IgG levels, which provided protection against H9N2 AIV challenge. More importantly, compared with oral administration of NC8-pSIP409-NP-M1-DCpep, intranasal administration induced stronger immune responses and provided effective protection against challenge with the H9N2 virus by reducing body weight loss, lung virus titers, and throat pathology. Taken together, these findings suggest that L. plantarum expressing NP-M1-DCpep has potential as a vaccine to combat H9N2 AIV infection.


Assuntos
Anticorpos Antivirais/biossíntese , Antígenos Virais/genética , Galinhas , Vírus da Influenza A Subtipo H9N2/imunologia , Vacinas contra Influenza/imunologia , Influenza Aviária/prevenção & controle , Lactobacillus plantarum/genética , Administração Intranasal , Administração Oral , Animais , Antígenos Virais/administração & dosagem , Antígenos Virais/imunologia , Imunidade nas Mucosas , Imunoglobulina A/biossíntese , Imunoglobulina A/imunologia , Imunoglobulina G/biossíntese , Imunoglobulina G/imunologia , Vacinas contra Influenza/administração & dosagem , Influenza Aviária/imunologia , Pulmão/virologia , Faringe/patologia , Faringe/virologia , Aves Domésticas , Linfócitos T/imunologia
17.
Biosci Biotechnol Biochem ; 81(8): 1489-1496, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28644752

RESUMO

Recombination activating gene 2 (RAG2) is necessary for immature B cell differentiation. Antibodies to human and rabbit RAG2 are currently commercially available, but antibodies to swine RAG remain unavailable to date. In this study, the swine RAG2 genes sequence was synthesized and then cloned into a pET-28a vector. The recombinant fusion protein was successfully expressed in E. coli, purified through nickel column chromatography, and further digested with Tobacco Etch Virus protease. The cleaved protein was purified by molecular-exclusion chromatography and named pRAG2. We used pRAG2 to immunize rabbits, collected the serum and purified rabbit anti-pRAG2 polyclonal antibodies. The rabbit anti-pRAG2 polyclonal antibodies were tested via immunofluorescence on eukaryotic cells overexpressing pRAG2 and also able to recognize pig natural RAG2 and human RAG2 protein in western blotting. These results indicated that the prepared rabbit anti-pRAG2 polyclonal antibodies may serve as a tool to detect immature B cell differentiation of swine.


Assuntos
Anticorpos/química , Proteínas de Ligação a DNA/biossíntese , Escherichia coli/genética , Expressão Gênica , Proteínas Nucleares/biossíntese , VDJ Recombinases/biossíntese , Animais , Anticorpos/isolamento & purificação , Anticorpos/metabolismo , Western Blotting , Clonagem Molecular , Proteínas de Ligação a DNA/genética , Proteínas de Ligação a DNA/imunologia , Endopeptidases/química , Escherichia coli/metabolismo , Imunofluorescência , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Humanos , Soros Imunes/química , Isoenzimas/biossíntese , Isoenzimas/genética , Isoenzimas/imunologia , Proteínas Nucleares/genética , Proteínas Nucleares/imunologia , Coelhos , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Suínos , VDJ Recombinases/genética , VDJ Recombinases/imunologia
18.
J Cell Mol Med ; 20(3): 526-36, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26805397

RESUMO

Gastric cancer is one of the most common malignancies worldwide. Interleukin-1-beta (IL-1ß) is a pro-inflammatory cytokine and potent inhibitor of gastric acid secretion. Some studies provided evidence of the association between IL-1B 31 polymorphism and gastric cancer risk while other studies did not. Therefore, we conducted a comprehensive meta-analysis to reassess the association. A systematic literature search of the PubMed and EMBASE databases identified 37 studies with 6108 cases and 8980 controls for this meta-analysis. The crude odd ratios (ORs) and the 95% confidence intervals (CIs) were calculated to evaluate the strength of the association. Meta-regression was used to determine the major source of heterogeneity across the studies. The pooled analysis did not suggest the significant association of IL-1B 31 C>T polymorphism with gastric cancer risk. Stratified analysis was performed by ethnicity, source of control, genotype method, and indicated a significantly increased gastric cancer risk associated with IL-1B 31T variant in the population-based subgroup (heterozygous model: OR = 1.22, 95% CI = 1.03-1.45). Moreover, stratified analysis by Helicobacter pylori infection status indicated that IL-1B 31 polymorphism increased gastric cancer risk in infection-positive subgroup (homozygous model: OR = 1.35, 95% CI = 1.02-1.78; heterozygous model: OR = 1.31, 95% CI = 1.04-1.66; recessive model: OR = 1.29, 95% CI = 1.04-1.61). The study suggested that IL-1B 31 polymorphism might confer susceptibility to gastric cancer in the presence of H. pylori infection, indicating a gene-environment interaction in gastric carcinogenesis.


Assuntos
Infecções por Helicobacter/genética , Helicobacter pylori/fisiologia , Interleucina-1beta/genética , Neoplasias Gástricas/genética , Estudos de Casos e Controles , Interação Gene-Ambiente , Estudos de Associação Genética , Predisposição Genética para Doença , Infecções por Helicobacter/microbiologia , Humanos , Polimorfismo de Nucleotídeo Único , Neoplasias Gástricas/microbiologia
19.
Circ J ; 80(4): 1024-33, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26911455

RESUMO

BACKGROUND: Previous work has demonstrated that the volume-regulated chloride channel is activated during foam cell formation, and inhibition of chloride movement prevents intracellular lipid accumulation. However, the mechanism explaining how chloride movement promotes foam cell formation is not clear. METHODS AND RESULTS: Foam cell formation was determined by Oil Red O staining. Western blotting and co-immunoprecipitation were used to examine protein expression and protein-protein interaction. [Cl(-)]iwas measured using 6-methoxy-N-ethylquinolinium iodide dye. The results showed that [Cl(-)]iwas decreased in monocytes/macrophages from patients with hypercholesterolemia and from apoE(-/-)mice fed with a high-fat diet. Lowering [Cl(-)]iupregulated scavenger receptor A (SR-A) expression, increased the binding and uptake of oxLDL, enhanced pro-inflammatory cytokine production and subsequently accelerated foam cell formation in macrophages from humans and mice. In addition, low Cl(-)solution stimulated the activation of JNK and p38 mitogen-activated protein kinases. Inhibition of JNK and p38 blocked Cl(-)reduced medium-induced SR-A expression and lipid accumulation. In contrast, reduction of [Cl(-)]ipromoted the interaction of SR-A with caveolin-1, thus facilitating caveolin-1-dependent SR-A endocytosis. Moreover, disruption of caveolae attenuated SR-A internalization, JNK and p38 activation, and ultimately prevented SR-A expression and foam cell formation stimulated by low Cl(-)medium. CONCLUSIONS: This data provide strong evidence that reduction of [Cl(-)]iis a critical contributor to intracellular lipid accumulation, suggesting that modulation of [Cl(-)]iis a novel avenue to prevent foam cell formation and atherosclerosis.


Assuntos
Cloretos/metabolismo , Células Espumosas/metabolismo , Hipercolesterolemia/metabolismo , Animais , Apolipoproteínas E/deficiência , Caveolina 1/genética , Caveolina 1/metabolismo , Gorduras na Dieta/efeitos adversos , Gorduras na Dieta/farmacologia , Ativação Enzimática/efeitos dos fármacos , Ativação Enzimática/genética , Células Espumosas/patologia , Hipercolesterolemia/induzido quimicamente , Hipercolesterolemia/genética , Hipercolesterolemia/patologia , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/metabolismo , Camundongos , Camundongos Knockout , Receptores Depuradores Classe A/genética , Receptores Depuradores Classe A/metabolismo , Regulação para Cima/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/genética , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
20.
Water Environ Res ; 88(4): 318-24, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27131055

RESUMO

The mesoporous graphitic carbon nitride (mpg-C3N4/r, r was defined as the initial silica/dicyandiamide mass ratio) was successfully synthesized by heating the mixture of silica and dicyandiamide in a nitrogen atmosphere. The morphology and structure of mpg-C3N4/r were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Brunauer-Emmett-Teller surface area measurement (BET), X-ray powder diffraction (XRD), and Fourier Transform Infrared spectroscopy (FT-IR). The adsorption performances of Ni (II) ions by mpg-C3N4/r were investigated. With increasing of r value, the BET specific surface area of the synthesized mpg-C3N4/r increased; the highest specific surface area of mpg-C3N4/1.5 increased up to 169.3 m2/g. This work shows that mpg-C3N4/1.5 is a promising, high-efficiency adsorbent that can be used to purify the water of a low Ni (II) ions concentration. The maximum adsorption capacity of Ni(II) ions by mpg-C3N4/1.5 was 15.26 mg/g. The adsorption properties of Ni (II) ions by mpg-C3N4/r complied well with pseudo-second-order kinetics and Langmuir isotherm model.


Assuntos
Níquel/química , Nitrilas/química , Poluentes Químicos da Água/química , Adsorção , Cinética , Purificação da Água/métodos
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