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PURPOSE: Vitamin D (VD) is a vitamin with anti-inflammatory and immunomodulatory effects. Several studies have shown that VD deficiency is associated with the incidence and severity of dry eye disease (DED) in humans. This meta-analysis compared signs and symptoms in patients with DED before and after VD supplementation. METHODS: Ten databases (PubMed, Cochrane, Web of science, Ovid, Embase, Scopus, CNKI, CSTJ, Wanfang, Sinomed) and four clinical trials registry websites (ISRCTN, CT, ICTRP, ICH GCP) were searched for studies on the efficacy of VD supplement in patients with DED that were published up to 2023 January. A total of 8 studies with 439 cases were included. Two reviewers independently assessed the quality of the literature and extracted data to calculate standard mean differences (SMD) for the Schirmer's test (SH test), tear film break-up time (TBUT), corneal fluorescein staining scores (CFSS, Oxford Scale, 0-5), lid hyperemia, ocular surface disease index (OSDI), and visual analogue pain scale (VAS). RESULTS: The analysis showed that VD supplementation improved tear production (SMD 1.43, 95% CI 0.81, 2.05, p = 0.000) and tear film stability (SMD 1.19, 95% CI 0.83, 1.55, p = 0.000) and reduced lid hyperemia (SMD -0.71, 95% CI -1.09, -0.32, p = 0.000) and OSDI (SMD -1.10, 95% CI -1.45, -0.74, p = 0.000), which is effective in alleviating the signs and symptoms of DED. CONCLUSIONS: VD supplementation could be an adjuvant treatment to alleviate the signs and symptoms of dry eye in the future. However, further studies regarding its most appropriate administration and dosage need to be conducted.
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3-Methylhistidine (3-MeHis) is increasingly used as an indicator of muscle protein breakdown. The development of a sensitive, simple, and non-invasive method for 3-MeHis assay is important in clinical practice. Herein, a sensitive, simple, and non-invasive electrogenerated chemiluminescence (ECL) method was proposed for the quantitation of 3-MeHis in urine by using an iridium(III) solvent complex ([Ir(dfppy)2(DMSO)Cl], dfppy = 2-(2,4-difluorophenyl)pyridine, Ir-DMSO) as a signal reagent. The photoluminescence (PL) and ECL responses of Ir-DMSO to 3-MeHis were studied. The ECL intensity of Ir-DMSO was enhanced in the presence of 3-MeHis because of the coordination recognition between Ir-DMSO and the imidazole group of 3-MeHis. Based on the enhancement of ECL intensity, 3-MeHis can be sensitively detected in the range of 5 to 25 µM. The detection limit was 0.4 µM. This is the first report of an ECL method for the quantitation of 3-MeHis. Further, to investigate the feasibility of the Ir-DMSO-based ECL method in practical applications, the developed ECL method was applied for 3-MeHis assay in urine samples of 28 healthy volunteers and 2 patients. The urine samples from patients hospitalized with obesity and kidney disease and healthy individuals were distinguished by the ECL responses of Ir-DMSO. The proposed ECL method based on the coordination recognition between iridium(III) solvent complex and the imidazole group of 3-MeHis allows inexpensive, fast, non-invasive, and sensitive detection of 3-MeHis in urine, which is promising for assessing large volumes of patients for routine analysis in clinical practices.
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Irídio , Limite de Detecção , Medições Luminescentes , Metilistidinas , Solventes , Humanos , Irídio/química , Medições Luminescentes/métodos , Metilistidinas/urina , Solventes/química , Complexos de Coordenação/química , Masculino , Técnicas Eletroquímicas/métodosRESUMO
Exosomes, as an emerging biomarker, have exhibited remarkable promise in early cancer diagnosis. Here, a highly sensitive, selective, and automatic electrochemiluminescence (ECL) method for the detection of cancerous exosomes was developed. Specific aptamer-(EK)4 peptide-tagged magnetic beads (MBs-(EK)4-aptamer) were designed as a magnetic capture probe in which the (EK)4 peptide was used to reduce the steric binding hindrance of cancerous exosomes with a specific aptamer. One new universal ECL signal nanoprobe (CD9 Ab-PEG@SiO2ϵRu(bpy)32+) was designed and synthesized by using microporous SiO2 nanoparticles as the carrier for loading ECL reagent Ru(bpy)32+, polyethylene glycol (PEG) layer, and anticluster of differentiation 9 antibody (CD9 Ab). A "sandwich" biocomplex was formed on the surface of the magnetic capture probe after mixing the capture probe, target exosomes, and ECL signal nanoprobe, and then it was introduced into an automated ECL analyzer for rapid and automatic ECL measurement. It was found that the designed signal nanoprobe shows a 270-fold improvement in the signal-to-noise ratio than that of the ruthenium complex-labeled CD9 antibody signal probe. The relative ECL intensity was proportional to MCF-7 exosomes as a model in the range of 102 to 104 particle/µL, with a detection limit of 11 particle/µL. Furthermore, the ECL method was employed to discriminate cancerous exosomes based on fingerprint responses using the designed multiple magnetic capture probes and the universal ECL signal nanoprobe. This work demonstrates that the utilization of a designed automated ECL tactic using the MBs-(EK)4-aptamer capture probe and the CD9 Ab-PEG@SiO2ϵRu(bpy)32+ signal nanoprobe will provide a unique and robust method for the detection and discrimination of cancerous exosomes.
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Aptâmeros de Nucleotídeos , Técnicas Eletroquímicas , Exossomos , Medições Luminescentes , Humanos , Exossomos/química , Aptâmeros de Nucleotídeos/química , Técnicas Eletroquímicas/métodos , Medições Luminescentes/métodos , Células MCF-7 , Dióxido de Silício/química , Técnicas Biossensoriais/métodos , Tetraspanina 29/análise , Polietilenoglicóis/químicaRESUMO
Herpes simplex keratitis (HSK) is a blinding disease caused by herpes simplex virus type 1 (HSV-1) infection, and rapid eradication of the virus from the affected cornea is imperative. Nod-like receptors (NLRs) are intracellular innate immune sensors closely associated with cell death, inflammation and immune responses. In this study, we investigated the role of NLRP12 in the antiviral immunology in HSK and the underlying mechanisms. We found that NLRP12 expression was significantly decreased in HSV-1-infected human corneal epithelial cells (HCE-Ts) and HSK mouse corneas. Overexpression of NLRP12 significantly reduced viral replication in infected HCE-Ts and functioned through inflammasome-mediated pyroptosis and downstream IL-18-IFN-γ axis. In HSK mouse models, overexpression of NLRP12 reduced viral replication in the cornea and alleviated HSK symptoms. This resulted from enhanced antiviral immune responses including the activation of specific immune cells in both the cornea and the draining lymph nodes. Specifically, the NLRP12-IL-18-IFN-γ axis regulated the interaction between infected corneal epithelial cells and macrophages. In conclusion, our study identified a role of NLRP12 in mediating pyroptosis and regulating antiviral immune responses. This novel finding opens the possibilities of NLRP12 as a viable target in the therapeutic strategies for HSV-1 infection.
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Herpesvirus Humano 1 , Interferon gama , Interleucina-18 , Ceratite Herpética , Camundongos Endogâmicos C57BL , Piroptose , Transdução de Sinais , Animais , Ceratite Herpética/imunologia , Ceratite Herpética/virologia , Humanos , Interleucina-18/metabolismo , Interleucina-18/imunologia , Interferon gama/metabolismo , Interferon gama/imunologia , Herpesvirus Humano 1/imunologia , Herpesvirus Humano 1/fisiologia , Camundongos , Córnea/virologia , Córnea/imunologia , Córnea/patologia , Feminino , Replicação Viral , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Peptídeos e Proteínas de Sinalização Intracelular/genética , Modelos Animais de Doenças , Inflamassomos/metabolismo , Inflamassomos/imunologia , Imunidade InataRESUMO
Cysteine (Cys), as one of the biological thiols, is related to many physiological and pathological processes in humans and plants. Therefore, it is necessary to develop a sensitive and selective method for the detection and imaging of Cys in biological organisms. In this work, a novel near-infrared (NIR) fluorescent probe, Probe-Cys, was designed by connecting furancarbonyl, as a new recognition moiety, with Fluorophore-OH via the decomposition of IR-806. The use of the furan moiety is anticipated to produce more effective fluorescence quenching because of the electron-donating ability of the O atom. Probe-Cys has outstanding properties, such as a new recognition group, an emission wavelength in the infrared region at 710 nm, a linear range (0-100 µM), a low detection limit of 0.035 µM, good water solubility, excellent sensitivity, and selectivity without the interference of Hcy, GSH, and HS-. More importantly, Probe-Cys could achieve the detection of endogenous Cys by reacting with the stimulant 1,4-dimercaptothreitol (DTT) and the inhibitor N-ethylmaleimide (NEM) in HepG2 cells and zebrafish. Ultimately, it was successfully applied to obtain images of Arabidopsis thaliana, revealing that the content of Cys in the meristematic zone was higher than that in the elongation zone, which was the first time that the NIR fluorescence probe was used to obtain images of Cys in A. thaliana. The superior properties of the probe exhibit its great potential for use in biosystems to explore the physiological and pathological processes associated with Cys.
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Arabidopsis , Perciformes , Humanos , Animais , Fluorescência , Peixe-Zebra , Cisteína , Células HeLa , Corantes Fluorescentes , GlutationaRESUMO
A highly sensitive and selective electrogenerated chemiluminescence (ECL) biosensor was developed for the determination of matrix metalloproteinase 3 (MMP-3) in serum via the target-induced cleavage of an oligopeptide. One ECL probe (named as Ir-peptide) was synthesized by covalently linking a new cyclometalated iridium(III) complex ([(3-pba)2Ir(bpy-COOH)](PF6)) (3-pba = 3-(2-pyridyl) benzaldehyde, bpy-COOH = 4'-methyl-2,2'-bipyridine-4-carboxylic acid) with an oligopeptide (CGVPLSLTMGKGGK). An ECL biosensor was fabricated by firstly casting Nafion and gold nanoparticles (AuNPs) on a glassy carbon electrode and then self-assembling both of the ECL probes, 6-mercapto-1-hexanol and zwitterionic peptide, on the electrode surface, from which the AuNPs could be used to amplify the ECL signal and Ir-peptide could serve as an ECL probe to detect the MMP-3. Thanks to the MMP-3-induced cleavage of the oligopeptide contributing to the decrease in ECL intensity and the amplification of the ECL signal using AuNPs, the ECL biosensor could selectively and sensitively quantify MMP-3 in the concentration range of 10-150 ng·mL-1 and with both a limit of quantification (26.7 ng·mL-1) and a limit of detection (8.0 ng·mL-1) via one-step recognition. In addition, the developed ECL biosensor showed good performance in the quantization of MMP-3 in serum samples, with a recovery of 92.6% ± 2.8%-105.6% ± 5.0%. An increased level of MMP-3 was found in the serum of rheumatoid arthritis patients compared with that of healthy people. This work provides a sensitive and selective biosensing method for the detection of MMP-3 in human serum, which is promising in the identification of patients with rheumatoid arthritis.
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Técnicas Biossensoriais , Ouro , Medições Luminescentes , Metaloproteinase 3 da Matriz , Nanopartículas Metálicas , Oligopeptídeos , Humanos , Metaloproteinase 3 da Matriz/sangue , Ouro/química , Nanopartículas Metálicas/química , Luminescência , Limite de Detecção , Eletrodos , Técnicas EletroquímicasRESUMO
An enzyme immunoassay was developed based on the coulometric measurement of immunoglobulin M (IgM) against Hantaan viruses (HTNV) by using virus-like particles (VLPs) as recognition molecules. The surface functionalization of screen-printed carbon electrodes (SPCEs) was achieved through paste-exfoliated graphene that was modified with a COOH group and a thionine mediator through supramolecular-covalent scaffolds, on SPCEs by using the binder contained in the ink. After the covalent immobilization of the antibody, the sensor was used for the sandwich enzyme immunoassay of IgM against HTNV. By using HTNV VLPs as the second recognization molecules, the resulting sensor efficiently monitored the reaction of IgM against HTNV and anti-IgM antibody with high specificity. By attaching HTNV nucleocapsid protein antibody conjugate with horseradish peroxidase (HRP) onto VLPs, the signal response of the assay was derived from the coulometric measurement of H2O2 reduction mediated by thionine on the electrode surface after the application of a potential (- 0.2 V vs. Ag/AgCl). The ratio of charges measured before or after H2O2 addition was used to quantify IgM because these charges could be used as background charges or total charges, respectively. The ratio exhibited good agreement with IgM concentration within a range 0.1 to 1000 pg mL-1, and a detection limit of 0.06 pg mL-1 was obtained. The assay demonstrated high sensitivity and specificity toward HTNV-specific IgM in serum.
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Técnicas Biossensoriais , Grafite , Fenotiazinas , Grafite/química , Carbono/química , Imunoensaio/métodos , Técnicas Biossensoriais/métodos , Peróxido de Hidrogênio/química , Imunoglobulina M , EletrodosRESUMO
A new fluorescence probe OHPD that could specifically identify acetylcholinesterase/butyrylcholinesterase has been developed and successfully applied to imaging in vivo. Probe OHPD shows significant color change, high selectivity, high sensitivity, and low detection limit for the detection of cholinesterase. Moreover, the real-time imaging in situ indicated that endogenous cholinesterase was mainly present in the yolk sac of zebrafish.
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Acetilcolinesterase , Butirilcolinesterase , Animais , Corantes Fluorescentes , Peixe-Zebra , Inibidores da Colinesterase/farmacologiaRESUMO
Histidine (His) and its metabolite analysis is significant due to their vital roles in the diagnosis of diseases. In practical applications, simple and effective detection and discrimination of these metabolic species are still a great challenge due to their highly similar structures. Herein, photoluminescence (PL)-electrochemiluminescence (ECL) dual-mode sensor arrays consisting of a series of sensing elements were proposed for simultaneous quantitation and accurate discrimination of His and its four key metabolites (including histamine, imidazole-4-acetic acid, N-acetylhistamine, and imidazole propionate). The sensing elements of these sensor arrays were constructed by employing two solvent iridium(III) complexes ([Ir(pbz)2(DMSO)Cl] and [Ir(ppy)2(DMSO)Cl], pbz = 3-(2-pyridyl)benzoic acid, ppy = 2-phenylpyridine) with excellent PL and ECL performances as cross-responsive sensing units. Based on diverse coordination abilities of the two complexes with the imidazole group of the five targets, PL and ECL responses of each sensing unit can be enhanced to various degrees, which generate unique fingerprint patterns for the corresponding targets. Through principal component analysis, the multifarious patterns (two-, three-, and four-element sensor arrays) can be transformed into simple visualization modes, from which His and its four key metabolites can be effectively discriminated against each other. Moreover, the quantitation of an individual metabolic species at different concentrations and the recognition of the mixtures with different ratios were also accurately achieved. Notably, His and its four key metabolites in urine can also be successfully discriminated by the as-fabricated sensor arrays, and the patients with kidney diseases can be identified clearly, providing a promising way for disease diagnosis.
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Dimetil Sulfóxido , Histidina , Humanos , Fotometria , Medições LuminescentesRESUMO
Numerous microorganisms, including bacteria and fungus, have been identified as capable of degrading rubber. Rubber biodegradation is still understudied due to its high stability and the lack of well-defined pathways and efficient enzymes involved in microorganism metabolism. However, rubber products manufacture and usage cause substantial environmental issues, and present physical-chemical methods involve dangerous chemical solvents, massive energy, and trash with health hazards. Eco-friendly solutions are required in this context, and biotechnological rubber treatment offers considerable promise. The structural and functional enzymes involved in poly (cis-1,4-isoprene) rubber and their cleavage mechanisms have been extensively studied. Similarly, novel bacterial strains capable of degrading polymers have been investigated. In contrast, relatively few studies have been conducted to establish natural rubber (NR) degrading bacterial consortia based on metagenomics, considering process optimization, cost effective approaches and larger scale experiments seeking practical and realistic applications. In light of the obstacles encountered during the constructing NR-degrading consortia, this study proposes the utilization of multi-omics tools to discern the underlying mechanisms and metabolites of rubber degradation, as well as associated enzymes and effective synthesized microbial consortia. In addition, the utilization of omics tool-based methods is suggested as a primary research direction for the development of synthesized microbial consortia in the future.
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A novel point-of-care testing (POCT) method for the determination of proteases was developed for the first time using a designed disposable capillary-fill device based on the cleavage of electrogenerated chemiluminescence (ECL)-label-tagged peptide probes and enabling elimination of the light-shielding from the magnetic beads (MBs). As a proof-of-principle, prostate-specific antigen (PSA) was taken as a model analyte, and streptavidin-coated magnetic beads bound with ruthenium-complex-tagged specific peptide (biotin-HSSKLQK) were utilized as MB ECL probes. The capillary-fill device was designed to be divided into a reaction zone and detection zone. In the reaction zone, the bio-cleavage reaction between the PSA analyte with the peptide on the surface of the MB ECL probes occurred, while in the detection zone, ECL emission was produced by a screen-printed carbon electrode, Ag/AgCl reference electrode and carbon counter electrode. When the analyte PSA was introduced into the suspension of MB ECL probes in the reaction zone of the device, biocleavage of the peptide occurred, and the cleaved Ru1 part was released from the surface of the MB ECL probes. The capillary-filled device was tilted 90°, and with the aid of gravity, the solution containing the released Ru1 part flowed to the surface of the working electrode in the detection region of the device, while the MB ECL probes were fixed in the reaction zone by an external magnet. PSA can be determined by the ECL emission from the released Ru1 part in the presence of the co-reactant tri-n-propylamine at the detection zone. Under the optimal conditions, the developed ECL method showed a low detection limit of 0.12 ng mL-1 for PSA. This work demonstrates that the developed ECL biosensing approach can eliminate the MB light-shielding effect and quantify proteases with high sensitivity and selectivity, which could be easily extended to POCT-based ECL biosensing for other proteases.
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Técnicas Biossensoriais , Peptídeo Hidrolases , Humanos , Masculino , Antígeno Prostático Específico , Luminescência , Peptídeos , Carbono , Medições Luminescentes/métodos , Técnicas Biossensoriais/métodosRESUMO
An electrochemiluminescence (ECL) bioassay with high sensitivity and anti-fouling ability was developed for determination of matrix metalloproteinase 9 (MMP-9) secreted from living cells under external stimulation. A peptide with sequence of CLGRMGLPGK and a new cyclometalated iridium(III) complex bearing carboxyl group, (pq)2Ir(dcbpy) (pq = 2-phenylquinoline, dcbpy = 2,2'-bipyridyl-4,4'-dicarboxyli acid, abbreviated as Ir) were employed as molecular recognition substrate and ECL emitter, respectively. The peptide was labelled with the Ir to form Ir-peptide as ECL probe. Ir-peptide was self-assembled onto Nafion and gold nanoparticles (AuNPs) modified glassy carbon electrode (AuNPs/Nafion/GCE) and then both of 6-mercapto-1-hexanol (MCH) and zwitterionic peptide as blocking reagents were co-assembled on Ir-peptide/AuNPs/Nafion/GCE to form an anti-fouling ECL peptide-based biosensor. MMP-9 can be quantified in the range 1.0-50 ng·mL-1 with a detection limit of 0.50 ng·mL-1 based on the decreased ECL intensity. Relative standard derivation was 2.3% for six fabricated anti-fouling ECL peptide-based biosensors after reaction with 50 ng·mL-1 MMP-9. The anti-fouling ECL peptide-based biosensor can be used to monitor MMP-9 secreted from living cells under external stimulation. 96.0%-108.0% of recoveries were obtained in 60-diluted cell culture media. This study demonstrates that the ECL biosensor by the combination of iridium(III) complex-based sensitive ECL method and the anti-fouling interface provides a promising way for the determination of MMP-9 in biological sample, which is viable in clinical diagnosis and point-of-care test of protease.
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Incrustação Biológica , Nanopartículas Metálicas , Ouro/química , Metaloproteinase 9 da Matriz , Irídio , Incrustação Biológica/prevenção & controle , Medições Luminescentes/métodos , Nanopartículas Metálicas/química , Peptídeos/químicaRESUMO
We propose a novel washing-free electrochemiluminescence (ECL) biosensor for the simultaneous detection of two types of N6 methyladenosines-RNAs (m6A-RNAs), which are potential cancer biomarkers, on the basis of binding-induced DNA strand displacement (BINSD). The biosensor integrated a tri-double resolution strategy that combined spatial and potential resolution, hybridization and antibody recognition, and ECL luminescence and quenching. The biosensor was fabricated by separately immobilizing two ECL reagents (gold nanoparticles/g-C3N4 nanosheets and ruthenium bipyridine derivative/gold nanoparticles/Nafion) and the capture DNA probe on the two sections of glassy carbon electrode. As a proof of concept, m6A-Let-7a-5p and m6A-miR-17-5p were chosen as model analytes, while m6A antibody-DNA3/ferrocene-DNA4/ferrocene-DNA5 was designed as an m6A-binding probe and DNA6/DNA7 was designed as a hybridization probe with DNA3 to release the quenching probes ferrocene-DNA4/ferrocene-DNA5. The recognition process led to the quenching of the ECL signals from both probes via BINSD. The proposed biosensor has the advantage of being washing-free. The ECL methods using the fabricated ECL biosensor with the designed probes exhibited a low detection limit of 0.03 pM for two m6A-RNAs and high selectivity. This work reveals that this strategy is promising for developing an ECL method for the simultaneous detection of two m6A-RNAs. The proposed strategy could be expanded to develop the analytical methods for the simultaneous detection of other RNA modifications by changing the antibody and hybridization probe sequences.
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Técnicas Biossensoriais , Nanopartículas Metálicas , Metalocenos , Ouro , Medições Luminescentes/métodos , Técnicas Biossensoriais/métodos , DNA/genética , AnticorposRESUMO
A sensitive and noninvasive cyclic peptide-based electrogenerated chemiluminescence biosensing method for the determination of sweat glucose was developed. Glucose can be quantified in sweat samples with a recovery of 93%-113% via one-step recognition, which is promising for the determination of sweat glucose.
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Técnicas Biossensoriais , Peptídeos Cíclicos , Suor , Glucose , Luminescência , Técnicas Biossensoriais/métodos , Medições Luminescentes/métodosRESUMO
A polyphasic taxonomic study was conducted on two Gram-negative, non-sporulating, non-motile bacterial strains, S2-20-2T and S2-21-1, isolated from a contaminated freshwater sediment in China. Comparative 16S rRNA gene sequence studies revealed a clear affiliation of two strains with Bacteroidetes, which showed the highest pairwise sequence similarities with Hymenobacter duratus BT646T (99.3%), Hymenobacter psychrotolerans Tibet-IIU11T (99.3%), Hymenobacter kanuolensis T-3T (97.6%), Hymenobacter swuensis DY53T (96.9%), Hymenobacter tenuis POB6T (96.8%), Hymenobacter seoulensis 16F7GT (96.7%), and Hymenobacter rigui KCTC 12533T (96.5%). The phylogenetic analysis based on 16S rRNA gene sequences showed that two strains formed a clear phylogenetic lineage with the genus Hymenobacter. Major fatty acids were identified as iso-C15:0, anteiso-C15:0, and summed feature 3 (C16:1 ω6c and/or C16:1 ω7c/t) and summed feature 4 (iso-C17:1 I and/or anteiso-C17:1 B). Major cellular polar lipids were identified as phosphatidylethanolamine, three unidentified aminolipids, an unidentified aminophosopholipid and an unidentified lipid. The respiratory quinone was detected as MK-7 and the genomic DNA G + C content was determined to be 57.9% (genome) for type strain S2-20-2T and 57.7 mol% (HPLC) for strain S2-21-1. The observed ANI and dDDH values between strain S2-20-2T and its closely related strains were 75.7-91.4% and 21.2-43.9%, respectively. Based on physiological, biochemical, genetic and genomic characteristics, we propose that strains S2-20-2T and S2-21-1 represent a novel species of the genus Hymenobacter, for which the name Hymenobacter sediminicola sp. nov. is proposed. The type strain is S2-20-2T (= CGMCC 1.18734T = JCM 35801T).
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Cytophagaceae , Ácidos Graxos , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ácidos Graxos/análise , DNA Bacteriano/genética , DNA Bacteriano/química , Técnicas de Tipagem Bacteriana , Vitamina K 2/químicaRESUMO
Numerous microorganisms and other invertebrates that are able to degrade polyethylene (PE) have been reported. However, studies on PE biodegradation are still limited due to its extreme stability and the lack of explicit insights into the mechanisms and efficient enzymes involved in its metabolism by microorganisms. In this review, current studies of PE biodegradation, including the fundamental stages, important microorganisms and enzymes, and functional microbial consortia, were examined. Considering the bottlenecks in the construction of PE-degrading consortia, a combination of top-down and bottom-up approaches is proposed to identify the mechanisms and metabolites of PE degradation, related enzymes, and efficient synthetic microbial consortia. In addition, the exploration of the plastisphere based on omics tools is proposed as a future principal research direction for the construction of synthetic microbial consortia for PE degradation. Combining chemical and biological upcycling processes for PE waste could be widely applied in various fields to promote a sustainable environment.
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The understanding of the dissolution processes of solids is important for the design and synthesis of solids in a controlled and precise manner and for predicting their fate in the aquatic environment. We report herein single-particle-based confocal laser scanning microscopy (CLSM) for tracking the dissolution surface kinetics of a single fluorescent cyclodextrin metal-organic framework (CD-MOF). As a proof of concept, CD-MOF containing fluorescein, named as CD-MOFâFL, was synthesized by encapsulating fluorescein into the interior of CD-MOF via a vapor diffusion method and used as a single-particle dissolution model because of its high FL efficiency and unique structure. The morphology of CD-MOFâFL and the distribution of fluorescein within CD-MOFâFL were characterized. The growth and dissolution processes of CD-MOFâFL at the single-particle level were visualized and quantified for the first time by recording the change of the fluorescence emission. Three processes, including nucleation, germination growth, and saturation stage, were found in the growth of CD-MOFâFL, and the growth kinetics followed Avrami's model. The dissolution rate at the face of a single CD-MOFâFL crystal was slower than that of its arris, and the dissolution rate of the CD-MOFâFL crystal was increased with the increase of the water amount in methanol solution. The dissolution process of the CD-MOFâFL crystal was a competitive process of erosion and diffusion in different methanol aqueous solutions, and the dissolution kinetics followed the Korsmeyer-Peppas model. These results offer new insights into the nature of dissolution kinetics of CD-MOFâFL and provide new venues for the quantitative analysis of solid dissolution and growth at the single-particle level.
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This work presents the role of commercial microfiltration membranes combined with single particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) in removing environmental matrix interference for model silver nanoparticles (AgNPs) determination. The filters with different pore sizes (0.22 µm, 0.45 µm, 0.8 µm) and materials (mixed cellulose ester, polyether sulfone, and nylon) were investigated to acquire the recovery of particle concentration and size of AgNPs spiked into different real aqueous solutions, including ultrapure water, tap water, surface water, and sewage effluent. The maximum recovery of nanoparticle concentration was 70.2% through the 0.8 µm polyether sulfone membrane. The heated filters were able to improve the recovery of AgNPs particle concentration in the real aqueous environment. Hence, the pretreatment method by SP-ICP-MS combined with filtration membrane was simple, fast, and low-cost to quantify AgNPs in natural water environments.
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This work reports the performance enhancement strategies on magnetic beads (MBs)-based electrochemiluminescence (ECL) platforms by using double magnetic field actuation of the ECL magnetic microbiosensors (MMbiosensors) for highly sensitive determination of cancer biomarker and exosomes. To obtain the high sensitivity and reproducibility of the ECL MMbiosensors, a series of strategies have been developed including replacing a conventional photomultiplier tube (PMT) with a diamagnetic PMT, replacing the stacked ring-disc magnets with circular-disc magnets lain-in glassy carbon electrode, adding a pre-concentration process of MBs using external magnet actuation. For fundamental research, the ECL MBs taken as the substitute of ECL MMbiosensors were prepared by binding biotinylated DNA tagged with Ru(bpy)32+ derivative (Ru1) to streptavidin-coated MB(MB@SA) were which showed that the developed strategies can enhance 45-fold sensitivity. Importantly, the developed MBs-based ECL platform was estimated by determination of prostate specific antigen (PSA) and exosomes. For PSA, MB@SAâ¢biotin-Ab1(PSA) was taken as the capture probe and Ru1-labeled Ab2 (PSA) was done as ECL probe, while for exosomes, MB@SAâ¢biotin-aptamer (CD63) was taken as the capture probe and Ru1-labeled Ab (CD9) was done as the ECL probe. The experiment results showed that the developed strategies can enhance 33-fold sensitivity of ECL MMbiosensors for PSA and exosomes. The detection limit is 0.28 ng mL-1 for PSA and 4.9 × 102 particle mL-1 for exosomes. This work demonstrated that a series of proposed magnetic field actuation strategies greatly increase the sensitivity of the ECL MMbiosensors. The developed strategies can be expanded to MBs-based ECL and electrochemical biosensors for clinical analysis with greater sensitivity.
Assuntos
Técnicas Biossensoriais , Exossomos , Neoplasias , Masculino , Humanos , Biomarcadores Tumorais , Antígeno Prostático Específico , Reprodutibilidade dos Testes , Técnicas Biossensoriais/métodos , Medições Luminescentes/métodos , Fenômenos Magnéticos , Campos Magnéticos , Neoplasias/diagnósticoRESUMO
This work reports washing-free electrogenerated chemiluminescence (ECL) magnetic microbiosensors based on target assistant proximity hybridization (TAPH) for multiple protein biomarkers for the first time. As a principle-of-proof, alpha-fetoprotein (AFP) was chosen as a model analyte, and biotin-DNA1 bound streptavidin-coated magnetic microbeads (MMB@SAâ biotin-DNA1) were designed as the universal capture MMB, while the corresponding two antibodies tagged with DNA2 or DNA3 were utilized as hybrid recognition probes, and ruthenium complex-tagged DNA4-10A was designed as a universal ECL signal probe. When the capture MMB was added into the mixture solution (containing the analyte, hybrid recognition probes, signal probe and tri-n-propylamine), biocomplexes were formed on the MMB. After the resulting MMB was efficiently brought to the surface of a magnetic glassy carbon electrode (MGCE), ECL measurement was performed without a washing step, resulting in an increase in the ECL intensity. A model for ECL measuring the second-order rate constants of hybridization reactions on MMB was derived. It was found that the rate constants for hybridization reactions on MMB in rotating mode are 1.6-fold higher than those in shaking mode, and a suitable DNA length of the signal probe can improve the signal-to-noise ratio. The washing-free ECL method was developed for the determination of AFP with a much lower detection limit (LOD) of 0.04 ng mL-1. The developed flexible strategy has been extended to determine D-dimer with an LOD of 0.1 ng mL-1 and myoglobinglobin with an LOD of 1.1 ng mL-1. This work demonstrated that the proposed strategy of ECL TAPH on MMB at MGCE is a washing-free and flexible promising strategy, and can be extended to qualify other multiple protein biomarkers in real clinical assays.