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1.
Chem Commun (Camb) ; 56(11): 1637-1640, 2020 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-31960844

RESUMO

We report a modularized sample purification system (MSPS) fabricated by 3D printing for rapid and high-throughput MALDI-MS analysis of various small-volume biological samples. The MSPS presents distinct advantages such as customizability, portability, expandability, simple operation and high throughput and also keeps the flexibility of being customized with more functional modules in future applications.


Assuntos
Impressão Tridimensional , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Água/análise , Líquido da Lavagem Broncoalveolar/química , Humanos , Soro/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/instrumentação
2.
Arch Virol ; 165(1): 127-135, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31741097

RESUMO

In clinical virome research, whole-genome/transcriptome amplification is required when starting material is limited. An improved method, named "template-dependent multiple displacement amplification" (tdMDA), has recently been developed in our lab (Wang et al. in BioTechniques 63:21-25. https://doi.org/10.2144/000114566, 2017). In combination with Illumina sequencing and bioinformatics pipelines, its application in virome sequencing was explored using a serum sample from a patient with chronic hepatitis C virus (HCV) infection. In comparison to an amplification-free procedure, virome sequencing via tdMDA showed a 9.47-fold enrichment for HCV-mapped reads and, accordingly, an increase in HCV genome coverage from 28.5% to 70.1%. Eight serum samples from acute patients liver failure (ALF) with or without known etiology were then used for virome sequencing with an average depth at 94,913x. Both similarity-based (mapping, NCBI BLASTn, BLASTp, and profile hidden Markov model analysis) and similarity-independent methods (machine-learning algorithms) identified viruses from multiple families, including Herpesviridae, Picornaviridae, Myoviridae, and Anelloviridae. However, their commensal nature and cross-detection ruled out an etiological interpretation. Together with a lack of detection of novel viruses in a comprehensive analysis at a resolution of single reads, these data indicate that viral agents might be rare in ALF cases with indeterminate etiology.


Assuntos
Biologia Computacional/métodos , Hepatite C Crônica/diagnóstico , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Falência Hepática Aguda/virologia , Soro/virologia , Anelloviridae/isolamento & purificação , Anelloviridae/fisiologia , Perfilação da Expressão Gênica/métodos , Hepacivirus/genética , Hepacivirus/isolamento & purificação , Hepatite C Crônica/sangue , Herpesviridae/isolamento & purificação , Herpesviridae/fisiologia , Humanos , Falência Hepática Aguda/sangue , Myoviridae/isolamento & purificação , Myoviridae/fisiologia , Picornaviridae/isolamento & purificação , Picornaviridae/fisiologia , Especificidade da Espécie , Simbiose , Sequenciamento Completo do Genoma/métodos
3.
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
4.
J Sci Food Agric ; 100(2): 874-884, 2020 Jan 30.
Artigo em Inglês | MEDLINE | ID: mdl-31680264

RESUMO

BACKGROUND: The low cost of aminoglycoside (AMG) antibiotics facilitates their excessive use in animal husbandry and the agriculture sector. This scenario has led to the occurrence of residues in the food chain. After several years of AMG use in antibacterial therapy, resistance to streptomycin has begun to appear. Most of the detection methods developed for AMG antibiotics lacks specificity. A broad target specific nanoprobe would be ideal for detecting the entire class of AMGs. A rapid and sensitive method for the detection of AMGs is urgently needed. RESULTS: Gallic acid-coated silver nanoparticles (AgNPs) were demonstrated as a nanoprobe for the colorimetric detection of AMGs (yellow to orange / red). A linear dynamic range of 50-650 pmol L-1 was achieved readily by ratiometric spectrophotometry (A560 /A400 ) with a limit of detection (LOD) as low as 36 pmol L-1 . The amine-groups of the AMGs function as molecular linkers, so that electrostatic coupling interactions between neighboring particles drive the formation of AgNP aggregates. The assay can also be applied for the determination of streptomycin residues in serum and milk samples. CONCLUSION: This study revealed the potential of an AgNP probe for the rapid and cost-effective detection of low-molecular-weight target analytes, such as the AMGs. A ligand-induced aggregation of AgNPs coated with gallic acid was reported to be a rapid and sensitive assay for AMGs. Analysis of streptomycin was demonstrated with excellent picomolar-level sensitivity. Thus, the validated method can find practical applications in the ultrasensitive detection of AMGs in complex and diagnostic settings. © 2019 Society of Chemical Industry.


Assuntos
Antibacterianos/análise , Colorimetria/métodos , Resíduos de Drogas/análise , Leite/química , Soro/química , Estreptomicina/análise , Água/química , Animais , Antibacterianos/farmacologia , Bovinos , Colorimetria/instrumentação , Limite de Detecção , Nanopartículas Metálicas/química , Prata/química
5.
Altern Lab Anim ; 47(3-4): 116-127, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31698922

RESUMO

Serum supplementation is crucial in in vitro cell culture to provide all the essential nutrients needed for cellular processes. Fetal bovine serum (FBS) is considered the 'gold standard', but its production raises serious ethical concerns. Human-derived alternatives to FBS exist in the form of human platelet lysates (hPLs) or human AB serum (ABS). However, these serum products are usually pooled from several donors, in order to have a standardised product without patient-specific deviations. Nevertheless, the use of patient-specific serum in cell culture might be the key to successful transplantation of the cultured cells in medical applications, particularly as it avoids the transmission of infectious components or xenogenic proteins. In addition, the production of non-pooled hPL from single donors is likely to be a cost-effective and time-saving method. The current study used hPL units isolated from single donors and tested their performance as medium supplements for cell culture in comparison with FBS or ABS. This proof-of-concept study aimed to assess the potential of non-pooled hPL for personalised serum supplementation, and thus optimise in vitro models by making them more relevant to human physiology. We showed that A549, HepG2 and Caco-2 human cell lines were generally able to adapt to the new culture conditions and maintain viability, morphology and certain cell-specific characteristics. These results indicate that non-pooled, single patient-derived hPL could be a suitable alternative for in vitro serum supplementation.


Assuntos
Técnicas de Cultura de Células , Soro , Células A549 , Células CACO-2 , Técnicas de Cultura de Células/métodos , Técnicas de Cultura de Células/normas , Proliferação de Células , Células Hep G2 , Humanos
6.
Klin Lab Diagn ; 64(11): 663-668, 2019.
Artigo em Russo | MEDLINE | ID: mdl-31747494

RESUMO

To study the concentration of vasculoendothelial growth factor (VEGF) in mixed saliva and serum of patients in normal conditions and with generalized periodontitis. The main group (n = 42) was represented by patients with generalized periodontitis. The comparison group (n = 36) consisted of patients without periodontal tissue diseases. The concentration of VEFR was determined by the method of enzyme-linked immunosorbent assay (ELISA) using a commercial test-system "VEGF - IFA - BEST" (A-8784) ("Vector - Best", Russia). The median values VEFR in saliva were 5.49 times higher than the values for serum in the main group (p = 0.000000) and 7.01 times in the comparison group (p = 0.000000). The concentration of VEFR in the saliva of the examined main group exceeded the similar values of the comparison group (p = 0,014857); the median and interquartile range for the main group was 1098.45 (925.5; 1291) pg/ml, and for the comparison group 1360.5 (998.9; 2062) pg/ml. There were no differences in the serum VEFR concentration (p = 0.775124). No significant correlation was found between the serum VEFR content and the mixed saliva. The Spearman's rank correlation coefficient for the main group was R = 0,0184358, and for the comparison group, respectively, R = 0.188932. The source of VEFR in saliva are the glands and cells of the oral mucosa, and not the process of exudation from blood serum. The high content of VEFR in the saliva of healthy people and a decrease in its level during periodontitis indicates the important role of this protein in the processes of maintaining the normal state of periodontal tissues and reparation of tissues of the oral mucosa.


Assuntos
Periodontite/diagnóstico , Saliva/química , Soro/química , Fator A de Crescimento do Endotélio Vascular/análise , Humanos , Federação Russa
7.
Zhongguo Zhong Yao Za Zhi ; 44(17): 3780-3785, 2019 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-31602953

RESUMO

The aim of this paper was to investigate the molecular mechanism of Calculus Bovis Sativus( CBS) in alleviating lipid accumulation in vitro by serum pharmacology. The CBS-containing serum of mice was obtained by serum pharmacology method to evaluate its effect on the proliferation of LO2 hepatocytes. The lipid reducing effects of CBS-containing serum through Nrf2 was evaluated by fructose-induced LO2 hepatocyte steatosis model,nuclear factor erythroid 2 related factor 2( Nrf2) agonist oltipraz combined intervention,cell oil red O staining and intracellular triglyceride( TG) content. The effects of CBS-containing serum on lipid peroxidation and hepatocytes apoptosis were evaluated by reactive oxygen species( ROS) and apoptosis assay,respectively. Real-time quantitative polymerase chain reaction( PCR) was used to detect the relative expression of lipid synthesis-related genes and apoptosis-related genes.RESULTS:: showed that CBS drug-containing serum had no significant effect on LO2 hepatocyte proliferation. As compared with the model group,CBS-containing serum could effectively reduce the formation of lipid droplets in fructose-induced LO2 hepatocytes,significantly reduce intracellular TG and ROS levels,and significantly reduce hepatocyte apoptosis rate( P < 0. 05). As compared with the model group,carbohydrate responsive element binding protein( ChREBP),sterol regulatory element binding protein-1 c( SREBP-1 c),fatty acid synthase( FAS),acetyl-CoA carboxylase 1( ACC1),stearoyl-CoA desaturase 1( SCD1),Bax and caspase-3 mRNA levels were significantly reduced in CBS drug-containing serum treatment group( P<0. 05). All of the above effects could be reversed by oltipraz.In conclusion,CBS-containing serum can significantly inhibit the fructose-induced LO2 liver fat deposition,and the mechanism may be related to reducing intracellular ROS level through the Nrf2 pathway and improving intracellular peroxidation state to reduce apoptosis.


Assuntos
Cálculos Biliares/química , Hepatócitos/citologia , Soro/química , Animais , Apoptose , Bovinos , Células Cultivadas , Fígado Gorduroso , Frutose , Hepatócitos/metabolismo , Metabolismo dos Lipídeos , Peroxidação de Lipídeos , Fígado , Medicina Tradicional Chinesa , Camundongos , Espécies Reativas de Oxigênio/metabolismo , Proteína de Ligação a Elemento Regulador de Esterol 1/metabolismo , Triglicerídeos
8.
J Biochem Mol Toxicol ; 33(12): e22407, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31581362

RESUMO

In this study, we investigated the effects of certain respiratory drugs, which are mainly used on human serum paraoxonase-1 (hPON1; EC 3.1.8.1). hPON1 was purified from human serum, with 354.91 fold and 45% yield by using two simple step procedures including, first, ammonium sulfate precipitation, then, Sepharose-4B-l-tyrosine-1-naphthylamine hydrophobic interaction chromatography. SDS-polyacrylamide gel electrophoresis showed a single protein band belonging to hPON1 with 43 kDa. All the pharmaceutical compounds inhibited the PON1 enzyme highly at the micromolar level. The obtained IC50 values for nine different pharmaceutics ranged from 0.219 µM (salbutamol sulfate) to 67.205 µM (montelukast sodium). So, all drugs could be considered as potent hPON1 inhibitors. Ki values and inhibition types were determined by Lineweaver-Burk graphs. While varenicline tartrate and moxifloxacin hydrochloride inhibited the enzyme in a noncompetitive manner, others inhibited it in a mixed manner.


Assuntos
Arildialquilfosfatase/química , Arildialquilfosfatase/isolamento & purificação , Broncodilatadores/química , Inibidores Enzimáticos/química , Soro/enzimologia , Arildialquilfosfatase/antagonistas & inibidores , Broncodilatadores/efeitos adversos , Broncodilatadores/uso terapêutico , Cromatografia em Gel/métodos , Eletroforese em Gel de Poliacrilamida , Inibidores Enzimáticos/efeitos adversos , Inibidores Enzimáticos/uso terapêutico , Humanos , Interações Hidrofóbicas e Hidrofílicas , Infecções Respiratórias/tratamento farmacológico
9.
Int J Nanomedicine ; 14: 7107-7121, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31564868

RESUMO

Background: Cervical cancer (CxCa) ranks as the fourth most prevalent women-related cancer worldwide. Therefore, there is a crucial need to develop newer treatment modalities. Ormeloxifene (ORM) is a non-steroidal, selective estrogen receptor modulator (SERM) that is used as an oral contraceptive in humans. Recent investigations suggest that ORM exhibits potent anti-cancer activity against various types of cancers. Nanoparticulates offer targeted delivery of anti-cancer drugs with minimal toxicity and promise newer approaches for cancer diagnosis and treatment. Therefore, the nanotherapy approach is superior compared to traditional chemotherapy, which is not site-specific and is often associated with various side effects. Methods: Pursuing this novel nanotherapy approach, our lab has recently developed ORM-loaded poly [lactic-co-glycolic acid] (PLGA), an FDA-approved biodegradable polymer, nanoparticles to achieve targeted drug delivery and improved bioavailability. Our optimized PLGA-ORM nanoformulation showed improved internalization in both dose- and energy-dependent manners, through endocytosis-mediated pathways in both Caski and SiHa cell lines. Additionally, we employed MTS and colony forming assays to determine the short- and long-term effects of PLGA-ORM on these cells. Results: Our results showed that this formulation demonstrated improved inhibition of cellular proliferation and clonogenic potential compared to free ORM. Furthermore, the PLGA-ORM nanoformulation exhibited superior anti-tumor activities in an orthotopic cervical cancer mouse model than free ORM. Conclusion: Collectively, our findings suggest that our novel nanoformulation has great potential for repurposing the drug and becoming a novel modality for CxCa management.


Assuntos
Benzopiranos/uso terapêutico , Nanopartículas/uso terapêutico , Neoplasias do Colo do Útero/tratamento farmacológico , Animais , Benzopiranos/farmacologia , Carcinogênese/efeitos dos fármacos , Carcinogênese/metabolismo , Carcinogênese/patologia , Linhagem Celular Tumoral , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Clonais , Modelos Animais de Doenças , Endocitose/efeitos dos fármacos , Eritrócitos/metabolismo , Feminino , Hemólise/efeitos dos fármacos , Humanos , Teste de Materiais , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Camundongos Nus , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Soro/química , Neoplasias do Colo do Útero/patologia
10.
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
11.
Parasit Vectors ; 12(1): 447, 2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31506092

RESUMO

BACKGROUND: Toxocara canis, a globally distributed roundworm, can cause debilitating disease in dogs and humans; however, little is known about the metabolomic response of the hosts to T. canis infection. There is an increasing need to understand the metabolic mechanisms underlying the pathogenesis of T. canis infection in dogs. Here, we examined the metabolomic changes in Beagle dogs' serum following T. canis infection using LC-MS/MS. RESULTS: The metabolic profiles of Beagle dogs' serum were determined at 12 h, 24 h, 10 d and 36 d after oral infection with 300 infectious T. canis eggs by LC-MS/MS. We tested whether the T. canis-associated differentially abundant metabolites could distinguish the serum of infected dogs from controls, as measured by the area under the receiver operating characteristic (ROC) curve (AUC). The differentially expressed metabolites were further evaluated by principal components analysis and pathway enrichment analysis. A total of 5756 and 5299 ions were detected in ESI+ and ESI- mode, respectively. ROC curve analysis revealed nine and five metabolite markers, at 12 hpi and 24 hpi to 36 dpi, respectively, with potential diagnostic value for toxocariasis. The levels of taurocholate, estradiol, prostaglandins and leukotriene were significantly changed. Primary bile acid biosynthesis pathway, steroid hormone biosynthesis pathway and biosynthesis of unsaturated fatty acids pathway were significantly altered by T. canis infection. CONCLUSIONS: These findings show that T. canis infection can induce several changes in the dog serum metabolome and that the metabolic signature associated with T. canis infection in dogs has potential for toxocariasis diagnosis.


Assuntos
Biomarcadores/sangue , Doenças do Cão/patologia , Metabolômica , Soro/química , Toxocara canis/crescimento & desenvolvimento , Toxocaríase/patologia , Animais , Cromatografia Líquida , Cães , Espectrometria de Massas em Tandem , Fatores de Tempo
12.
Artigo em Inglês | MEDLINE | ID: mdl-31536838

RESUMO

In recent decades cryogels as monolithic materials have gained interest as stationary phase in chromatography for purification of biomolecules. In this study, polyacrylamide-alginate (PAAm-Alg) monolithic cryogels were prepared by cryo-copolymerization of acrylamide and alginate monomers and methylene-bisacrylamide as crosslinker to be used as a matrix in affinity chromatography for purification of proteins. Ortho-phospho-L-tyrosine (P-Tyr) was covalently attached onto PAAm-Alg cryogels via bisoxirane-activation (PAAm-Alg-Bix-P-Tyr) and both derivatized and non-derivatized cryogels were utilized for the purification of immunoglobulin G (IgG) from human serum. Cryogels were characterized by scanning electron microscopy, swelling tests, elemental analysis, FTIR, and flow dynamics. The effects of buffer systems, conductivity, and pH on IgG adsorption were studied. Through breakthrough curve analysis a dynamic capacity of 9.2 mg IgG/mL with an IgG purity of 94% was obtained (based on ELISA analysis of IgG and albumin) for PAAm-Alg-Bix-P-Tyr cryogel when human serum was diluted in 10 mmol/L NaP buffer at pH 6.0. The adsorption isotherm data were well described by the Langmuir model with value of maximum adsorption capacity of 36.12 ±â€¯3.63 mg of IgG/g for PAAm-Alg-Bix-P-Tyr. The PAAm-Alg-Bix-P-Tyr cryogel provides an attractive alternative for adsorption of IgG from human serum.


Assuntos
Resinas Acrílicas/química , Alginatos/química , Criogéis/química , Imunoglobulina G/isolamento & purificação , Soro/química , Tirosina/química , Adsorção , Tampões (Química) , Cromatografia de Afinidade/métodos , Reagentes para Ligações Cruzadas/química , Condutividade Elétrica , Humanos , Concentração de Íons de Hidrogênio , Imunoglobulina G/análise , Fosfitos/química , Polimerização , Porosidade , Propriedades de Superfície
13.
Fa Yi Xue Za Zhi ; 35(4): 396-401, 2019 Aug.
Artigo em Inglês, Chinês | MEDLINE | ID: mdl-31532145

RESUMO

Abstract: Objective To study the protein expression of cluster of differentiation 63 (CD63) in lung tissues of guinea pigs that died of anaphylactic shock and discuss the diagnostic value of CD63 for death from anaphylactic shock. Methods Twenty guinea pigs were randomly divided into control group, anaphylactic shock immediate death group, cold storage group (4 ℃ for 48 h) and frozen group (-20 ℃ for 7 d). The animal model of guinea pigs that died of anaphylactic shock was established with human mixed serum injection. The expression changes of CD63 protein and CD63 mRNA in lung tissues were detected by hematoxylin-eosin (HE) staining, immunohistochemical staining, Western blotting, enzyme-linked immunosorbent assay (ELISA) and real-time RT-PCR. Results HE staining results showed congestion, and edema of lung tissues, and eosinophil infiltration in the anaphylactic shock groups. Western blotting analysis results showed that the expression of CD63 protein in the lung tissues of guinea pigs that died of anaphylactic shock was significantly higher than that in the control group (P<0.05). Comparison between the anaphylactic shock groups was made, and the differences had no statistical significance. The results of immunohistochemical staining and real-time RT-PCR were consistent with that of Western blotting. ELISA results showed that CD63 protein expression in the immediate death group was higher than that in the control group (P<0.05). Conclusion The expression of CD63 protein and CD63 mRNA in the lung tissues of guinea pigs that died of anaphylactic shock is significantly enhanced. Animal carcasses which were put in cold storage for 48 h and frozen for 7 d do not affect the examination of the above indicators. CD63 protein is expected to become an auxiliary diagnostic indicator of death from anaphylactic shock.


Assuntos
Anafilaxia/metabolismo , Pulmão/metabolismo , Tetraspanina 30/metabolismo , Anafilaxia/mortalidade , Animais , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Cobaias , Humanos , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Soro
14.
J Steroid Biochem Mol Biol ; 195: 105472, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31550504

RESUMO

Blood for determining 25-hydroxyvitamin D3 [25(OH)D3] is usually obtained through venipuncture although, as an alternative for serum, dried blood spot (DBS) can be considered. The aim of this proof-of-concept study was to investigate levels of agreement between measurements of 25(OH)D3 obtained with DBS compared with serum. 301 Chinese participants were included who completed 25(OH)D3 measurement from DBS and from simultaneously collected blood samples obtained by venipuncture. Measurements of both DBS and serum 25(OH)D3 were performed using liquid chromatography followed by tandem mass spectrometry. Agreement between the two methods was assessed with Passing and Bablok regression analysis and Bland-Altman plot. Measurements showed a good correlation (Pearson's correlation coefficient r = 0.929, P < 0.001) between the two methods. After recalculating for a 13% difference, a regression equation of DBS 25(OH)D3 = -1.91 + 1.00 serum 25(OH)D3 was found in Passing and Bablok regression analysis. Bland-Altman analysis showed a fixed bias of 1.7 nmol/L; upper and lower limit of agreement was 24.1 nmol/L and -20.7 nmol/L, respectively. Sensitivity of recalculated DBS for 25(OH)D3 concentrations <30 and <50 nmol/L was 87.8% and 91.1%, respectively, and specificity was 89.2% and 83.1%, respectively. In conclusion, a good agreement was found between the measurement of 25(OH)D3 obtained with DBS compared with serum. DBS may possibly be used in a future screening program, but it is less suitable for individualized vitamin D status assessment.


Assuntos
Calcifediol/sangue , Soro/química , Vitaminas/sangue , Adulto , Idoso , Grupo com Ancestrais do Continente Asiático , Cromatografia Líquida , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Países Baixos , Reprodutibilidade dos Testes , Espectrometria de Massas em Tandem
15.
Int J Med Sci ; 16(8): 1102-1106, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31523172

RESUMO

Mesenchymal stem cells are an ideal source for regenerative medicine. For clinical use, cell culture should be done at stable conditions, thus the use of serum should be avoided because of the batch-to-batch variations of serum. Although several kinds of serum-free media are available, a method to confirm whether they contain serum has not been established yet. During studies on effect of adipocyte mesenchymal stem cells (Ad-MSCs) on pain using a human pain gene array, we noticed that BDKRB1 gene was constantly upregulated when serum was used in the culture medium. In this study, we attempted to establish further the potential of this gene as a new marker indicative of the presence of serum in media. Using a real-time quantitative PCR gene array screening containing 84 functional genes, we verified BDKRB1 as a specific gene upregulated in the presence of serum. The expression of BDKRB1 in Ad-MSCs was induced not only by bovine serum but also by human serum. The BDKRB1 expression was induced even when Ad-MSCs was cultured with 0.1% serum in the medium. We concluded that BDKRB1 is a valuable marker to detect traces of both human and animal serum in Ad-MSCs cultures. Our study provides a new method to confirm the absence of serum in media and ensure a stable cell culture condition.


Assuntos
Meios de Cultura/análise , Células-Tronco Mesenquimais/citologia , Receptor B1 da Bradicinina/genética , Soro , Animais , Células Cultivadas , Meios de Cultura/química , Meios de Cultura/farmacologia , Meios de Cultura Livres de Soro/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/fisiologia , Receptor A2A de Adenosina/genética
16.
Analyst ; 144(19): 5700-5705, 2019 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-31486454

RESUMO

To analyze multiple analytes in trace samples, low-dosage and high efficiency are crucial in many common cases. Herein, we developed a facile method using a single-channel surface plasmon resonance (SPR)-based biosensor for the simultaneous detection of gentamicin (GEN) and melamine (MEL) in milk and serum with only one sample injection. Based on a sandwich immunoassay, non-interfering antibodies against GEN from mouse (AbGEN) and against MEL from rabbit (AbMEL) were chosen to capture the analytes. Secondary antibodies against mouse (AbM) and rabbit (AbR) were used to bind with AbGEN and AbMEL to determine the concentrations of GEN and MEL on a single channel of an SPR sensor. All of the detection process could be done in 10 min with 50 µL of sample injection. According to the response shifts of AbM and AbR, two standard curves for GEN and MEL were obtained successively, with the limit of detection (LOD) values at 4.4 ng mL-1 and 1.3 ng mL-1, respectively. Moreover, the feasibility was determined by spiking milk and serum samples with GEN and MEL, with recoveries in the range of 81.6%-118.0%. Importantly, the analytes can be substituted by others for much more applications. This method is also expected to multiply the detection efficiency of multi-channel SPR biosensors with low-dosage samples in the future.


Assuntos
Técnicas Biossensoriais/métodos , Imunoensaio/métodos , Ressonância de Plasmônio de Superfície/métodos , Animais , Anticorpos/imunologia , Técnicas Biossensoriais/instrumentação , Gentamicinas/análise , Gentamicinas/imunologia , Humanos , Limite de Detecção , Camundongos , Leite/química , Coelhos , Reprodutibilidade dos Testes , Soro , Ressonância de Plasmônio de Superfície/instrumentação , Triazinas/análise
17.
Environ Sci Technol ; 53(19): 11447-11457, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31476116

RESUMO

We investigated associations between serum perfluoroalkyl acid (PFAA) concentrations in children aged 4, 8, and 12 years (sampled in 2008-2015; n = 57, 55, and 119, respectively) and exposure via placental transfer, breastfeeding, and ingestion of PFAA-contaminated drinking water. Sampling took place in Uppsala County, Sweden, where the drinking water has been historically contaminated with perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHxS), perfluorooctanesulfonate (PFOS), perfluoroheptanoate (PFHpA), and perfluorooctanoate (PFOA). PFOS showed the highest median concentrations in serum (3.8-5.3 ng g-1 serum), followed by PFHxS (1.6-5.0 ng g-1 serum), PFOA (2.0-2.5 ng g-1 serum), and perfluorononanoate (PFNA) (0.59-0.69 ng g-1 serum) in children. Including all children, serum PFOA, PFHxS, and PFOS concentrations in children increased 10, 10, and 1.3% (adjusted mean), respectively, per unit (ng g-1 serum) of increase in the maternal serum level (at delivery), the associations being strongest for 4 year-old children. PFHxS and PFOS significantly increased 3.9 and 3.8%, respectively, per month of nursing, with the highest increase for 4 year-olds. PFOA, PFBS, PFHxS, and PFOS increased 1.2, 207, 7.4, and 0.93%, respectively, per month of cumulative drinking water exposure. Early life exposure to PFOA, PFHxS, and PFOS is an important determinant of serum concentrations in children, with the strongest influence on younger ages. Drinking water with low to moderate PFBS, PFHxS, PFOS, and PFOA contamination is an important source of exposure for children with background exposure from other sources.


Assuntos
Ácidos Alcanossulfônicos , Água Potável , Fluorcarbonetos , Caprilatos , Criança , Pré-Escolar , Ingestão de Líquidos , Feminino , Humanos , Gravidez , Soro , Suécia , Poluição da Água
18.
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
19.
J Pharm Pharmacol ; 71(10): 1497-1507, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31385295

RESUMO

OBJECTIVE: To evaluate the impact of PEG content on poly(lactic-co-glycolic acid) (PLGA) NP physicochemical properties, hydrophobic drug release (rifampicin as a model drug) and human serum protein binding. METHODS: Rifampicin loaded and unloaded nanoparticles with PEG content of 0-17% (w/w) were prepared by an emulsification-evaporation technique. Nanoparticles were characterized for size, zeta potential and morphology. PEGlyation was confirmed using proton nuclear magnetic resonance (1H NMR). Fluorescence spectroscopy and dynamic light scattering were used to determine nanoparticle-protein binding, binding constants and stability of nanoparticles in human serum, respectively. Drug loading and release were determined by UV-VIS spectroscopy and drug release data was mathematically modelled. KEY FINDINGS: A NP PEG content of 17% w/w significantly retarded release of rifampicin from PLGA NPs and altered kinetics of drug release. Stern-Volmer (Ksv) protein binding constants decreased upon PEG incorporation. A 2% w/w PEG was sufficient to significantly reduce protein binding extent to PLGA NPs and maintain particle size distributions. CONCLUSION: The ability to fine tune drug release and formation of protein corona around nanoparticles is crucial to formulation scientists. This study suggests that PLGA NPs with low PEG content might be suitable for extended circulation and rapid drug release and that higher PEG content retards hydrophobic drug release.


Assuntos
Nanopartículas/química , Polietilenoglicóis/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Albumina Sérica Humana/química , Soro/química , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Espectroscopia de Ressonância Magnética/métodos , Tamanho da Partícula , Ácido Poliglicólico/química
20.
Med Sci Monit ; 25: 5850-5855, 2019 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-31385574

RESUMO

BACKGROUND The aim of this study was to detect the expression levels of chemokines (CX3CL1, CXCL-11, CXCL-12, CCL3, CCL4, and CCL20) in the serum of esophageal cancer patients and a normal control group, and to explore the correlations of those expression levels with the pathological type, progression, and metastasis of esophageal cancer. MATERIAL AND METHODS A total of 50 normal people and 50 untreated patients initially diagnosed with esophageal cancer (including 17 cases of non-metastatic esophageal cancer, 33 cases of metastatic esophageal cancer, 36 cases of esophageal squamous cell carcinoma and 14 cases of esophageal adenocarcinoma) were collected. The liquid chip (Luminex) technology was applied to detect the expression levels of the above-mentioned serum chemokines in the two groups. The results were analyzed using Statistical Product and Service Solution 20.0 software. RESULTS The expression levels of CX3CL1, CXCL-12, and CCL20 in esophageal cancer group were evidently higher than those in normal control group (P<0.001, P<0.001 and P=0.003, respectively). There were no statistically significant differences in chemokine expressions between metastatic esophageal cancer group and non-metastatic esophageal cancer group (P>0.05). The expression level of serum CCL4 in esophageal adenocarcinoma group was remarkably higher than that in esophageal squamous cell carcinoma group [18.45 (11.94) versus 13.37 (9.29), Z=-2.039, P=0.031]. In esophageal cancer group and normal control group, the serum CX3CL1 was positively correlated with CCL20 (r=0.649, P<0.001, r=0.758, P<0.001). CONCLUSIONS The expressions of serum CX3CL1, CXCL-12, and CCL20 are increased markedly in the patients, which may promote the occurrence, development and metastasis of esophageal cancer.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias Esofágicas/patologia , Soro/química , Adulto , Idoso , Carcinoma de Células Escamosas/patologia , Quimiocinas/sangue , China , Neoplasias Esofágicas/sangue , Neoplasias Esofágicas/metabolismo , Carcinoma de Células Escamosas do Esôfago/patologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
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