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Lytic polysaccharide monooxygenases (LPMOs) oxidatively depolymerize recalcitrant polysaccharides, which is important for biomass conversion. The catalytic domains of many LPMOs are linked to carbohydrate-binding modules (CBMs) through flexible linkers, but the function of these CBMs in LPMO catalysis is not well understood. In this study, we utilized MtLPMO9L and MtLPMO9G derived from Myceliophthora thermophila to investigate the impact of CBMs on LPMO activity, with particular emphasis on their influence on H2O2 tolerance. Using truncated forms of MtLPMO9G generated by removing the CBM, we found reduced substrate binding affinity and enzymatic activity. Conversely, when the CBM was fused to the C terminus of the single-domain MtLPMO9L to create MtLPMO9L-CBM, we observed a substantial improvement in substrate binding affinity, enzymatic activity, and notably, H2O2 tolerance. Furthermore, molecular dynamics simulations confirmed that the CBM fusion enhances the proximity of the active site to the substrate, thereby promoting multilocal cleavage and impacting the exposure of the copper active site to H2O2. Importantly, the fusion of CBM resulted in more efficient consumption of H2O2 by LPMO, leading to improved enzymatic activity and reduced auto-oxidative damage of the copper active center.
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Dominio Catalítico , Peróxido de Hidrógeno , Oxigenasas de Función Mixta , Polisacáridos , Sordariales , Cobre/metabolismo , Peróxido de Hidrógeno/efectos adversos , Peróxido de Hidrógeno/metabolismo , Oxigenasas de Función Mixta/metabolismo , Polisacáridos/metabolismo , Sordariales/enzimología , Sordariales/metabolismo , Simulación de Dinámica MolecularRESUMEN
BACKGROUND: Haemonchus contortus (H. contortus) is the most common parasitic nematode in ruminants and is prevalent worldwide. H. contortus resistance to albendazole (ABZ) hinders the efficacy of anthelmintic drugs, but little is known about the molecular mechanisms that regulate this of drug resistance. Recent research has demonstrated that long noncoding RNAs (lncRNAs) can exert significant influence as pivotal regulators of the emergence of drug resistance. RESULTS: In this study, transcriptome sequencing was conducted on both albendazole-sensitive (ABZ-sensitive) and albendazole-resistant (ABZ-resistant) H. contortus strains, with three biological replicates for each group. The analysis of lncRNA in the transcriptomic data revealed that there were 276 differentially expressed lncRNA (DElncRNA) between strains with ABZ-sensitive and ABZ-resistant according to the criteria of |log2Foldchange|≥ 1 and FDR < 0.05. Notably, MSTRG.12969.2 and MSTRG.9827.1 exhibited the most significant upregulation and downregulation, respectively, in the resistant strains. The potential roles of the DElncRNAs included catalytic activity, stimulus response, regulation of drug metabolism, and modulation of the immune response. Moreover, we investigated the interactions between DElncRNAs and other RNAs, specifically MSTRG.12741.1, MSTRG.11848.1, MSTRG.5895.1, and MSTRG.14070.1, involved in regulating drug stimulation through cis/trans/antisense/lncRNAâmiRNA-mRNA interaction networks. This regulation leads to a decrease (or increase) in the expression of relevant genes, consequently enhancing the resistance of H. contortus to albendazole. Furthermore, through comprehensive analysis of competitive endogenous RNAs (ceRNAs) involved in drug resistance-related pathways, such as the mTOR signalling pathway and ABC transporter signalling pathway, the relevance of the MSTRG.2499.1-novel-m0062-3p-HCON_00099610 interaction was identified to mainly involve the regulation of catalytic activity, metabolism, ubiquitination and transcriptional regulation of gene promoters. Additionally, quantitative real-time polymerase chain reaction (qRT-PCR) validation indicated that the transcription profiles of six DElncRNAs and six DEmRNAs were consistent with those obtained by RNA-seq. CONCLUSIONS: The results of the present study allowed us to better understand the changes in the lncRNA expression profile of ABZ-resistant H. contortus. In total, these results suggest that the lncRNAs MSTRG.963.1, MSTRG.12741.1, MSTRG.11848.1 and MSTRG.2499.1 play important roles in the development of ABZ resistance and can serve as promising biomarkers for further study.
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Antihelmínticos , Haemonchus , ARN Largo no Codificante , Animales , Albendazol/farmacología , Albendazol/análisis , Albendazol/metabolismo , Haemonchus/genética , ARN Largo no Codificante/genética , ARN Largo no Codificante/metabolismo , Transcriptoma , Antihelmínticos/farmacología , Antihelmínticos/metabolismo , Antihelmínticos/uso terapéuticoRESUMEN
HnRNP A/B belongs to the heterogeneous nuclear ribonucleoprotein (hnRNP) family and plays an important role in regulating viral protein translation and genome replication. Here, we found that overexpression of hnRNP A/B promoted spring viremia of carp virus (SVCV) and cyprinid herpesvirus 3 (CyHV3) replication. Further, hnRNP A/B was shown to act as a negative regulator of type I interferon (IFN) response. Mechanistically, hnRNP A/B interacted with MITA, TBK1 and IRF3 to initiate their degradation. In addition, hnRNP A/B bound to the kinase domain of TBK1, the C terminal domain of MITA and IAD domain of IRF3, and the RRM1 domain of hnRNP A/B bound to TBK1, RRM2 domain bound to IRF3 and MITA. Our study provides novel insights into the functions of hnRNP A/B in regulating host antiviral response.
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Enfermedades de los Peces , Proteínas de Peces , Proteínas Serina-Treonina Quinasas , Infecciones por Rhabdoviridae , Rhabdoviridae , Animales , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/virología , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Proteínas de Peces/metabolismo , Rhabdoviridae/fisiología , Infecciones por Rhabdoviridae/inmunología , Infecciones por Rhabdoviridae/veterinaria , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Serina-Treonina Quinasas/inmunología , Inmunidad Innata/genética , Factor 3 Regulador del Interferón/genética , Factor 3 Regulador del Interferón/metabolismo , Factor 3 Regulador del Interferón/inmunología , Carpas/inmunología , Carpas/genética , Herpesviridae/fisiología , Infecciones por Herpesviridae/veterinaria , Infecciones por Herpesviridae/inmunología , Interferón Tipo I/inmunología , Interferón Tipo I/genética , Interferón Tipo I/metabolismo , Proteínas de Pez CebraRESUMEN
DExD/H-box helicase (DDX) 5 belongs to the DExD/H-box helicase family. DDX family members play differential roles in the regulation of innate antiviral immune response. However, whether DDX5 is involved in antiviral immunity remains unclear. In this study, we found that DDX5 serves as a negative regulator of type I interferon (IFN) response. Overexpression of DDX5 inhibited IFN production induced by Spring viremia of carp virus (SVCV) and poly(I:C) and enhanced virus replication by targeting key elements of the RLR signaling pathway (MAVS, MITA, TBK1, IRF3 and IRF7). Mechanistically, DDX5 directly interacted with TBK1 to promote its autophagy-mediated degradation. Moreover, DDX5 was shown to block the interaction between TRAF3 and TBK1, hence preventing nuclear translocation of IRF3. Together, these data shed light on the roles of DDX5 in regulating IFN response.
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Interferón Tipo I , Proteínas Serina-Treonina Quinasas , Proteínas Serina-Treonina Quinasas/genética , Proteínas Serina-Treonina Quinasas/metabolismo , Factor 3 Asociado a Receptor de TNF/genética , Factor 3 Asociado a Receptor de TNF/metabolismo , Fosforilación , Diclorodifenil Dicloroetileno , Inmunidad Innata , Interferón Tipo I/metabolismo , Factor 3 Regulador del Interferón/genética , Factor 3 Regulador del Interferón/metabolismo , AntiviralesRESUMEN
In this study, 858 novel long non-coding RNAs (lncRNAs) were predicted as sensitive and resistant strains of Haemonchus contortus to ivermectin. These lncRNAs underwent bioinformatic analysis. In total, 205 lncRNAs significantly differed using log2 (difference multiplicity) > 1 or log2 (difference multiplicity) < - 1 and FDR < 0.05 as the threshold for significant difference analysis. We selected five lncRNAs based on significant differences in expression, cis-regulation, and their association with the Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathways. These expressions of lncRNAs, namely MSTRG.12610.1, MSTRG.8169.1, MSTRG.6355.1, MSTRG.980.1, and MSTRG.9045.1, were significantly downregulated. These findings were consistent with the results of transcriptomic sequencing. We further investigated the relative expression of target gene mRNAs and the regulation of mRNA and miRNA, starting with lncRNA cis-regulation of mRNA, and constructed a lncRNA-mRNA-miRNA network regulation. After a series of statistical analyses, we finally screened out UGT8, Unc-116, Fer-related kinase-1, GGPP synthase 1, and sart3, which may be involved in developing drug resistance under the regulation of their corresponding lncRNAs. The findings of this study provide a novel direction for future studies on drug resistance targets.
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Resistencia a Medicamentos , Haemonchus , Ivermectina , ARN Largo no Codificante , Animales , Haemonchus/genética , Haemonchus/efectos de los fármacos , ARN Largo no Codificante/genética , Ivermectina/farmacología , Resistencia a Medicamentos/genética , Hemoncosis/parasitología , Hemoncosis/veterinaria , Antihelmínticos/farmacología , MicroARNs/genética , Biología Computacional , Perfilación de la Expresión Génica , Regulación de la Expresión Génica/efectos de los fármacosRESUMEN
One-unit-cell, single-crystal, hexagonal CuInP2 S6 atomically thin sheets of≈0.81â nm in thickness was successfully synthesized for photocatalytic reduction of CO2 . Exciting ethene (C2 H4 ) as the main product was dominantly generated with the yield-based selectivity reaching ≈56.4 %, and the electron-based selectivity as high as ≈74.6 %. The tandem synergistic effect of charge-enriched Cu-In dual sites confined on the lateral edge of the CuInP2 S6 monolayer (ML) is mainly responsible for efficient conversion and high selectivity of the C2 H4 product as the basal surface site of the ML, exposing S atoms, can not derive the CO2 photoreduction due to the high energy barrier for the proton-coupled electron transfer of CO2 into *COOH. The marginal In site of the ML preeminently targets CO2 conversion to *CO under light illumination, and the *CO then migrates to the neighbor Cu sites for the subsequent C-C coupling reaction into C2 H4 with thermodynamic and kinetic feasibility. Moreover, ultrathin structure of the ML also allows to shorten the transfer distance of charge carriers from the interior onto the surface, thus inhibiting electron-hole recombination and enabling more electrons to survive and accumulate on the exposed active sites for CO2 reduction.
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OBJECTIVE: The study aimed to observe the effects of noise exposure on the pericytes of the cochlear stria vascularis (SV) in mice and to investigate its molecular mechanism. METHOD: Male C57BL/6J mice aged 6-8 weeks were used as the subjects. Auditory Brainstem Response (ABR) was used to assess hearing loss. Hematoxylin and Eosin (HE) staining was conducted to observe morphological alterations in the SV. Immunofluorescence combined with transmission electron microscopy (TEM) was used to scrutinize changes in pericytes following acoustic injury. Western blotting (WB) was used to assess the expression variations of the migration-related protein Osteopontin (OPN). Evans Blue assay was performed to evaluate the permeability of the blood labyrinth barrier (BLB). 4-Hydroxynonenal (4-HNE) staining, in conjunction with measurements of Superoxide Dismutase (SOD), Malondialdehyde (MDA), and Catalase (CAT) content, was used to ascertain whether oxidative stress injury occurred in the SV. WB, combined with immunofluorescence, was used to examine alterations in the expression of proliferator-activated receptor-gamma coactivator 1α (PGC-1α) in the SV and pericytes. RESULTS: Noise exposure resulted in permanent hearing loss in C57BL/6J mice, accompanied by SV swelling, migration of pericytes from their vascular attachments, BLB leakage, elevated oxidative stress levels in the SV, and reduced expression of PGC-1α on both the SV and migrating pericytes. CONCLUSION: Noise exposure may potentially increase oxidative stress levels in the SV, downregulate the expression levels of PGC-1α, promote pericytes migration, and subsequently lead to an elevation in BLB permeability.
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Sordera , Oído Interno , Pérdida Auditiva Provocada por Ruido , Animales , Humanos , Masculino , Ratones , Cóclea/metabolismo , Sordera/metabolismo , Oído Interno/metabolismo , Pérdida Auditiva Provocada por Ruido/metabolismo , Ratones Endogámicos C57BL , Pericitos/metabolismo , Coactivador 1-alfa del Receptor Activado por Proliferadores de Peroxisomas gamma/metabolismoRESUMEN
OBJECTIVE: We aimed to summarize the laboratory findings and clinical features of hemolytic disease of the fetus and newborn (HDFN). METHODS: We retrospectively analyzed the data for 17 infants with anti-M-induced HDFN (anti-M-HDFN) diagnosed between June 2013 and May 2019. Their maternal history, neonatal diagnosis on admission, and laboratory test results were compared with those of 15 infants with HDFN involving the ABO blood group system, 15 infants with HDFN involving the Rh system, and 15 premature infants. RESULTS: In the anti-M-HDFN group, 94.12% (16/17), 35.29% (6/17), and 17.65% (3/17) had free antibodies in plasma, a positive direct antiglobulin test, and a positive elution test, respectively. In 12 infants, free antibody reactions were stronger at 4°C than at 37°C, and the antibody titer at 4°C ranged from 1 to 512. All 17 infants with anti-M-HDFN developed anemia: 14 were treated with blood transfusion and 1 with neonatal exchange transfusion. Sixteen infants improved, and one died. Anti-M-HDFN had a higher rate of maternal stillbirth, lower gestational age, lower birthweight, and higher incidence of respiratory distress than other HDFN types. CONCLUSION: Anti-M may cause HDFN. It may present with varying degrees of anemia, low regenerative anemia, and low bilirubin levels. In addition, infants with anti-M-HDFN may have a negative elution test and direct antiglobulin test. These tests are helpful in examining antibody responses at a low temperature of 4°C.
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Anemia , Eritroblastosis Fetal , Femenino , Recién Nacido , Humanos , Estudios Retrospectivos , Isoanticuerpos , Sistema del Grupo Sanguíneo ABO , Feto , Anemia/complicacionesRESUMEN
Infections of Cyprinid herpesvirus 2 in goldfish and farmed crucian carp (Carassius auratus gibelio) are still an urgent problem worldwide. Detection and prevention are necessary for the control of haematopoietic necrosis disease caused by CyHV-2. Although many sensitive molecular diagnostic methods have been developed, effective immunodiagnosis and neutralization approaches based on monoclonal antibodies (MAbs) against CyHV-2 are still important to CyHV-2 study. In this experiment, purified CyHV-2 was used as antigens to produce monoclonal antibodies (Mabs). Six Mabs bound to different proteins were selected by Dot-blot screening and Western-blot analysis, and no one had cross-reactivity with closely related koi herpesvirus. Among them, Mabs 2E1-B10, 1F5-A3 and 4C4-A7 belonged to IgG1 isotype, while other three Mabs 3G9-B11, 3B4-G5 and 4F4-B7 belonged to IgM isotype. These six Mabs all could specifically detect CyHV-2 in CyHV-2 infected caudal fin of Carassius auratus gibelio (GiCF) cells by immunofluorescence assays. Then, the neutralization ability was tested in vitro, and the result showed that all six Mabs can attenuate CPE by CyHV-2 in vitro among which 2E1-B10 had the best neutralization ability. The virus proteins recognized by these six Mabs were identified by mass spectrometry identification, and the result showed they probably were ORF88, ORF55R, ORF115 and ORF151R. This study is the first to prepare Mabs by purifying CyHV-2, which will provide a practical basis for the in-depth study of CyHV-2 virus.
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Enfermedades de los Peces , Infecciones por Herpesviridae , Herpesviridae , Animales , Anticuerpos Monoclonales , Carpa Dorada , Infecciones por Herpesviridae/diagnóstico , Infecciones por Herpesviridae/veterinaria , Inmunoglobulina G , Inmunoglobulina MRESUMEN
Cyprinid herpesvirus 2 (CyHV-2) is a viral pathogen worldwide and causing high mortality on goldfish and silver crucian carp (Carassius auratus gibelio). In order to establish a stable and sensitive immunological diagnostic approach, the recombinant ORF121 protein encoded by the CyHV-2 ORF121 gene, was selected as a capture antigen to identify cells and tissues infected with CyHV-2 by immunological methods in this study. Firstly, the open reading frame of CyHV-2 ORF121 was cloned into the PGEX-4T-3 vector and expressed in Escherichia coli. Purified recombinant ORF121 protein was then used as an antigen to prepare monoclonal antibodies, and an efficient hybridoma cell line was selected by dot-blot assay. The resulting mAb-3D9 was applied to detect CyHV-2 in infected caudal fin of Carassius auratus gibelio (GiCF) cells and fish tissues by western blotting, immunofluorescence assays and immunohistological asays. The monoclonal antibody could specifically identify CyHV-2 in infected GiCF cells and the gills, the kidney and the spleen tissues, and it could attenuate CPE by CyHV-2 in vitro, suggesting it can be applied for CyHV-2 detection in the crucian carp and ORF121 may be a candidate vaccine against CyHV-2.
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Enfermedades de los Peces , Infecciones por Herpesviridae , Herpesviridae , Animales , Anticuerpos Monoclonales , Enfermedades de los Peces/diagnóstico , Carpa Dorada , Herpesviridae/genética , Infecciones por Herpesviridae/diagnóstico , Infecciones por Herpesviridae/veterinariaRESUMEN
Objective To observe the patients after transvaginal mesh(TVM)implantation surgery by using transperineal ultrasound(TPUS),compare the diagnosis of pelvic organ prolapse(POP)by TPUS and clinical examination[according to the Pelvic Organ Prolapse Quantification(POP-Q)system published by the International Continence Society],and to explore the role of ultrasound in postoperative evaluation as well as the high-risk factors of post-surgery POP recurrence. Methods This is a retrospective study based on the POP-Q records and TPUS data sets of patients within 6 months after TVM surgery during September 2013 and November 2019.The diagnostic results of TPUS and POP-Q were compared.The incidences of hiatal ballooning and levator avulsion were separately compared between the TPUS group and the control group. Results A total of 147 patients were enrolled.The Kappa values between TPUS and POP-Q in the diagnosis of anterior and posterior compartment POP were 0.268(P=0.000)and 0.235(P=0.001),respectively.There were altogether 37 cases diagnosed inconsistently between TPUS and POP-Q,including 14(9.52%)cases of anterior compartment prolapse and 23(15.65%)cases of posterior compartment prolapse.TPUS diagnosed 32 more cases of prolapse than POP-Q,which included 13(8.84%)cases of anterior compartment prolapse and 19(12.93%)cases of posterior compartment prolapse.The incidence of hiatal ballooning in the TPUS prolapse group was significantly higher than control group(51.35% vs.33.94%,χ2=3.950,P=0.047).The incidence of levator avulsion showed no significant difference between the two groups(P=1.000). Conclusions TPUS diagnosis of POP after TVM surgery has consistency with the POP-Q diagnosis of International Continence Society.TPUS can detect more POP cases,and thus it may act as a supplement for clinical diagnosis.Hiatal ballooning is associated with POP recurrence after TVM surgery.
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Prolapso de Órgano Pélvico , Mallas Quirúrgicas , Humanos , Diafragma Pélvico/diagnóstico por imagen , Prolapso de Órgano Pélvico/diagnóstico por imagen , Estudios Retrospectivos , Mallas Quirúrgicas/efectos adversos , UltrasonografíaRESUMEN
BACKGROUND: Enterovirus 71 (EV71) is one of the major causative agents of hand, foot, and mouth disease (HFMD), which is sometimes associated with severe central nervous system disease in children. There is currently no specific medication for EV71 infection. Quercetin, one of the most widely distributed flavonoids in plants, has been demonstrated to inhibit various viral infections. However, investigation of the anti-EV71 mechanism has not been reported to date. METHODS: The anti-EV71 activity of quercetin was evaluated by phenotype screening, determining the cytopathic effect (CPE) and EV71-induced cells apoptosis. The effects on EV71 replication were evaluated further by determining virus yield, viral RNA synthesis and protein expression, respectively. The mechanism of action against EV71 was determined from the effective stage and time-of-addition assays. The possible inhibitory functions of quercetin via viral 2Apro, 3Cpro or 3Dpol were tested. The interaction between EV71 3Cpro and quercetin was predicted and calculated by molecular docking. RESULTS: Quercetin inhibited EV71-mediated cytopathogenic effects, reduced EV71 progeny yields, and prevented EV71-induced apoptosis with low cytotoxicity. Investigation of the underlying mechanism of action revealed that quercetin exhibited a preventive effect against EV71 infection and inhibited viral adsorption. Moreover, quercetin mediated its powerful therapeutic effects primarily by blocking the early post-attachment stage of viral infection. Further experiments demonstrated that quercetin potently inhibited the activity of the EV71 protease, 3Cpro, blocking viral replication, but not the activity of the protease, 2Apro, or the RNA polymerase, 3Dpol. Modeling of the molecular binding of the 3Cpro-quercetin complex revealed that quercetin was predicted to insert into the substrate-binding pocket of EV71 3Cpro, blocking substrate recognition and thereby inhibiting EV71 3Cpro activity. CONCLUSIONS: Quercetin can effectively prevent EV71-induced cell injury with low toxicity to host cells. Quercetin may act in more than one way to deter viral infection, exhibiting some preventive and a powerful therapeutic effect against EV71. Further, quercetin potently inhibits EV71 3Cpro activity, thereby blocking EV71 replication.
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Enterovirus Humano A/efectos de los fármacos , Infecciones por Enterovirus/prevención & control , Quercetina/farmacología , Proteínas Virales/antagonistas & inhibidores , Proteasas Virales 3C , Animales , Antivirales/química , Antivirales/farmacología , Línea Celular Tumoral , Chlorocebus aethiops , Cisteína Endopeptidasas/metabolismo , Efecto Citopatogénico Viral/efectos de los fármacos , Enterovirus Humano A/fisiología , Infecciones por Enterovirus/virología , Humanos , Simulación del Acoplamiento Molecular , Unión Proteica , Quercetina/química , Quercetina/metabolismo , ARN Viral/biosíntesis , ARN Viral/efectos de los fármacos , Células Vero , Proteínas Virales/biosíntesis , Proteínas Virales/efectos de los fármacos , Proteínas Virales/metabolismo , Acoplamiento Viral/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
RATIONALE: Rhizoma coptidis extract and its alkaloids show various pharmacological activities, but its metabolic profile in human plasma has not been thoroughly investigated. In the present research, the metabolism of Rhizoma coptidis at a clinical dose (5 g/60 kg/day) was systematically analyzed to determine its biotransformation processes in human plasma. METHODS: In this research, the metabolites of Rhizoma coptidis in human plasma after oral administration of Rhizoma coptidis extract at a clinical dose were investigated using ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution LTQ-Orbitrap mass spectrometry. The structural elucidation of the constituents was confirmed by comparing their retention times (tR ) and MSn fragments with those of standards and literature reports. RESULTS: In total, two prototypes and twelve metabolites were detected in human plasma. The two prototypes were confidently identified using reference standards. Of the compounds detected, M7 (berberrubinen-9-O-glucuronide) was the most abundant based on its peak area, which indicates that this compound might be a pharmacokinetic marker for Rhizoma coptidis alkaloids in humans. Based on the metabolites detected in human plasma, a possible metabolic pathway for Rhizoma coptidis in vivo was proposed. CONCLUSIONS: The results indicated that the alkaloids in Rhizoma coptidis were extensively biotransformed in vivo mainly via conjugation with glucuronic acid (GluA) or sulfuric acid (SulA) to form phase II metabolites, and the GluA metabolites are likely the dominant form in human plasma. To the best of our knowledge, this is the first in vivo evaluation of the metabolic profile of the whole Rhizoma coptidis extract in human plasma, which is essential for determining the chemicals responsible for the pharmacological activities of Rhizoma coptidis in vivo. Moreover, it would be beneficial for us to further systematically study the pharmacokinetic behavior of Rhizoma coptidis in humans.
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Medicamentos Herbarios Chinos/química , Rizoma/química , Adulto , Alcaloides/sangre , Alcaloides/química , Biotransformación , Cromatografía Líquida de Alta Presión/métodos , Coptis chinensis , Medicamentos Herbarios Chinos/administración & dosificación , Medicamentos Herbarios Chinos/metabolismo , Femenino , Humanos , Masculino , Espectrometría de Masas/métodos , Metaboloma , Estructura Molecular , Adulto JovenRESUMEN
OBJECTIVE: The aim of this study was to establish a rapid and accurate method for the detection of the Streptococcus agalactiae antibody (SA-Ab) to determine the presence of the bovine mastitis (BM)-causative pathogen. METHODS: The multi-subunit fusion protein rSip-Pgk-FbsA was prokaryotically expressed and purified. The triple activities of the membrane surface-associated proteins Sip, phosphoglycerate kinase (Pgk), and fibronectin (FbsA) were used as the diagnostic antigens to establish an indirect enzyme-linked immunosorbent assay (ELISA) method for the detection of SA-Ab in BM. RESULTS: The optimal antigen coating concentration was 2 µg/mL, the optimal serum dilution was 1:160, and the optimal dilution of the enzyme-labeled secondary antibody was 1:6000. The sensitivity, specificity, and repeatability tests showed that the method established in this study had no cross-reaction with antibodies to Streptococcus pyogenes, Escherichia coli, Staphylococcus aureus, and Staphylococcus epidermidis in the sera. The results of the sensitivity test showed that a positive result could be obtained even if the serum dilution reached 1:12,800, indicating the high sensitivity and good repeatability of the method. The positive coincidence rate of this method was 98.6%, which is higher than that of previous tests established with the Sip or Pgk mono-antigen fusion protein, respectively, demonstrating the relatively higher sensitivity of this newly established method. The detection rate for 389 clinical samples was 46.53%. CONCLUSIONS: The indirect ELISA method established in this study could provide a more accurate and reliable serological method for the rapid detection of S. agalactiae in cases of BM.
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Proteínas Bacterianas/metabolismo , Ensayo de Inmunoadsorción Enzimática/veterinaria , Mastitis Bovina/microbiología , Infecciones Estreptocócicas/veterinaria , Streptococcus agalactiae/inmunología , Streptococcus agalactiae/metabolismo , Animales , Proteínas Bacterianas/genética , Bovinos , Femenino , Mastitis Bovina/diagnóstico , Proteínas Recombinantes , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Infecciones Estreptocócicas/diagnóstico , Infecciones Estreptocócicas/microbiologíaRESUMEN
1. In China, Fructus Gardeniae was used as a traditional Chinese medicine (TCM) with a wide array of biological activities. The bioactive components identified in Fructus Gardeniae mainly included iridoids, flavonids, pigments, and so on. Among them, iridoids were regarded as important compounds in Fructus Gardeniae. Though analyses of the constituents in biological samples after oral administration of Fructus Gardeniae effective fraction or its active compounds have been reported, few efforts have been made to investigate the metabolic profile of Fructus Gardeniae in humans. In this study, the constituents and metabolites of Fructus Gardeniae in human blood and urine after oral administration of Fructus Gardeniae were investigated using ultra high-performance liquid chromatography (UHPLC) coupled with high-resolution LTQ-Orbitrap mass spectrometery. 2. Totally, 14 constituents (two parent compounds and 12 metabolites) of Fructus Gardeniae were identified in human plasma and urine either by comparing the retention time and mass spectrometry data with that of reference compounds or by the accurate high-resolution MS/MS data of the chemicals. The compounds identified were mainly iridoid glycosides such as geniposide and the derivatives of genipin-O-glucuronide. Among them, 11 metabolites were detected in human plasma and urine while the other three metabolites including geniposidic acid (M1), demethylation derivative of genipin-O-glucuronide (M2), and dehydration product of mono-hydroxylated genipin-O-glucuronide (M9) were only discovered in human urine. Further, the possible metabolic pathways of Fructus Gardeniae in vivo were proposed and the peak area-time curve of the most abundant metabolite genipin-O-glucuronide (M13) in human plasma after oral administration of Fructus Gardeniae was depicted. The results suggested that a metabolic difference existed between rats and humans. 3. The results obtained in the present research would provide basic information to understand the metabolic profile of Fructus Gardeniae in humans and explore the chemicals responsible for the hepatotoxicity of Fructus Gardeniae in vivo. Moreover, it would be beneficial for us to further study the pharmacokinetic behavior of Fructus Gardeniae in humans systematically.
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Medicamentos Herbarios Chinos/metabolismo , Gardenia , Animales , Cromatografía Líquida de Alta Presión , Humanos , Ratas , Espectrometría de Masas en TándemRESUMEN
A transparent SERS platform was fabricated via the gel-trapping method coupled with a liquid/liquid interface self-assembly technique. We employed gold nanorods as the building blocks for interface self-assembly because of their strong localized surface plasmons upon excitation by infrared radiation. Based on a "top cover" configuration, this transparent SERS platform endows high signal reproducibility for directly detecting liquid samples by confining the sample droplet into a regular shape. The Au NR PDMS platform was able to directly detect crystal violet in aqueous solutions down to 10 ppb level with high enhancement factor (0.87 × 10(5)) and signal uniformity (RSD = 3.9%). Furthermore, SERS-based anti-fungal agent detection on a fish scale was demonstrated by simply covering the fish scale with a tailored GNRs PDMS film. The experimental results clearly show that the Au NR PDMS SERS platform has great potential for on-site real time detection of contaminants in water as well as on curved surfaces.
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Accurate acoustic source localization at a low sampling rate (less than 10 kHz) is still a challenging problem for small portable systems, especially for a multitasking micro-embedded system. A modification of the generalized cross-correlation (GCC) method with the up-sampling (US) theory is proposed and defined as the US-GCC method, which can improve the accuracy of the time delay of arrival (TDOA) and source location at a low sampling rate. In this work, through the US operation, an input signal with a certain sampling rate can be converted into another signal with a higher frequency. Furthermore, the optimal interpolation factor for the US operation is derived according to localization computation time and the standard deviation (SD) of target location estimations. On the one hand, simulation results show that absolute errors of the source locations based on the US-GCC method with an interpolation factor of 15 are approximately from 1/15- to 1/12-times those based on the GCC method, when the initial same sampling rates of both methods are 8 kHz. On the other hand, a simple and small portable passive acoustic source localization platform composed of a five-element cross microphone array has been designed and set up in this paper. The experiments on the established platform, which accurately locates a three-dimensional (3D) near-field target at a low sampling rate demonstrate that the proposed method is workable.
RESUMEN
Microalgal-bacterial (MB) consortia create an excellent eco-system for simultaneous COD/BOD and nutrients (N and P) removals in a single step with significant reduction in or complete elimination of aeration and carbonation in the biological wastewater treatment processes. The integration of membrane separation technology with the MB processes has created a new paradigm for research and development. This paper focuses on a comprehensive and critical literature review of recent advances in these emerging processes. Novel membrane process configurations and process conditions affecting the biological performance of these novel systems have been systematically reviewed and discussed. Membrane fouling issues and control of MB biofilm formation and thickness associated with these emerging suspended growth or immobilized biofilm processes are addressed and discussed. The research gaps, challenges, outlooks of these emerging processes are identified and discussed in-depth. The findings from the literature suggest that the membrane-based MB processes are advanced biotechnologies with a significant reduction in energy consumption and process simplification and high process efficiency that are not achievable with current technologies in wastewater treatment. There are endless opportunities for research and development of these novel and emerging membrane-based MB processes.
Asunto(s)
Membranas Artificiales , Microalgas , Eliminación de Residuos Líquidos , Aguas Residuales , Microalgas/fisiología , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/microbiología , Biopelículas , Bacterias , Reactores Biológicos , Purificación del Agua/métodosRESUMEN
Photocatalytic CO2 reduction captures solar energy to convert CO2 into hydrocarbon fuels, thus shifting the dependence on rapidly depleting fossil fuels. Among the various proposed photocatalysts, systems containing metal active sites (MASs) possess obvious advantages, such as effective photogenerated carrier separation, suitable adsorption and activation of intermediates, and achievable C-C coupling to generate multi-carbon (C2+) products. The present review aims to summarize the typical photocatalytic materials with MAS, highlighting the critical role of different formulations of MAS in CO2 photoreduction, especially for C2+ product generation. State-of-the-art progress in the characterization and theoretical calculations for MAS-containing photocatalysts is also emphasized. Finally, the challenges and prospects of catalytic systems involving MAS for solar-driven CO2 conversion are outlined, providing inspiration for the future design of materials for efficient photocatalytic energy conversion.
RESUMEN
Fenton or Fenton-like reactions have been widely used in various fields, including solar energy conversion to generate hydroxyl radicals, environmental remediation, biology, and life science. However, the slow Fe3+/Fe2+ cycle and narrow applicable pH range still present significant challenges. Here, a heterostructured CoFe-layered double hydroxide/MoS2 nanocomposite (CoFe-LDH/MoS2) was prepared via simple electrostatic interactions. The heterostructure establishes a robust interfacial contact, leading to an abundance of exposed Mo6+ sites. Consequently, the developed CoFe-LDH/MoS2+H2O2 system exhibited superior performance in the degradation of tetracycline (>85%) within 60 min across a wide pH range from acidic to basic. Moreover, the CoFe-LDH/MoS2 heterojunction catalysts exhibited exceptional resistance to common anions and efficiently degraded various organic pollutants. The mechanism study verified that the CoFe-LDH/MoS2 had high efficiency in producing 1O2 and â§OH to degrade various organic pollutants. The present study will serve as a foundation for creating efficient catalyst systems for related environmental remediation.