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1.
Biotechnol J ; 19(6): e2400074, 2024 Jun.
Article de Anglais | MEDLINE | ID: mdl-38896409

RÉSUMÉ

The ELISA is the most worldwide method for immunoassay. However, the ELISA is losing ground due to low reproducibility of manual experimental processes in both R&D and IVD areas. An automated platform is a good solution, but there are still limitations owning to extremely high cost and requiring large space to set up especially for a small size laboratory. Here, we present a novel all-in-one platform called "VEUS" settable on the laboratory table that offers comprehensive automation of the entire multiplex immunoassay process by exploiting antibody conjugated magnetic particles, quality control and then immunoanalytical reaction, thereby enhancing detection sensitivity and high reproducibility. As a proof of concept, the system exhibits a sensitive LOD of 0.6 and 3.1 pg mL-1 within 1 h run, comparable precision that of molecular diagnostic systems based on PCR method, enabling rapid multiplex diagnosis of Influenza A, Influenza B, and COVID-19 viruses with similar symptoms. Through automation by the all-in-one system, it can be used by novice users, something innovative for immunoassays, relying heavily on user experience. Furthermore, it can contribute to streamline entire immunoassay processes of diverse biomarkers with high reproducibility and convenience in laboratories.


Sujet(s)
SARS-CoV-2 , Humains , Dosage immunologique/méthodes , SARS-CoV-2/immunologie , SARS-CoV-2/isolement et purification , Anticorps immobilisés/immunologie , Anticorps immobilisés/composition chimique , Reproductibilité des résultats , COVID-19/diagnostic , COVID-19/virologie , Test ELISA/méthodes , Virus de la grippe A/immunologie , Virus influenza B/immunologie , Laboratoire automatique/méthodes , Limite de détection
2.
Int J Mol Sci ; 25(9)2024 May 02.
Article de Anglais | MEDLINE | ID: mdl-38732183

RÉSUMÉ

The impact of microplastics (MPs) on the metabolic functions of the liver is currently unclear and not completely understood. To investigate the effects of the administration of MPs on the hepatic metabolism of normal and obese mice, alterations in the lipid, glucose (Glu), and amino acid regulation pathways were analyzed in the liver and adipose tissues of C57BL/6Korl (wild type, WT) or C57BL/6-Lepem1hwl/Korl mice (leptin knockout, Lep KO) orally administered polystyrene (PS) MPs for 9 weeks. Significant alterations in the lipid accumulation, adipogenesis, lipogenesis, and lipolysis pathways were detected in the liver tissue of MP-treated WT and Lep KO mice compared to the vehicle-treated group. These alterations in their liver tissues were accompanied by an upregulation of the serum lipid profile, as well as alterations in the adipogenesis, lipogenesis, and lipolysis pathways in the adipose tissues of MP-treated WT and Lep KO mice. Specifically, the level of leptin was increased in the adipose tissues of MP-treated WT mice without any change in their food intake. Also, MP-induced disruptions in the glycogenolysis, Glu transporter type 4 (GLUT4)-5' AMP-activated protein kinase (AMPK) signaling pathway, levels of lipid intermediates, and the insulin resistance of the liver tissues of WT and Lep KO mice were observed. Furthermore, the levels of seven endogenous metabolites were remarkably changed in the serum of WT and Lep KO mice after MP administrations. Finally, the impact of the MP administration observed in both types of mice was further verified in differentiated 3T3-L1 adipocytes and HepG2 cells. Thus, these results suggest that the oral administration of MPs for 9 weeks may be associated with the disruption of lipid, Glu, and amino acid metabolism in the liver tissue of obese WT and Lep KO mice.


Sujet(s)
Acides aminés , Glucose , Métabolisme lipidique , Foie , Souris de lignée C57BL , Souris knockout , Microplastiques , Polystyrènes , Animaux , Foie/métabolisme , Foie/effets des médicaments et des substances chimiques , Souris , Glucose/métabolisme , Métabolisme lipidique/effets des médicaments et des substances chimiques , Acides aminés/métabolisme , Administration par voie orale , Leptine/métabolisme , Tissu adipeux/métabolisme , Tissu adipeux/effets des médicaments et des substances chimiques , Adipogenèse/effets des médicaments et des substances chimiques , Mâle , Lipogenèse/effets des médicaments et des substances chimiques , Obésité/métabolisme , Obésité/étiologie , Obésité/génétique , Humains , Lipolyse/effets des médicaments et des substances chimiques
3.
Sci Rep ; 6: 32647, 2016 09 06.
Article de Anglais | MEDLINE | ID: mdl-27596274

RÉSUMÉ

High field magnetic resonance imaging (MRI)-based delineation of the substantia nigra (SN) and visualization of its inner cellular organization are promising methods for the evaluation of morphological changes associated with neurodegenerative diseases; however, corresponding MR contrasts must be matched and validated with quantitative histological information. Slices from two postmortem SN samples were imaged with a 7 Tesla (7T) MRI with T1 and T2* imaging protocols and then stained with Perl's Prussian blue, Kluver-Barrera, tyrosine hydroxylase, and calbindin immunohistochemistry in a serial manner. The association between T2* values and quantitative histology was investigated with a co-registration method that accounts for histology slice preparation. The ventral T2* hypointense layers between the SNr and the crus cerebri extended anteriorly to the posterior part of the crus cerebri, which demonstrates the difficulty with an MRI-based delineation of the SN. We found that the paramagnetic hypointense areas within the dorsolateral SN corresponded to clusters of neuromelanin (NM). These NM-rich zones were distinct from the hypointense ventromedial regions with high iron pigments. Nigral T2* imaging at 7T can reflect the density of NM-containing neurons as the metal-bound NM macromolecules may decrease T2* values and cause hypointense signalling in T2* imaging at 7T.


Sujet(s)
Produits de contraste/composition chimique , Imagerie par résonance magnétique , Mélanines/métabolisme , Modifications postmortem , Substantia nigra/métabolisme , Substantia nigra/anatomopathologie , Adulte , Femelle , Humains , Fer/métabolisme , Mâle , Adulte d'âge moyen
4.
Xenobiotica ; 41(6): 501-10, 2011 Jun.
Article de Anglais | MEDLINE | ID: mdl-21341987

RÉSUMÉ

Genetic variants of Na(+)-taurocholate co-transporting polypeptide (NTCP; SLC10A1) and ileal apical sodium-dependent bile acid transporter (ASBT; SLC10A2), which greatly contribute to bile acid homeostasis, were extensively explored in the Korean population and functional variants of NTCP were compared among Asian populations. From direct DNA sequencing, six SNPs were identified in the SLC10A1 gene and 14 SNPs in the SLC10A2 gene. Three of seven coding variants were non-synonymous SNPs: two variants from SLC10A1 (A64T, S267F) and one from SLC10A2 (A171S). No linkage was analysed in the SLC10A1 gene because of low frequencies of genetic variants, and the SLC10A2 gene was composed of two separated linkage disequilibrium blocks contrary to the white population. The stably transfected NTCP-A64T variant showed significantly decreased uptakes of taurocholate and rosuvastatin compared with wild-type NTCP. The decreased taurocholate uptake and increased rosuvastatin uptake were shown in the NTCP-S267F variant. The allele frequencies of these functional variants were 1.0% and 3.1%, respectively, in a Korean population. However, NTCP-A64T was not found in Chinese and Vietnamese subjects. The frequency distribution of NTCP-S267F in Koreans was significantly lower than those in Chinese and Vietnamese populations. Our data suggest that NTCP-A64T and -S267F variants cause substrate-dependent functional change in vitro, and show ethnic difference in their allelic frequencies among Asian populations although the clinical relevance of these variants is remained to be evaluated.


Sujet(s)
Transporteurs d'anions organiques sodium-dépendants/génétique , Polymorphisme de nucléotide simple , Symporteurs/génétique , Asiatiques , Bile/métabolisme , Fluorobenzènes/métabolisme , Fréquence d'allèle , Variation structurale du génome , Humains , Déséquilibre de liaison , Transporteurs d'anions organiques sodium-dépendants/métabolisme , Pyrimidines/métabolisme , Rosuvastatine de calcium , Sulfonamides/métabolisme , Symporteurs/métabolisme , Acide taurocholique/métabolisme
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