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1.
J Agric Food Chem ; 68(2): 686-696, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31877248

RESUMO

Metabolites of serum and milk from genetically modified (GM) cows and contrast check (CK) cows were comparatively investigated. Serum and milk were collected from genetically modified (GM) cows and contrast check (CK) cows, and then, they were analyzed using ultraperformance liquid chromatography-mass spectrometry (UPLC-MS) and gas chromatography-mass spectrometry (GC-MS). Although the level of some blood biochemical indexes for GM cows was shifted up or down, they were generally in normal physiological condition. Serum samples from lactoferrin GM cows exhibited reduced levels of amino acids and elevated levels of indoleacetate, α-keto acids, long-chain fatty acids, etc. GM milk possessed elevated levels of pentose and amino sugar metabolites, including arabitol, xylulose, glucuronate, and N-acetylgalactosamine. Interestingly, some essential nutrients, such as certain unsaturated fatty acids (e.g., eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA)), and some necessary rare sugars were significantly upregulated. Compared to the CK group, a Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis was conducted based on the increased or decreased metabolites identified in the serum and milk samples of the GM group. The results showed that the GM cows were in healthy condition and their milk has improved benefits for customers. The milk from genetically modified cows was found to be a promising milk source for producing recombinant human lactoferrin (rhLF) for human beings.


Assuntos
Animais Geneticamente Modificados/metabolismo , Lactoferrina/genética , Leite/química , Soro/química , Animais , Animais Geneticamente Modificados/genética , Bovinos/genética , Bovinos/metabolismo , Ácidos Graxos/química , Ácidos Graxos/metabolismo , Feminino , Ácidos Indolacéticos/sangue , Cetoácidos/sangue , Lactoferrina/metabolismo , Metabolômica , Leite/metabolismo , Soro/metabolismo , Açúcares/sangue
2.
Int J Nanomedicine ; 14: 8285-8302, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31802866

RESUMO

Background: Curcumin has been widely used owing to its various medicinal properties including antitumor effects. However, its clinical application is limited by its instability, poor solubility and low bioavailability. Folic acid (FA)-functionalized nanoformulations may enhance the sustained release of an anticancer drug (curcumin) by tumor-specific targeting to improve therapeutic benefit. This study aims to design a nanoconjugate (NC) comprised of folate-curcumin-loaded gold-polyvinylpyrrolidone nanoparticles (FA-CurAu-PVP NPs) for targeted delivery in breast cancer model systems. Methods: We developed curcumin-loaded FA-functionalized Au-PVP NCs by layer-by-layer assembly. The folic acid-curcumin Au-PVP NCs (FA-CurAu-PVP NCs) were characterized by ultraviolet-visible spectra, Fourier transform infrared spectroscopy, X-ray powder diffraction and thermogravimetric analysis. In vitro anticancer and antimigratory effects of NCs were examined by performing MTT and wound migration assays. The in vivo antitumor efficacy of NCs was investigated using a preclinical breast cancer orthotopic mouse model. Results: Curcumin (40 µg/mL) was loaded along with conjugation of folate onto Au-PVP NPs to form FA-CurAu-PVP NCs. The size and charge of the NCs were increased gradually through layer-by-layer assembly and showed 80% release of curcumin at acidic pH. The NC did not show aggregation when incubated with human serum and mimicked an intrinsic peroxidase-like property in the presence of 3,3',5,5'-tetramethylbenzidine substrate. The MTT data using these NCs showed efficient anticancer activity at lower doses in estrogen/progesterone receptor (ER/PR)-negative cells compared with ER/PR-positive cells. Furthermore, the NCs did not show cytotoxicity at the investigated concentration in human breast epithelial and mouse fibroblast cell lines. They showed inhibitory effects on cell migration and high antitumor efficacy in in vivo analysis. Conclusion: These results suggest that folate-based tumor targeting using CurAu-PVP NCs is a promising approach for tumor-specific therapy of breast cancer without harming normal cells.


Assuntos
Neoplasias da Mama/tratamento farmacológico , Sistemas de Liberação de Medicamentos , Ácido Fólico/química , Ouro/química , Nanopartículas Metálicas/química , Polímeros/química , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Curcumina/farmacologia , Curcumina/uso terapêutico , Portadores de Fármacos/química , Liberação Controlada de Fármacos , Feminino , Humanos , Concentração de Íons de Hidrogênio , Nanopartículas Metálicas/ultraestrutura , Camundongos , Nanoconjugados/química , Povidona/química , Soro/metabolismo
3.
Exp Oncol ; 41(4): 318-322, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31868335

RESUMO

This study aimed to investigate the effects of hypoxia and serum deprivation on regulation of fucosyltransferase-3 (FUT3) expression in breast cancer cells. MATERIALS AND METHODS: FUT3 expression was evaluated in T47D and MCF7 cells. Transcriptional and protein analysis was performed under hypoxia and serum deprivation conditions after 6 and 24 hours; and after 24 and 48 hours, respectively. RESULTS: In T47D cells, experimental conditions induced a significant decrease in FUT3 expression at both, transcriptional and protein levels, while in MCF7 cells the same conditions induced a significant increase of FUT3 expression. CONCLUSIONS: Regulation of FUT3 expression under hypoxic and serum deprivation conditions may be involved in the acquisition of advantages related to apoptosis resistance and metastasis promotion, according to the intrinsic differences presented by T47D and MCF7 cells.


Assuntos
Neoplasias da Mama/genética , Fucosiltransferases/genética , Regulação Neoplásica da Expressão Gênica , Apoptose , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Feminino , Humanos , Células MCF-7 , Soro/metabolismo , Hipóxia Tumoral
4.
Pharm Res ; 37(1): 10, 2019 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-31872347

RESUMO

PURPOSE: As nanoparticles (NPs) are intravenously entering the bloodstream, proteins in the plasma can recognize and bind them to form a protein corona that affects how NPs are perceived by biological systems. The complement is an essential part of the innate immunity that contributes to non-specific host defense. How complement recognizes NPs has not been elucidated. Here, we developed a proteomics and biochemical approach to understand the applied risk of activated complement by NPs. METHODS: Complement proteins absorbed on Hydroxyapatite Nanoparticles (HAP-NPs) and Silicon dioxide Nanoparticles (SiO2-NPs) were analyzed by proteomics with LC-MS. The effect of complement activation was studied by iC3b/Sc5b-9/C3a/C4a/C5a with ELISA. An inhibitory model was established via EDTA and EGTA to confirm the selective pathway activation of both NPs. Finally, the regulation of complement by NPs was analyzed by western blot. RESULTS: The results indicate that HAP-NPs start the activation of the complement through the classical pathway because of the absorption of C1q and the release of C1r/C1s. Meanwhile, the soluble regulatory molecules such as CFI, C4bp, and CFH tried to resist the complement system activation by the cleavage of C3 convertase. In contrast, SiO2-NPs can activate the alternative pathway of the complement through the absorption of CFD and CFB. CONCLUSION: It was clarified that HAP-NPs and SiO2-NPs activate complement through different mechanisms. These studies provide a scientific basis for the design and modification of nano-drug carriers for delaying their recognition and clearance by the mononuclear phagocytic system and simultaneously reducing the immunotoxicity of NPs. The understanding of protein corona is conducive to innovation in the field of "immune-safe-by-design" nanomedicines.


Assuntos
Complemento C1q/metabolismo , Portadores de Fármacos/química , Nanopartículas/química , Coroa de Proteína/química , Coroa de Proteína/metabolismo , Ativação do Complemento/efeitos dos fármacos , Proteínas do Sistema Complemento/metabolismo , Composição de Medicamentos/métodos , Liberação Controlada de Fármacos , Durapatita/química , Humanos , Proteômica , Soro/metabolismo , Transdução de Sinais , Dióxido de Silício/química , Manejo de Espécimes , Espectrometria de Massas em Tandem/métodos
5.
Immunohorizons ; 3(12): 585-592, 2019 12 16.
Artigo em Inglês | MEDLINE | ID: mdl-31843785

RESUMO

Chemically defined serum-free media are increasingly used as a tool to help standardize experiments by eliminating the potential variability contributed by pooled serum. These media are formulated for the culture and expansion of specific cell types, maintaining cell viability without the need for exogenous animal proteins. Formulated serum-free media could thus help improve viability and reduce variability during sample preparation for flow cytometry, yet a thorough analysis of how such media impact fluorochrome-Ab conjugates has not been performed. In this study, we expose fluorescent Ab-labeled cells or Ab capture beads to white light in the presence of various hematopoietic cell culture media and provide evidence that formulated serum-free media permit rapid light-initiated fluorescent dye degradation in a cell-independent manner. We observed fluorescence signal loss of several dyes, which included fluorescence spillover into adjacent detectors. Finally, photostability of Ab-fluorochrome conjugates in formulated serum-free media is partially restored in the presence of either serum or vitamin C, implicating reactive oxygen species in the observed signal loss. Thus, our data indicate that formulated serum-free media designed to standardize cell culture are not currently optimized for use with fluorochrome-Ab conjugates, and thus, extreme caution should be exercised when using these media in cytometric experiments.


Assuntos
Meios de Cultura Livres de Soro/metabolismo , Corantes Fluorescentes/metabolismo , Luz , Proteólise/efeitos da radiação , Anticorpos/metabolismo , Ácido Ascórbico/metabolismo , Doadores de Sangue , Linfócitos T CD4-Positivos/metabolismo , Sobrevivência Celular , Citometria de Fluxo/métodos , Humanos , Soro/metabolismo
6.
Res Vet Sci ; 127: 57-64, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31678454

RESUMO

The objective of this study was to evaluate the effects of in-feed clinoptilolite (CPL) on serum metabolic and antioxidative biomarkers, acute phase proteins and reproductive performance in cows during pregnancy and lactation. A total of 78 Holstein-Friesian cows were randomly assigned into two groups: the treatment group, cows fed CPL (n = 38) which received 50 g of powdered CPL twice a day from day 180 before parturition to day 60 postpartum; and the control group (n = 40). Blood samples were taken on days 180, 90, 60, 30 and 10 before parturition, on day of calving and on days 5, 12, 19, 26, 33, 40 and 60 postpartum, and were analysed for metabolic biomarkers: glucose, triglycerides, total cholesterol, high density lipoprotein cholesterol, non-esterified fatty acids, beta-hydroxybutyrate (BHB), antioxidative biomarkers and acute phase proteins: paraoxonase-1 (PON1), apolipoprotein A-I, haptoglobin (Hp) and serum amyloid A (SAA). CPL supplementation increased concentration of glucose and significantly decreased (P < .05) level of BHB during puerperium. The SAA concentration in CPL-fed cows was significantly decreased (P < .05) on days 33, 40 and 60 postpartum as well as Hp concentration on days 0 and 12 postpartum. The results of this study suggest that the CPL-fed cows may have improved metabolic status due to the tendency of greater glucose levels and decreased BHB values during early lactation. In addition, acute phase response was lower (P < .05) in CPL-fed cows. Such an outcome might be attributed to the effect of dietary CPL on intensity and severity of the negative energy balance and inflammatory response in dairy cows.


Assuntos
Reação de Fase Aguda/veterinária , Antioxidantes/metabolismo , Biomarcadores/metabolismo , Doenças dos Bovinos/tratamento farmacológico , Zeolitas/metabolismo , Reação de Fase Aguda/tratamento farmacológico , Reação de Fase Aguda/metabolismo , Ração Animal/análise , Animais , Bovinos , Doenças dos Bovinos/metabolismo , Dieta/veterinária , Suplementos Nutricionais/análise , Feminino , Lactação/fisiologia , Gravidez , Distribuição Aleatória , Soro/metabolismo , Zeolitas/administração & dosagem
7.
Ann Clin Lab Sci ; 49(5): 678-681, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31611215

RESUMO

Recently, the removal of false reaction from beads 10 (DRB1 *04:04), 30 (DRB1 *16:01), and 31 (DRB1 *16:02) by fetal bovine serum (FBS) treatment in LABScreen Single Antigen Class II Assay was reported. We aimed to confirm the reaction in many cases. Fifty-nine sera showed positivity on at least two among beads 10, 30 and 31 from Nov 2017 to Oct 2018 in Seoul National University Hospital were included. FBS treatment was performed on 59 sera, and Single Antigen Class II Assay was repeated. Among 59 cases, the negative conversion rates of DR16 (57/59, 96.6%), DR4 (37/39, 94.9%), and accompanied other antibodies, e.g.) DP19 (39/45, 86.7%) were very high. The prior use of intravenous immunoglobulin in a non-specific binding group was significantly higher than the sex, age-matched control group (P=0.005).


Assuntos
Feto/metabolismo , Antígenos de Histocompatibilidade Classe II/metabolismo , Imunoensaio/métodos , Soro/metabolismo , Adulto , Animais , Estudos de Casos e Controles , Bovinos , Feminino , Humanos , Masculino
8.
Life Sci ; 235: 116840, 2019 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-31494171

RESUMO

AIMS: Ovarian ischemia as a consequence of torsion constitutes a gynecologic emergency affecting females during reproductive age. Its management by detorsion results in ovarian ischemia-reperfusion (IR) injury. Thus, a conservative treatment with detorsion is highly recommended. Therefore, we attempted to investigate the effect and underlying mechanisms of angiotensin 1-7 (Ang-(1-7)) treatment against ovarian IR injury. MAIN METHODS: Female rats were included into: Sham group; Ang-(1-7) (300 µg/kg, i.p.) group; ovarian IR groups with and without Ang-(1-7) treatment. We determined ovarian Ang-(1-7), malondialdehyde (MDA) and nitric oxide (NO) in addition to serum total anti-oxidant capacity (TAC) levels. Ovarian gene expressions of angiotensin converting enzyme 2 (ACE2), Mas receptor, tumor necrosis factor alpha (TNF-α) and B-cell leukemia/lymphoma-2 (BCL-2) were estimated. Furthermore, histopathological changes and ovarian expressions of nuclear factor kappa B (NF-κB), inducible and endothelial nitric oxide synthases (iNOS and eNOS) were done. KEY FINDINGS: Treatment of ovarian IR rats with Ang-(1-7) led to marked improvement of ovarian damage through histological examination which was accompanied with marked increase in ovarian Ang-(1-7) level and expressions of ACE2 and Mas receptor, decrease in MDA and NO levels and expressions of NF-kB, iNOS and TNF-α with increase in serum TAC levels and ovarian expressions of eNOS and BCL-2. SIGNIFICANCE: Our results proved the protective effect of Ang-(1-7) against ovarian IR injury in rats and this may be attributed to ACE2/Ang (1-7)/Mas axis which showed anti-oxidant, anti-inflammatory and anti-apoptotic effects. Therefore, Ang-(1-7) can be used in the future for treatment of ovarian IR injury.


Assuntos
Angiotensina I/farmacologia , Ovário/efeitos dos fármacos , Fragmentos de Peptídeos/farmacologia , Traumatismo por Reperfusão/prevenção & controle , Animais , Antioxidantes/metabolismo , Feminino , Expressão Gênica/efeitos dos fármacos , Malondialdeído/metabolismo , NF-kappa B/biossíntese , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/biossíntese , Óxido Nítrico Sintase Tipo III/biossíntese , Ovário/lesões , Ovário/metabolismo , Peptidil Dipeptidase A/biossíntese , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Ratos , Receptores Acoplados a Proteínas-G/biossíntese , Soro/metabolismo , Fator de Necrose Tumoral alfa/biossíntese
9.
Acta Trop ; 200: 105186, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31542371

RESUMO

The small blood flukes of genus Schistosoma, which cause one of the most prevalent and serious parasitic zoonosis schistosomiasis, are dependent on immune-related factors of their mammalian host to facilitate their growth and development, and the formation of granulomatous pathology caused by eggs deposited in host's liver and intestinal wall. Schistosome development is hampered in the mice lacking just T cells, and is even more heavily retarded in the severe combined immunodeficient (SCID) mice lacking both T and B lymphocytes. Nevertheless, it's still not clear about the underlying regulatory molecular mechanisms of schistosome growth and development by host's immune system. This study, therefore, detected and compared the serum metabolic profiles between the immunodeficient mice and immunocompetent mice (SCID mice vs. BALB/c mice) before and after S. japonicum infection (on the thirty-fifth day post infection using liquid chromatography-mass spectrometry (LC-MS). Totally, 705 ion features in electrospray ionization in positive-ion mode (ESI+) and 242 ion features in ESI- mode were identified, respectively. First, distinct serum metabolic profiles were identified between SCID mice and BALB/c mice without S. japonicum worms infection. Second, uniquely perturbed serum metabolites and their enriched pathways were also obtained between SCID mice and BALB/c mice after S. japonicum infection, which included differential metabolites due to both species differences and differential responses to S. japonicum infection. The metabolic pathways analysis revealed that arachidonic acid metabolism, biosynthesis of unsaturated fatty acids, linoleic acid metabolism, glycosylphosphatidylinositol (GPI)-anchor biosynthesis, alpha-linolenic acid metabolism, glycerophospholipid metabolism, sphingolipid metabolism and purine metabolism were enriched based on the differential serum metabolites between SCID mice and BALB/c mice after S. japonicum infection, which was addressed to be related to the retarded growth and development of S. japonicum in SCID mice. These findings provide new clues to the underlying molecular events of host's systemic metabolic changes on the growth and development of S. japonicum worms, and also provide quite promising candidates for exploitation of drugs or vaccines against schistosome and schistosomiasis.


Assuntos
Metabolômica , Camundongos Endogâmicos BALB C/crescimento & desenvolvimento , Camundongos SCID/crescimento & desenvolvimento , Schistosoma japonicum/imunologia , Esquistossomose Japônica/imunologia , Soro/imunologia , Soro/metabolismo , Animais , Feminino , Camundongos , Camundongos Endogâmicos BALB C/metabolismo , Camundongos SCID/metabolismo
10.
Neurology ; 93(9): e917-e926, 2019 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-31366722

RESUMO

OBJECTIVE: To determine the long-term association of hemoglobin levels and anemia with risk of dementia, and explore underlying substrates on brain MRI in the general population. METHODS: Serum hemoglobin was measured in 12,305 participants without dementia of the population-based Rotterdam Study (mean age 64.6 years, 57.7% women). We determined risk of dementia and Alzheimer disease (AD) (until 2016) in relation to hemoglobin and anemia. Among 5,267 participants without dementia with brain MRI, we assessed hemoglobin in relation to vascular brain disease, structural connectivity, and global cerebral perfusion. RESULTS: During a mean follow-up of 12.1 years, 1,520 individuals developed dementia, 1,194 of whom had AD. We observed a U-shaped association between hemoglobin levels and dementia (p = 0.005), such that both low and high hemoglobin levels were associated with increased dementia risk (hazard ratio [95% confidence interval (CI)], lowest vs middle quintile 1.29 [1.09-1.52]; highest vs middle quintile 1.20 [1.00-1.44]). Overall prevalence of anemia was 6.1%, and anemia was associated with a 34% increased risk of dementia (95% CI 11%-62%) and 41% (15%-74%) for AD. Among individuals without dementia with brain MRI, similar U-shaped associations were seen of hemoglobin with white matter hyperintensity volume (p = 0.03), and structural connectivity (for mean diffusivity, p < 0.0001), but not with presence of cortical and lacunar infarcts. Cerebral microbleeds were more common with anemia. Hemoglobin levels inversely correlated to cerebral perfusion (p < 0.0001). CONCLUSION: Low and high levels of hemoglobin are associated with an increased risk of dementia, including AD, which may relate to differences in white matter integrity and cerebral perfusion.


Assuntos
Doença de Alzheimer/epidemiologia , Anemia/epidemiologia , Infarto Encefálico/epidemiologia , Encéfalo/irrigação sanguínea , Demência/epidemiologia , Hemoglobinas/metabolismo , Doença de Alzheimer/sangue , Doença de Alzheimer/patologia , Anemia/patologia , Encéfalo/patologia , Infarto Encefálico/patologia , Comorbidade , Demência/sangue , Demência/patologia , Feminino , Humanos , Imagem por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Vias Neurais/patologia , Neuroimagem , Prevalência , Fatores de Risco , Soro/metabolismo , Substância Branca/patologia
11.
Cell Host Microbe ; 26(2): 252-264.e10, 2019 08 14.
Artigo em Inglês | MEDLINE | ID: mdl-31399369

RESUMO

Obesity and type 2 diabetes (T2D) are metabolic disorders that are linked to microbiome alterations. However, their co-occurrence poses challenges in disentangling microbial features unique to each condition. We analyzed gut microbiomes of lean non-diabetic (n = 633), obese non-diabetic (n = 494), and obese individuals with T2D (n = 153) from German population and metabolic disease cohorts. Microbial taxonomic and functional profiles were analyzed along with medical histories, serum metabolomics, biometrics, and dietary data. Obesity was associated with alterations in microbiome composition, individual taxa, and functions with notable changes in Akkermansia, Faecalibacterium, Oscillibacter, and Alistipes, as well as in serum metabolites that correlated with gut microbial patterns. However, microbiome associations were modest for T2D, with nominal increases in Escherichia/Shigella. Medications, including antihypertensives and antidiabetics, along with dietary supplements including iron, were significantly associated with microbiome variation. These results differentiate microbial components of these interrelated metabolic diseases and identify dietary and medication exposures to consider in future studies.


Assuntos
Diabetes Mellitus Tipo 2/complicações , Microbioma Gastrointestinal/fisiologia , Obesidade/complicações , Obesidade/microbiologia , Animais , Bactérias/classificação , Bactérias/genética , Bactérias/isolamento & purificação , Biodiversidade , Dieta , Suplementos Nutricionais , Fezes/microbiologia , Feminino , Alemanha , Humanos , Ferro/metabolismo , Magnésio/metabolismo , Masculino , Doenças Metabólicas/complicações , Metagenômica , Camundongos , Camundongos Endogâmicos C57BL , Análise Multivariada , Avaliação Nutricional , Soro/metabolismo
12.
Anal Bioanal Chem ; 411(26): 6983-6994, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31463516

RESUMO

This study investigated the optimal inter-batch normalization method for gas chromatography/tandem mass spectrometry (GC/MS/MS)-based targeted metabolome analysis of rodent blood samples. The effect of centrifugal concentration on inter-batch variation was also investigated. Six serum samples prepared from a mouse and 2 quality control (QC) samples from pooled mouse serum were assigned to each batch, and the 3 batches were analyzed by GC/MS/MS at different days. The following inter-batch normalization methods were applied to metabolome data: QC-based methods with quadratic (QUAD)- or cubic spline (CS)-fitting, total signal intensity (TI)-based method, median signal intensity (MI)-based method, and isotope labeled internal standard (IS)-based method. We revealed that centrifugal concentration was a critical factor to cause inter-batch variation. Unexpectedly, neither the QC-based normalization methods nor the IS-based method was able to normalize inter-batch variation, though MI- or TI-based normalization methods were effective in normalizing inter-batch variation. For further validation, 6 disease model rat and 6 control rat plasma were evenly divided into 3 batches, and analyzed as different batches. Same as the results above, MI- or TI-based methods were able to normalize inter-batch variation. In particular, the data normalized by TI-based method showed similar metabolic profiles obtained from their intra-batch analysis. In conclusion, the TI-based normalization method is the most effective to normalize inter-batch variation for GC/MS/MS-based metabolome analysis. Graphical abstract.


Assuntos
Metaboloma , Metabolômica/métodos , Plasma/metabolismo , Soro/metabolismo , Animais , Centrifugação/métodos , Cromatografia Gasosa-Espectrometria de Massas/métodos , Masculino , Camundongos Endogâmicos ICR , Controle de Qualidade , Ratos , Síndrome da Serotonina/sangue , Síndrome da Serotonina/metabolismo , Espectrometria de Massas em Tandem/métodos
13.
J Chromatogr A ; 1603: 83-91, 2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31288928

RESUMO

In the acidic environment of the stomach, nitrosatable pesticide residues may react with nitrite to form potentially carcinogenic pesticide-associated N-nitroso compounds (PANOCs). The objective of this study was to develop a method for the analysis of 10 nitrosatable pesticides and breakdown products in human serum and urine. Three sample preparation methods were evaluated for extraction of target analytes from the biomatrices. Deproteinization by methanol for 300-µL aliquots of serum with a final extract volume of 225 µL resulted in excessive ion enhancement of some analytes and suppression of others. Three types of solid-phase extraction cartridges were tested for optimal analyte retention from 200-µL aliquots of serum with a final extract volume of 400 µL; this approach resulted in significant analyte loss for some compounds. The Quick, Easy, Cheap, Effective, Rugged, and Safe approach resulted in a suitable method for extraction of the analytes from each biomatrix. Biofluid samples (500 µL) were spiked to 100 µg L-1 with analytical standards and extracted using 500 µL of acetonitrile (ACN) with 4% acetic acid (AcOH) for serum and 0.1% AcOH in ACN for urine. For extraction, 200 mg magnesium sulfate (MgSO4) and 50 mg sodium acetate were added for serum and 200 mg MgSO4 and 50 mg sodium chloride were added for urine. Final extract volumes for both biomatrices using the QuEChERS method was 400 µL after dilution. Samples were analyzed via ultra-high pressure liquid chromatography/high-resolution accurate mass orbital ion trap mass spectrometry. Mean recoveries for target analytes in serum and urine ranged between 74 and 120% (%RSD < 12) and 96 to 116% (%RSD ≤ 10), respectively. These methods may be used in large-scale biomonitoring studies to analyze PANNs and their parent compounds in human serum and urine.


Assuntos
Cromatografia Líquida/métodos , Compostos Nitrosos/análise , Resíduos de Praguicidas/análise , Soro/metabolismo , Extração em Fase Sólida/métodos , Manejo de Espécimes/métodos , Espectrometria de Massas em Tandem/métodos , Humanos , Soro/química , Urinálise/métodos
14.
Am J Chin Med ; 47(5): 1025-1041, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31327237

RESUMO

Myocardial infarction (MI), the most common symptom is chest pain, occurs when blood flow decreases or stops to a part of the heart, causing damage to the heart muscle. Electroacupuncture pretreatment (EP) is a recent observation which has been shown to induce ischemic tolerance like the ischemia preconditioning, suggesting that EP may be a promising preventive strategy for individual susceptibility to MI. This study investigated mechanisms that underlie the effect of EP on MI through the use of gas chromatography-mass spectrometry (GC-MS)-based metabolic profiling. Male Sprague-Dawley rats were randomly divided to receive or not receive three days of EP at PC6 (Neiguan). Then on the fourth day, each group was further divided to undergo mock surgery or MI, induced by ligation of the left anterior descending coronary artery. After 24h, the blood samples and hearts were collected for the follow-up research. The results showed that treatment by EP significantly reduced the levels of CK-MB, cTnT, AST, and MDH in serum and decreased myocardial infarction area. According to GC-MS-based serum metabolic profiling and analysis, a total of 636 characteristic peaks were identified, including 158 known and 478 unknown metabolites. MI caused comprehensive metabolic changes in glycolysis-related metabolites, malate-aspartate shuttle (MAS) metabolites, and purine metabolites with anti-oxidant functions, while EP reversed more than half of the differential metabolic changes, mainly affecting amino acid and energy metabolism, especially the glutamate metabolism and MAS. In a word, our findings suggest that EP exerts its cardioprotective effect on MI by regulating amino acid and energy metabolisms. Meanwhile, GC-MS-based metabolomics provided a powerful way to characterize the metabolic features of MI, with and without EP, and thereby improved our understanding of the effect and mechanisms of EP.


Assuntos
Eletroacupuntura , Isquemia Miocárdica/terapia , Soro/química , Pontos de Acupuntura , Animais , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Masculino , Metaboloma , Metabolômica , Isquemia Miocárdica/sangue , Ratos , Ratos Sprague-Dawley , Soro/metabolismo
15.
Nat Commun ; 10(1): 2961, 2019 07 04.
Artigo em Inglês | MEDLINE | ID: mdl-31273197

RESUMO

Persistent inflammation is a hallmark of many human diseases, including anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV) and atherosclerosis. Here, we describe a dominant trigger of inflammation: human serum factor H-related protein FHR1. In vitro, this protein selectively binds to necrotic cells via its N-terminus; in addition, it binds near necrotic glomerular sites of AAV patients and necrotic areas in atherosclerotic plaques. FHR1, but not factor H, FHR2 or FHR3 strongly induces inflammasome NLRP3 in blood-derived human monocytes, which subsequently secrete IL-1ß, TNFα, IL-18 and IL-6. FHR1 triggers the phospholipase C-pathway via the G-protein coupled receptor EMR2 independent of complement. Moreover, FHR1 concentrations of AAV patients negatively correlate with glomerular filtration rates and associate with the levels of inflammation and progressive disease. These data highlight an unexpected role for FHR1 during sterile inflammation, may explain why FHR1-deficiency protects against certain diseases, and identifies potential targets for treatment of auto-inflammatory diseases.


Assuntos
Proteínas Inativadoras do Complemento C3b/metabolismo , Inflamassomos/metabolismo , Monócitos/metabolismo , Monócitos/patologia , Doenças Vasculares/metabolismo , Doenças Vasculares/patologia , Proteína C-Reativa/metabolismo , Proteínas do Sistema Complemento/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Proteínas Imobilizadas/metabolismo , Inflamação/metabolismo , Inflamação/patologia , Mediadores da Inflamação/metabolismo , Interleucina-1beta/metabolismo , Lipoproteínas LDL/metabolismo , Malondialdeído/metabolismo , Modelos Biológicos , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Necrose , Ligação Proteica , Receptores Acoplados a Proteínas-G/metabolismo , Soro/metabolismo , Fosfolipases Tipo C/metabolismo
16.
Med Sci Monit ; 25: 5518-5524, 2019 Jul 25.
Artigo em Inglês | MEDLINE | ID: mdl-31342946

RESUMO

BACKGROUND The aim of this study was to evaluate the efficacy of RASSF1A promoter hypermethylation of serum or sputum in diagnosis of non-small cell lung cancer (NSCLC) by pooling open published data. MATERIAL AND METHODS Open-published studies relevant to RASSF1A promoter hypermethylation and NSCLC diagnosis were screened through Medline, EMBASE, the Cochrane Library, Web of Science, Google Scholar, and CBM. Number of cases of true positive (tp), false positive (fp), false negative (fn), and true negative (tn) by RASSF1A gene promoter hypermethylation was extracted from each of the include original studies. The combined diagnostic sensitivity, specificity, and symmetric receiver operating characteristic curve (SROC) were calculated, as was the effect size. RESULTS Twelve studies with 826 NSCLC and 598 controls were included in the present work. The combined sensitivity and specificity were 0.45 (95%CI: 0.41-0.48) (random effects) and 0.99(95%CI: 0.98-1.00) (fixed effects) respectively. The pooled positive likelihood ratio (+LR) and negative likelihood ratio (-LR) were 20.27 (9.64-42.61) and 0.53 (0.42-0.66), respectively, through the random effects model. The combined DOR was 46.63 (95%CI: 17.30-125.65) through the fixed effects model. The AUC of the SROC was 0.9989, calculated through Moses's model for RASSF1A promoter hypermethylation as a biomarker in diagnosis of NSCLC. CONCLUSIONS The low diagnostic sensitivity for RASSF1A gene promoter hypermethylation indicated that it is not suitable for NSCLC screening. However, the high specificity made it effective for NSCLC confirmation diagnosis, which could be used instead of pathological diagnosis.


Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Metilação de DNA , Neoplasias Pulmonares/genética , Proteínas Supressoras de Tumor/genética , Área Sob a Curva , Biomarcadores Tumorais/genética , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Humanos , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , Regiões Promotoras Genéticas , Curva ROC , Sensibilidade e Especificidade , Soro/metabolismo , Escarro/metabolismo , Transcriptoma , Proteínas Supressoras de Tumor/metabolismo
17.
Mol Biol Rep ; 46(4): 4437-4441, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31154604

RESUMO

Human mesenchymal stem cells (MSCs) are presently on the top of hierarchy in the field of stem cell therapy, due to their miraculous therapeutic abilities in diminishing the symptoms of many chronic diseases and initiating regeneration and repair for various damaged tissues and organs. The foremost initial step to reach high success rate in any MSCs based study is the optimization of culture growth media by establishing a suitable fetal bovine serum (FBS) percentage that suits the purpose of MSCs based experiment. Choosing the suitable FBS percentage is a controversial issue and merely depends on the researchers experience and suggested recommendations by the suppliers. Despite the huge improvements in overall MSCs investigating approaches, there are no definite protocols that set up a range of FBS percentages that can be followed. Toward achieving this objective, we evaluate in the present report the effect of using various FBS percentages (5-20%) added to DMEM low glucose media, on the biological behaviour of MSCs. Growing MSCs in high FBS percentages containing culture media (15% and 20% FBS) increase the proliferation and expansion rate of MSCs, although it decreases the immunosuppressive properties. On the other hand, adding low FBS percentage (7% FBS) to MSCs culture media enhanced the immunosuppression characteristics of MSCs, even though the proliferation rate was moderately reduced. 7% FBS is the cut off percentage that can be used without negatively altering major MSCs biological properties in which using 5% FBS will cause a tremendous decrease in the proliferation capacity and immunosuppressive properties. This report may assist other researchers in choosing appropriate FBS percentage when preparing MSCs culture media that serve the purpose of their MSCs based studies.


Assuntos
Técnicas de Cultura de Células/métodos , Células-Tronco Mesenquimais/metabolismo , Animais , Células da Medula Óssea/citologia , Bovinos/sangue , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas/metabolismo , Meios de Cultura/análise , Sangue Fetal/metabolismo , Humanos , Soro/metabolismo
18.
Ann Afr Med ; 18(2): 103-107, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31070153

RESUMO

Background: Exacerbations in Chronic obstructive pulmonary disease (COPD) have a considerable impact on morbidity, mortality, and quality of life. Procalcitonin (PCT) a polypeptide normally produced in neuroendocrine cells of the thyroid and lungs is a marker of systemic inflammation and bacterial infection. Objectives: The aim of this study was to determine the levels of PCT in serum of acute exacerbation of COPD patients (AECOPD) and stable COPD patients in North Indian population. Materials and Methods: The study was conducted on 80 AECOPD and 80 stable COPD patients in respiratory medicine department at tertiary care hospital in north India. PCT levels were measured in serum by ELISA kit. GraphPad Prism version 6.01 (GraphPad software Inc.; La, Jolla, CA, USA) was used for analysis of data. Results: The present study showed that mean serum PCT levels were significantly higher in AECOPD group (1.31 ± 0.79) as compared to stable COPD group (0.1 ± 0.09) (P < 0.001). Conclusion: The study confirms that PCT levels were higher in AECOPD patients as compared to stable COPD patients. PCT could be used as a biomarker of exacerbations of COPD and can be used to target management and guiding the treatment in patients with acute exacerbations of COPD.


Assuntos
Pró-Calcitonina/sangue , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Qualidade de Vida , Soro/metabolismo , Adulto , Idoso , Biomarcadores/sangue , Feminino , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/sangue , Doença Pulmonar Obstrutiva Crônica/psicologia
19.
Thromb Haemost ; 119(7): 1058-1071, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31055798

RESUMO

Interaction between the transcription factors, hypoxia-inducible factor (HIF1α and HIF2α) and Sp1, mediates hypoxia-driven expression of FVII gene encoding coagulation factor VII (fVII) in ovarian clear cell carcinoma (CCC) cells. This mechanism is synergistically enhanced in response to serum starvation, a condition possibly associated with tumor hypoxia. This transcriptional response potentially results in venous thromboembolism, a common complication in cancer patients by producing procoagulant extracellular vesicles (EVs). However, which deficient serum factors are responsible for this characteristic transcriptional mechanism is unknown. Here, we report that cholesterol deficiency mediates synergistic FVII expression under serum starvation and hypoxia (SSH) via novel sterol regulatory element binding protein-1 (SREBP1)-driven mechanisms. Unlike conventional mechanisms, SREBP1 indirectly enhances FVII transcription through the induction of a new target, glucocorticoid-induced leucine zipper (GILZ) protein. GILZ expression induced in response to hypoxia by a HIF1α-dependent mechanism activates SREBP1 under SSH, suggesting reciprocal regulation between SREBP1 and GILZ. Furthermore, GILZ binds to the FVII locus. Xenograft tumor samples analyzed by chromatin immunoprecipitation confirmed that HIF1α-aryl hydrocarbon nuclear translocator and GILZ bind to the TSC22D3 (GILZ) and FVII gene loci, respectively, thereby potentially modulating chromatin function to augment FVII transcription. Thus, deficiency of both O2 and cholesterol, followed by interplay between HIFs, Sp1, and SREBP1-GILZ pathways synergistically induce fVII synthesis, resulting in the shedding of procoagulant EVs.


Assuntos
Micropartículas Derivadas de Células/metabolismo , Coagulantes/metabolismo , Fator VII/genética , Hipóxia/metabolismo , Neoplasias Ovarianas/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Fatores de Transcrição/metabolismo , Animais , Linhagem Celular Tumoral , Colesterol/metabolismo , Montagem e Desmontagem da Cromatina , Fator VII/metabolismo , Feminino , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Camundongos , Soro/metabolismo , Transdução de Sinais , Proteína de Ligação a Elemento Regulador de Esterol 1/genética , Fatores de Transcrição/genética , Ensaios Antitumorais Modelo de Xenoenxerto
20.
Int J Mol Sci ; 20(10)2019 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-31130623

RESUMO

In an earlier study, signs of commencing degeneration of spinal motor neurons were induced in mice with short-term intraperitoneal injections of immunoglobulin G (IgG) taken from patients with amyotrophic lateral sclerosis (ALS). Since in that study, neither weakness nor loss of motor neurons was noted, to test whether the ALS IgG in this paradigm has the potential to evoke relentless degeneration of motor neurons, treatment with repeated injections over a longer period was carried out. Mice were systematically injected intraperitoneally with serum taken from ALS patients over a 75-day period. At selected time points, the isometric force of the limbs, number of spinal motor neurons and their intracellular calcium levels were determined. Furthermore, markers of glial activation and the motoneuronal uptake of human IgG were monitored. During this period, gliosis and progressive motoneuronal degeneration developed, which led to gradual loss of spinal motor neurons, more than 40% at day 21, along with decreasing muscle strength in the limbs. The inclusion-like accumulation of IgG appeared in the perikarya with the increase of intracellular calcium in the cell bodies and motor nerve terminals. Our results demonstrate that ALS serum can transfer motor neuron disease to mice.


Assuntos
Esclerose Amiotrófica Lateral/metabolismo , Doença dos Neurônios Motores/metabolismo , Neurônios Motores/patologia , Soro/metabolismo , Esclerose Amiotrófica Lateral/sangue , Animais , Cálcio/metabolismo , Modelos Animais de Doenças , Feminino , Humanos , Imunoglobulina G/administração & dosagem , Imunoglobulina G/sangue , Imunoglobulina G/metabolismo , Injeções Intraperitoneais , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Doença dos Neurônios Motores/patologia , Neurônios Motores/metabolismo
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