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1.
Clin Med (Lond) ; 20(5): 463-467, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32934038

RESUMO

OBJECTIVE: The objective was to study hospitalised COVID-19 patients' mortality and intensive care unit (ICU) admission with covariates of interest (age, gender, ethnicity, clinical presentation, comorbidities and admission laboratory findings). METHODS: Logistic regression analyses were performed for patients admitted to University Hospital, University Hospitals Coventry and Warwickshire NHS Trust, between 24 January 2020 - 13 April 2020. RESULTS: There were 321 patients hospitalised. Median age was 73 years and 189 (59%) were male. Ethnicity was divided between Caucasian (77%), and black, Asian, and minority ethnic (BAME) groups (23%). Commonest symptoms were dyspnoea (62.9%), fever (59.1%) and cough (56%). Gastrointestinal symptoms amounted to 11.8%.Forty-four patients (13.7%) received ICU care. ICU male to female ratio was 3:1 (p=0.027; odds ratio (OR) 2.3; 95% confidence interval (CI) 1.1-4.9), BAME (p=0.008; OR 2.5; 95% CI 1.3-4.9), age >65 years (p=0.026; OR 0.28; 95% CI 0.09-0.93), heart disease (p=0.009; OR 0.2; 95% CI 0.1-0.6) and elevated C-reactive protein (CRP; p<0.001; OR 1.004; 95% CI 1.002-1.008) were associated with ICU admission.One-hundred and four patients (32.4%) died. Age >65 years (p=0.011; OR 5; 95% CI 1.6-21.9), neutrophils (p=0.047), neutrophil:lymphocyte ratio (NLR; p=0.028), CRP (p<0.001) and albumin (p=0.002) were associated with mortality. When analysis adjusted for age, CRP (p<0.001; OR 1.006; 95% CI 1.004-1.008) and albumin (p=0.005; OR 0.94; 95% CI 0.90-0.98) remained associated with mortality. CONCLUSIONS: COVID-19 has high mortality. BAME and male patients were associated with ICU admission. High CRP and low albumin (after correcting for age) were associated with mortality.


Assuntos
Albuminas/metabolismo , Proteína C-Reativa/metabolismo , Causas de Morte , Infecções por Coronavirus/sangue , Infecções por Coronavirus/mortalidade , Mortalidade Hospitalar/tendências , Pneumonia Viral/sangue , Pneumonia Viral/mortalidade , Idoso , Idoso de 80 Anos ou mais , Infecções por Coronavirus/fisiopatologia , Feminino , Avaliação Geriátrica , Hospitalização/estatística & dados numéricos , Hospitais Universitários , Humanos , Unidades de Terapia Intensiva/estatística & dados numéricos , Modelos Logísticos , Masculino , Razão de Chances , Pandemias , Pneumonia Viral/fisiopatologia , Estudos Retrospectivos , Medição de Risco , Índice de Gravidade de Doença , Centros de Atenção Terciária , Reino Unido
2.
Medicine (Baltimore) ; 99(28): e20654, 2020 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-32664063

RESUMO

Albumin-bilirubin (ALBI) showed its prognostic and predictive value in hepatobiliary disease like hepatocellular carcinoma. However, little has been known about its role in pancreatic cancer.In this retrospective study, 149 patients with advanced pancreatic cancer (APC) treated in the Shanghai General Hospital from January 2009 to December 2014 were enrolled as the training cohort and 120 patients treated from January 2015 to December 2018 were taken as the validation cohort. We generated the ALBI score according previous studies. The correlations between ALBI and clinicopathological parameters were evaluated with the Pearson Chi-square test. Kaplan-Meier method and log-rank test were conducted to determine the correlation between ALBI and overall survival (OS). Then we used Cox regression model to investigate the prognostic significance of ALBI. We further assessed retrospectively whether ALBI score could be used to identify combination therapy candidates for APC.Eastern Cooperative Oncology Group Performance Status, hemoglobin, aspartate aminotransferase, and alanine aminotransferase were found to be significantly correlated with ALBI. Kaplan-Meier analysis showed that the median OS in patients with a pretreatment ALBI ≥-2.6 was 7.0 months, which was significantly shorter than OS of patients with a ALBI <-2.6 (13.0 months, P = .001). ALBI was independently correlated with OS in multivariate analysis. In the subgroup analysis, ALBI showed significant prognostic value in patients with liver metastasis but not those without liver metastasis in all 3 cohorts. In addition, only in the group with ALBI <-2.6, patients receiving combination therapy showed better prognosis than those receiving monotherapy.In conclusion, ALBI was a promising prognostic biomarker in APC with liver metastasis. ALBI also showed predictive value in identifying combination therapy candidates for patients with APC.


Assuntos
Albuminas/metabolismo , Bilirrubina/sangue , Carcinoma/sangue , Neoplasias Pancreáticas/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/sangue , Carcinoma/diagnóstico , Carcinoma/secundário , Feminino , Humanos , Neoplasias Hepáticas/secundário , Masculino , Pessoa de Meia-Idade , Neoplasias Pancreáticas/diagnóstico , Neoplasias Pancreáticas/patologia , Prognóstico , Estudos Retrospectivos
3.
Yakugaku Zasshi ; 140(10): 1269-1274, 2020 Oct 01.
Artigo em Japonês | MEDLINE | ID: mdl-32684555

RESUMO

We previously reported that tolvaptan may influence warfarin pharmacodynamics in vivo; however, the mechanism responsible for this influence was not clear. In this study, we investigated the drug-drug interactions between warfarin and tolvaptan by measuring warfarin blood concentrations in 18 patients who received warfarin therapy and in 24 who received warfarin+tolvaptan therapy. The free warfarin concentrations significantly increased in patients who were also receiving oral tolvaptan (p=0.04). In vitro albumin-binding experiments showed that the free warfarin concentrations significantly increased with the addition of tolvaptan, in a dose-dependent manner, through albumin-binding substitution (approximately 2.5 times). Both clinical and in vitro data showed that tolvaptan increased the unbound warfarin serum concentration. The prothrombin time-international normalized ratio (PT-INR) tended to increase within 2 weeks when tolvaptan was added at clinically used doses (p=0.14). Special attention is warranted in cases with a serum tolvaptan concentration of ≥125 ng/mL (≥7.5 mg/d) for at least 2 weeks following oral tolvaptan administration.


Assuntos
Anticoagulantes/sangue , Interações Medicamentosas , Coeficiente Internacional Normatizado , Tempo de Protrombina , Tolvaptan/farmacologia , Varfarina/sangue , Varfarina/farmacologia , Administração Oral , Idoso , Idoso de 80 Anos ou mais , Albuminas/metabolismo , Anticoagulantes/administração & dosagem , Relação Dose-Resposta a Droga , Feminino , Humanos , Técnicas In Vitro , Masculino , Pessoa de Meia-Idade , Ligação Proteica , Fatores de Tempo , Tolvaptan/administração & dosagem , Varfarina/administração & dosagem , Varfarina/metabolismo
4.
J Biosci Bioeng ; 130(2): 212-216, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32312490

RESUMO

To clinically apply bioartificial livers (BALs), an effective liver cell cryopreservation method is required for a stable cell supply. In this study, we performed tissue-engineered construct (TEC) cryopreservation of fetal liver cells (FLCs) in which FLCs cultured within a porous polymer scaffold were cryopreserved. Growth and albumin secretion in TEC-cryopreserved FLCs after thawing were compared to freshly isolated FLCs (control experiments). The effect of preculture duration prior to cryopreservation (0-3 weeks) on these functions was also examined. In the three-dimensional cultures, the TEC-cryopreserved FLCs with preculturing showed constant growth, and this growth was comparable to controls. On the contrary, the TEC-cryopreserved FLCs without preculturing did not proliferate after thawing. Albumin secretion of TEC-cryopreserved FLCs with preculturing rapidly increased up to day 12 and high secretory activity comparable to controls was maintained thereafter in FLCs with 1- or 2-week preculturing, suggesting this as an appropriate preculture duration. Compared to conventionally cryopreserved FLCs, growth and albumin secretion in the TEC-cryopreserved FLCs were significantly higher, indicating their usefulness as a potent cell source for BALs.


Assuntos
Albuminas/metabolismo , Hepatócitos/citologia , Fígado Artificial , Polímeros/química , Engenharia Tecidual/métodos , Tecidos Suporte/química , Animais , Sobrevivência Celular , Células Cultivadas , Criopreservação/normas , Feto , Fígado/citologia , Camundongos , Polímeros/farmacologia , Porosidade
5.
Nucleic Acids Res ; 48(9): 4658-4671, 2020 05 21.
Artigo em Inglês | MEDLINE | ID: mdl-32313951

RESUMO

Erythropoietic protoporphyria (EPP) is a rare genetic disease in which patients experience acute phototoxic reactions after sunlight exposure. It is caused by a deficiency in ferrochelatase (FECH) in the heme biosynthesis pathway. Most patients exhibit a loss-of-function mutation in trans to an allele bearing a SNP that favors aberrant splicing of transcripts. One viable strategy for EPP is to deploy splice-switching oligonucleotides (SSOs) to increase FECH synthesis, whereby an increase of a few percent would provide therapeutic benefit. However, successful application of SSOs in bone marrow cells is not described. Here, we show that SSOs comprising methoxyethyl-chemistry increase FECH levels in cells. We conjugated one SSO to three prototypical targeting groups and administered them to a mouse model of EPP in order to study their biodistribution, their metabolic stability and their FECH splice-switching ability. The SSOs exhibited distinct distribution profiles, with increased accumulation in liver, kidney, bone marrow and lung. However, they also underwent substantial metabolism, mainly at their linker groups. An SSO bearing a cholesteryl group increased levels of correctly spliced FECH transcript by 80% in the bone marrow. The results provide a promising approach to treat EPP and other disorders originating from splicing dysregulation in the bone marrow.


Assuntos
Ferroquelatase/genética , Oligonucleotídeos/administração & dosagem , Protoporfiria Eritropoética/metabolismo , Processamento de RNA , Albuminas/metabolismo , Animais , Medula Óssea/metabolismo , Células COS , Chlorocebus aethiops , Modelos Animais de Doenças , Ferroquelatase/metabolismo , Humanos , Células K562 , Camundongos , Oligonucleotídeos/sangue , Oligonucleotídeos/química , Oligonucleotídeos/farmacocinética , Polimorfismo de Nucleotídeo Único , Protoporfiria Eritropoética/genética , Protoporfiria Eritropoética/terapia , Sítios de Splice de RNA , Distribuição Tecidual
6.
Life Sci ; 248: 117475, 2020 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-32119963

RESUMO

AIMS: Liver fibrosis is a crucial pathological feature which could result in cirrhosis and hepatocarcinoma. But until now, there is no favourable treatment for it. Apigenin (APG) is a flavonoid, which exhibits efficient anti-liver fibrosis activity, but its underlying mechanisms were rarely studied. So this work aims to estimate the potential therapeutic action of APG on liver fibrosis rats and to gain insight into its system-level mechanisms. MAIN METHODS: Hepatic fibrosis was induced by CCl4 in Wistar rats, and APG was given in the light of the regimen. Biochemical indexes, histopathological change and immunohistochemistry of liver were evaluated. The optimal effect group of APG was selected for further transcriptomic and proteomic analysis. KEY FINDINGS: APG ameliorated liver fibrosis via reducing the levels of AST, ALT, ALP, LDH, Hyp, TP, TB, DB, HA, LN, PCIII and IV-C, mitigating fibrosis and inflammation of liver in H&E and Masson staining. Mechanistically, APG elevated the activity of ALB, SOD and GSH-PX with reducing the level of MDA. The results of microarray and TMT revealed that 4919 genes and 4876 proteins were differentially expressed in the APG and model groups. Besides, transcriptomics and proteomics analyses unfolded 120 overlapped proteins, enriched in 111 GO terms containing apoptotic process, angiogenesis, cell migration and proliferation, etc. Meanwhile, KEGG pathway analysis showed that 26 pathways containing HIF-1/MAPK/eNOS/VEGF/PI3K/Akt signaling pathway, regulation of actin cytoskeleton and focal adhesion mostly. SIGNIFICANCE: APG can ameliorate CCl4-induced liver fibrosis via VEGF-mediated FAK phosphorylation through the MAPKs, PI3K/Akt, HIF-1, ROS, and eNOS pathways, which may hopefully become the anti-liver fibrosis activity of natural product.


Assuntos
Anti-Inflamatórios/farmacologia , Apigenina/farmacologia , Doença Hepática Induzida por Substâncias e Drogas/prevenção & controle , Cirrose Hepática/prevenção & controle , Fígado/efeitos dos fármacos , Transcriptoma/efeitos dos fármacos , Alanina Transaminase/genética , Alanina Transaminase/metabolismo , Albuminas/metabolismo , Fosfatase Alcalina/genética , Fosfatase Alcalina/metabolismo , Animais , Aspartato Aminotransferases/genética , Aspartato Aminotransferases/metabolismo , Bilirrubina/sangue , Tetracloreto de Carbono/administração & dosagem , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/genética , Doença Hepática Induzida por Substâncias e Drogas/patologia , Quinase 1 de Adesão Focal/genética , Quinase 1 de Adesão Focal/metabolismo , Glutationa Peroxidase/genética , Glutationa Peroxidase/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , L-Lactato Desidrogenase/genética , L-Lactato Desidrogenase/metabolismo , Fígado/metabolismo , Fígado/patologia , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/genética , Cirrose Hepática/patologia , Masculino , Malondialdeído/antagonistas & inibidores , Malondialdeído/metabolismo , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Óxido Nítrico Sintase Tipo III/genética , Óxido Nítrico Sintase Tipo III/metabolismo , Proteômica/métodos , Ratos , Ratos Wistar , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo
7.
Nat Commun ; 11(1): 1573, 2020 03 26.
Artigo em Inglês | MEDLINE | ID: mdl-32218438

RESUMO

Near-infrared (NIR) fluorescent probes are among the most attractive chemical tools for biomedical imaging. However, their in vivo applications are hindered by albumin binding, generating unspecific fluorescence that masks the specific signal from the analyte. Here, combining experimental and docking methods, we elucidate that the reason for this problem is an acceptor (A) group-mediated capture of the dyes into hydrophobic pockets of albumin. This pocket-capturing phenomenon commonly applies to dyes designed under the twisted intramolecular charge-transfer (TICT) principle and, therefore, represents a generic but previously unidentified backdoor problem. Accordingly, we create a new A group that avoids being trapped into the albumin pockets (pocket-escaping) and thereby construct a NIR probe, BNLBN, which effectively prevents this backdoor problem with increased imaging accuracy for liver fibrosis in vivo. Overall, our study explains and overcomes a fundamental problem for the in vivo application of a broad class of bioimaging tools.


Assuntos
Corantes Fluorescentes/química , Raios Infravermelhos , Albuminas/metabolismo , Animais , Feminino , Cirrose Hepática/patologia , Camundongos Endogâmicos C57BL , Ligação Proteica , Reprodutibilidade dos Testes
8.
Nucleic Acids Res ; 48(8): 4382-4395, 2020 05 07.
Artigo em Inglês | MEDLINE | ID: mdl-32182359

RESUMO

Conjugation of antisense oligonucleotide (ASO) with a variety of distinct lipophilic moieties like fatty acids and cholesterol increases ASO accumulation and activity in multiple tissues. While lipid conjugation increases tissue exposure in mice and reduces excretion of ASO in urine, histological review of skeletal and cardiac muscle indicates that the increased tissue accumulation of lipid conjugated ASO is isolated to the interstitium. Administration of palmitic acid-conjugated ASO (Palm-ASO) in mice results in a rapid and substantial accumulation in the interstitium of muscle tissue followed by relatively rapid clearance and only slight increases in intracellular accumulation in myocytes. We propose a model whereby increased affinity for lipid particles, albumin, and other plasma proteins by lipid-conjugation facilitates ASO transport across endothelial barriers into tissue interstitium. However, this increased affinity for lipid particles and plasma proteins also facilitates the transport of ASO from the interstitium to the lymph and back into circulation. The cumulative effect is only a slight (∼2-fold) increase in tissue accumulation and similar increase in ASO activity. To support this proposal, we demonstrate that the activity of lipid conjugated ASO was reduced in two mouse models with defects in endothelial transport of macromolecules: caveolin-1 knockout (Cav1-/-) and FcRn knockout (FcRn-/-).


Assuntos
Oligonucleotídeos Antissenso/farmacocinética , Ácido Palmítico , Albuminas/genética , Albuminas/metabolismo , Animais , Proteínas Sanguíneas/metabolismo , Caveolina 1/genética , Feminino , Coração , Células Hep G2 , Antígenos de Histocompatibilidade Classe I/genética , Humanos , Lipoproteínas HDL/metabolismo , Lipoproteínas LDL/metabolismo , Sistema Linfático/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Miocárdio/metabolismo , Oligonucleotídeos Antissenso/química , Músculo Quadríceps/metabolismo , Receptores Fc/genética , Distribuição Tecidual
9.
Angiology ; 71(4): 366-371, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32000500

RESUMO

Contrast-induced nephropathy (CIN) accounts for about 10% of all hospital-acquired acute kidney injury. We aimed to assess the role of the combination of 2 inflammatory biomarkers, the C-reactive protein (CRP)/albumin ratio (CAR), in the development of CIN after percutaneous coronary intervention (PCI) in patients with non-ST-elevation myocardial infarction (NSTEMI). Patients with NSTEMI (n = 205) treated by PCI were classified according to the development of CIN. Both groups were compared according to clinical, laboratory, and demographic characteristics, including inflammatory biomarkers and specifically, CAR. Contrast-induced nephropathy was observed in 10.2% of patients. More advanced age, the presence of diabetes and dyslipidemia, left ventricular ejection fraction, and CAR correlated with the development of CIN. Analysis also showed a significant association between CAR and the development of CIN (CAR in CIN (+): 8.54 ± 8.48, range: 0.7-32, median: 7.13 vs CAR in CIN (-): 2.36 ± 3.01, range: 0.1-24, median: 1.33, P < .001). Multivariate logistic regression analysis showed the impact of CAR on the development of CIN (odds ratio: 1.244, 95% confidence interval: 1.102; 1.392, P < .01). We conclude that CAR, as a combination of 2 inflammatory biomarkers, is a more accurate predictor of CIN development compared with the single-marker assessment of albumin and CRP in the context of NSTEMI.


Assuntos
Lesão Renal Aguda/induzido quimicamente , Albuminas/metabolismo , Proteína C-Reativa/metabolismo , Meios de Contraste/efeitos adversos , Infarto do Miocárdio sem Supradesnível do Segmento ST/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores/metabolismo , Estudos Transversais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Infarto do Miocárdio sem Supradesnível do Segmento ST/cirurgia , Intervenção Coronária Percutânea , Valor Preditivo dos Testes , Estudos Prospectivos
10.
Food Chem ; 317: 126423, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-32097824

RESUMO

The impact of pH (6-9) and NaCl concentration (0-0.5 mol.L-1) on sunflower protein extraction was studied through design of experiments. The considered criteria were protein extraction yield (total proteins, helianthinin and albumins), chlorogenic acids covalently bound to proteins, and free chlorogenic acid concentration in the aqueous extract. Statistical analysis showed that the obtained by design of experiments the polynomial models of each extraction criteria were reliable for predicting the responses. They were employed in an original multi-objective optimization methodology. The optimal conditions revealed to be pH 7.3/0.3 mol.L-1 NaCl yielded 46.83% and 59.16% of total protein and albumin extraction yield, 1.730 and 1.998 mg.g-1 of chlorogenic acids covalently bound to helianthinin and albumins in aqueous extract, respectively. The sunflower protein isolate obtained after extraction in this condition had good solubility (40-80% at pH 5-8), functional properties (foaming and emulsifying) and a satisfying color.


Assuntos
Helianthus/metabolismo , Extração Líquido-Líquido/métodos , Proteínas de Plantas/isolamento & purificação , Extração em Fase Sólida/métodos , Albuminas/análise , Albuminas/isolamento & purificação , Albuminas/metabolismo , Ácido Clorogênico/química , Ácido Clorogênico/metabolismo , Concentração de Íons de Hidrogênio , Isomerismo , Extração Líquido-Líquido/instrumentação , Proteínas de Plantas/análise , Proteínas de Plantas/metabolismo , Polifenóis/análise , Polifenóis/metabolismo , Ligação Proteica , Cloreto de Sódio/química , Extração em Fase Sólida/instrumentação
11.
Food Chem ; 316: 126350, 2020 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-32045819

RESUMO

This work characterises the peptide and protein profiles of Theobroma cacao beans of the genotype IMC 67 at different fermentation stages, using the Styrofoam-box fermentation method and employing UHPLC-ESI MS/MS for the analysis of peptides and proteins extracted from the beans. A total of 1058 endogenous peptides were identified and quantified over four fermentation time points. The majority of these peptides were formed after 2 and 4 days of fermentation, and originated predominantly from the proteolysis of two storage proteins - vicilin and a 21 kDa albumin. The changes in the peptide profile over fermentation were subsequently evaluated, and potential markers for assessing the degree of fermentation were identified. In particular, changes of the relative abundance of the major cocoa proteins detected can be proposed as potential markers for the fermentation stage. Furthermore, PCA of both the peptidomic and proteomic data has allowed differentiation of beans at different fermentation stages.


Assuntos
Cacau/metabolismo , Peptídeos/metabolismo , Proteômica , Albuminas/metabolismo , Reatores Biológicos , Cromatografia Líquida de Alta Pressão , Fermentação , Poliestirenos , Proteínas de Armazenamento de Sementes/metabolismo , Espectrometria de Massas em Tandem
12.
PLoS One ; 15(2): e0229654, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32106262

RESUMO

Human hepatocytes are essential materials in pharmaceutical researches. Not only primary human hepatocytes (PHH) but also human iPS cell-derived hepatocyte-like cells (human iPS-HLCs) are expected to be applied as materials for pharmaceutical researches. To date, several culture media have been developed for culturing human hepatocytes. However, there have been no reports comparing these media to determine which is most suitable for culturing human hepatocytes. In this study, we compared five commercial media (Hepatocyte Culture Medium (HCM), HepatoZYME-SFM, Cellartis Power Primary HEP Medium, DMEM/F12, and William's E Medium (WEM)) to determine which is most suitable for culturing PHH and human iPS-HLCs. In hepatic differentiation of human iPS cells (day 14-25 of differentiation), albumin (ALB) and urea secretion abilities and CYP2C9, CYP2C19, and CYP3A4 activities were the highest when using HCM or WEM. During maintenance of human iPS-HLCs, ALB and urea producing abilities and CYP2C9, CYP2C19, and CYP3A4 activities were the highest when using HCM. Importantly, we found that human iPS-HLCs cultured in HCM were maintained for 3 weeks or more without impairment of their hepatic functions. These results suggest that it is necessary to select an optimal medium for hepatic differentiation and maintenance of human iPS-HLCs. In the case of PHH culture, there was little difference in hepatic functions among the five media. However, the CYP2C9, CYP2C19, and CYP3A4 activities were the highest when using HCM and WEM. In conclusion, it is important to select the optimal medium for specific application when carrying out pharmaceutical researches using human hepatocytes.


Assuntos
Técnicas de Cultura de Células/métodos , Meios de Cultura , Hepatócitos/citologia , Hepatócitos/metabolismo , Albuminas/metabolismo , Diferenciação Celular , Linhagem Celular , Células Cultivadas , Citocromo P-450 CYP2C19/metabolismo , Citocromo P-450 CYP2C9/metabolismo , Citocromo P-450 CYP3A/metabolismo , Humanos , Células-Tronco Pluripotentes Induzidas/citologia , Ureia/metabolismo
13.
Am J Physiol Renal Physiol ; 318(3): F851-F859, 2020 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-32068462

RESUMO

Albuminuria is frequently associated with proximal tubule (PT) cytotoxicity that can feed back to cause glomerular damage and exacerbate kidney disease. PT cells express megalin and cubilin receptors that bind to and internalize albumin over a broad concentration range. How the exposure to high concentrations of albumin leads to PT cytotoxicity remains unclear. Fatty acids and other ligands bound to albumin are known to trigger production of reactive oxygen species (ROS) that impair PT function. Alternatively or in addition, uptake of high concentrations of albumin may overload the endocytic pathway and elicit downstream responses. Here, we used a well-differentiated PT cell culture model with high endocytic capacity to dissect the effects of albumin versus its ligands on endocytic uptake and degradation of albumin, production of ROS, and cell viability. Cellular responses differed dramatically, depending on the preparation of albumin tested. Knockdown of megalin or cubilin failed to prevent ROS production mediated by albumin ligands, suggesting that receptor-mediated internalization of albumin was not necessary to trigger cellular responses to albumin ligands. Moreover, albumin induced cytotoxic responses when added to the basolateral surface of PT cells. Whereas overnight incubation with high concentrations of fatty acid-free albumin had no overt effects on cell function or viability, lysosomal degradation kinetics were slowed upon longer exposure, consistent with overload of the PT endocytic/degradative pathway. Together, the results of our study demonstrate that the PT responds independently to albumin and to its ligands and suggest that the consequences of albumin overload in vivo may be dependent on metabolic state.


Assuntos
Albuminas/metabolismo , Aconitato Hidratase/metabolismo , Albuminas/administração & dosagem , Animais , Linhagem Celular , Técnicas de Silenciamento de Genes , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Proteína-2 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Estresse Oxidativo , Interferência de RNA , Espécies Reativas de Oxigênio , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo
14.
Ann Biol Clin (Paris) ; 78(1): 93-107, 2020 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-32108587

RESUMO

The measurement performance of 13 biochemistry parameters (CEA, CA 19-9, amylase, lipase, sodium, potassium, chloride, creatinine, glucose, protein, albumin, LDH, triglycerides) was tested in a panel of biological fluids other than blood and urine (peritoneal, pleural, pancreatic fluids ...). Our protocol, based on a risk analysis, allowed us to justify our choices and compare the performance obtained with those of the serum or plasma matrix already validated. Thus, the coefficients of variation obtained in body fluids are comparable. The assessment of accuracy (spiking and dilution tests) shows the absence of bias, which is consistent with the absence of matrix effect. The linearity studied by dilution tests shows that the upper limits of the measurement interval communicated by the supplier are applicable to body fluids. The absence of contamination and stability have been also confirmed. All analytes are stable for 3 days at room temperature, 7 days between 2 and 8̊C, and 6 months at -20̊C; except LDH and lipase. For most analytes, at least one interference (hemolysis, icterus, lipemia) was found. Finally, a bibliographical study, confronted with the experience of prescribers, led us to define optimal thresholds to help interpret patients' results. In conclusion, this work has allowed us to validate analytical methods for body fluids testing after relying on their comparability to the blood matrix. We have also been able to adapt our practices and finally be accredited according to the standard NF IN ISO 15189.


Assuntos
Biomarcadores/análise , Líquidos Corporais/química , Técnicas de Laboratório Clínico , Albuminas/análise , Albuminas/metabolismo , Amilases/análise , Amilases/metabolismo , Biomarcadores/metabolismo , Líquidos Corporais/metabolismo , Antígeno CA-19-9/análise , Antígeno CA-19-9/metabolismo , Antígeno Carcinoembrionário/análise , Antígeno Carcinoembrionário/metabolismo , Cloretos/análise , Cloretos/metabolismo , Técnicas de Laboratório Clínico/métodos , Técnicas de Laboratório Clínico/normas , Creatinina/análise , Creatinina/metabolismo , Glucose/análise , Glucose/metabolismo , Humanos , L-Lactato Desidrogenase/análise , L-Lactato Desidrogenase/metabolismo , Lipase/análise , Lipase/metabolismo , Potássio/análise , Potássio/metabolismo , Proteínas/análise , Proteínas/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Sódio/análise , Sódio/metabolismo , Temperatura , Triglicerídeos/análise , Triglicerídeos/metabolismo
15.
Proc Natl Acad Sci U S A ; 117(4): 1976-1987, 2020 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-31924745

RESUMO

Upon activation, fibrinogen forms large fibrin biopolymers that coalesce into clots which assist in wound healing. Limited insights into their molecular architecture, due to the sheer size and the insoluble character of fibrin clots, have restricted our ability to develop novel treatments for clotting diseases. The, so far resolved, disparate structural details have provided insights into linear elongation; however, molecular details like the C-terminal domain of the α-chain, the heparin-binding domain on the ß-chain, and other functional domains remain elusive. To illuminate these dark areas, we applied cross-linking mass spectrometry (XL-MS) to obtain biochemical evidence in the form of over 300 distance constraints and combined this with structural modeling. These restraints additionally define the interaction network of the clots and provide molecular details for the interaction with human serum albumin (HSA). We were able to construct the structural models of the fibrinogen α-chain (excluding two highly flexible regions) and the N termini of the ß-chain, confirm these models with known structural arrangements, and map how the structure laterally aggregates to form intricate lattices together with the γ-chain. We validate the final model by mapping mutations leading to impaired clot formation. From a list of 22 mutations, we uncovered structural features for all, including a crucial role for ßArg'169 (UniProt: 196) in lateral aggregation. The resulting model can potentially serve for research on dysfibrinogenemia and amyloidosis as it provides insights into the molecular mechanisms of thrombosis and bleeding disorders related to fibrinogen variants. The structure is provided in the PDB-DEV repository (PDBDEV_00000030).


Assuntos
Albuminas/metabolismo , Reagentes para Ligações Cruzadas/metabolismo , Fibrina/química , Fibrina/metabolismo , Espectrometria de Massas/métodos , Modelos Estruturais , Trombose/fisiopatologia , Albuminas/química , Fibrina/genética , Humanos , Mutação , Conformação Proteica
16.
PLoS One ; 15(1): e0227980, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31978133

RESUMO

INTRODUCTION: Particles in exhaled air (PEx) provide samples of respiratory tract lining fluid from small airways containing, for example, Surfactant protein A (SP-A) and albumin, potential biomarkers of small airway disease. We hypothesized that there are differences between morning, noon, and afternoon measurements and that the variability of repeated measurements is larger between days than within days. METHODS: PEx was obtained in sixteen healthy non-smoking adults on 11 occasions, within one day and between days. SP-A and albumin were quantified by ELISA. The coefficient of repeatability (CR), intraclass correlation coefficient (ICC), and coefficient of variation (CV) were used to assess the variation of repeated measurements. RESULTS: SP-A and albumin increased significantly from morning towards the noon and afternoon by 13% and 25% on average, respectively, whereas PEx number concentration and particle mean mass did not differ significantly between the morning, noon and afternoon. Between-day CRs were not larger than within-day CRs. CONCLUSIONS: Time of the day influences the contents of SP-A and albumin in exhaled particles. The variation of repeated measurements was rather high but was not influenced by the time intervals between measurements.


Assuntos
Albuminas/isolamento & purificação , Doença Pulmonar Obstrutiva Crônica/diagnóstico , Proteína A Associada a Surfactante Pulmonar/isolamento & purificação , Sistema Respiratório/química , Adulto , Idoso , Ar/análise , Albuminas/metabolismo , Biomarcadores/química , Testes Respiratórios , Expiração/fisiologia , Feminino , Voluntários Saudáveis , Humanos , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/metabolismo , Doença Pulmonar Obstrutiva Crônica/patologia , Proteína A Associada a Surfactante Pulmonar/metabolismo , Sistema Respiratório/metabolismo , Espirometria/métodos
17.
BMC Plant Biol ; 20(1): 45, 2020 Jan 29.
Artigo em Inglês | MEDLINE | ID: mdl-31996140

RESUMO

BACKGROUND: Wheat grain avenin-like proteins (ALPs) belong to a recently discovered class of wheat grain storage protein. ALPs in wheat grains not only have beneficial effects on dough quality but also display antifungal activities, which is a novel observation for wheat storage proteins. Previous studies have shown that ALPs are likely present in the albumin/globulin fractions of total protein extract from wheat flour. However, the accumulation characteristics of these ALPs in the mature wheat grain remains unknown. RESULTS: In the present study, a total of 13 ALPs homologs were isolated and characterized in the albumin/globulin fractions of the wheat protein extract. A combination of multiple techniques including RP-HPLC, SDS-PAGE, MALDI-TOF and peptide sequencing were used for accurate separation and identification of individual ALP homolog. The C-terminal TaALP-by-4AL/7DS, TaALP-by-4AL/7AS/7DS, TaALP-bx/4AL/7AS/7DS, TaALP-ay-7DS, TaALP-ay-4AL, TaALP-ax-4AL, TaALP-ax-7AS, and TaALP-ax-7DS, were separated as individual protein bands from wheat flour for the first time. These unique ALPs peptides were mapped to the latest wheat genome assembly in the IWGSC database. The characteristic defence related proteins present in albumin and globulin fractions, such as protein disulfide-isomerase (PDI), grain softness protein (GSP), alpha-amylase inhibitors (AAIs) and endogenous alpha-amylase/subtilisin inhibitor were also found to co-segregate with these identified ALPs, avenin-3 and α-gliadins. The molecular weight range and the electrophoresis segregation properties of ALPs were characterised in comparison with the proteins containing the tryp_alpha_amyl domain (PF00234) and the gliadin domain (PF13016), which play a role in plant immunity and grain quality. We examined the phylogenetic relationships of the AAIs, GSP, avenin-3, α-gliadins and ALPs, based on the alignment of their functional domains. MALDI-TOF profiling indicated the occurrence of certain post-translations modifications (PTMs) in some ALP subunits. CONCLUSIONS: We reported for the first time the complete profiling of ALPs present in the albumin/globulin fractions of wheat grain protein extracts. We concluded that majority of the ALPs homologs are expressed in wheat grains. We found clear evidence of PTMs in several ALPs peptides. The identification of both gliadin domain (PF13016) and Tryp_alpha_amyl domain (PF00234) in the mature forms of ALPs highlighted the multiple functional properties of ALPs in grain quality and disease resistance.


Assuntos
Grão Comestível/metabolismo , Prolaminas/metabolismo , Triticum/metabolismo , Albuminas/metabolismo , Cromatografia Líquida de Alta Pressão , Eletroforese em Gel de Poliacrilamida , Globulinas/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Processamento de Proteína Pós-Traducional , Espectrometria de Massas em Tandem
18.
Angiology ; 71(4): 360-365, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31888345

RESUMO

Several laboratory parameters have been used to assess inflammatory process and determine cardiovascular risk. The C-reactive protein to albumin ratio (CAR) is a novel marker of inflammation and its clinical importance has not been clearly elucidated in coronary artery disease (CAD). We compared the diagnostic value of CAR with other inflammatory parameters in detecting significant CAD. Patients (n = 421) with stable angina pectoris who underwent coronary angiography for the suspected CAD were included. Neutrophil to lymphocyte ratio (NLR), monocyte to lymphocyte ratio (MLR), platelet to lymphocyte ratio, uric acid, monocyte to high-density cholesterol (HDL-C) ratio, mean platelet volume to lymphocyte ratio (MPVLR), and platelet to mean corpuscular volume (MCV) ratio were measured. Patients with significant CAD had a significantly higher NLR (P = .043), MLR (P = .004), uric acid (P < .001), monocyte to HDL-C ratio (P = .004), and CAR (P < .001) compared to patients without significant CAD. However, MPVLR and platelet to MCV ratio weren't different between 2 groups. The area under the curve (AUC) of CAR was the highest AUC among all inflammatory parameters for predicting significant CAD. Multivariate analysis showed that age (odds ratio [OR]: 1.046, 95% confidence interval [CI], 1.020-1.072, P < .001) and CAR (OR: 1.175, 95% CI, 1.126-1.226, P < .001) were the only independent predictors of significant CAD. In conclusion, CAR had the strongest diagnostic value in detecting significant CAD among the inflammatory parameters evaluated in this study.


Assuntos
Albuminas/metabolismo , Angina Estável/diagnóstico , Angina Estável/metabolismo , Proteína C-Reativa/metabolismo , Doença da Artéria Coronariana/metabolismo , Biomarcadores/metabolismo , Angiografia Coronária , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade
19.
World J Surg Oncol ; 18(1): 9, 2020 Jan 13.
Artigo em Inglês | MEDLINE | ID: mdl-31931816

RESUMO

OBJECTIVE: Provide an updated and comprehensive evaluation of the prognostic value of the albumin-fibrinogen ratio (AFR) and the fibrinogen-prealbumin ratio (FPR) for patients with cancer. MATERIALS AND METHODS: Four databases (PubMed, Web of Science, Cochrane Library, and WanFang) were searched. The primary endpoints were overall survival (OS), disease-free survival (DFS), and progression-free survival (PFS). Pooled data were synthesized using StataMP 14 and expressed as hazard ratios (HRs) and 95% confidence intervals (CIs). RESULTS: This update examined 19 studies (7282 cases) that assessed the correlation of AFR with cancer prognosis. Pooled univariate and multivariate analyses indicated significant correlations of low AFR with poor OS (HR 2.18, 95%CI 1.87-2.55 and HR 1.75, 95%CI 1.54-2.00, respectively), poor DFS (HR 1.89, 95%CI 1.54-2.32 and HR 1.51, 95%CI 1.29-1.76, respectively), and poor PFS (HR 1.68, 95%CI 1.42-1.99 and HR 1.48, 95%CI 1.16-1.88, respectively). Pooled univariate and multivariate analyses of 6 studies (2232 cases) indicated high FPR significantly correlated with poor OS (HR 2.37, 95%CI 2.03-2.77 and HR 1.97, 95%CI 1.41-2.77, respectively). One study reported that high FPR correlated with poor DFS (univariate analysis: HR 2.20, 95%CI 1.35-3.57; multivariate analysis: HR 1.77, 95%CI 1.04-2.99) and one study reported a correlation of high FPR with poor PFS in univariate analysis alone (HR 1.79, 95%CI 1.11-2.88). CONCLUSION: A low AFR and a high FPR correlated with increased risk of cancer mortality and recurrence. AFR and FPR may be promising prognostic markers for cancers.


Assuntos
Albuminas/metabolismo , Fibrinogênio/metabolismo , Neoplasias/sangue , Neoplasias/patologia , Pré-Albumina/metabolismo , Biomarcadores Tumorais/sangue , Humanos , Prognóstico
20.
Eur J Med Chem ; 186: 111863, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31740050

RESUMO

In recent years, drug discovery paradigm has been shifted from conventional single target inhibition toward multitarget design concept. In current research, we have reported synthesis, in-vitro, in-vivo and acute toxicity determination of N-substituted pyrrolidine-2,5-dione derivatives as multitarget anti-inflammatory agents. We synthesized cycloalkyl, alkyl and aryl carbonyl derivatives by the Michael addition of ketones to N-substituted maleimides using self-assembled three component system as an organocatalyst. Anti-inflammatory potential of the compounds was determined by using different in-vitro assays, like cyclooxygenase-1, cyclooxygenase-2 and 5-lipoxygenase, albumin denaturation and anti-protease assays. Amongst the synthesized compounds, 13a-e series of compounds showed inhibition in low micromolar to submicromolar ranges. These compounds also demonstrated COX-2 selectivity. Compound 13e with IC50 value 0.98 µM and SI of 31.5 emerged as the most potent inhibitor of COX-2. Based on in-vitro results, in-vivo anti-inflammatory investigations were performed on compounds 3b and 13evia carrageenan induced paw edema test. The possible mode of action of compounds 3b and 13e were ascertained with various mediators like histamine, bradykinin, prostaglandin and leukotriene. In-vivo acute toxicity study showed the safety of synthesized compounds up to 1000 mg/kg dose. The selectivity of the compounds against cyclooxygenase isoforms was supported by docking simulations. Selective COX-2 inhibitors showed significant interactions with the amino acid residues present in additional secondary COX-2 enzyme pocket. Furthermore, in-silico pharmacokinetic predictions confer the drug-like characteristics.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Desenho de Fármacos , Inibidores Enzimáticos/farmacologia , Simulação de Acoplamento Molecular , Pirrolidinas/farmacologia , Albuminas/antagonistas & inibidores , Albuminas/metabolismo , Animais , Anti-Inflamatórios não Esteroides/síntese química , Anti-Inflamatórios não Esteroides/química , Araquidonato 5-Lipoxigenase/metabolismo , Carragenina , Ciclo-Oxigenase 2/metabolismo , Relação Dose-Resposta a Droga , Edema/induzido quimicamente , Edema/tratamento farmacológico , Edema/metabolismo , Inibidores Enzimáticos/síntese química , Inibidores Enzimáticos/química , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Pirrolidinas/síntese química , Pirrolidinas/química , Relação Estrutura-Atividade
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