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1.
Biochem Med (Zagreb) ; 30(1): 010502, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31839720

RESUMO

Extravascular body fluids (EBF) analysis can provide useful information in the differential diagnosis of conditions that caused their accumulation. Their unique nature and particular requirements accompanying EBF analysis need to be recognized in order to minimize possible negative implications on patient safety. This recommendation was prepared by the members of the Working group for extravascular body fluid samples (WG EBFS). It is designed to address the total testing process and clinical significance of tests used in EBF analysis. The recommendation begins with a chapter addressing validation of methods used in EBF analysis, and continues with specific recommendations for serous fluids analysis. It is organized in sections referring to the preanalytical, analytical and postanalytical phase with specific recommendations presented in boxes. Its main goal is to assist in the attainment of national harmonization of serous fluid analysis and ultimately improve patient safety and healthcare outcomes. This recommendation is intended to all laboratory professionals performing EBF analysis and healthcare professionals involved in EBF collection and processing. Cytological and microbiological evaluations of EBF are beyond the scope of this document.


Assuntos
Líquidos Corporais/química , Técnicas de Laboratório Clínico/normas , Líquidos Corporais/metabolismo , Exsudatos e Transudatos/química , Exsudatos e Transudatos/metabolismo , Guias como Assunto , Humanos , Segurança do Paciente , Derrame Pleural/diagnóstico , Garantia da Qualidade dos Cuidados de Saúde , Sociedades Médicas , Manejo de Espécimes/normas
2.
Artigo em Inglês | MEDLINE | ID: mdl-31610480

RESUMO

Naoshuantong capsule (NSTC) is an oral traditional Chinese medicine formula used widely in the clinic for ischemic stroke. The absorbed ingredients and metabolites of NSTC have never been reported before. In this study, a method incorporating rapid resolution liquid chromatography with quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was used to identify absorbed ingredients and metabolites after oral administration of NSTC. A total of 15 constituents were detected and identified as prototypes of NSTC. 109 metabolites related to catechin, gallic acid, paeoniflorin, chlorogenic acid, protocatechuate, typhaneoside, ß-elemene, calycosin were identified in serum, urine and brain. 19 metabolites of typhaneoside, 3 metabolites of ß-elemene, 12 metabolites of calycosin were reported for the first time. This is the first time to explore the absorption and metabolism of NSTC. The work will provide helpful information for further research of the mechanism and application of NSTC.


Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Medicamentos de Ervas Chinesas/metabolismo , Espectrometria de Massas em Tandem/métodos , Animais , Líquidos Corporais/metabolismo , Encéfalo/metabolismo , Catequina/sangue , Ácido Clorogênico/sangue , Cromatografia Líquida de Alta Pressão/métodos , Ácido Gálico/sangue , Glucosídeos/sangue , Glicosídeos/metabolismo , Hidroxibenzoatos/sangue , Isoflavonas/sangue , Masculino , Medicina Tradicional Chinesa/métodos , Metaboloma , Camundongos , Camundongos Endogâmicos C57BL , Monoterpenos/sangue , Sesquiterpenos/sangue
3.
Adv Clin Chem ; 93: 115-167, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31655729

RESUMO

The qualitative and quantitative determination of insulin and its related substances (e. g., C-peptide) is of great importance in many different areas of analytical chemistry. In particular, due to the steadily increasing prevalence of metabolic disorders such as diabetes mellitus, an adequate control of the circulating amount of insulin is desirable. In addition, also in forensics and doping control analysis, the determination of insulin in blood, urine or other biological matrices plays a major role. However, in order to establish general reference values for insulin and C-peptide for diabetology, the comparability of measured concentrations is indispensable. This has not yet been fully implemented, although enormous progress has been made in recent years, and the search for a "gold standard" method is still ongoing. In addition to established ligand-binding assays, an increasing number of mass-spectrometric methods have been developed and employed as the to-date available systems (for example, high-resolution/high accuracy mass spectrometers) provide the sensitivity required to determine analyte concentrations in the sub-ng/mL (sub-100pmol/L) level. Meanwhile, also high-throughput measurements have been realized to meet the requirement of testing a high number of samples in a short period of time. Further developments aim at enabling the online measurement of insulin in the blood with the help of an insulin sensor and, in the following, in addition to a brief review, today's state of the art testing developments are summarized.


Assuntos
Líquidos Corporais/metabolismo , Insulina/metabolismo , Sequência de Aminoácidos , Peptídeo C/metabolismo , Medicina Legal , Humanos , Insulina/sangue , Insulina/normas , Insulina/urina , Limite de Detecção , Espectrometria de Massas
4.
J Clin Pathol ; 72(12): 785-799, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31611285

RESUMO

Progresses in liquid-based assays may provide novel useful non-invasive indicators of cardiovascular (CV) diseases. By analysing circulating cells or their products in blood, saliva and urine samples, we can investigate molecular changes present at specific time points in each patient allowing sequential monitoring of disease evolution. For example, an increased number of circulating endothelial cells may be a diagnostic biomarker for diabetic nephropathy and heart failure with preserved ejection fraction. The assessment of circulating cell-free DNA (cfDNA) levels may be useful to predict severity of acute myocardial infarction, as well as diagnose heart graft rejection. Remarkably, circulating epigenetic biomarkers, including DNA methylation, histone modifications and non-coding RNAs are key pathogenic determinants of CV diseases representing putative useful biomarkers and drug targets. For example, the unmethylated FAM101A gene may specifically trace cfDNA derived from cardiomyocyte death providing a powerful diagnostic biomarker of apoptosis during ischaemia. Moreover, changes in plasma levels of circulating miR-92 may predict acute coronary syndrome onset in patients with diabetes. Now, network medicine provides a framework to analyse a huge amount of big data by describing a CV disease as a result of a chain of molecular perturbations rather than a single defect (reductionism). We outline advantages and challenges of liquid biopsy with respect to traditional tissue biopsy and summarise the main completed and ongoing clinical trials in CV diseases. Furthermore, we discuss the importance of combining fluid-based assays, big data and network medicine to improve precision medicine and personalised therapy in this field.


Assuntos
Líquidos Corporais/metabolismo , Doenças Cardiovasculares/diagnóstico , Técnicas de Diagnóstico Molecular , Medicina de Precisão , Biomarcadores/metabolismo , Doenças Cardiovasculares/tratamento farmacológico , Doenças Cardiovasculares/genética , Doenças Cardiovasculares/metabolismo , Ácidos Nucleicos Livres/sangue , Tomada de Decisão Clínica , Humanos , Biópsia Líquida , Seleção de Pacientes , Valor Preditivo dos Testes , Prognóstico , Reprodutibilidade dos Testes
5.
Eur J Pharm Biopharm ; 144: 252-265, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31563633

RESUMO

Nanoscale cerium dioxide (nanoceria) has industrial applications, capitalizing on its catalytic, abrasive, and energy storage properties. It auto-catalytically cycles between Ce3+ and Ce4+, giving it pro-and anti-oxidative properties. The latter mediates beneficial effects in models of diseases that have oxidative stress/inflammation components. Engineered nanoparticles become coated after body fluid exposure, creating a corona, which can greatly influence their fate and effects. Very little has been reported about nanoceria surface changes and biological effects after pulmonary or gastrointestinal fluid exposure. The study objective was to address the hypothesis that simulated biological fluid (SBF) exposure changes nanoceria's surface properties and biological activity. This was investigated by measuring the physicochemical properties of nanoceria with a citric acid coating (size; morphology; crystal structure; surface elemental composition, charge, and functional groups; and weight) before and after exposure to simulated lung, gastric, and intestinal fluids. SBF-exposed nanoceria biological effect was assessed as A549 or Caco-2 cell resazurin metabolism and mitochondrial oxygen consumption rate. SBF exposure resulted in loss or overcoating of nanoceria's surface citrate, greater nanoceria agglomeration, deposition of some SBF components on nanoceria's surface, and small changes in its zeta potential. The engineered nanoceria and SBF-exposed nanoceria produced no statistically significant changes in cell viability or cellular oxygen consumption rates.


Assuntos
Líquidos Corporais/química , Líquidos Corporais/metabolismo , Cério/química , Cério/metabolismo , Nanopartículas/metabolismo , Propriedades de Superfície/efeitos dos fármacos , Células A549 , Células CACO-2 , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Humanos , Mitocôndrias/efeitos dos fármacos , Nanopartículas/química , Oxirredução/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Consumo de Oxigênio/efeitos dos fármacos
7.
J Pharm Biomed Anal ; 175: 112791, 2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31398629

RESUMO

The anti-rheumatic fraction (ARF), is responsible for the anti-inflammatory and analgesic effects of Dianbaizhu derived from the aerial part of Gaultheria leucocarpa var. yunnanensis (Ericaceae). The gastrointestinal metabolism of ARF was investigated in vitro through simulating a series of models-gastric juice, intestinal juice, and human intestinal bacteria, analyzed by HPLC-DAD and UPLC-LTQ-Orbitrap-MSn. ARF includes three categories: methyl salicylate glycosides, organic acids and the others. The primordial and metabolic components of ARF bio-transformed by simulated gastric fluid (36 and 13), intestinal fluid (29 and 7) and two human fecal bacteria (34 and 34, 40 and 25) were characterized, respectively. The methyl salicylate glycosides, MSTG-B, MSTG-A and gaultherin, with terminal-xylosyl-moiety in sugar chain were always being found in the whole gastrointestinal incubation processing. The metabolites were formed through hydrolysis of ester and glucosidic bond, as well as methylation, hydroxylation, acetylation, sulfation, reduction, decarboxylation, deglycosylation and glucuronidation. The metabolic conversion effect of the four index compounds, MSTG-B, MSTG-A, gaultherin, and chlorogenic acid by human intestinal bacteria exhibited much stronger. Those markers' variation in content-time curve in volunteer A gut flora were faster than that in volunteer B's. These results indicate that ARF is relatively stable in the gastrointestinal tract.


Assuntos
Antirreumáticos/metabolismo , Líquidos Corporais/metabolismo , Suco Gástrico/metabolismo , Microbioma Gastrointestinal/fisiologia , Trato Gastrointestinal/metabolismo , Gaultheria/metabolismo , Intestinos/fisiologia , Anti-Inflamatórios/metabolismo , Bactérias/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Ericaceae/metabolismo , Fezes/química , Fezes/microbiologia , Suco Gástrico/microbiologia , Trato Gastrointestinal/microbiologia , Glucosídeos/metabolismo , Glicosídeos/metabolismo , Humanos , Intestinos/microbiologia
8.
BMC Womens Health ; 19(1): 109, 2019 08 12.
Artigo em Inglês | MEDLINE | ID: mdl-31405377

RESUMO

BACKGROUND: This study aims to investigate the difference in vaginal microecology, local immunity and HPV infection among childbearing-age women with different degrees of cervical lesions. METHODS: A total of 432 patients were included in this study. Among these patients, 136 patients had LSIL, 263 patients had HSIL and 33 patients had CSCC. These patients were assigned as the research groups. In addition, 100 healthy females were enrolled and assigned as the control group. RESULTS: The microbiological indexes of vaginal secretions were evaluated. Furthermore, the concentrations of SIgA, IgG, IL-2 and IL-10 in vaginal lavage fluid, as well as the presence of HPV, mycoplasma and Chlamydia in cervical secretions, were detected. The results is that: (1) Differences in evaluation indexes of vaginal microecology among all research groups and the control group were statistically significant (P < 0.0001). As the degree of cervical lesions increased, the number of Lactobacillus decreased, and there was an increase in prevalence of bacterial imbalance, and the diversity, density and normal proportion of bacteria was reduced. Furthermore, the incidence of HPV, trichomonads, clue cell and Chlamydia infection increased. Moreover, the positive rate of H2O2 decreased, while the positive rates of SNa and GADP increased. (2) Differences in the ratio of IL-2 and IL-10 in the female genital tract among all research groups and the control group were statistically significant (P < 0.0001). CONCLUSIONS: As the degree of cervical lesions increased, IL-2 decreased, IL-10 increased and IL-2/IL-10 decreased, while SIgA and IgG were elevated. The reduction of dominant Lactobacillus in the vagina, impairment of H2O2 function, flora ratio imbalance, pathogen infections, reduction in IL-2/IL-10 ratio, and changes in SIgA and IgG levels could all be potential factors that influenced the pathogenicity of HPV infection and the occurrence and development of cervical lesions.


Assuntos
Infecções por Chlamydia/epidemiologia , Infecções por Papillomavirus/epidemiologia , Lesões Intraepiteliais Escamosas Cervicais/epidemiologia , Vagina/imunologia , Vagina/microbiologia , Adulto , Líquidos Corporais/metabolismo , China/epidemiologia , Chlamydia trachomatis/isolamento & purificação , Coagulase/metabolismo , Feminino , Humanos , Peróxido de Hidrogênio/metabolismo , Imunoglobulina A/metabolismo , Imunoglobulina G/metabolismo , Interleucina-10/metabolismo , Interleucina-2/metabolismo , Mycoplasma/isolamento & purificação , Neuraminidase/metabolismo , Papillomaviridae/isolamento & purificação , Esfregaço Vaginal , Adulto Jovem
9.
Environ Sci Pollut Res Int ; 26(29): 29763-29779, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31407264

RESUMO

Dibutyl phthalate (DBP), a persistent environmental pollutant, can induce neural tube abnormal development in animals. The possible effects of DBP exposure on human neural tube defects (NTDs) remain elusive. In this study, the distribution of DBP in the body fluid of human NTDs was detected by GC-MS. Then, chick embryos were used to investigate the effects of DBP on early embryonic development. Oxidative stress indicators in chick embryos and the body fluid of human NTDs were detected by ELISA. The cell apoptosis and total reactive oxygen species (ROS) level in chick embryos were detected by whole-mount TUNEL and oxidized DCFDA, respectively. The study found that the detection ratio of positive DBP and its metabolites in maternal urine was higher in the NTD population than that in normal controls. 8-hydroxy-2 deoxyguanosine (8-OHDG) and malondialdehyde (MDA) were evidently upregulated and superoxide dismutase (SOD) was observably downregulated in amniotic fluid and urine. Animal experiments indicated that DBP treatment induced developmental toxicity in chick embryos by enhancing the levels of oxidative stress and cell apoptosis. MDA was increased and SOD was decreased in DBP-treated embryos. Interestingly, the supplement of high-dose choline (100 µg/µL), not folic acid, could partially restore the teratogenic effects of DBP. Our data collectively suggest that the incidence of NTDs is closely associated with DBP exposure. This study may provide new insight for NTD prevention.


Assuntos
Galinhas/metabolismo , Colina/metabolismo , Dibutilftalato/toxicidade , Desenvolvimento Embrionário/efeitos dos fármacos , Poluentes Ambientais/toxicidade , Defeitos do Tubo Neural/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Líquidos Corporais/metabolismo , Embrião de Galinha , Galinhas/crescimento & desenvolvimento , Dibutilftalato/urina , Poluentes Ambientais/urina , Feminino , Ácido Fólico/metabolismo , Humanos , Exposição Materna/efeitos adversos , Teratogênese/efeitos dos fármacos
10.
Environ Pollut ; 254(Pt A): 113007, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31421570

RESUMO

Parabens are extensively applied in cosmetics, drugs or food as preservatives and have become common pollutants in environmental media. However, data on human exposure to these chemicals is still limited, especially for children. This study aimed to investigate parabens in urine samples of children and to evaluate the cumulative risk of paraben exposure. Five short-chain parabens were measured in 255 urine samples collected from children in a kindergarten and elementary schools from South China. Methyl paraben (MeP), ethyl paraben (EtP) and n-propyl paraben (PrP) were widely detected in urine samples (detection rates > 94.9%), indicating their widespread exposure. The urinary median concentrations of MeP, EtP and PrP were 2.25, 0.33 and 0.50 µg/L, respectively. Significantly positive correlations (p < 0.01) were observed between MeP and PrP in urine, suggesting similar sources and/or metabolic pathways of these two chemicals. The median estimated daily intakes (EDIs) of parabens were determined to be 18.1 and 9.79 µg/kg-bw/day for kindergarten children and elementary school students, respectively. Estimation of human intake and exposure risks indicated potential risks of PrP exposure for elementary school students. This is the first study addressing paraben exposure in South China children.


Assuntos
Exposição Ambiental/estatística & dados numéricos , Poluentes Ambientais/urina , Parabenos/metabolismo , Líquidos Corporais/metabolismo , Criança , China , Cosméticos/análise , Exposição Ambiental/análise , Alimentos , Humanos , Parabenos/análise , Fatores de Risco , Instituições Acadêmicas , Estudantes
11.
Int J Mol Sci ; 20(14)2019 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-31337156

RESUMO

Exosomes and other classes of extracellular vesicles (EVs) have gained interest due to their role in cell-to-cell communication. Knowledge of the molecular content of EVs may provide important information on features of parental cells and mechanisms of cross-talk between cells. To study functions of EVs it is essential to know their composition, that includes proteins, nucleic acids, and other classes biomolecules. The metabolome, set of molecules the most directly related to the cell phenotype, is the least researched component of EVs. However, the metabolome of EVs circulating in human blood and other bio-fluids is of particular interest because of its potential diagnostic value in cancer and other health conditions. On the other hand, the metabolome of EVs released to culture media in controlled conditions in vitro could shed light on important aspects of communication between cells in model systems. This paper summarizes the most common approaches implemented in EV metabolomics and integrates currently available data on the composition of the metabolome of EVs obtained in different models with particular focus on human body fluids and cancer cells.


Assuntos
Exossomos/metabolismo , Metaboloma , Metabolômica , Líquidos Corporais/metabolismo , Comunicação Celular , Vesículas Extracelulares/metabolismo , Humanos , Metabolismo dos Lipídeos , Biópsia Líquida/métodos , Espectrometria de Massas/métodos , Metabolômica/métodos , Neoplasias/metabolismo
12.
Int J Mol Sci ; 20(14)2019 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-31330872

RESUMO

Sphingolipids (SL) modulate several cellular processes including cell death, proliferation and autophagy. The conversion of sphingomyelin (SM) to ceramide and the balance between ceramide and sphingosine-1-phosphate (S1P), also known as the SL rheostat, have been associated with oxidative stress and neurodegeneration. Research in the last decade has focused on the possibility of targeting the SL metabolism as a therapeutic option; and SL levels in biofluids, including serum, plasma, and cerebrospinal fluid (CSF), have been measured in several neurodegenerative diseases with the aim of finding a diagnostic or prognostic marker. Previous reviews focused on results from diseases such as Alzheimer's Disease (AD), evaluated total SL or species levels in human biofluids, post-mortem tissues and/or animal models. However, a comprehensive review of SL alterations comparing results from several neurodegenerative diseases is lacking. The present work compiles data from circulating sphingolipidomic studies and attempts to elucidate a possible connection between certain SL species and neurodegeneration processes. Furthermore, the effects of ceramide species according to their acyl-chain length in cellular pathways such as apoptosis and proliferation are discussed in order to understand the impact of the level alteration in specific species. Finally, enzymatic regulations and the possible influence of insulin resistance in the level alteration of SL are evaluated.


Assuntos
Líquidos Corporais/metabolismo , Doenças Neurodegenerativas/metabolismo , Esfingolipídeos/metabolismo , Animais , Apoptose , Biomarcadores , Vias Biossintéticas , Ceramidas/metabolismo , Cromatografia Líquida de Alta Pressão , Ácidos Graxos/metabolismo , Humanos , Incidência , Resistência à Insulina , Metabolismo dos Lipídeos , Doenças Neurodegenerativas/diagnóstico , Doenças Neurodegenerativas/epidemiologia , Doenças Neurodegenerativas/etiologia , Fenótipo , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem
13.
J Steroid Biochem Mol Biol ; 193: 105426, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31301352

RESUMO

Oxysterols are oxidized derivatives of cholesterol that are formed enzymatically or via reactive oxygen species or both. Cholesterol or oxysterols ingested as food are absorbed and packed into lipoproteins that are taken up by hepatic cells. Within hepatic cells, excess cholesterol is metabolised to form bile acids. The endoplasmic reticulum acts as the main organelle in the bile acid synthesis pathway. Metabolised sterols originating from this pathway are distributed within other organelles and in the cell membrane. The alterations to membrane oxysterol:sterol ratio affects the integrity of the cell membrane. The presence of oxysterols changes membrane fluidity and receptor orientation. It is well documented that hydroxylase enzymes located in mitochondria facilitate oxysterol production via an acidic pathway. More recently, the presence of oxysterols was also reported in lysosomes. Peroxisomal deficiencies favour intracellular oxysterols accumulation. Despite the low abundance of oxysterols compared to cholesterol, the biological actions of oxysterols are numerous and important. Oxysterol levels are implicated in the pathogenesis of multiple diseases ranging from chronic inflammatory diseases (atherosclerosis, Alzheimer's disease and bowel disease), cancer and numerous neurodegenerative diseases. In this article, we review the distribution of oxysterols in sub-cellular organelles and in biological fluids.


Assuntos
Líquidos Corporais/metabolismo , Oxisteróis/metabolismo , Animais , Humanos , Organelas/metabolismo
14.
Soft Matter ; 15(31): 6392-6399, 2019 Aug 21.
Artigo em Inglês | MEDLINE | ID: mdl-31312830

RESUMO

Bumblebees and some other tiny animals feed on nectar by visiting flowers in their neighborhood. Some bee species appear to be highly specialized, their tongue being adapted to specific flowers. Bombus terrestris in contrast is able to feed on a wide variety of flowers and can thus be considered as a kind of universal nectar catcher. Since plant nectars show highly variable sugar content, Bombus terrestris have developed a capture mechanism that works for almost any fluid viscosity. Their tongues are decorated with very elongated papillae forming a hairy coating surrounding a rod-like main stalk. When settled on a flower, Bombus rapidly dip their tongue into the inflorescence to catch the highly sought-after nectar. To determine the physical mechanism at the origin of this outstanding ability, the capture dynamics was followed from videos recorded during viscous fluid ingestion. Surprisingly, the volume per lap and the lapping frequency are independent of the fluid viscosity over three orders of magnitude. To explain this observation, we designed a physical model of viscous dipping with structured rods. Predictions of the model compared to observations for bees showed that the nectar is not captured with the help of viscous drag, as proposed in the Landau-Levich-Derjaguin model, but thanks to the hairy structure that traps the viscous fluid, capillary forces drastically limiting the drainage. Our approach can be transposed to others nectar foragers such as bats and hummingbirds.


Assuntos
Abelhas , Líquidos Corporais/metabolismo , Flores/química , Néctar de Plantas/metabolismo , Língua/metabolismo , Animais , Comportamento Animal , Fenômenos Biomecânicos , Carboidratos/química , Modelos Biológicos , Língua/citologia , Gravação em Vídeo
15.
Eur J Pharm Biopharm ; 142: 387-395, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31306752

RESUMO

Oral administration of drug products is the preferred administration route. In recent decades there has been an increase in drug candidates with low solubility and/or low permeability. To increase the possibility of oral administration for the poorly permeating drugs, the use of absorption modifying excipients (AMEs) has been proposed. These types of AMEs may also affect the regulatory assessment of a novel drug delivery system if they affect the absorption of a drug from any of the four BCS classes. The effects of AMEs have previously been investigated in various animal models, including the single-pass intestinal perfusion (SPIP) in rats. To further improve the biorelevance and the in vivo predictiveness of the SPIP model, four compounds (atenolol, enalaprilat, ketoprofen, metoprolol) were perfused in fasted or fed state simulated intestinal fluid (FaSSIF or FeSSIF) together with the AMEs N-acetyl-cysteine, caprate, or sodium dodecyl sulfate. For the highly soluble and poorly permeating compounds enalaprilat and atenolol (BCS class III), the flux was increased the most by the addition of SDS in both FaSSIF and FeSSIF. For ketoprofen (BCS class II), the flux decreased in the presence of all AMEs in at least one of the perfusion media. The flux of metoprolol (BCS class I) was not affected by any of the excipients in none of simulated prandial states. The changes in magnitude in the absorption of the compounds were in general smaller in FeSSIF than in FaSSIF. This may be explained by a reduced free concentration AMEs in FeSSIF. Further, the results in FeSSIF were similar to those from intrajejunal bolus administration in rat in a previous study. This suggests that the biorelevance of the SPIP method may be increased when investigating the effects of AMEs, by the addition of intraluminal constituents representative to fasted and/or fed state to the inlet perfusate.


Assuntos
Excipientes/química , Jejum/metabolismo , Absorção Intestinal/efeitos dos fármacos , Jejuno/metabolismo , Preparações Farmacêuticas/química , Preparações Farmacêuticas/metabolismo , Administração Oral , Animais , Disponibilidade Biológica , Líquidos Corporais/metabolismo , Mucosa Intestinal/metabolismo , Intestinos/fisiologia , Masculino , Permeabilidade/efeitos dos fármacos , Ratos , Ratos Wistar , Dodecilsulfato de Sódio/química , Solubilidade/efeitos dos fármacos
16.
Eur J Pharm Biopharm ; 142: 307-314, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31288077

RESUMO

The influence of physiological factors on the solubility of drug compounds has been thoroughly investigated in humans. However, as these factors vary between species and since many in vivo studies are carried out in rats or mice, it has been difficult to establish sufficient in vitro in vivo relations. The aim of this study was to develop a physiologically relevant in vitro dissolution model simulating the gastrointestinal (GI) fluids of fasted rats and compare it to previously published in vitro and in vivo data. To develop the in vitro model, the pH was measured in situ in six segments of the GI tract of anesthetised rats, then the fluids from the stomach, the proximal and the distal small intestine were collected and characterized with regard to osmolality, and bile acid and phospholipid concentration. The pH and osmolality were found to increase throughout the GI tract. The bile acids and phospholipids were present in high concentrations in the proximal small intestine, and the bile acid concentration doubled in the distal part, where the phospholipid concentration decreased. Matrix-assisted laser desorption ionisation mass spectrometry imaging was applied on a cross section of the small intestine, to study which bile acids and phospholipid classes were present in the small intestine of rats. Both cholic acid, taurocholic acid and glycocholic acid were detected, and phosphatidylcholine (34:2) was found to be mainly present in the intestinal wall or mucus, whereas lysophosphatidylcholine (16:0) was also detected in the lumen. Based on these observations, biorelevant media were developed to simulate fluids in the stomach and the proximal part of the small intestine in fasted rats. The media were implemented in a two-step in vitro dissolution model, which was found to better predict the in vivo performance of furosemide, when compared to previously published in vitro and in vivo data.


Assuntos
Líquidos Corporais/metabolismo , Líquidos Corporais/fisiologia , Mucosa Gástrica/metabolismo , Intestino Delgado/metabolismo , Intestino Delgado/fisiologia , Preparações Farmacêuticas/metabolismo , Estômago/fisiologia , Animais , Humanos , Concentração de Íons de Hidrogênio , Masculino , Camundongos , Concentração Osmolar , Ratos , Ratos Sprague-Dawley , Solubilidade
17.
Int J Mol Sci ; 20(11)2019 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-31195629

RESUMO

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that has no effective treatment. The lack of any specific biomarker that can help in the diagnosis or prognosis of ALS has made the identification of biomarkers an urgent challenge. Multiple panels have shown alterations in levels of numerous cytokines in ALS, supporting the contribution of neuroinflammation to the progressive motor neuron loss. However, none of them is fully sensitive and specific enough to become a universal biomarker for ALS. This review gathers the numerous circulating cytokines that have been found dysregulated in both ALS animal models and patients. Particularly, it highlights the opposing results found in the literature to date, and points out another potential application of inflammatory cytokines as therapeutic targets.


Assuntos
Esclerose Amiotrófica Lateral/sangue , Biomarcadores/sangue , Citocinas/sangue , Esclerose Amiotrófica Lateral/imunologia , Esclerose Amiotrófica Lateral/terapia , Animais , Líquidos Corporais/metabolismo , Humanos , Sistema Imunitário/metabolismo , Inflamação/patologia
18.
Mol Med Rep ; 20(2): 1103-1112, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173186

RESUMO

DL0410, a dual­action cholinesterase inhibitor and histamine­3 receptor antagonist with a novel structural scaffold, may be a potential candidate for the treatment of Alzheimer's disease (AD). To the best of the authors' knowledge, this is the first study to demonstrate a reliable method for the measurement of DL0410 in rat plasma, brain, bile, urine and feces samples, and identification of its primary metabolites. The pharmacokinetic properties of DL0410 were analyzed by liquid chromatography­mass spectrometry at oral doses of 25, 50 and 100 mg/kg and intravenous dose of 5 mg/kg. The investigation of the excretion and metabolism of DL0410 was determined following liquid­liquid extraction for biliary, urinary and fecal samples. Finally, the cytochrome (CY)P450 isoforms involved in the production of DL0410 metabolites with recombinant human cytochrome P450 enzymes were characterized. The results suggested that DL0410 was not well absorbed; however, was distributed to the entorhinal cortex and hippocampus of the brain. A total of two common metabolites of the reduction of DL0140 in the bile, urine and feces were identified and CYP2D6 was involved in this reaction. The pharmacokinetic results of DL0410 provided information for the illustration of its pharmacodynamic properties, mechanism of action and promoted its continued evaluation as a therapeutic agent for AD treatment.


Assuntos
Compostos de Bifenilo/farmacocinética , Inibidores da Colinesterase/farmacocinética , Citocromo P-450 CYP2D6/metabolismo , Antagonistas dos Receptores Histamínicos H3/farmacocinética , Piperidinas/farmacocinética , Doença de Alzheimer/tratamento farmacológico , Animais , Bile/metabolismo , Compostos de Bifenilo/análise , Compostos de Bifenilo/uso terapêutico , Líquidos Corporais/metabolismo , Encéfalo/metabolismo , Inibidores da Colinesterase/análise , Inibidores da Colinesterase/uso terapêutico , Fezes/química , Feminino , Antagonistas dos Receptores Histamínicos H3/análise , Antagonistas dos Receptores Histamínicos H3/uso terapêutico , Humanos , Masculino , Piperidinas/análise , Piperidinas/uso terapêutico , Ratos , Ratos Sprague-Dawley
19.
Molecules ; 24(12)2019 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-31234484

RESUMO

Improvement in high-throughput MALDI-TOF MS analysis requires practical and efficient sample preparation protocols for high acquisition rates. The use of hexagonal mesoporous silica (HMS) sorbents in combination with MALDI-TOF MS was explored as a versatile tool for peptidomic profiling of clinical specimens difficult to process, but considered important sources of disease biomarkers: synovial fluid and sputum. A rapid and efficient procedure, based on dispersive solid-phase extraction of peptides using commercially available wormhole mesostructured HMS, was tested for: a) pre-concentration of standard peptides in serially diluted solution up to the sub-nanomolar range; b) peptidome profiling of sputum and synovial fluid. The use of HMS, as dispersed sponges, significantly amplified the peptidic repertoire of sputum and synovial fluid by excluding from the adsorptive process large size proteins, which mask and/or suppress peptidome signals. The protocol proposed, as dispersive solid phase extraction, ensures good analytical performances. Moreover, it is economical and rapid, as it avoids the use of less reproducible and prolonged sample preparation procedures, such as the use of ultrafiltration filter devices. These findings may contribute to defining a high-throughput screening MS-based platform for monitoring key peptidic features of difficult to analyse bodily fluids in a clinical setting.


Assuntos
Peptídeos/análise , Proteômica , Dióxido de Silício/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Biomarcadores , Líquidos Corporais/química , Líquidos Corporais/metabolismo , Humanos , Projetos Piloto , Proteômica/métodos , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Escarro
20.
Cell Mol Neurobiol ; 39(7): 901-915, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31190159

RESUMO

Parkinson's disease (PD) is an age-related, threatening neurodegenerative disorder with no reliable treatment till date. Identification of specific and reliable biomarker is a major challenge for disease diagnosis and designing effective therapeutic strategy against it. PD pathology at molecular level involves abnormal expression and function of several proteins, including alpha-synuclein. These proteins affect the normal functioning of neurons through various post-translational modifications and interaction with other cellular components. The role of protein anomalies during PD pathogenesis can be better understood by the application of proteomics approach. A number of proteomic studies conducted on brain tissue, blood, and cerebrospinal fluid of PD patients have identified a wide array of protein alterations underlying disease pathogenesis. However, these studies are limited by the types of brain regions or biofluids utilized in the research. For a complete understanding of PD mechanism and discovery of reliable protein biomarkers, it is essential to analyze the proteome of different PD-associated brain regions and easily accessible biofluids such as saliva and urine. The present review summarizes the major advances in the field of PD research in humans utilizing proteomic techniques. Moreover, potential samples for proteomic analysis and limitations associated with the analyses of different types of samples have also been discussed.


Assuntos
Doença de Parkinson/metabolismo , Proteômica , Líquidos Corporais/metabolismo , Encéfalo/metabolismo , Encéfalo/patologia , Humanos , Especificidade de Órgãos , Doença de Parkinson/sangue , Doença de Parkinson/líquido cefalorraquidiano
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