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1.
J Antimicrob Chemother ; 79(9): 2292-2297, 2024 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-38997220

RESUMO

OBJECTIVES: Carbapenem-resistant Klebsiella pneumoniae (CRKP) is a major nosocomial infectious pathogen with rapidly increasing prevalence. The genomic epidemiological characteristics of CRKP nationwide, especially the evolving trends within the predominant clones, should be evaluated clearly. METHODS: We collected 3415 K. pneumoniae strains from 28 hospitals across China. Antimicrobial susceptibility testing and WGS were performed. Subsequent genomic analyses, including sequence typing, K-locus (KL) identification, antimicrobial resistance gene screening, and virulence score assessment were performed. The phylogenetic relationship of clonal group 11 was determined based on core-genome analysis, and the presence of the pLVPK-like virulence plasmid in ST11 isolates was confirmed using plasmid core-gene analysis. Additionally, the trends of the ST11 lineage with different KL types on a global scale were investigated using Beast2. RESULTS: Of the K. pneumoniae strains, 708 were identified as CRKP isolates (20.7%), of which 97.7% were MDR. ST11 was the predominant clone, and KPC-2 was the prevalent carbapenemase in China, although the prevalence of specific clones and carbapenemases varied by geographic region. Among ST11 isolates, KL47 and KL64 were the predominant KL types, and KL64 gradually replaced KL47, with a higher percentage of KL64 isolates harbouring the pLVPK-like plasmid. Global genome data showed a significant increase in the effective population size of KL64 over the last 5 years. CONCLUSIONS: The prevalence of CRKP was very high in certain regions in China. The increasing convergence of virulence and resistance, particularly in ST11-KL64 isolates, should be given more attention and further investigation.


Assuntos
Antibacterianos , Enterobacteriáceas Resistentes a Carbapenêmicos , Infecções por Klebsiella , Klebsiella pneumoniae , Testes de Sensibilidade Microbiana , Filogenia , Plasmídeos , Sequenciamento Completo do Genoma , China/epidemiologia , Klebsiella pneumoniae/genética , Klebsiella pneumoniae/efeitos dos fármacos , Klebsiella pneumoniae/isolamento & purificação , Klebsiella pneumoniae/classificação , Infecções por Klebsiella/epidemiologia , Infecções por Klebsiella/microbiologia , Humanos , Plasmídeos/genética , Antibacterianos/farmacologia , Enterobacteriáceas Resistentes a Carbapenêmicos/genética , Enterobacteriáceas Resistentes a Carbapenêmicos/efeitos dos fármacos , Enterobacteriáceas Resistentes a Carbapenêmicos/isolamento & purificação , beta-Lactamases/genética , Genoma Bacteriano , Carbapenêmicos/farmacologia , Proteínas de Bactérias/genética , Tipagem de Sequências Multilocus , Virulência/genética , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/microbiologia , Epidemiologia Molecular , Prevalência , Farmacorresistência Bacteriana Múltipla/genética
2.
Int J Mol Sci ; 25(3)2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-38338731

RESUMO

CTP synthase (CTPS), the rate-limiting enzyme in the de novo synthesis of CTP, assembles into a filamentous structure termed the cytoophidium. The Hippo pathway regulates cell proliferation and apoptosis. The relationship of the nucleotide metabolism with the Hippo pathway is little known. Here, we study the impact of the Hippo pathway on the cytoophidium in Drosophila melanogaster posterior follicle cells (PFCs). We find that the inactivation of the Hippo pathway correlates with reduced cytoophidium length and number within PFCs. During the overexpression of CTPS, the presence of Hippo mutations also reduces the length of cytoophidia in PFCs. In addition, we observe that knocking down CTPS mitigates hpo (Hippo)-associated over-proliferation. In summary, our results suggest that there is a connection between the Hippo pathway and the nucleotide biosynthesis enzyme CTPS in PFCs.


Assuntos
Proteínas de Drosophila , Drosophila , Animais , Drosophila/metabolismo , Drosophila melanogaster/metabolismo , Via de Sinalização Hippo , Citoesqueleto/metabolismo , Nucleotídeos/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas de Drosophila/genética , Proteínas de Drosophila/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo
3.
Glycobiology ; 33(8): 673-682, 2023 10 06.
Artigo em Inglês | MEDLINE | ID: mdl-37504513

RESUMO

Nonalcoholic fatty liver disease is caused by an imbalance in lipid metabolism and immune response to pose a risk factor for liver fibrosis. Recent evidence indicates that M2 macrophages secrete transforming growth factor-ß1, which contributes to liver fibrosis. Galectin-12 has been demonstrated to regulate lipid metabolism and macrophage polarization. The purpose of this study is to investigate the role of galectin-12 in the development of nonalcoholic fatty liver disease and fibrosis. Liver tissue from wild-type C57BL/6 mice fed with a high-fat diet containing cholesterol and cholic acid for 4-12 weeks was used to examine galectin-12 expression and its correlation with nonalcoholic fatty liver disease. Furthermore, the effects of galectin-12 on M2 macrophages during the progression of nonalcoholic fatty liver disease were investigated by studying Kupffer cells from galectin-12 knockout mice and doxycycline-inducible Gal12-/-THP-1 cells. Ablation of galectin-12 promoted M2 polarization of Kupffer cells, as indicated by higher levels of M2 markers, such as arginase I and chitinase 3-like protein 3. Furthermore, the activation of signal transducer and activator of transcription 6 was significantly higher in Gal12-/- macrophages activated by interleukin-4, which was correlated with higher levels of transforming growth factor-ß1. Moreover, Gal12-/- macrophage-conditioned medium promoted hepatic stellate cells myofibroblast differentiation, which was indicated by higher α-smooth muscle actin expression levels compared with those treated with LacZ control medium. Finally, we demonstrated that galectin-12 knockdown negatively regulated the suppressor of cytokine signaling 3 levels. These findings suggested that galectin-12 balances M1/M2 polarization of Kupffer cells to prevent nonalcoholic fatty liver disease progression.


Assuntos
Células de Kupffer , Hepatopatia Gordurosa não Alcoólica , Animais , Camundongos , Células de Kupffer/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Fator de Crescimento Transformador beta1 , Camundongos Endogâmicos C57BL , Fígado/metabolismo , Cirrose Hepática , Dieta Hiperlipídica/efeitos adversos , Galectinas/metabolismo
4.
Mol Pain ; 19: 17448069221149834, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36550612

RESUMO

Irritable bowel syndrome (IBS) related chronic visceral pain affects 20% of people worldwide. The treatment options are very limited. Although the scholarly reviews have appraised the potential effects of the intestinal microbiota on intestinal motility and sensation, the exact mechanism of intestinal microbiota in IBS-like chronic visceral pain remains largely unclear. The purpose of this study is to investigate whether Folic Acid (FA) attenuated visceral pain and its possible mechanisms. Chronic visceral hyperalgesia was induced in rats by neonatal colonic inflammation (NCI). 16S rDNA analysis of fecal samples from human subjects and rats was performed. Patch clamp recording was used to determine synaptic transmission of colonic-related spinal dorsal horn. Alpha diversity of intestinal flora was increased in patients with IBS, as well as the obviously increased abundance of Clostridiales order (a main bacteria producing hydrogen sulfide). The hydrogen sulfide content was positive correlation with visceral pain score in patients with IBS. Consistently, NCI increased Clostridiales frequency and hydrogen sulfide content in feces of adult rats. Notably, the concentration of FA was markedly decreased in peripheral blood of IBS patients compared with non-IBS human subjects. FA supplement alleviated chronic visceral pain and normalized the Clostridiales frequency in NCI rats. In addition, FA supplement significantly reduced the frequency of sEPSCs of neurons in the spinal dorsal horn of NCI rats. Folic Acid treatment attenuated chronic visceral pain of NCI rats through reducing hydrogen sulfide production from Clostridiales in intestine.


Assuntos
Sulfeto de Hidrogênio , Síndrome do Intestino Irritável , Dor Visceral , Humanos , Adulto , Ratos , Animais , Sulfeto de Hidrogênio/farmacologia , Sulfeto de Hidrogênio/uso terapêutico , Ratos Sprague-Dawley , Clostridiales , Ácido Fólico/farmacologia , Ácido Fólico/uso terapêutico , Hidrogênio , Dor Visceral/tratamento farmacológico , Inflamação , Sulfetos
5.
Am J Physiol Gastrointest Liver Physiol ; 325(4): G356-G367, 2023 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-37529842

RESUMO

Chronic visceral pain is a common symptom of irritable bowel syndrome (IBS). Exosomes are involved in the development of pain. Rab27a can mediate the release of exosomes. The purpose of this study is to investigate how Rab27a-mediated exosome secretion in the anterior cingulate cortex (ACC) regulates visceral hyperalgesia induced with neonatal maternal deprivation (NMD) in adult mice. The colorectal distension method was adopted to measure visceral pain. The BCA protein assay kit was applied to detect the exosome protein concentration. Western blotting, quantitative PCR, and immunofluorescence technique were adopted to detect the expression of Rab27a and the markers of exosomes. Exosomes extracted from ACC were more in NMD mice than in control (CON) mice. Injection of the exosome-specific inhibitor GW4869 in ACC attenuated colorectal visceral pain of NMD mice. Injection of NMD-derived exosomes produced colorectal visceral pain in CON mice. Rab27a was upregulated in ACC of NMD mice. Rab27a was highly expressed in ACC neurons of NMD mice, rather than astrocytes and microglia. Injection of Rab27a-siRNA reduced the release of exosomes and attenuated the colorectal visceral pain in NMD mice. This study suggested that overexpression of Rab27a increased exosome secretion in ACC neurons, thus contributing to visceral hyperalgesia in NMD mice.NEW & NOTEWORTHY This work demonstrated that the expression of Rab27a in the anterior cingulate cortex was upregulated, which mediated multivesicular bodies trafficking to the plasma membrane and led to the increased release of neuronal exosomes, thus contributing to colorectal visceral pain in neonatal maternal deprivation (NMD) mice. Blocking the release of exosomes or downregulation of Rab27a could alleviate colorectal visceral pain in NMD mice. These data may provide a promising strategy for the treatment of visceral pain in irritable bowel syndrome patients.


Assuntos
Neoplasias Colorretais , Exossomos , Síndrome do Intestino Irritável , Dor Visceral , Camundongos , Animais , Giro do Cíngulo , Dor Visceral/metabolismo , Hiperalgesia/etiologia , Privação Materna , Exossomos/metabolismo , Proteínas rab27 de Ligação ao GTP/genética , Proteínas rab27 de Ligação ao GTP/metabolismo
6.
BMC Musculoskelet Disord ; 24(1): 407, 2023 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-37217909

RESUMO

BACKGROUND: Intermediate vertebral collapse is a newly discovered complication of consecutive two-level anterior cervical discectomy and fusion (ACDF). There have been no analytical studies related to the effects of endplate defects on the biomechanics of the intermediate vertebral bone after ACDF. This study aimed to compare the effects of endplate defects on the intermediate vertebral bone biomechanics in the zero-profile (ZP) and cage-and-plate (CP) methods of consecutive 2-level ACDF and to determine whether collapse of the intermediate vertebra is more likely to occur using ZP. METHODS: A three-dimensional finite element (FE) model of the intact cervical spine (C2-T1) was constructed and validated. The intact FE model was then modified to build ACDF models and imitate the situation of endplate injury, establishing two groups of models (ZP, IM-ZP and CP, IM-ZP). We simulated cervical motion, such as flexion, extension, lateral bending and axial rotation, and compared the range of motion (ROM), upper and lower endplate stress, fusion fixation device stress, C5 vertebral body stress, intervertebral disc internal pressure (intradiscal pressure, or IDP) and the ROM of adjacent segments in the models. RESULTS: There was no significant difference between the IM-CP model and the CP model in the ROM of the surgical segment, upper and lower endplate stress, fusion fixation device stress, C5 vertebral body stress, IDP, or ROM of the adjacent segments. Compared with the CP model, the endplate stress of the ZP model is significantly higher in the flexion, extension, lateral bending and axial rotation conditions. Compared with the ZP model, endplate stress, screw stress, C5 vertebral stress and IDP in IM-ZP were significantly increased under flexion, extension, lateral bending and axial rotation conditions. CONCLUSIONS: Compared to consecutive 2-level ACDF using CP, collapse of the intermediate vertebra is more likely to occur using ZP due to its mechanical characteristics. Intraoperative endplate defects of the anterior lower margin of the middle vertebra are a risk factor leading to collapse of the middle vertebra after consecutive 2-level ACDF using ZP.


Assuntos
Disco Intervertebral , Fusão Vertebral , Humanos , Análise de Elementos Finitos , Fusão Vertebral/métodos , Vértebras Cervicais/diagnóstico por imagem , Vértebras Cervicais/cirurgia , Discotomia/efeitos adversos , Discotomia/métodos , Disco Intervertebral/diagnóstico por imagem , Disco Intervertebral/cirurgia , Amplitude de Movimento Articular , Fenômenos Biomecânicos
7.
Beijing Da Xue Xue Bao Yi Xue Ban ; 55(6): 1022-1027, 2023 Dec 18.
Artigo em Zh | MEDLINE | ID: mdl-38101783

RESUMO

OBJECTIVE: To detect the expression of plasma exosomal microRNA (miRNA) in systemic sclerosis (SSc), and to investigate its clinical significance. METHODS: A total of 20 patients who were initially diagnosed with SSc and did not receive medication in Department of Rheumatology and Immunology of Meizhou People' s Hospital from January 2020 to January 2022 were recruited, as well as 15 healthy individuals whose gender and age matched with those of the SSc patients. Plasma exosomes were isolated using ultracentrifugation method. The expression levels of exosomal miR-34-5p, miR-92-3p and miR-142-3p were detected by quantative real-time polymerase chain reaction (qRT-PCR). Correlations between the expression levels of exosomal miRNAs and clinical characteristic were analyzed by Spearman's rank correlation coefficient test. RESULTS: The mean age of 20 patients with SSc was (52.6±12.6) years, including 7 males and 13 females. Among the 20 SSc patients, 13 cases were diagnosed as limited cutaneous systemic sclerosis (lcSSc) and 7 cases were diagnosed as diffuse cutaneous systemic sclerosis (dcSSc) according to the extent of skin involvement. According to the findings of high resolution chest CT, 7 of 20 SSc patients were diagnosed with interstitial lung disease (ILD) and 13 SSc patients were diagnosed with non-ILD. The expression levels of exosomal miR-34-5p, miR-92-3p and miR-142-3p were significantly elevated in the SSc patients compared with those in the healthy controls group (P=0.003, P=0.000 1, and P=0.016, respectively). Compared with the SSc patients without ILD, the expression levels of miR-34-5p and miR-142-3p were significantly lower in the SSc patients with ILD (P=0.037 and P=0.015, respectively). The expression levels of exosomal miR-34-5p and miR-142-3p showed negative correlation with ILD (r=-0.48, P=0.031 and r=-0.55, P=0.011, respectively), and arthritis (r=-0.46, P=0.040 and r=-0.48, P=0.032, respectively). The expression levels of exosomal miR-142-3p showed a negative correlation with erythrocyte sedimentation rate (ESR) (r=-0.55, P=0.012). CONCLUSION: Plasma exosomal miR-34-5p, miR-92-3p and miR-142-3p were dysregulated in SSc. The dyregulation of exosomal miR-34-5p and miR-142-3p showed correlation with SSc associated ILD (SSc-ILD).


Assuntos
Doenças Pulmonares Intersticiais , MicroRNAs , Escleroderma Sistêmico , Masculino , Feminino , Humanos , Adulto Jovem , Adulto , Relevância Clínica , MicroRNAs/genética , Escleroderma Sistêmico/genética
8.
Microb Ecol ; 81(2): 314-322, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32935184

RESUMO

In this study, the joint use of high throughput sequencing, real-time quantitative PCR, and ammonia-oxidizing bacteria (AOB)-inhibiting allylthiourea was used to differentiate between the contributions of ammonia-oxidizing archaea (AOA) vs AOB to ammonia oxidation and ascertain how AOA and AOB responded to two widely used river remediation techniques (aeration and Ca(NO3)2 injection). Results showed that ammonia oxidation was largely attributed to ATU-sensitive AOB rather than AOA and Nitrosomonas was the predominant AOB-related genus (53.86%) in the malodorous river. The contribution of AOB to ammonia oxidation in the context of aeration and Ca(NO3)2 injection was 75.51 ± 2.77% and 60.19 ± 10.44%, respectively. The peak of AOB/AOA ratio and the marked increase of relative abundances of Nitrosomonas and Nitrosospira in aeration runs further demonstrated aeration favored the ammonia oxidation of AOB. Comparatively, Ca(NO3)2 injection could increase the ammonia oxidation contribution of AOA from 31.32 ± 6.06 to 39.81 ± 10.44% and was significantly correlated with Nitrosococcus of AOB (r = 0.796, p < 0.05), Candidatus_Nitrosopelagicus of AOA (r = 0.986, p < 0.01), and AOA Simpson diversity (r = - 0.791, p < 0.05). Moreover, Candidatus_Nitrosopelagicus was only present in Ca(NO3)2 runs. Taken together, Ca(NO3)2 was recognized as an important factor in mediating the growth and ecological niches of ammonia oxidizers.Graphical abstract.


Assuntos
Amônia/metabolismo , Archaea/metabolismo , Bactérias/metabolismo , Recuperação e Remediação Ambiental/métodos , Sedimentos Geológicos/microbiologia , Rios/microbiologia , Amônia/análise , Archaea/classificação , Archaea/genética , Archaea/isolamento & purificação , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Compostos de Cálcio/análise , Sedimentos Geológicos/química , Nitratos/análise , Oxirredução , Oxigênio/análise , Rios/química , Especificidade da Espécie , Poluentes Químicos da Água/metabolismo
9.
J Anat ; 236(5): 916-922, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31961950

RESUMO

Many studies have demonstrated the association between facet tropism and disc herniation in the lumbar spine. Some of them found that lumbar disc herniation was on the side of the more sagittal facet joint interface. However, little is understood about the association of facet tropism with disc herniation in the cervical spine. As the relationship between the facet orientation and the side of cervical disc herniation (CDH) is unclear, the purpose of this study is to investigate that relationship. Ninety-six patients with single-level CDH (C4-C5, C5-C6 or C6-C7) were included in the CDH group of this study. Another 50 age-matched and gender-matched healthy participants who accepted physical examinations were enrolled as the control group. The cervical facet angles of two sides were measured using axial computed tomography (CT). The intersection angle of the midsagittal line of the vertebra to the facet line represents the facet angle. Facet tropism was defined as the angular difference of 7º between the left and the right sides. Facet tropism angle was recorded as the absolute value of the difference of facet angles between two sides. There were 20 herniations at C4-C5 level, 50 herniations at C5-C6 level and 26 herniations at C6-C7 level. The present study showed that more cases in the CDH group had facet tropism than did those in the control group at C4-C5, C5-C6 and C6-C7 level (p = .021, p = .001, p = .015, respectively). The facet tropism angles in the CDH group were significantly bigger than those in the control group at C4-C5, C5-C6 and C6-C7 level (p = .001, p = .002, p = .028, respectively). In the CDH group, the facet angles on the herniated side were found to be significantly bigger than those on the healthy side at C4-C5, C5-C6 and C6-C7 level (p = .000, p = .000, p = .037, respectively). The findings of this present study suggest that facet tropism is associated with the disc herniation in the cervical spine. We also found that cervical disc herniates towards the side of the bigger facet angle with respect to the sagittal plane. There is a need for future studies to verify the biomechanical impact of facet tropism on CDH.


Assuntos
Vértebras Cervicais/diagnóstico por imagem , Deslocamento do Disco Intervertebral/diagnóstico por imagem , Disco Intervertebral/diagnóstico por imagem , Articulação Zigapofisária/diagnóstico por imagem , Feminino , Humanos , Imageamento por Ressonância Magnética , Masculino , Estudos Retrospectivos , Tomografia Computadorizada por Raios X
10.
Anal Chem ; 91(2): 1424-1431, 2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30562003

RESUMO

Aptamers have emerged as promising molecular tools for small-molecule analyte sensing. However, the performance of such aptasensors is generally limited by leakage since it has been difficult to completely suppress signal in the absence of analyte, resulting in a compromise between sensitivity and specificity. Here, we describe a methodology for the ultrasensitive detection of analytes combining aptasensors with single-molecule kinetic fingerprinting. A short, fluorescently labeled DNA probe is utilized to detect the structural changes upon ligand binding to the designed hairpin-shaped aptasensor probe. The Poisson statistics of binding and dissociation events of the DNA probe to single surface-immobilized aptasensor molecules is monitored by total internal reflection fluorescence microscopy, permitting the high-accuracy discrimination of the ligand bound and ligand-free states, resulting in zero background. The programmable dynamics of the hairpin enables fine-tuning of the hybridization kinetics of the fluorescent probe, rendering the acquisition time sufficiently flexible to optimize discrimination. Remarkable detection limits are achieved for a diverse set of analytes when spiked into chicken meat extract: the nucleotide adenosine (0.3 pM), the insecticide acetamiprid (0.35 pM), and the dioxin-like toxin PCB-77 (0.72 pM), which is superior to recently reported aptasensors. Our generalizable method significantly improves the performance of aptasensors, with the potential to extend to other molecular biomarkers.


Assuntos
Aptâmeros de Nucleotídeos/metabolismo , Técnicas Biossensoriais/métodos , Limite de Detecção , Bibliotecas de Moléculas Pequenas/análise , Animais , DNA de Cadeia Simples/genética , DNA de Cadeia Simples/metabolismo , Corantes Fluorescentes/metabolismo , Sequências Repetidas Invertidas , Cinética , Bibliotecas de Moléculas Pequenas/metabolismo
11.
Anal Chem ; 90(21): 13044-13050, 2018 11 06.
Artigo em Inglês | MEDLINE | ID: mdl-30289245

RESUMO

Rapid quantification of permanganate (MnO4-) in aqueous solution with a convenient and sensitive single-particle-detection (SPD) method is demonstrated by dark-field optical microscopy. The design is based on the selective etching of the Ag shell of a glucose-protected GNPs@Ag nanoparticle by MnO4-. In the presence of MnO4-, a noticeable red-shift of localized surface-plasmon resonance (LSPR, from blue to green) together with a tremendous decrease in the extinction coefficient from individual GNPs@Ag nanoparticles is observed. MnO4- can then be quantified by calculating the ratio between the number of green and blue particles on the cover glass surface after the etching process. A linear dynamic range of 0-6 µM and a limit of detection (LOD) as low as 46 nM were readily achieved, which are much lower than those of spectroscopic measurements in bulk solution. In tap water, a comparable LOD (50 nM) and satisfactory recovery efficiency are demonstrated. As a consequence of these merits, the method demonstrated herein will find promising applications for the ultrasensitive detection of MnO4- under complex milieu in the future.

12.
Anal Chem ; 90(2): 1177-1185, 2018 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-29243478

RESUMO

Exploring the diffusion dynamics of a viral capsid proteins (VCP)-functionalized nanocarrier on a living cell membrane could provide much kinetic information for the better understanding of their biological functionality. Gold nanoparticles are an excellent core material of nanocarriers because of the good biocompatibility as well as versatile surface chemistry. However, due to the strong scattering background from subcellular organelles, it is a grand challenge to selectively image an individual nanocarrier on a living cell membrane. In this work, we demonstrated a convenient strategy to effectively screen the scattering background from living cells for single-particle imaging with a polarization-resolved dual-channel imaging module. By taking advantage of the polarization of anisotropic gold nanoparticles (gold nanorods, GNRs), the signals from cell components could be counteracted after subtracting the sequential images one by one, while those transiently rotating GNRs on the cell membrane still exist in the processed image. In contrast to the previously reported methods, this method does not require a complicated optical setup alignment and sophisticated digital image analysis process. According to the single-particle imaging results, the majority of VCP-GNRs were anchoring on the cell membrane with confined diffusion. Interestingly, on further inspection of the diffusion trajectories, the particles displayed anomalous confined diffusion with randomly distributed large walking steps during the whole track. Non-Gaussian step distribution was noted, indicating heterogeneous binding and desorption processes on the cell membrane. As a consequence of the robust background screening capability, this approach would find broad applications for single-particle imaging under a noisy environment, e.g., living cells.


Assuntos
Proteínas do Capsídeo/análise , Infecções por Circoviridae/virologia , Circovirus/química , Ouro/química , Hepatócitos/virologia , Nanopartículas Metálicas/química , Imagem Óptica/métodos , Anisotropia , Infecções por Circoviridae/patologia , Desenho de Equipamento , Células Hep G2 , Hepatócitos/patologia , Humanos , Microscopia/instrumentação , Microscopia/métodos , Imagem Óptica/instrumentação
13.
Sensors (Basel) ; 18(2)2018 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-29385105

RESUMO

Stable rotor levitation is a challenge for rotational gyroscopes (magnetically suspended gyroscopes (MSG) and electrostatically suspended gyroscopes (ESG)) with a ring- or disk-shaped rotor, which restricts further improvement of gyroscope performance. In addition, complicated pick-up circuits and feedback control electronics propose high requirement on fabrication technology. In the proposed gyroscope, a ball-disk shaped rotor is supported by a water-film bearing, formed by centrifugal force to deionized water at the cavity of the lower supporting pillar. Water-film bearing provides stable mechanical support, without the need for complicated electronics and control system for rotor suspension. To decrease sliding friction between the rotor ball and the water-film bearing, a supherhydrophobic surface (SHS) with nano-structures is fabricated on the rotor ball, resulting in a rated spinning speed increase of 12.4% (under the same driving current). Rotor is actuated by the driving scheme of brushless direct current motor (BLDCM). Interaction between the magnetized rotor and the magnetic-conducted stator produces a sinusoidal rotor restoring torque, amplitude of which is proportional to the rotor deflection angle inherently. Utilization of this magnetic restoring effect avoids adding of a high amplitude voltage for electrostatic feedback, which may cause air breakdown. Two differential capacitance pairs are utilized to measure input angular speeds at perpendicular directions of the rotor plane. The bias stability of the fabricated gyroscope is as low as 0.5°/h.

14.
Anal Bioanal Chem ; 408(11): 2963-73, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26780709

RESUMO

Serotonin is an important neurotransmitter that regulates a wide range of physiological, neuropsychological, and behavioral processes. Consequently, serotonin deficiency is involved in a wide variety of neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, schizophrenia, and depression. The pathophysiological mechanisms underlying serotonin deficiency, particularly from a lipidomics perspective, remain poorly understood. This study therefore aimed to identify novel lipid biomarkers associated with serotonin deficiency by lipidomic profiling of tryptophan hydroxylase 2 knockout (Tph2-/-) mice. Using a high-throughput normal-/reversed-phase two-dimensional liquid chromatography-quadrupole time-of-flight mass spectrometry (NP/RP 2D LC-QToF-MS) method, 59 lipid biomarkers encompassing glycerophospholipids (glycerophosphocholines, lysoglycerophosphocholines, glycerophosphoethanolamines, lysoglycerophosphoethanolamines glycerophosphoinositols, and lysoglycerophosphoinositols), sphingolipids (sphingomyelins, ceramides, galactosylceramides, glucosylceramides, and lactosylceramides) and free fatty acids were identified. Systemic oxidative stress in the Tph2-/- mice was significantly elevated, and a corresponding mechanism that relates the lipidomic findings has been proposed. In summary, this work provides preliminary findings that lipid metabolism is implicated in serotonin deficiency.


Assuntos
Biomarcadores/metabolismo , Lipídeos/química , Serotonina/metabolismo , Triptofano Hidroxilase/metabolismo , Animais , Camundongos , Camundongos Knockout , Estresse Oxidativo , Triptofano Hidroxilase/genética
15.
J Sep Sci ; 39(20): 3964-3972, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27542351

RESUMO

Fatty acids, which are well-known for their influence on human metabolism and signal transduction, are also a substantial component of cellular membranes and regulate the basic properties and functions of membranes. Owing to their multiple functions, fatty acid profiles of cell membranes are of great interest to those who are studying the relationship between membrane biochemical compositions and functions. A HCl-catalyzed derivation method and a gas chromatography with mass spectrometry analysis method were developed to accurately profile the fatty acids in cell membranes of erythrocytes, leukocytes, and platelets. The detection limits of all 35 fatty acids ranged from 0.58 to 22 ng/mL and the limits of quantitation were between 2.1 and 72 ng/mL. Finally, the established method was used to profile the membrane fatty acids of 44 healthy volunteers from the north and south of China. Results revealed significant differences in the fatty acid profiles from the two regions, particularly those of the erythrocytes. This technique may be applied to cell membrane studies to generate new biological hypotheses concerning fatty acid composition and membrane functions as well as to construct related disease profiles.


Assuntos
Membrana Eritrocítica/química , Ácidos Graxos/análise , Cromatografia Gasosa-Espectrometria de Massas , China , Humanos
17.
Rapid Commun Mass Spectrom ; 29(16): 1491-500, 2015 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-26212164

RESUMO

RATIONALE: Although serotonin deficiency is involved with various physiological disorders such as Alzheimer's disease, Parkinson's disease, schizophrenia and depression, the serotonin-dependent pathomechanisms remain poorly understood, particularly from a lipidomics perspective. METHODS: This study therefore aimed to identify novel lipid biomarkers associated with serotonin deficiency by lipid profiling of p-chlorophenylalanine (pCPA)-treated, serotonin-deficient mice using continuous-flow normal-phase/reversed-phase two-dimensional liquid chromatography/quadrupole time-of-flight mass spectrometry (NP/RP 2D LC/QTOFMS). Principal component analysis (PCA) was performed to distinguish significantly altered lipids between the pCPA-treated mice and control mice. RESULTS: Eighteen lipid biomarkers were associated with pCPA-induced serotonin deficiency. Specifically, lipid species of lysophosphatidylethanolamine (LPE), phosphatidylethanolamine (PE), sphingomyelin (SM), galactosylceramide (GalCer), glucotosylceramide (GluCer), lactosylceramide (LacCer) and triacylglycerol (TG) were down-regulated whereas glycerophosphocholine (PC) and phosphatidylinositol (PI) were up-regulated in the pCPA-treated mice compared with control mice. CONCLUSIONS: This work demonstrates the significant effects of serotonin deficiency on lipid metabolisms and will facilitate improved understanding of pathomechanisms in serotonin deficiency, particularly from a lipidomics perspective.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Fenclonina , Metabolismo dos Lipídeos/efeitos dos fármacos , Lipídeos/sangue , Serotonina , Espectrometria de Massas em Tandem/métodos , Animais , Biomarcadores/análise , Fenclonina/efeitos adversos , Fenclonina/farmacologia , Lipídeos/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Análise de Componente Principal , Serotonina/deficiência , Serotonina/metabolismo
18.
J Agric Food Chem ; 72(17): 10117-10126, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38631034

RESUMO

Storage is important for the garlic cloves industry because it is critical to enabling a year-round supply. This study aimed to investigate the changes in biochemical and metabolic profiles in garlic cloves in terms of different temperatures and cultivars during storage using nontargeted and targeted metabolomics. The results showed that the storage temperatures and times were important factors affecting the composition and metabolite content of garlic cloves. In detail, the metabolic profiling of garlic cloves changed significantly at 22 °C, which was mainly related to sprouting. Furthermore, γ-glutamyl peptide was converted into the corresponding flavor precursors or free amino acids, leading to the fluctuation in the amount of nutrients in garlic cloves. In contrast, the quality of garlic cloves remained stable for 290 days at 0 °C though metabolism still occurred, which indicated that the slight chemical changes did not impact the quality significantly and low temperature could prolong their dormancy.


Assuntos
Armazenamento de Alimentos , Alho , Alho/química , Alho/metabolismo , Temperatura , Aminoácidos/metabolismo , Aminoácidos/análise , Metabolômica
19.
Environ Pollut ; 342: 123066, 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38048871

RESUMO

While traditional culture-dependent methods can effectively detect certain microorganisms, the comprehensive composition of the municipal drinking water (DW) microbiome, including bacteria, archaea, and viruses, remains unknown. Metagenomic sequencing has opened the door to accurately determine and analyze the entire microbial community of DW, providing a comprehensive understanding of DW species diversity, especially in the context of public health concerns during the COVID-19 era. In this study, we found that most of the culturable bacteria and some fecal indicator bacteria, such as Escherichia coli and Pseudomonas aeruginosa, were non-culturable using culture-dependent methods in all samples. However, metagenomic analysis showed that the predominant bacterial species in the DW samples belonged to the phyla Proteobacteria and Planctomycetes. Notably, the genus Methylobacterium was the most abundant in all water samples, followed by Sphingomonas, Gemmata, and Azospirilum. While low levels of virulence-associated factors, such as the Esx-5 type VII secretion system (T7SS) and DevR/S, were detected, only the erythromycin resistance gene erm(X), an rRNA methyltransferase, was identified at low abundance in one sample. Hosts corresponding to virulence and resistance genes were identified in some samples, including Mycobacterium spp. Archaeal DNA (Euryarchaeota, Crenarchaeota) was found in trace amounts in some DW samples. Viruses such as rotavirus, coxsackievirus, human enterovirus, and SARS-CoV-2 were negative in all DW samples using colloidal gold and real-time reverse transcription polymerase chain reaction (RT‒PCR) methods. However, DNA encoding a new order of reverse-transcribing viruses (Ortervirales) and Herpesvirales was found in some DW samples. The metabolic pathways of the entire microbial community involve cell‒cell communication and signal secretion, contributing to cooperation between different microbial populations in the water. This study provides insight into the microbial community and metabolic process of DW in Hangzhou, China, utilizing both culture-dependent methods and metagenomic sequencing combined with bioinformatics tools during the COVID-19 pandemic era.


Assuntos
Água Potável , Microbiota , Humanos , Água Potável/microbiologia , Pandemias , Bactérias/genética , Archaea , Microbiota/genética , RNA Ribossômico 16S/genética
20.
Chemosphere ; 364: 142999, 2024 Aug 02.
Artigo em Inglês | MEDLINE | ID: mdl-39097107

RESUMO

Organochlorine pesticides (OCPs) are still occurring in various foodstuffs after the ban on their use. However, it remains unclear concerning the contamination source of OCPs in livestock and poultry food products and associated health risks. To fill this gap, we characterized the residual levels of 19 OCPs in multiple types of meats and eggs, which were sampled across China within the same period. Dichlorodiphenyltrichloroethanes (DDTs) were dominant in eggs, with the mean levels being 0.76 and 2.03 µg/kg for chicken eggs and duck eggs, respectively. By contrast, hexachlorocyclohexanes (HCHs) were the top one OCP in beef and lamb, with its mean levels being 0.51 and 0.65 µg/kg, respectively. Hexachlorobenzene (HCB) was rather detected in the poultry products. The componential ratio analysis implicated recent inputs of several banned OCPs including technical HCH and DDT, HCB and aldrin in multiple regions, which may origin from local industrial activities or possible illegal use. Risk assessment based on the risk quotient method suggested that daily consumption of cooked meats and eggs contaminated by dieldrin may pose a carcinogenic risk in adult residents of Jiangsu province. We concluded that OCPs remain present in meats and eggs at levels of health concern regionally in China.

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