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1.
Molecules ; 27(3)2022 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-35164316

RESUMO

In this study, temperature-responsive polymer-protein conjugate was synthesized using a "grafting from" concept by introducing a chain transfer agent (CTA) into bovine serum albumin (BSA). The BSA-CTA was used as a starting point for poly(N-isopropylacrylamide) (PNIPAAm) through reversible addition-fragmentation chain transfer polymerization. The research investigations suggest that the thermally responsive behavior of PNIPAAm was controlled by the monomer ratio to CTA, as well as the amount of CTA introduced to BSA. The study further synthesized the human serum albumin (HSA)-PNIPAAm conjugate, taking the advantage that HSA can specifically adsorb indoxyl sulfate (IS) as a uremic toxin. The HSA-PNIPAAm conjugate could capture IS and decreased the concentration by about 40% by thermal precipitation. It was also revealed that the protein activity was not impaired by the conjugation with PNIPAAm. The proposed strategy is promising in not only removal of uremic toxins but also enrichment of biomarkers for early diagnostic applications.


Assuntos
Resinas Acrílicas/química , Soroalbumina Bovina/química , Albumina Sérica Humana/química , Toxinas Urêmicas/isolamento & purificação , Resinas Acrílicas/síntese química , Adsorção , Animais , Bovinos , Humanos , Indicã/isolamento & purificação , Soroalbumina Bovina/síntese química , Albumina Sérica Humana/síntese química , Temperatura
2.
Blood Purif ; 51(1): 38-46, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-33789268

RESUMO

INTRODUCTION: Haemodialysis (HD) allow depuration of uraemic toxins by diffusion, convection, and adsorption. Online haemodiafiltration (HDF) treatments add high convection to enhance removal. There are no prior studies on the relationship between convection and adsorption in HD membranes. The possible benefits conferred by intrinsic adsorption on protein-bound uraemic toxins (PBUTs) removal are unknown. METHODS: Twenty-two patients underwent their second 3-days per week HD sessions with randomly selected haemodialysers (polysulfone, polymethylmethacrylate, cellulose triacetate, and polyamide copolymer) in high-flux HD and HDF. Blood samples were taken at the beginning and at the end of the treatment to assess the reduction ratio (RR) in a wide range of molecular weight uraemic toxins. A mid-range removal score (GRS) was also calculated. An elution protocol was implemented to quantify the amount of adsorbed mass (Mads) for each molecule in every dialyser. RESULTS: All synthetic membranes achieved higher RR for all toxins when used in HDF, specially the polysulfone haemodialyser, resulting in a GRS = 0.66 ± 0.06 (p < 0.001 vs. cellulose triacetate and polyamide membranes). Adsorption was slightly enhanced by convection for all membranes. The polymethylmethacrylate membrane showed expected substantial adsorption of ß2-microglobulin (MadsHDF = 3.5 ± 2.1 mg vs. MadsHD = 2.1 ± 0.9 mg, p = 0.511), whereas total protein adsorption was pronounced in the cellulose triacetate membrane (MadsHDF = 427.2 ± 207.9 mg vs. MadsHD = 274.7 ± 138.3 mg, p = 0.586) without enhanced PBUT removal. DISCUSSION/CONCLUSION: Convection improves removal and slightly increases adsorption. Adsorbed proteins do not lead to enhanced PBUTs depuration and limit membrane efficiency due to fouling. Selection of the correct membrane for convective therapies is mandatory to optimize removal efficiency.


Assuntos
Hemodiafiltração/instrumentação , Membranas Artificiais , Toxinas Urêmicas/isolamento & purificação , Adsorção , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Uremia/terapia , Toxinas Urêmicas/sangue
3.
ACS Appl Mater Interfaces ; 13(38): 45149-45160, 2021 Sep 29.
Artigo em Inglês | MEDLINE | ID: mdl-34520182

RESUMO

The removal of uremic toxins from patients with acute kidney injury is a key issue in improving the quality of life for people requiring peritoneal dialysis. The currently utilized method for the removal of uremic toxins from the human organism is hemodialysis, performed on semipermeable membranes where the uremic toxins, along with small molecules, are separated from proteins and blood cells. In this study, we describe a mixed-linker modulated synthesis of zirconium-based metal-organic frameworks for efficient removal of uremic toxins. We determined that the efficient adsorption of uremic toxins is achieved by optimizing the ratio between -amino functionalization of the UiO-66 structure with 75% of -NH2 groups within organic linker structure. The maximum adsorption of hippuric acid and 3-indoloacetic acid was achieved by UiO-66-NH2 (75%) and by UiO-66-NH2 (75%) 12.5% HCl prepared by modulated synthesis. Furthermore, UiO-66-NH2 (75%) almost completely adsorbs 3-indoloacetic acid bound to bovine serum albumin, which was used as a model protein to which uremic toxins bind in the human body. The high adsorption capacity was confirmed in recyclability test, which showed almost 80% removal of 3-indoloacetic acid after the third adsorption cycle. Furthermore, in vitro cytotoxicity tests as well as hemolytic activity assay have proven that the UiO-66-based materials can be considered as potentially safe for hemodialytic purposes in living organisms.


Assuntos
Hipuratos/isolamento & purificação , Ácidos Indolacéticos/isolamento & purificação , Rins Artificiais , Estruturas Metalorgânicas/química , Ácidos Ftálicos/química , Toxinas Urêmicas/isolamento & purificação , Adsorção , Animais , Chlorocebus aethiops , Eritrócitos/efeitos dos fármacos , Células HEK293 , Hipuratos/química , Humanos , Ácidos Indolacéticos/química , Estruturas Metalorgânicas/síntese química , Estruturas Metalorgânicas/toxicidade , Ácidos Ftálicos/síntese química , Ácidos Ftálicos/toxicidade , Toxinas Urêmicas/química , Células Vero , Zircônio/química
4.
J Mol Recognit ; 34(10): e2901, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-33975380

RESUMO

The last 5 years have seen a series of advances in the application of isothermal titration microcalorimetry (ITC) and interpretation of ITC data. ITC has played an invaluable role in understanding multiprotein complex formation including proteolysis-targeting chimeras (PROTACS), and mitochondrial autophagy receptor Nix interaction with LC3 and GABARAP. It has also helped elucidate complex allosteric communication in protein complexes like trp RNA-binding attenuation protein (TRAP) complex. Advances in kinetics analysis have enabled the calculation of kinetic rate constants from pre-existing ITC data sets. Diverse strategies have also been developed to study enzyme kinetics and enzyme-inhibitor interactions. ITC has also been applied to study small molecule solvent and solute interactions involved in extraction, separation, and purification applications including liquid-liquid separation and extractive distillation. Diverse applications of ITC have been developed from the analysis of protein instability at different temperatures, determination of enzyme kinetics in suspensions of living cells to the adsorption of uremic toxins from aqueous streams.


Assuntos
Calorimetria/métodos , Descoberta de Drogas/métodos , Enzimas/química , Proteínas/química , Animais , Pesquisa Biomédica/métodos , Calorimetria/instrumentação , Catálise , Entropia , Enzimas/metabolismo , Humanos , Extração Líquido-Líquido/métodos , Minerais/química , Minerais/isolamento & purificação , Toxinas Urêmicas/química , Toxinas Urêmicas/isolamento & purificação
5.
Blood Purif ; 50(4-5): 492-498, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33291102

RESUMO

BACKGROUND: A new medium cut-off (MCO) membranes has been designed to achieve better removal capacities for middle and large middle molecules in hemodialysis (HD) treatment. AIM: The aim of this study was to evaluate the removal efficacy of Theranova® in standard HD in comparison with standard high-flux HD. METHODS: Four HD patients (M/F 1/4) were included in 12-week observational pilot study in HD with Theranova® 400 and Theranova® 500 dialyzers. Each patient was assessed 4 times, T0 with high-flux dialyzers, T1 at 1 month, T2 at second month, and T3 at third month, by measuring pre- and post-HD samples of urea, Cr, ß2-microglobilin (ß2M), myoglobin, albumin, free light chains kappa (FLC-k), and free light chains lambda (FLC-λ). RESULTS: The data showed a higher average removal rate for all the uremic toxins with Theranova® dialyzers for ß2M, myoglobin, FLC-k, and FLC-λ (62.7, 56.9, 63.5, and 54.6%, respectively) during the 3 months. Albumin retention was observed and did not change between T0 and T3 (p = 0.379). CONCLUSION: Compared to high-flux membranes, MCO membranes show greater permeability for middle molecules in midterm report.


Assuntos
Diálise Renal/instrumentação , Adulto , Idoso , Feminino , Humanos , Cadeias kappa de Imunoglobulina/sangue , Cadeias kappa de Imunoglobulina/isolamento & purificação , Masculino , Pessoa de Meia-Idade , Permeabilidade , Projetos Piloto , Diálise Renal/métodos , Ureia/sangue , Ureia/isolamento & purificação , Toxinas Urêmicas/sangue , Toxinas Urêmicas/isolamento & purificação , Microglobulina beta-2/sangue , Microglobulina beta-2/isolamento & purificação
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