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1.
Cancer Immunol Immunother ; 68(9): 1537-1545, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31482306

RESUMO

PURPOSE: To evaluate the clinical-pathological and prognostic significance of the circulating PD-L1 level in patients with surgically treated NSCLC, by combining data for PD-L1 expression with other immune-related markers and tumor metabolism. METHODS: Overall, 40 patients with resected NSCLC (stage Ia-IIIa) who had preoperative blood storage and underwent staging PET/CT were enrolled for the study. In all cases, we determined plasma levels of PD-L1 (pg/ml), immune-reactive areas (IRA %) covered by CD3, CD68, CD20, CD8, PD-1, and PD-L1 in the tumor specimen, and metabolic parameters on PET, i.e., SUVmax, SUVpeak, metabolic tumor volume (MTV), and total lesion glycolysis (TLG). Variables were statistically analyzed to establish their association with disease-free survival (DFS). RESULTS: The circulating levels of PD-L1 in the bloodstream could be determined in 38/40 (95%) samples. The mean and median expression levels were 34.86 pg/ml and 24.83 pg/ml, respectively. We did not find any statistically significant correlation between circulating PD-L1 and tissue expression of PD-L1/PD-1. Some mild degree of positive correlation was determined between tissue PD-L1 and SUVmax (ρ = 0.390; p = 0.0148). Hierarchical clustering combining circulating, tissue, and metabolic parameters identified clusters with high metabolic tumor burden or high expression of plasma PD-L1 levels (Z score ≥ 2) as having a poor DFS (p = 0.033). The multivariate analysis detected stage and metabolism (i.e., SUVmax and SUVpeak) as independent prognostic factors for DFS. CONCLUSION: Plasma levels of PD-L1 are independent of the expression of PD-1/PD-L1 in NSCLC tumor tissue and, when combined with other clinical-pathological parameters, allow for the identification of clusters with different outcomes.


Assuntos
Antígeno B7-H1/metabolismo , Proteínas Sanguíneas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno B7-H1/genética , Proteínas Sanguíneas/genética , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Feminino , Regulação Neoplásica da Expressão Gênica , Glicólise , Humanos , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/mortalidade , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Prognóstico , Análise de Sobrevida , Resultado do Tratamento , Carga Tumoral
2.
Eur J Pharm Biopharm ; 143: 44-50, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31421208

RESUMO

Today, a growing number of nanotherapeutics is utilized to deliver poorly soluble compounds using the intravenous route of administration. The drug release and the direct transfer of the active pharmaceutical ingredient to serum proteins plays an important role in bioavailability and accumulation of the drug at the target site. It is closely related to the formation of a protein corona as well as the plasma protein binding of the compound. In the present study, two in vitro drug release methods, the flow-through cell and the dispersion releaser technology, were evaluated with regards to their capability to measure a time-resolved profile of the serum protein binding. In this context, the photosensitizer temoporfin and temoporfin-loaded liposomes were tested. While in the fine capillaries of the flow-through cell a rapid agglomeration of proteins occurred, the dispersion releaser technology in combination with the four-step model enabled the measurement of the transfer of drugs from liposomes to proteins. In presence of 10% of fetal calf serum approximately 20% of the model compound temoporfin were bound to serum proteins within the first 3 h. At higher serum concentration this binding remained stable for approximately 10 h.


Assuntos
Proteínas Sanguíneas/metabolismo , Lipossomos/química , Mesoporfirinas/química , Mesoporfirinas/metabolismo , Animais , Disponibilidade Biológica , Bovinos , Portadores de Fármacos/química , Liberação Controlada de Fármacos/efeitos dos fármacos , Cinética , Tamanho da Partícula , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/metabolismo , Ligação Proteica/efeitos dos fármacos
3.
Clin Biochem ; 73: 90-97, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31401122

RESUMO

BACKGROUND: Patients treated for human immunodeficiency virus (HIV) infection are prone to developing chronic kidney disease (CKD). Current methods used in assessing kidney function suffer inaccuracy in HIV-infected patients. This study aims to identify biomarkers that could complement existing methods of kidney assessment among HIV-infected subjects. METHODS: Plasma protein profiling was performed for HIV patients with CKD presented with negative/trace proteinuria (non-proteinuric) (n = 8) and their matched non-CKD controls, using two-dimensional gel electrophoresis (2DE); selected protein candidates were identified using mass spectrometry. Subsequently, altered plasma abundance of protein candidates were verified using Western blotting in HIV-infected subjects with non-proteinuric CKD (n = 8), proteinuric CKD (n = 5), and their matched non-CKD controls, as well as in HIV-uninfected subjects with impaired kidney function (n = 3) and their matched controls. RESULTS: Analysis of 2DE found significantly altered abundance of five protein candidates between HIV-infected patients with non-proteinuric CKD and without CKD: alpha-1-microglobulin (A1M), serum albumin (ALB), zinc-alpha-2-glycoprotein (AZGP1), haptoglobin (HP), and retinol binding protein (RBP4). Western blotting showed an increased abundance of A1M and HP in HIV-infected patients with non-proteinuric CKD compared to their non-CKD controls, whereas A1M, AZGP1, and RBP4 were significantly increased in HIV-infected patients with proteinuric CKD compared to their non-CKD controls. Such pattern was not found in HIV-uninfected subjects with impaired kidney function. CONCLUSION: The data suggests four proteins that may be used as biomarkers of CKD in HIV-infected patients. Further validation in a larger cohort of HIV-infected patients is necessary for assessing the clinical use of these proposed biomarkers for CKD.


Assuntos
Proteínas Sanguíneas/metabolismo , Infecções por HIV/sangue , HIV-1 , Proteinúria/sangue , Proteômica , Insuficiência Renal Crônica/sangue , Idoso , Biomarcadores/sangue , Feminino , Infecções por HIV/complicações , Humanos , Masculino , Pessoa de Meia-Idade , Proteinúria/complicações , Insuficiência Renal Crônica/complicações
4.
BMC Bioinformatics ; 20(1): 333, 2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31195980

RESUMO

BACKGROUND: Parametric feature selection methods for machine learning and association studies based on genetic data are not robust with respect to outliers or influential observations. While rank-based, distribution-free statistics offer a robust alternative to parametric methods, their practical utility can be limited, as they demand significant computational resources when analyzing high-dimensional data. For genetic studies that seek to identify variants, the hypothesis is constrained, since it is typically assumed that the effect of the genotype on the phenotype is monotone (e.g., an additive genetic effect). Similarly, predictors for machine learning applications may have natural ordering constraints. Cross-validation for feature selection in these high-dimensional contexts necessitates highly efficient computational algorithms for the robust evaluation of many features. RESULTS: We have developed an R extension package, fastJT, for conducting genome-wide association studies and feature selection for machine learning using the Jonckheere-Terpstra statistic for constrained hypotheses. The kernel of the package features an efficient algorithm for calculating the statistics, replacing the pairwise comparison and counting processes with a data sorting and searching procedure, reducing computational complexity from O(n2) to O(n log(n)). The computational efficiency is demonstrated through extensive benchmarking, and example applications to real data are presented. CONCLUSIONS: fastJT is an open-source R extension package, applying the Jonckheere-Terpstra statistic for robust feature selection for machine learning and association studies. The package implements an efficient algorithm which leverages internal information among the samples to avoid unnecessary computations, and incorporates shared-memory parallel programming to further boost performance on multi-core machines.


Assuntos
Algoritmos , Estudo de Associação Genômica Ampla , Aprendizado de Máquina , Proteínas Sanguíneas/metabolismo , Simulação por Computador , Polimorfismo de Nucleotídeo Único/genética , Característica Quantitativa Herdável
5.
Ann Hematol ; 98(8): 1827-1834, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31190133

RESUMO

In this study, we aimed to investigate the pattern and association of genetic mutations occurring within the alpha hemoglobin-stabilizing protein (AHSP) gene among HbE beta thalassemia patients with varying phenotypic expressions. Fifty-four diagnosed cases of HbE beta thalassemia (transfusion dependent and independent) were included in the study. Among them, 38 patients with similar genotypes (IVS 1-5, alpha gene deletion and triplication, Xmn polymorphism) were selected for further analysis. AHSP gene sequencing was done for these 38 samples to study associated mutations in AHSP gene. HbE beta thalassemia patients with similar genotypes but different phenotypic expressions were found to have mutations in the AHSP gene. There were five mutations found most prevalent among the samples analyzed for AHSP gene sequencing. Among these, two mutations were from intron 1 region of AHSP and three mutations were found in exon 3. The most prevalent mutation was found at the Oct binding site at intron 1 of AHSP. The mutations in exon 3 were more prevalent among the TDT groups. A mutation in exon 3 changing the amino acid (33rd) from serine to phenylalanine was found to be associated with only TDT group. This study documents that among the HbE beta thalassemia patients with varying severity, an exon mutation in AHSP is significantly prevalent only among the TDT group. Further understanding of the mechanism will shed light upon the impact of AHSP in modifying the disease severity in thalassemia.


Assuntos
Proteínas Sanguíneas/genética , Deleção de Genes , Duplicação Gênica , Hemoglobina E/genética , Chaperonas Moleculares/genética , Talassemia beta/genética , Adolescente , Adulto , Sequência de Bases , Proteínas Sanguíneas/metabolismo , Transfusão de Sangue/estatística & dados numéricos , Criança , Pré-Escolar , Análise Mutacional de DNA , Eritrócitos/metabolismo , Eritrócitos/patologia , Éxons , Feminino , Expressão Gênica , Genótipo , Hemoglobina E/metabolismo , Humanos , Íntrons , Masculino , Pessoa de Meia-Idade , Modelos Moleculares , Chaperonas Moleculares/metabolismo , Fenótipo , Estrutura Secundária de Proteína , Índice de Gravidade de Doença , Talassemia beta/metabolismo , Talassemia beta/patologia , Talassemia beta/terapia
6.
Cell Mol Life Sci ; 76(18): 3515-3523, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31089746

RESUMO

Cytokine-like protein 1 (Cytl1), also named Protein C17 or C4orf4 is located on human chromosome 4p15-p16 and encodes a polypeptide of 126 amino acid residues that displays characteristics of a secretory protein. Cytl1 is expressed by a sub-population of CD34+ human mononuclear cells from bone marrow and cord blood, and by chondrocytes (cartilage-forming cells). In this review, we explore evidence suggesting that Cytl1 may be involved in the regulation of chondrogenesis, cartilage homeostasis and osteoarthritis progression, accompanied by the modulation of Sox9 and insulin-like growth factor 1 expression. In addition, Cytl1 exhibits chemotactic and pro-angiogenic biological effects. Interestingly, CCR2 (C-C chemokine receptor type 2) has been identified as a likely receptor for Cytl1, which mediates the ERK signalling pathway. Cytl1 also appears to mediate the TGF-beta-Smad signalling pathway, which is hypothetically independent of the CCR2 receptor. More recently, studies have also potentially linked Cytl1 with a variety of conditions including cardiac fibrosis, smoking, alcohol dependence risk, and tumours such as benign prostatic hypertrophy, lung squamous cell carcinoma, neuroblastoma and familial colorectal cancer. Defining the molecular structure of Cytl1 and its role in disease pathogenesis will help us to design therapeutic approaches for Cytl1-associated pathological conditions.


Assuntos
Proteínas Sanguíneas/metabolismo , Cartilagem/metabolismo , Citocinas/metabolismo , Proteínas Sanguíneas/química , Proteínas Sanguíneas/genética , Condrócitos/citologia , Condrócitos/metabolismo , Condrogênese , Citocinas/química , Citocinas/genética , Humanos , Osteoartrite/metabolismo , Osteoartrite/patologia , Receptores CCR2/metabolismo , Transdução de Sinais
7.
Zhongguo Zhong Yao Za Zhi ; 44(7): 1475-1484, 2019 Apr.
Artigo em Chinês | MEDLINE | ID: mdl-31090307

RESUMO

To determine the plasma protein binding rate of the nine compounds in Inula cappa extraction by the method of equilibrium dialysis. The proteins in plasma samples were precipitated by methanol, and the ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS) was developed for determination of the concentrations of the nine active compounds, namely chlorogenic acid, scopolin, neochlorogenic acid, cryptochlorogenic acid, 1,3-O-dicaffeoylquinic acid, galuteolin, 3,4-O-dicaffeoylquinic acid, 3,5-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, with the internal standard of puerarin. We found that all components have a good linearity(r≥0.999), and accuracy, precision, extraction recovery and stability conformed to the requirements of determination, without endogenous compounds disturbing within the range of optimum concentration. This suggested that the method was stable and reliable, and could be used for the determination of the plasma protein binding rates of the nine active compounds in rat and human plasma of I. cappa. The plasma protein binding rates of the nine active compounds in rat and human plasma respectively were(41.07±0.046)%-(94.95±0.008)%, and(37.66±0.043)%-(97.46±0.013)%. According to the results, there were differences in the plasma protein binding rates of the nine compounds in I. cappa extraction between rat and human.


Assuntos
Proteínas Sanguíneas/metabolismo , Inula/química , Compostos Fitoquímicos/metabolismo , Extratos Vegetais/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Humanos , Ligação Proteica , Ratos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem
8.
Folia Histochem Cytobiol ; 57(2): 43-55, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31099889

RESUMO

Biological membranes are organized in various microdomains, one of the best known being called membrane rafts. The major function of these is thought to organize signaling partners into functional complexes. An important protein found in membrane raft microdomains of erythroid and other blood cells is MPP1 (membrane palmitoylated protein 1)/p55. MPP1 (p55) belongs to the MAGUK (membrane-associated guanylate kinase homolog) family and it is a major target of palmitoylation in the red blood cells (RBCs) membrane. The well-known function of this protein is to participate in formation of the junctional complex of the erythrocyte mem-brane skeleton. However, its function as a "raft organizer" is not well understood. In this review we focus on recent reports concerning MPP1 participation in membrane rafts organization in erythroid cells, including its role in signal transduction. Currently it is not known whether MPP1 could have a similar role in cell types other than erythroid lineage. We present also preliminary data regarding the expression level of MPP1 gene in several non-erythroid cell lines.


Assuntos
Proteínas Sanguíneas/metabolismo , Membrana Celular/metabolismo , Eritrócitos/metabolismo , Microdomínios da Membrana/metabolismo , Proteínas de Membrana/metabolismo , Proteínas Sanguíneas/genética , Colesterol/metabolismo , Humanos , Fluidez de Membrana/fisiologia , Lipídeos de Membrana/metabolismo , Proteínas de Membrana/genética , Ligação Proteica
9.
Biomed Res Int ; 2019: 2181370, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31032337

RESUMO

Introduction: Oxidative stress is a state of imbalance between the production of reactive oxygen species and antioxidant defenses. It results in the oxidation of all cellular elements and, to a large extent, proteins, causing inter alia the formation of carbonyl groups in their structures. The study focused on assessment of changes in the plasma protein-bound carbonyls in police horses after combat training and after rest and the applicability of infrared spectroscopy with a Fourier transform, utilizing the attenuated total reflectance (FTIR-ATR) in detecting plasma protein oxidation. Methods: We evaluated the influence of both the different concentrations of hydrogen peroxide and combat training on protein carbonylation in horse blood plasma. The oxidation of plasma proteins was assessed using a spectrophotometric method based on the carbonyl groups derivatization with 2,4-dinitrophenylhydrazine (DNPH). The measured values were correlated with the carbonyl groups concentrations determined by means of the FTIR-ATR method. Results: The linear correlation between the DNPH and FTIR-ATR methods was shown. The concentration of plasma protein-bound carbonyls significantly deceased in police horses after one-day rest when compared to the values measured directly after the combat training (a drop by 23%, p<0.05 and 29%, p<0.01 measured by DNPH and FTIR-ATR methods, respectively). These results were consistent with the proteins phosphorylation analysis. Conclusion: The FTIR-ATR method may be applied to measure the level of plasma proteins peroxidation.


Assuntos
Antioxidantes/metabolismo , Proteínas Sanguíneas/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Carbonilação Proteica/efeitos dos fármacos , Animais , Antioxidantes/química , Proteínas Sanguíneas/química , Proteínas Sanguíneas/efeitos dos fármacos , Cavalos , Humanos , Hidrazinas/química , Hidrazinas/metabolismo , Peróxido de Hidrogênio/farmacologia , Oxirredução , Espécies Reativas de Oxigênio/sangue , Espectroscopia de Infravermelho com Transformada de Fourier
10.
In Vivo ; 33(3): 771-776, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31028196

RESUMO

BACKGROUND/AIM: Hemostatic system components contribute to cancer progression independently from their roles in hemostasis. It has been shown that protein Z (PZ)/protein Z-dependent protease inhibitor (ZPI) inhibit coagulation factor X (FX). The aim of the study was to analyze the expression of PZ/ZPI in relation to the main coagulation factor - FX in human endometrial cancer tissue. MATERIALS AND METHODS: Immunohistochemical analysis was performed on 21 endometrial cancer specimens employing antibodies against ZPI, PZ and FX. RESULTS: Endometrial cancer cells showed a strong expression of ZPI and PZ and medium expression of FX. Normal endometrial tissue showed no expression of ZPI, PZ or FX. CONCLUSION: Strong expression of PZ and ZPI in endometrial cancer cells suggests a role of these proteins in endometrial cancer.


Assuntos
Proteínas Sanguíneas/metabolismo , Neoplasias do Endométrio/metabolismo , Fator X/metabolismo , Biomarcadores , Neoplasias do Endométrio/patologia , Feminino , Humanos , Imuno-Histoquímica , Ligação Proteica , Transporte Proteico
11.
Gen Comp Endocrinol ; 280: 97-103, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31002824

RESUMO

Continuous light or dark photoperiods are the leading cause of disruption in the circadian rhythm of day-night cycle. The purpose of this study was to understand the cellular redox balance in a model of circadian disrupted rat model and determine the effect of melatonin supplementation. Young male Wistar rats were randomly divided into five groups (n = 4). Group (I): normal day-night (12 h:12 h) cycle, Group (II): normal rats treated with melatonin, Group (III): rats subjected to continuous light exposure (CLE), Group (IV): CLE rats treated with melatonin, and Group (V) Rats subjected to continuous dark. Melatonin (10 mg/kg) was administered orally at dusk to the Group (II) & (IV). Rats were sacrificed after 10 days of treatment and biomarkers of oxidative stress were evaluated. Results demonstrated significant (p < 0.05) increase of malondialdehyde (MDA), plasma membrane redox system (PMRS), protein carbonyl oxidation (PCO), advanced oxidation protein products (AOPPs), and advanced glycation end products (AGEs) during CLE. A significantly (p < 0.05) decreased level of reduced glutathione (GSH) and ferric reducing antioxidant potential in plasma (FRAP) was also observed during CLE. Treatment with melatonin in CLE rats showed reduced level of MDA, PMRS, PCO, AOPPs and AGEs while GSH and FRAP activity were increased. During continuous dark exposure (CDE) the biomarkers of oxidative stress were attenuated compared to control. Supplementation of melatonin could be a promising strategy to maintain redox homeostasis during prolonged condition of light exposure and other conditions of redox imbalance.


Assuntos
Ritmo Circadiano/fisiologia , Suplementos Nutricionais , Homeostase , Melatonina/farmacologia , Animais , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Proteínas Sanguíneas/metabolismo , Peso Corporal/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Eritrócitos/efeitos dos fármacos , Eritrócitos/metabolismo , Glutationa/metabolismo , Homeostase/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Malondialdeído/metabolismo , Melatonina/administração & dosagem , Modelos Animais , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Ratos Wistar , Espécies Reativas de Oxigênio/metabolismo
12.
Nat Med ; 25(5): 805-813, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31011203

RESUMO

Chronic inflammation is postulated to be involved in the development of end-stage renal disease in diabetes, but which specific circulating inflammatory proteins contribute to this risk remain unknown. To study this, we examined 194 circulating inflammatory proteins in subjects from three independent cohorts with type 1 and type 2 diabetes. In each cohort, we identified an extremely robust kidney risk inflammatory signature (KRIS), consisting of 17 proteins enriched in tumor necrosis factor-receptor superfamily members, that was associated with a 10-year risk of end-stage renal disease. All these proteins had a systemic, non-kidney source. Our prospective study findings provide strong evidence that KRIS proteins contribute to the inflammatory process underlying end-stage renal disease development in both types of diabetes. These proteins point to new therapeutic targets and new prognostic tests to identify subjects at risk of end-stage renal disease, as well as biomarkers to measure responses to treatment of diabetic kidney disease.


Assuntos
Nefropatias Diabéticas/sangue , Nefropatias Diabéticas/etiologia , Falência Renal Crônica/sangue , Falência Renal Crônica/etiologia , Adulto , Idoso , Biomarcadores/sangue , Proteínas Sanguíneas/genética , Proteínas Sanguíneas/metabolismo , Estudos de Coortes , Diabetes Mellitus Tipo 1/sangue , Diabetes Mellitus Tipo 1/complicações , Diabetes Mellitus Tipo 1/genética , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/genética , Nefropatias Diabéticas/genética , Progressão da Doença , Feminino , Humanos , Mediadores da Inflamação/sangue , Falência Renal Crônica/genética , Masculino , Pessoa de Meia-Idade , Prognóstico , Estudos Prospectivos , Proteômica , Receptores do Fator de Necrose Tumoral/sangue , Receptores do Fator de Necrose Tumoral/genética , Fatores de Risco
13.
Int J Nanomedicine ; 14: 2055-2067, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30988608

RESUMO

Background: Understanding of iron oxide nanoparticles (IONP) interaction with the body milieu is crucial to guarantee their efficiency and biocompatibility in nanomedicine. Polymer coating to IONP, with polyethyleneglycol (PEG) and polyvinylpyrrolidone (PVP), is an accepted strategy to prevent toxicity and excessive protein binding. Aim: The aim of this study was to investigate the feature of IONP adsorption of complement proteins, their activation and consequent inflammatory response as a strategy to further elucidate their biocompatibility. Methods: Three types of IONP with different surface characteristics were used: bare (IONP-bare), coated with PVP (IONP-PVP) and PEG-coated (IONP-PEG). IONPs were incubated with human plasma and adsorbed proteins were identified. BALB/c mice were intravenously exposed to IONP to evaluate complement activation and proinflammatory response. Results: Protein corona fingerprinting showed that PEG surface around IONP promoted a selective adsorption of complement recognition molecules which would be responsible for the complement system activation. Furthermore, IONP-PEG activated in vitro, the complement system and induced a substantial increment of C3a and C4a anaphylatoxins while IONP-bare and IONP-PVP did not. In vivo IONP-PEG induced an increment in complement activation markers (C5a and C5b-9), and proinflammatory cytokines (IL-1ß, IL-6, TNF-α). Conclusion: The engineering of nanoparticles must incorporate the association between complement proteins and nanomedicines, which will regulate the immunostimulatory effects through a selective adsorption of plasma proteins and will enable a safer application of IONP in human therapy.


Assuntos
Proteínas Sanguíneas/metabolismo , Proteínas do Sistema Complemento/metabolismo , Compostos Férricos/química , Inflamação/patologia , Nanopartículas/química , Polietilenoglicóis/química , Adsorção , Anafilatoxinas/metabolismo , Animais , Ativação do Complemento , Humanos , Interleucina-1beta/metabolismo , Masculino , Camundongos Endogâmicos BALB C , Nanopartículas/ultraestrutura , Povidona/química , Coroa de Proteína/metabolismo , Fator de Necrose Tumoral alfa/metabolismo
14.
Bioanalysis ; 11(5): 393-406, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30874444

RESUMO

AIM: Develop LC-MS/MS-based assays to measure total and free complement C5 in cynomolgus monkey serum as a target engagement biomarker for pharmacokinetic/pharmacodynamic correlation study. Materials & methods/results: The C5-specific signature peptide derived from pellet digestion of serum proteins with and without prior immunodepletion of the drug-bound C5 by protein A beads was quantified to assess free and total C5 levels, respectively. Conditions for immunodepletion by protein A were optimized to ensure complete depletion of IgGs (and drug-bound C5). The effect of sample dilution on drug-target dissociation and thus free C5 measurement was evaluated by applying a mathematical simulation. CONCLUSION: The procedure described here allows for the assessment of protein target engagement, aiding in pharmacokinetic/pharmacodynamic correlation analysis and human dose projection.


Assuntos
Proteínas Sanguíneas/metabolismo , Cromatografia Líquida/métodos , Espectrometria de Massas em Tandem/métodos , Animais , Macaca fascicularis
16.
Anal Bioanal Chem ; 411(11): 2383-2394, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30820631

RESUMO

Solid-phase microextraction (SPME) is an alternative method to dialysis and ultrafiltration for the determination of plasma protein binding (PPB) of drugs. It is particularly advantageous for complicated analytes where standard methods are not applicable. Di-2-pyridylketone 4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC) is a lead compound of novel thiosemicarbazone anti-cancer drugs, which entered clinical trials in 2016. However, this agent exhibited non-specific binding on filtration membranes and had intrinsic chelation activity, which precluded standard PPB methods. In this study, using a simple and fast procedure, we prepared novel SPME fibers for extraction of DpC based on a metal-free, silicon string support, covered with C18 sorbent. Reproducibility of the preparation process was demonstrated by the percent relative standard deviation (RSD) of ≤ 9.2% of the amount of DpC extracted from PBS by several independently prepared fibers. The SPME procedure was optimized by evaluating extraction and desorption time profiles. Suitability of the optimized protocol was verified by examining reproducibility, linearity, and recovery of DpC extracted from PBS or plasma. All samples extracted by SPME were analyzed using an optimized and validated UHPLC-MS/MS method. The developed procedure was applied to the in vitro determination of PPB of DpC at two clinically relevant concentrations (500 and 1000 ng/mL). These studies showed that DpC is highly bound to plasma proteins (PPB ≥ 88%) and this did not differ significantly between both concentrations tested. This investigation provides novel data in the applicability of SPME for the determination of PPB of chelators, as well as useful information for the clinical development of DpC. Graphical abstract.


Assuntos
Antineoplásicos/metabolismo , Proteínas Sanguíneas/metabolismo , Piridinas/metabolismo , Microextração em Fase Sólida/instrumentação , Tiossemicarbazonas/metabolismo , Adsorção , Animais , Bovinos , Cromatografia Líquida de Alta Pressão/métodos , Desenho de Equipamento , Ligação Proteica , Ratos , Silício/química , Microextração em Fase Sólida/métodos , Espectrometria de Massas em Tandem/métodos
17.
Nutrients ; 11(3)2019 Feb 27.
Artigo em Inglês | MEDLINE | ID: mdl-30818777

RESUMO

To better understand the variability of the type and level of serum proteins in human milk, the milk serum proteome of Chinese mothers during lactation was investigated using proteomic techniques and compared to the milk serum proteome of Dutch mothers. This showed that total milk serum protein concentrations in Chinese human milk decreased over a 20-week lactation period, although with variation between mothers in the rate of decrease. Variation was also found in the composition of serum proteins in both colostrum and mature milk, although immune-active proteins, enzymes, and transport proteins were the most abundant for all mothers. These three protein groups account for many of the 15 most abundant proteins, with these 15 proteins covering more than 95% of the total protein concentrations, in both the Chinese and Dutch milk serum proteome. The Dutch and Chinese milk serum proteome were also compared based on 166 common milk serum proteins, which showed that 22% of the 166 serum proteins differed in level. These differences were observed mainly in colostrum and concern several highly abundant proteins. This study also showed that protease inhibitors, which are highly correlated to immune-active proteins, are present in variable amounts in human milk and could be relevant during digestion.


Assuntos
Proteínas Sanguíneas/química , Lactação/fisiologia , Leite Humano/química , Proteínas Sanguíneas/metabolismo , China , Feminino , Humanos , Países Baixos
18.
Environ Toxicol Pharmacol ; 68: 148-154, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30903934

RESUMO

The interactions between human serum albumin (HSA) and α1- acid glycoprotein (AGP), the main plasma proteins binding drugs/xenobiotics, and some endocrine disrupting chemicals (EDCs), such as bisphenol A (BPA) and some of its structural analogues, bisphenol S (BPS), bisphenol F (BPF), bisphenol E (BPE), bisphenol B (BPB), bisphenol AF (BPAF), bisphenol A diglycidyl ether (BADGE) and bisphenol M (BPM), were characterized by biomimetic liquid chromatography (LC). The interactions between bisphenols (BPs) and either HSA or AGP protein was found to be non-specific and essentially lipophilicity-driven. To get more information on the binding of BPs and plasma proteins, in silico predictions and molecular docking simulations were exploited, and the results achieved in silico were compared to those observed in vitro. BPM was the one exhibiting the highest affinity on both plasma proteins according to these data. Our findings clarified the binding of these EDCs to plasma proteins and offered insights into the biodistribution and bioaccumulation processes underlying their toxicity.


Assuntos
Compostos Benzidrílicos/toxicidade , Proteínas Sanguíneas/metabolismo , Disruptores Endócrinos/toxicidade , Fenóis/toxicidade , Biomimética , Cromatografia Líquida , Simulação por Computador , Simulação de Acoplamento Molecular
19.
Aust Vet J ; 97(3): 75-80, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30809814

RESUMO

BACKGROUND: Cefovecin has a long duration of antibiotic activity in cats and dogs, somewhat attributable to its high plasma protein binding. AIMS: To determine the cefovecin binding to plasma proteins in vitro in selected Australian marsupials and to quantify the change in cetovecin concentration over time following subcutaneous injection in koalas. METHODS AND RESULTS: Various cefovecin concentrations were incubated with plasma and quantified using HPLC. The median (range) bound percentages when 10 µg/mL of cefovecin was incubated with plasma were 11.1 (4.1-20.4) in the plasma of the Tasmanian devil, 12.7 (5.8-17.3) in the koala, 18.9 (14.6-38.0) in the eastern grey kangaroo, 16.9 (15.7-30.2) in the common brush-tailed possum, 37.6 (25.3-42.3) in the eastern ring-tailed possum and 36.4 (35.0-38.3) in the red kangaroo, suggesting that cefovecin may have a shorter duration of action in these species than in cats and dogs. Cefovecin binding to plasma proteins in thawed, frozen equine plasma was also undertaken for assay quality control and the median (range) plasma protein binding (at 10 µg/mL) was 95.6% (94.9-96.6%). Cefovecin was also administered to six koalas at 8 mg/kg subcutaneously and serial blood samples were collected at 3, 6, 24, 48, 72, 96 h thereafter. Cefovecin plasma concentrations were not quantifiable in four koalas and in the other two, the mean plasma concentration at t = 3 h was 1.04 ± 0.01 µg/mL. CONCLUSION: Because of the limited pharmacokinetic data generated, no further pharmacokinetic analysis was performed; however, a single injected bolus of cefovecin is likely to have a short duration of action in koalas (hours, rather than days).


Assuntos
Antibacterianos/metabolismo , Proteínas Sanguíneas/metabolismo , Cefalosporinas/metabolismo , Marsupiais/sangue , Animais , Antibacterianos/administração & dosagem , Antibacterianos/farmacocinética , Austrália , Cefalosporinas/administração & dosagem , Cefalosporinas/farmacocinética , Cromatografia Líquida de Alta Pressão/veterinária , Feminino , Técnicas In Vitro , Injeções Subcutâneas/veterinária , Masculino , Marsupiais/metabolismo , Phascolarctidae/sangue , Phascolarctidae/metabolismo
20.
J Therm Biol ; 80: 133-140, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30784477

RESUMO

The present study evaluates whether increased water temperature induces reproduction by Lophiosilurus alexandri under controlled conditions, and investigates the effects of this procedure on sexual steroids, hematological profile and behavior. A 44-week experiment was performed with four wild males and 12 wild females that had been acclimatized to captive conditions. Water temperature was maintained at 24.4 ±â€¯1.0 °C for weeks 1-22, and then at 29.0 ±â€¯1.1 °C for weeks 22-44. Spawn weight, number of eggs/spawn and hatching rate were satisfactory and ranged 27.5-127.5 g, 1209-5183 and 83-89%, respectively. Hematocrit, leukocytes and glucose were not influenced by increased water temperature, while higher values for erythrocytes were observed for both sexes. The lowest value for plasma protein was for females maintained at 29.0 °C, while the lowest value for testosterone was obtained at the end of the study period at a temperature in 29.0 °C. Serum values of 17ß-estradiol were higher in females than in males, however, there was no evidence of variation as a function of experimental temperature or interaction with sex. The reproductive behavior of L. alexandri in captivity is described for the first time. The present study demonstrates that adult individuals are able to maintain a stable hematological profile during an increase in mean water temperature from 24.4 °C to 29.0 °C, even during the reproductive period, and still produce good quality larvae. Nonetheless, whether spawning was associated with increased 17ß-estradiol levels could not be determined.


Assuntos
Peixes-Gato/fisiologia , Reprodução , Temperatura Ambiente , Aclimatação , Animais , Comportamento Animal , Proteínas Sanguíneas/metabolismo , Contagem de Eritrócitos , Estradiol/sangue , Feminino , Masculino , Testosterona/sangue
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