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1.
Psychophysiology ; 61(7): e14564, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38487932

RESUMO

Anxiety is a common psychological disorder associated with other mental disorders, with depression being the most common comorbidity. Few studies have examined the neural mechanisms underlying anxiety after controlling for depression. This study aimed to explore whether there are differences in cortical activation in anxiety patients with different severities whose depression are normal. In the current study, depression levels were normal for 366 subjects-139 healthy subjects, 117 with mild anxiety, and 110 with major anxiety. Using the Hospital Anxiety and Depression Scale (HADS) and a verbal fluency task (VFT) to test subjects' anxiety and depression and cognitive function, respectively. A 53-channel guided near-infrared spectroscopic imaging technology (fNIRS) detected the concentration of oxyhemoglobin (oxy-Hb). Correlation analysis between anxiety severity and oxy-Hb concentration in the brain cortex was performed, as well as ANOVA analysis of oxy-Hb concentration among the three anxiety severity groups. The results showed that anxiety severity was significantly and negatively correlated with oxy-Hb concentrations in the left frontal eye field (lFEF) and in the right dorsolateral prefrontal area (rDLPFC). The oxy-Hb concentration in the lFEF and the rDLPFC were significantly lower in the major anxiety disorder group than that in the control group. This suggests that decreased cortical activity of the lFEF and rDLPFC may be neural markers of anxiety symptoms after controlling for depression. Anxiety symptoms without depression may be result from the dysfunction of the cognitive control network (CCN) which includes the lFEF and rDLPFC.


Assuntos
Espectroscopia de Luz Próxima ao Infravermelho , Humanos , Masculino , Feminino , Adulto , Adulto Jovem , Ansiedade/fisiopatologia , Oxiemoglobinas/metabolismo , Depressão/fisiopatologia , Pessoa de Meia-Idade , Função Executiva/fisiologia , Transtornos de Ansiedade/fisiopatologia , Córtex Cerebral/diagnóstico por imagem , Córtex Cerebral/fisiopatologia
2.
Environ Res ; 234: 116505, 2023 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-37406724

RESUMO

The worldwide spreading of severe acute respiratory syndrome SARS-CoV2 pandemic, a massive setback to every human being. In response to strategies actions against Covid-19 spreading many detection, prevention, and post-measures are being studied in large capacities. Association of SARS-CoV2 with ACE2 is well acknowledged and used for developing point-of-care detection kits. Recently, cases and studies have surfaced showing relation of ACE I/D polymorphism with spreading of SARS-CoV2 and highlighted a slip section towards detection and these studies show specificity with older males, high diabetes, and hypertension. To address the raised concern, we report synthesis of unique SnO2-xNx tpod nanostructure, showing affirmative attachment to both ACE1 and ACE2 efficiently. The attachment is examined in different ratios and studied with µ-Raman spectroscopy. The tpod nanostructure has served with its signature raman signals and used as probe for detection of SARS-CoV2 spike protein (S1). The linearity response for tpod raman signal at 630.4 cm-1 shows R2 0.9705, comparatively peak 1219.13 cm-1 show R2 0.9865 and calculated limit of detection of 35 nM.


Assuntos
COVID-19 , Masculino , Humanos , SARS-CoV-2/genética , RNA Viral , Peptidil Dipeptidase A , Enzima de Conversão de Angiotensina 2 , Glicoproteína da Espícula de Coronavírus/genética
3.
Luminescence ; 38(7): 954-998, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35929140

RESUMO

This review briefly emphasizes the different detection approaches (electrochemical sensors, chemiluminescence, surface-enhanced Raman scattering), functional nanostructure materials (quantum dots, metal nanoparticles, metal nanoclusters, magnetic nanomaterials, metal oxide nanoparticles, polymer-based nanomaterials, and carbonaceous nanomaterials) and detection mechanisms. Furthermore, the emphasis of this review is on the integration of functional nanomaterials with optical spectroscopic techniques for the identification of various biomarkers (nucleic acids, glucose, uric acid, oxytocin, dopamine, ascorbic acid, bilirubin, spermine, serotonin, thiocyanate, Pb2+ , Cu2+ , Hg2+ , F- , peptides), and cancer biomarkers (mucin 1, prostate specific antigen, carcinoembryonic antigen, CA15-3, human epidermal growth factor receptor 2, C-reactive protein, and interleukin-6). Analytical characteristics of nanomaterials-based optical sensors are summarized in the tables, providing the insights of nanomaterials-based optical sensors for biomarkers detection. Finally, the opportunities and challenges of nanomaterials-based optical analytical approaches for the detection of various biomarkers (inorganic, organic, biomolecules, peptides and proteins) are discussed.


Assuntos
Técnicas Biossensoriais , Nanoestruturas , Humanos , Técnicas Eletroquímicas/métodos , Nanoestruturas/química , Biomarcadores Tumorais , Polímeros/química , Metais , Técnicas Biossensoriais/métodos
4.
Analyst ; 145(21): 7072, 2020 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-32901639

RESUMO

Correction for 'Progress of electrospray ionization and rapid evaporative ionization mass spectrometric techniques for the broad-range identification of microorganisms' by Suresh Kumar Kailasa et al., Analyst, 2019, 144, 1073-1103, DOI: 10.1039/C8AN02034E.

5.
Anal Bioanal Chem ; 412(26): 7141-7154, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32876723

RESUMO

Lysozyme (LYZ) sensors have attracted increased attention because rapid and sensitive detection of LYZ is highly desirable for various diseases associated with humans. In this research, we designed L-cysteine-protected ultra small photoluminescent (PL) copper nanoclusters (CuNCs) conjugated with ß-cyclodextrin (ß-CD) for rapid detection of LYZ in human serum samples at room temperature. The proposed ß-CD-CuNCs exhibited excellent water solubility, ultrafine size, good dispersion, bright photoluminescence, and good photostability. The ß-CD-CuNCs exhibit an excitation and emission maximum at 370 nm and 492 nm, respectively, with an absolute quantum yield (QY) of 54.6%. ß-CD-CuNCs showed a fluorescence lifetime of 12.7 ns. The addition of LYZ would result in PL quenching from ß-CD-CuNCs. The lowest detectable LYZ concentration was 50 nM for the naked eye and the limit of detection (LOD) and limit of quantification (LOQ) were 0.36 nM and 1.21 nM, respectively, by emission measurement observed in the LYZ concentration range from 30 to 100 nM. It is important to note that the PL ß-CD-CuNC strategy possessed great selectivity toward LYZ relative to other biomolecules. The proposed nanosensor was efficiently applied to determine the LYZ level in human serum sample average recoveries from 96.15 to 104.05% and relative standard deviation (RSD) values lower than 3.0%. Moreover, the proposed sensing system showed many advantages, including high speed, high sensitivity, high selectivity, low cost, and simple preparation.


Assuntos
Cobre/química , Ciclodextrinas/química , Cisteína/química , Luminescência , Nanopartículas Metálicas/química , Muramidase/sangue , Humanos , Interações Hidrofóbicas e Hidrofílicas , Limite de Detecção , Reprodutibilidade dos Testes , Espectroscopia de Infravermelho com Transformada de Fourier/métodos
6.
Analyst ; 144(4): 1073-1103, 2019 Feb 11.
Artigo em Inglês | MEDLINE | ID: mdl-30698588

RESUMO

Several non-culture molecular (multiplex polymerase chain reaction assays, DNA microarrays, massive parallel DNA sequencing, in situ hybridization, microbiome profiling, and molecular typing of pathogens) and analytical (electrophoresis, gel electrophoresis, surface-enhanced Raman scattering, and mass spectrometry) tools have been developed in recent years for the identification of bacteria and diagnosis of bacterial infections from clinical samples. Among mass spectrometric techniques, electrospray ionization (ESI) and rapid evaporative ionization (REI) mass spectrometric (MS) techniques have attracted much attention in the identification of microorganisms (bacteria, fungi, and viruses), and in the diagnosis of various bacterial infections. This review highlights the developed ESI-MS-based methods, including polymerase chain reaction (PCR) combined with ESI-MS and capillary electrophoresis (CE) and liquid chromatography (LC)-ESI-MS, for the identification of microorganisms (pathogenic bacteria, fungi, and viruses) in various samples. Recent applications of ESI- and REI-MS in identifying pathogenic bacteria are depicted in tables, and some significant findings are summarized.


Assuntos
Bactérias/isolamento & purificação , Espectrometria de Massas por Ionização por Electrospray/métodos , Bactérias/química , Humanos , Volatilização
8.
Mikrochim Acta ; 186(3): 130, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30707295

RESUMO

A method is reported for the synthesis of highly luminescent copper/molybdenum bimetallic nanoclusters (Cu/Mo NCs) using cysteine as both a capping and reducing agent. The nanoclusters display bluish-green luminescence (excitation/emission peaks at 370/490 nm) and a relative quantum yield of 26%. The capped Cu/Mo NCs were used as a fluorescent probe for determination of the antineoplastic drug methotrexate (MTX) via an inner filter effect. Fluorescence intensity decreases linearly in the 50 nM to 100 µM MTX concentration range. The limit of detection is 13.7 nM. This approach has been successfully applied to the determination of MTX in spiked human urine with a typical recovery of 99%. Graphical abstract Schematic of a fluorometric method for the determination of methotrexate (MTX) which exerts a strong inner filter effect on the fluorescence of cysteine-capped copper/molybdenum nanoclusters (CuMo NCs) at the wavelength of excitation (370 nm).


Assuntos
Cobre/química , Cisteína/química , Nanopartículas Metálicas/química , Metotrexato/urina , Molibdênio/química , Corantes Fluorescentes/química , Humanos , Concentração de Íons de Hidrogênio , Limite de Detecção , Tamanho da Partícula , Espectrometria de Fluorescência , Propriedades de Superfície
9.
Chemistry ; 24(29): 7417-7427, 2018 May 23.
Artigo em Inglês | MEDLINE | ID: mdl-29493025

RESUMO

The present study deals with photothermal therapy of solid tumors using different forms of oxygen-deficient sub-stoichiometric two-dimensional (2D) molybdenum oxide nanoflakes (α-MoO3-x ). Upon exfoliation of molybdenum oxide power using fine gridding followed by ultrasonication, bluish green molybdenum oxide (BG α-MoO3 ) was obtained. Oxygen vacancies in BG were generated upon irradiation with an intense xenon lamp. Irradiating the BG for 3 and 5 h, deep blue (B) and olive green (G) oxygen-deficient nanoflakes were obtained respectively. All exhibited high NIR absorption, making these nanomaterials suitable for photothermal therapy. All three forms were functionalized with polypyrrole (PPy@BG, PPy@B, PPy@G) to boost the photothermal stability and transduction efficiency. After functionalization and irradiation with 808 nm laser, the enhancement of temperature for BG, B, G was 50, 65, 52 °C respectively and the corresponding photothermal transduction efficiencies (PTE) were 29.32, 44.42 and 42.00 %. Each of the nanoflakes were found to be highly biocompatible and photostable both in vitro and in vivo. There was substantial decrease in the size of tumors after seven days of treatment on tumor-bearing experimental mice models.

10.
Anal Bioanal Chem ; 409(21): 4943-4950, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28744559

RESUMO

In most applications of quantum dots (QDs) for surface-assisted laser desorption/ionization mass spectrometry (SALDI-MS), one side of QDs is supported by a solid substrate (stainless - steel plate), whereas the other side is in contact with the target analytes. Therefore, the surface capping agent of QDs is a key parameter for laser desorption/ionization mass spectrometry (LDI-MS). Cadmium telluride quantum dots (CdTe QDs) modified with different capping agents are synthesized, characterized, and applied for surface tuning laser desorption/ionization mass spectrometry (STLDI-MS). Data shows that CdTe quantum dot modified cysteine (cys@CdTe QDs) has an absorption that matches with the wavelength of the N2 laser (337 nm). The synergistic effect of large surface area and absorption of the laser irradiation of cys@CdTe QDs enhances the LDI-MS process for small - molecule analysis, including α-, ß-, and γ-cyclodextrin, gramicidin D, perylene, pyrene, and triphenylphosphine. Cys@CdTe QDs are also applied using Al foils as substrates. Aluminum foil combined with cys@CdTe QDs enhances the ionization efficiency and is cheap compared to traditional matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) with a stainless - steel plate.

11.
Mikrochim Acta ; 185(1): 65, 2017 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-29594588

RESUMO

An optical method is described for the quantitation of glucose by using oxygen-deficient α-MoO3-x nanoflakes. It is based on the use of glucose oxidase (GOx) which produces hydrogen peroxide on oxidation of glucose. Hydrogen peroxide then oxidizes the α-MoO3-x nanoflakes, and this results in a visible color change from blue to colorless. The color change can be measured photometrically at 740 nm. The method has a 68 nM detection limit. Graphical Abstract Mechanism of glucose detection using blue colored oxygen deficient 2D α-MoO3-x nanoflakes. Hydrogen peroxide (H2O2) is formed as a by-product in the conversion of glucose to glucono-1,5-lactone by glucose oxidase (GOx). In the presence of H2O2, the oxygen vacancies in α-MoO3-x nanoflakes are filled up, and this leads to the loss of blue color of the nanoflakes because they are converted back to colorless bulk α-MoO3.


Assuntos
Técnicas Biossensoriais/métodos , Glucose Oxidase/metabolismo , Glucose/análise , Molibdênio/química , Nanoestruturas/química , Óxidos/química , Oxigênio/química , Biocatálise , Cor , Glucose Oxidase/química , Peróxido de Hidrogênio/metabolismo , Limite de Detecção , Modelos Moleculares , Oxirredução , Conformação Proteica
12.
Rapid Commun Mass Spectrom ; 30(13): 1443-53, 2016 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-27321831

RESUMO

RATIONALE: Phosphorylation is a post-translational modification of proteins that plays very important role in a large number of biological processes. However, despite recent advancements in phosphoproteome research, large-scale detection and characterization of phosphopeptides by mass spectrometry (MS) is still a challenging task due to the low abundance of phosphopeptides and sub-stoichiometric nature of phosphorylation sites. On-particle microwave-assisted trypsin digestion of phosphoproteins and enrichment of phosphopeptides is an effective method for identification/characterization of phosphopeptides. Magnetic nanoparticles typically can absorb microwave radiation and generate heat in order to resolve complex phosphproteins and to enhance the digestion rate and capture the phosphopeptides on their modified surfaces. METHODS: In this study, we used a cheap and efficient method for rapid microwave-assisted tryptic digestion of phosphoproteins and simultaneous enrichment of phosphopeptides using CoFe2 O4 -ZnO magnetic nanoparticles. Using this technique, the digestion time of phosphoproteins can be reduced and the phosphopeptides can be quickly analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). For the first time, we have applied CoFe2 O4 -ZnO magnetic nanoparticles for enrichment of phosphopeptides from standard phosphoproteins (ß-casein and ovalbumin), complex samples (human serum and egg white) and a protein mixture of ß-casein and BSA (1:100). RESULTS: Our results demonstrate that the capture efficiency of CoFe2 O4 -ZnO nanoparticles for ß-casein and ovalbumin in MALDI-TOFMS is very high (detection limits 0.2 fmol and 20 fmol, respectively). The CoFe2 O4 -ZnO nanoparticles have high affinity for phosphopeptide enrichment for ß-casein in complex mixtures with BSA at 1:10 and 1:100 molar ratios in the microwave within 30 s. CONCLUSIONS: Compared with other reported magnetic nanoparticles, the CoFe2 O4 -ZnO nanoparticles are easy to prepare and handle, and can save time in the phosphopeptide enrichment procedure, making these nanoparticle a good choice for highly sensitive phosphopeptide enrichment. Copyright © 2016 John Wiley & Sons, Ltd.


Assuntos
Micro-Ondas , Nanopartículas , Fosfopeptídeos/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Caseínas , Humanos , Fosfoproteínas
13.
Rapid Commun Mass Spectrom ; 30(12): 1403-12, 2016 06 30.
Artigo em Inglês | MEDLINE | ID: mdl-27197033

RESUMO

RATIONALE: Investigation of nanoparticles for laser desorption/ionization mass spectrometry (LDI-MS) is routinely reported. However, the effect of surface capping of nanomaterials for LDI-MS is not well studied. METHODS: Different capping agents of quantum dots (CdTe) affect the spectra quality and sensitivity of protein analysis and protein digestion using trypsin enzyme assisted by microwave. Surface modification of CdTe quantum dots with different capping agents, namely 3-mercaptopropionic acid (3-MPA), 4-aminothiophenol (4-ATP), 4-mercaptobenzoic acid (4-MBA), 11-mercaptoundecanoic acid (11-MUA), cysteine (Cys) and thioglycolic acid (TG), were investigated for quantum dots (QDs)-assisted trypsin protease followed by analysis using mass spectrometry. RESULTS: CdTe QDs were used as a surface to assist trypsin protease and laser desorption/ionization mass spectrometry (surface-assisted laser desorption/ionization mass spectrometry, SALDI-MS). The MS profiles for the investigated analytes (bovine serum albumin (BSA), lysozyme, cytochrome c, α-casein, transferrin and myoglobin) revealed almost the absence of degradation that implies the softness of the present technique. QDs-assisted LDI-MS offered high sensitivity and high resolution. QDs showed significant enhancement of microwave-assisted trypsin digestion of the investigated proteins and these improvements boosted the identifications of fragments with a database. CONCLUSIONS: A capping agent of quantum dots affects the analysis of proteins and peptides using LDI-MS. CdTe QDs offer sensitive, high-resolution and simple analysis of proteins. QDs improved the protein digestion using the microwave-assisted trypsin digestion. Copyright © 2016 John Wiley & Sons, Ltd.


Assuntos
Proteômica , Pontos Quânticos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Compostos de Cádmio , Compostos de Sulfidrila
14.
Anal Bioanal Chem ; 408(17): 4485-502, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26973236

RESUMO

Gold nanoparticles (AuNPs) assisted laser desorption/ionization mass spectrometry (GALDI-MS) provided new horizons and offered many functions for various applications. This review summarized AuNPs applications for analytical, biotechnology and proteomics. AuNPs efficiently absorbed the laser radiation and transferred the energy to the analyte for the desorption/ionization process. The unique features of AuNPs such as large surface area and high absorption coefficient lead not only to high resolution, low interference and low limit of detection, but also offered selective detection for certain species. AuNPs provided an excellent surface for the analysis of several species such as small molecules, biomarkers, proteins and cells (pathogenic bacteria or cancer cells). AuNPs played many roles such as surface for LDI-MS, probe and stationary phase for separation or preconcentration. AuNPs modified various surface chemistry was applied for a wide range of different wavelength. AuNPs severed as a source of Au(+) ions that were suitable for analyte cationisation. Characterization of Au nanoclusters (AuNCs) by mass spectrometry, pros and cons were also highlighted. Graphical Abstract Schematic representation of the analysis by Gold Nanoparticles Assisted Laser Desorption/Ionization Mass Spectrometry (GALDI-MS).


Assuntos
Ouro/química , Nanopartículas Metálicas/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos
15.
Anal Bioanal Chem ; 408(23): 6269-81, 2016 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-27565791

RESUMO

Bacteria or their protein and peptide entity enrichment using biomolecules-functionalized magnetic nanoparticles, and analysis by matrix assisted laser desorption/ionization mass spectrometry (MALDI MS) is a promising technique to analyze microorganisms. High and low molecular weight proteins like penicillin-binding proteins are responsible for final step synthesis of peptidoglycan biosynthesis; those are the target of lactam antibiotics. In this paper, we synthesized magnetic nanoparticles (mag-NPs) and further modified them with 3-aminopropyltriethoxysilane, and then the ß-lactam antibiotic amoxicillin was covalently linked to their surface. ß-Lactam group attributes as penicillin binding proteins (PBPs) in bacteria. Staphylococcus aureus and Escherichia coli were used as model bacteria for enrichment based on the ß-lactam affinity of magnetic nanoparticles, and then the bacteria were easily separated by an external magnet. Several high molecular weight penicillin binding proteins (PBPs) were detected by MALDI MS containing 10(4) and 10(3) colony-forming unit (cfu) per milileter (mL) of S. aureus and E. coli, respectively. In the case of E. coli, higher molecular weight PBPs were observed at 20 to 55 kDa in MALDI mass spectra. However, S. aureus bacteria resulted with femAB operon-based proteins, with molecular weight of 49570.4 Da, by MALDI MS after using amoxicillin functionalized-mag-NPs. The current approach provides an effective bacteria detection and preconcentration method that has high potential in the near future for fast and sensitive diagnosis of pathogenic bacteria infection. Graphical Abstract Schematic for large proteins analysis by MALDI TOF MS (a) mag-NPs and bacterial interaction (b) Penicillin binding proteins trapping by Amox-mag-NPs.


Assuntos
Amoxicilina/química , Proteínas de Bactérias/análise , Escherichia coli/química , Nanopartículas de Magnetita/química , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Staphylococcus aureus/química , Proteínas de Bactérias/isolamento & purificação , Escherichia coli/isolamento & purificação , Infecções por Escherichia coli/microbiologia , Humanos , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/isolamento & purificação
16.
Analyst ; 140(5): 1555-65, 2015 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-25587827

RESUMO

Graphene oxide (GO)-modified sinapinic acid (3,5-dimethoxy-4-hydroxycinnamic acid, SA) (SA@GO) was synthesized and characterized; it was then investigated as a new surface assisted laser desorption/ionization mass spectrometry (SALDI-MS) for proteomics and pathogenic bacteria biosensing. SA@GO could effectively decrease the time necessary for sweet spotting searching, reducing the amount of organic matrix and solvent and enhance the sensitivity. SA@GO shows high performance as a matrix alone without the need to add trifluoroacetic acid (TFA). However, the analysis of the intact bacteria cells shows improvement in the signal intensity (2-5 fold) and offers a low limit of detection. All these analyses could be performed with low concentrations (1-10 fmol) and tiny volumes (0.5-1 µL). This study demonstrated that the exploration of new hybrid materials is pivotal to achieve high performance and high ionization. Because of the plane of GO, it assists protein-protein interactions that make it undergo softer ionization.


Assuntos
Técnicas Biossensoriais/métodos , Ácidos Cumáricos/química , Grafite/química , Proteômica/métodos , Pseudomonas aeruginosa/isolamento & purificação , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Staphylococcus aureus/isolamento & purificação , Celulase/metabolismo , Fluorescência , Humanos , Lactalbumina/metabolismo , Lasers , Muramidase/metabolismo , Nanocompostos/química , Infecções por Pseudomonas/diagnóstico , Infecções por Pseudomonas/metabolismo , Infecções por Pseudomonas/microbiologia , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier , Infecções Estafilocócicas/diagnóstico , Infecções Estafilocócicas/metabolismo , Infecções Estafilocócicas/microbiologia , Tripsina/metabolismo
17.
Anal Bioanal Chem ; 406(22): 5447-54, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24994666

RESUMO

This is the first capillary electrophoresis (CE) analysis for diagnosis of hemophilia A (HA). The intron 22 inversion of factor VIII gene (F8) causes 40-50 % of severe bleeding disorder of HA in all human populations. Consequently, identification of the disease-causing mutations is becoming increasingly important for accurate genetic counseling and prenatal diagnosis. In this study, the key steps of inverse-shifting polymerase chain reaction (IS-PCR) and of short-end injection capillary electrophoresis were used for more specific and rapid genotyping of intron 22 inversion of F8. In IS-PCR, three specific primers were used to amplify 512-bp amplicon for wild type and 584-bp amplicon for patients with intron 22 inversion. The capillary gel electrophoresis (CGE) system was performed using 1× Tris-borate-EDTA (TBE) buffer containing 0.3 % (w/v) polyethylene oxide (PEO). The PCR amplicons were electrokinetically injected at 10 kV for 10 s at a temperature of 25 °C. The optimal short-end injection CGE was applied to detect the F8 gene of HA patients and carriers within 5 min. Intron 22 inversion was indeed found on some HA patients (13/35, 37.1 %). All genotyping results showed good agreement with DNA sequencing method and long-distance polymerase chain reaction (LD-PCR). The IS-PCR combined with short-end injection CGE method was feasible and efficient for intron 22 inversion screening of F8 in the HA populations.


Assuntos
Eletroforese Capilar , Fator VIII/genética , Genótipo , Hemofilia A/genética , Íntrons , Reação em Cadeia da Polimerase , Soluções Tampão , DNA/química , Análise Mutacional de DNA , Ácido Edético/química , Feminino , Hemofilia A/diagnóstico , Heterozigoto , Humanos , Masculino , Mutação , Polietilenoglicóis/química , Polímeros/química , Manejo de Espécimes
18.
J Nanosci Nanotechnol ; 14(1): 447-59, 2014 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24730274

RESUMO

The development of novel nanomaterial-based analytical methods for biological analysis in proteomics and clinical diagnosis has been making significant progress. In the long-lasting efforts to improve the detection sensitivity of analytical instruments, functional nanomaterials have been significantly applied as effective probes by integrating various analytical tools for bioanalysis. Among these nanomaterials, quantum dots (QDs) have been recently proved as highly potential materials as fluorescent sensors for biomolecules assays due to their high quantum yield, narrow and tunable emission spectrum and excellent photostability. In this review, we introduce the use of surface modified QDs as fluorescence probes for detecting proteins, peptides and other biomolecules based on fluorescence quenching and fluorescence resonance energy transfer (FRET).


Assuntos
Biopolímeros/análise , Transferência Ressonante de Energia de Fluorescência/métodos , Corantes Fluorescentes/síntese química , Técnicas de Sonda Molecular , Pontos Quânticos , Espectrometria de Fluorescência/métodos , Propriedades de Superfície
19.
Proc Natl Acad Sci U S A ; 108(4): 1579-84, 2011 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-21205894

RESUMO

IL-25 (IL-17E) is a T-helper cell type 2 (Th2) cytokine best described as a potentiator of Th2 memory responses. Reports of expression of its receptor, IL-25R, on airways structural cells suggest a wider role for IL-25 in remodeling. We hypothesized that IL-25 stimulates local angiogenesis in the asthmatic bronchial mucosa. Immunoreactive IL-25(+), IL-25R(+), and CD31(+) (endothelial) cells in sections of bronchial biopsies from asthmatics and controls were detected by immunohistochemistry. The effect of IL-25 on angiogenesis was examined using an in vitro assay. Real-time PCR was used to detect expression of IL-25R and VEGF mRNA in cultured human vascular endothelial cells (HUVEC), and a cell proliferation kit (WST-8) was used to measure the effect of IL-25 on HUVEC proliferation. Immunostaining showed that IL-25(+), IL-25R(+), and CD31(+)/IL-25R(+) cells were significantly elevated in the bronchial mucosa of asthmatics compared with controls (P < 0.003). In asthmatics, the numbers of IL-25(+) cells correlated inversely with the forced expiratory volume in 1 s (r = -0.639; P = 0.01). In vitro, HUVEC constitutively expressed IL-25R, which was up-regulated further by TNF-α. IL-25 and TNF-α also increased expression of VEGF and VEGF receptors. IL-25 increased HUVEC proliferation and the number, length, and area of microvessel structures in a concentration-dependent manner in vitro. VEGF blockade, the PI3K-specific inhibitor LY294002, and the MAPK/ERK1/2 (MEK1/2)-specific inhibitor U0126 all markedly attenuated IL-25-induced angiogenesis, and the inhibitors also reduced IL-25-induced proliferation and VEGF expression. Our findings suggest that IL-25 is elevated in asthma and contributes to angiogenesis, at least partly by increasing endothelial cell VEGF/VEGF receptor expression through PI3K/Akt and Erk/MAPK pathways.


Assuntos
Asma/imunologia , Brônquios/irrigação sanguínea , Interleucina-17/imunologia , Células Th2/imunologia , Adulto , Asma/metabolismo , Asma/fisiopatologia , Western Blotting , Brônquios/efeitos dos fármacos , Brônquios/patologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Humanos , Imuno-Histoquímica , Memória Imunológica/imunologia , Interleucina-17/metabolismo , Interleucina-17/farmacologia , Pessoa de Meia-Idade , Fosfatidilinositol 3-Quinases/metabolismo , Fosforilação/efeitos dos fármacos , Molécula-1 de Adesão Celular Endotelial a Plaquetas/genética , Molécula-1 de Adesão Celular Endotelial a Plaquetas/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT6/metabolismo , Células Th2/metabolismo , Fator A de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto Jovem
20.
J Affect Disord ; 350: 521-530, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38237870

RESUMO

BACKGROUND: Previous studies proposed that functional near-infrared spectroscopy (fNIRS) can be used to distinguish between not only different severities of depressive symptoms but also different subgroups of depression, such as anxious and non-anxious depression, bipolar and unipolar depression, and melancholia and non-melancholia depression. However, the differences in brain haemodynamic activation between depression subgroups (such as confirmed depression [CD] and suspected depression [SD]) with different symptom severities and the possible correlation between symptom severity and haemodynamic activation in specific brain regions using fNIRS have yet to be clarified. METHODS: The severity of depression symptoms was classified using the Hospital Anxiety and Depression scale (HADS) and the Mini International Neuropsychiatric Interview by psychiatrists. We recruited 654 patients with depression who had varying severities of depressive symptoms, including 276 with SD and 378 with CD, and 317 with HCs from among Chinese college students. The 53-channel fNIRS was used to detect the cerebral hemodynamic difference of the three groups during the VFT (verbal fluency task). RESULTS: Compared with the HC, region-specific fNIRS leads indicate CD patients had significant lower haemodynamic activation in three particular prefrontal regions: 1) right dorsolateral prefrontal cortex (DLPFC), 2) bilateral frontopolar cortex (FPC), and 3) right Broca's area (BA). SD vs. HC comparisons revealed only significant lower haemodynamic activation in the right FPC area. Compared to SD patients, CD patients exhibited decreased hemodynamic activation changes in the right DLPFC and the right BA. Correlation analysis established a significant negative correlation between the hemodynamic changes in the bilateral FPC and the severity of depressive symptoms. CONCLUSIONS: The right DLPFC and right BA are expected to be physiological mechanisms to distinguish depression subgroups (CD, SD) with different symptom severities. The haemodynamic changes in the bilateral FPC was nagatively associated with the symptom severity of depression.


Assuntos
Depressão , Transtorno Depressivo , Humanos , Depressão/diagnóstico por imagem , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Córtex Pré-Frontal/diagnóstico por imagem , Transtorno Depressivo/diagnóstico por imagem , Área de Broca
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