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1.
Spectrochim Acta A Mol Biomol Spectrosc ; 265: 120385, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34536885

RESUMO

In this work, a strong blue-emitting fluorescent biosensor based on graphite carbon nitride nanoparticles (GCNNs) (Ex = 340 nm and Em = 435 nm) was synthesized by a facile one-step hydrothermal method. With the aid of hydrogen peroxide and horseradish peroxidase, pyrocatechol structure of dopamine (DA) was oxidized to o-quinone structure of polydopamine (PDA) by hydroxyl radical. PDA was able to rapidly and significantly quench fluorescence of GCNNs. In the meanwhile, oxidative self-polymerization from DA to PDA would be blocked by antioxidants, such as glutathione (GSH) and ascorbic acid (AA). Thus, the fluorescence of GCNNs@DA sensor would be recovered owing to the decrease of o-quinone. Based on above-mentioned dual recognition strategy of "turn-off" and "turn off-on", a fast, simple and ultrasensitive method was developed to measure DA and antioxidants. Under the optimal experimental conditions, the detection limits of DA, GSH and AA were 0.064 µmol L-1, 0.11 µmol L-1 and 0.16 µmol L-1 with relative standard deviations of 1.7%, 9.3% and 8.0%, respectively. As one of metal-free quantum dots, our GCNNs-based sensors were also successfully applied to the determination of DA as well as GSH and AA in human serum. The recoveries for the spiked samples were in the range of 93.8%-109% and 95.0%-110% of DA and antioxidants, which shows great promise to clinicalapplication.


Assuntos
Grafite , Pontos Quânticos , Antioxidantes , Carbono , Dopamina , Humanos , Limite de Detecção , Nitrilas , Soro
2.
Talanta ; 237: 122916, 2022 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-34736654

RESUMO

Herein, we show differences in blood serum of asymptomatic and symptomatic pregnant women infected with COVID-19 and correlate them with laboratory indexes, ATR FTIR and multivariate machine learning methods. We collected the sera of COVID-19 diagnosed pregnant women, in the second trimester (n = 12), third-trimester (n = 7), and second-trimester with severe symptoms (n = 7) compared to the healthy pregnant (n = 11) women, which makes a total of 37 participants. To assign the accuracy of FTIR spectra regions where peak shifts occurred, the Random Forest algorithm, traditional C5.0 single decision tree algorithm and deep neural network approach were used. We verified the correspondence between the FTIR results and the laboratory indexes such as: the count of peripheral blood cells, biochemical parameters, and coagulation indicators of pregnant women. CH2 scissoring, amide II, amide I vibrations could be used to differentiate the groups. The accuracy calculated by machine learning methods was higher than 90%. We also developed a method based on the dynamics of the absorbance spectra allowing to determine the differences between the spectra of healthy and COVID-19 patients. Laboratory indexes of biochemical parameters associated with COVID-19 validate changes in the total amount of proteins, albumin and lipase.


Assuntos
COVID-19 , Feminino , Humanos , Laboratórios , Aprendizado de Máquina , Gravidez , Gestantes , SARS-CoV-2 , Soro , Análise Espectral , Vibração
3.
Spectrochim Acta A Mol Biomol Spectrosc ; 264: 120246, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34371315

RESUMO

Endometriomas are typically an advanced form of endometriosis that leads to the formation of scar tissue, adhesions, and an inflammatory reaction. There is no certain serum marker for the diagnosis of endometriosis. This study aims to research the correlation between the amount of peaks corresponding to proteins and lipids with the volume of endometrioma and determine the chemical structure of blood serum collected from women suffering from endometriosis patients with endometrioma and healthy subjects using Fourier Transform Infrared (FTIR) spectroscopy. FTIR spectroscopy is used as a non-invasive diagnostic technique for the discrimination of endometriosis women with endometrioma and control blood sera. The FTIR spectra of 100 serum samples acquired from 50 patients and 50 healthy individuals were used for this study. For this purpose, multivariate analyses such as Principal Component Analysis (PCA), Partial Last Square analysis (PLS) with Variables Importance in Projection (VIP), and probability models, were performed. Our results showed that FTIR range 1500 cm-1 and 1700 cm-1 and around 2700 cm-1 - 3000 cm-1, regions may be used for the diagnosis of endometriosis. Also, we find that proteins and lipids fraction increase with the volume of endometrioma. Moreover, PLS and VIP analysis suggested that lipids could be helpful in the diagnosis of endometriosis women with endometrioma.


Assuntos
Endometriose , Endometriose/diagnóstico , Feminino , Humanos , Análise de Componente Principal , Soro , Espectroscopia de Infravermelho com Transformada de Fourier
4.
Spectrochim Acta A Mol Biomol Spectrosc ; 264: 120305, 2022 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-34455381

RESUMO

In order to improve the accuracy of spectral analysis of complex solutions, based on the "M + N" theory, this paper proposes to use single (365 nm ± 5 nm) ultraviolet LED as emission light to detect transmission spectrum and excited fluorescence spectrum. Taking the total bilirubin in serum as the measurement object, the dual-mode spectrum of transmission and fluorescence about serum is collected, which increases the amount of information. In order to verify the effectiveness of the method proposed in this paper, the transmission and fluorescence spectra of the samples were also collected. Then three models of total bilirubin concentration are established by transmission spectrum, fluorescence spectrum and dual-mode spectrum of transmission and fluorescence respectively. Through the comparison of the parameters of the three models, the model established by dual-mode spectrum of transmission and fluorescence is good. The Rc of the model is 0.91 and the RMSEC is 3.00 (µmol/L). The Rp is 0.92, the RMSEP is 3.53 (µmol/L). Compared with transmission spectrum modeling and fluorescence spectrum modeling, the RMSEP of dual-mode spectrum modeling was reduced by 34.8% and 22.6% respectively. The experimental results show that the measurement method of dual-mode spectrum of transmission and fluorescence by using a single ultraviolet light source proposed in this paper based on the "M + N" theory increases the information of solution composition, which provides a new method for the analysis of the same characteristic components.


Assuntos
Bilirrubina , Raios Ultravioleta , Soro , Espectrometria de Fluorescência
5.
Chemosphere ; 287(Pt 3): 132300, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34563784

RESUMO

Though many persistent organic pollutants (POPs) are closely regulated the human population is still exposed to these ubiquitous chemicals from the environment and diet. Safe management and human biomonitoring of POPs is necessary to understand the risk of exposure. Within human biomonitoring the mass of sample is often limited, therefore robust methods using smaller sample amounts are necessary. This study developed a 96-well plate solid phase extraction (SPE) method for determination of selected POPs: polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs), polybrominated diphenyl ethers (PBDEs), hexabromocyclododecane (HBCD) and non-persistent novel flame retardants (NFRs) in low volume blood serum. Non-destructive clean-up coupling Oasis HLB extraction plate with Phree phospholipid removal plate was employed. Extraction efficiency was determined at low and high concentrations in certified reference materials NIST SRM 1957 and 1958, respectively. Target compounds deviated from certified values on average by 15% and 21% for SRM 1957 and SRM 1958, respectively. Observed limit of detections (LODs) ranged from 0.36 pg/mL (PCB 180) to 66.07 pg/mL (δ-HCH). The applicability for real samples is demonstrated on 48 samples from pregnant women enrolled in the pilot phase of the CELSPAC: TNG study. In total, 30 target compounds were detected in at least one sample. The method developed here provides a fast and reliable analysis of human blood serum with possibility to introduce automation for the sample preparation procedure.


Assuntos
Poluentes Ambientais , Hidrocarbonetos Clorados , Praguicidas , Bifenilos Policlorados , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Éteres Difenil Halogenados/análise , Humanos , Hidrocarbonetos Clorados/análise , Poluentes Orgânicos Persistentes , Praguicidas/análise , Bifenilos Policlorados/análise , Gravidez , Soro/química , Extração em Fase Sólida
6.
Sci Rep ; 11(1): 21633, 2021 11 04.
Artigo em Inglês | MEDLINE | ID: mdl-34737330

RESUMO

Although the serum lipidome is markedly affected by COVID-19, two unresolved issues remain: how the severity of the disease affects the level and the composition of serum lipids and whether serum lipidome analysis may identify specific lipids impairment linked to the patients' outcome. Sera from 49 COVID-19 patients were analyzed by untargeted lipidomics. Patients were clustered according to: inflammation (C-reactive protein), hypoxia (Horowitz Index), coagulation state (D-dimer), kidney function (creatinine) and age. COVID-19 patients exhibited remarkable and distinctive dyslipidemia for each prognostic factor associated with reduced defense against oxidative stress. When patients were clustered by outcome (7 days), a peculiar lipidome signature was detected with an overall increase of 29 lipid species, including-among others-four ceramide and three sulfatide species, univocally related to this analysis. Considering the lipids that were affected by all the prognostic factors, we found one sphingomyelin related to inflammation and viral infection of the respiratory tract and two sphingomyelins, that are independently related to patients' age, and they appear as candidate biomarkers to monitor disease progression and severity. Although preliminary and needing validation, this report pioneers the translation of lipidome signatures to link the effects of five critical clinical prognostic factors with the patients' outcomes.


Assuntos
COVID-19/metabolismo , Lipídeos/sangue , Soro/química , Adulto , Idoso , Biomarcadores/sangue , COVID-19/sangue , Dislipidemias/metabolismo , Feminino , Humanos , Itália , Lipidômica/métodos , Lipídeos/análise , Masculino , Pessoa de Meia-Idade , Estresse Oxidativo/fisiologia , Prognóstico , SARS-CoV-2/metabolismo , SARS-CoV-2/patogenicidade , Esfingomielinas/sangue
7.
Mol Biol (Mosk) ; 55(6): 1011-1020, 2021.
Artigo em Russo | MEDLINE | ID: mdl-34837705

RESUMO

Expression levels of matrix metalloproteinases, in particular MT1-MMP, are elevated in pancreatic cancer (PC) cells, and this is associated with increased tumor proliferation, invasion, and migration. MT1-MMP is considered a promising target for drug therapy of PC, but the use of inhibitors and therapeutic antibodies to MT1-MMP is limited because maximal efficiency is only observed in a narrow time interval, at the early asymptomatic stages of the disease. This problem could be solved by immunization to MPs at the moment of detection of the primary tumor. This therapeutic effect could be provided by specific antibodies that can be re-produced in case of relapses. Here, we selected the optimal mode for immunization of mice with MT1-MMP fragments that allows us to obtain a high titer of specific antibodies in the blood serum. The obtained antiserums effectively inhibited MT1-MMP enzymatic activity, migration of PANC-02 PC cells through the collagen matrix, and activation of the main inducers of epithelial -mesenchymal transition, TGF-ß and MMP-2. These results maybe useful in the development of drugs for PC treatment, and the approach we propose might form the basis for design of antitumor drugs with prolonged action.


Assuntos
Metaloproteinase 14 da Matriz , Neoplasias Pancreáticas , Animais , Movimento Celular , Metaloproteinase 14 da Matriz/genética , Metaloproteinases da Matriz , Camundongos , Neoplasias Pancreáticas/tratamento farmacológico , Neoplasias Pancreáticas/genética , Soro
8.
Arch Razi Inst ; 76(3): 445-452, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34824738

RESUMO

Brucellosis is an anthropozoonotic disease. Infection of livestock with Brucella is endemic in most parts of Iran. Sistan-Baluchestan is bordered on the east by the countries of Afghanistan and Pakistan. The high prevalence of brucellosis in livestock in the eastern neighboring countries results in transmission of the disease to this province. The present research aimed to determine the prevalence of brucellosis in small ruminants in the Sistan region of Iran and to compare serological and molecular tests for the detection of brucellosis. Blood samples were taken from 150 randomly selected sheep and goats, and sera were separated. All sera were analyzed by serological (Wright and 2-ME) and molecular (Polymerase Chain Reaction (PCR)) tests. Serological tests were carried out according to the instructions of the Iranian Veterinary Organization The degree of agreement between serological tests and PCR was determined by kappa value. In this study, 17 cases (11.3%) were identified as positive by the PCR method. Wright and 2-ME tests had the highest agreement with PCR in titers ≥2/80 and ≥2/40, respectively. The results of this study show that the brucellosis in sheep and goats has a greater prevalence in the Sistan region than in most other parts of Iran, and this is important in terms of public health. It is suggested that brucellosis vaccination coverage in livestock be increased in this area and that the people in Sistan region must be notified about methods for preventing brucellosis. Also, further studies to compare conventional serologic tests with the gold standard test are recommended.


Assuntos
Brucella , Brucelose , Doenças das Cabras , Animais , Brucelose/epidemiologia , Brucelose/veterinária , Doenças das Cabras/epidemiologia , Cabras , Irã (Geográfico)/epidemiologia , Reação em Cadeia da Polimerase/veterinária , Soro , Ovinos
10.
J Med Life ; 14(4): 549-556, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34621381

RESUMO

Vitamin D is known to alter immune regulation. It binds to the vitamin D receptors (VDR) expressed on T lymphocytes and macrophages. In individuals with Hashimoto's thyroiditis, serum vitamin D levels were found to be lower compared to healthy controls. The study's objective was to investigate the association between VDR gene polymorphism (rs2228570) with blood serum levels of 25-OH vitamin D in patients with thyroid pathology from western Ukraine. The study involved a total of 153 patients with various forms of thyroid pathology. 25-OH vitamin D levels in the serum of the patients and healthy individuals were quantified with ELISA using the 25-OH vitamin D Total (Vit D-Direct) Test System ELISA Kit (Monobind Inc.®, United States, Product Code: 9425-300) on the EIA Reader Sirio S (Seac, Italy). Genotyping of the VDR (rs2228570) gene polymorphism was performed using TaqMan probes and TaqMan Genotyping Master Mix (4371355) on CFX96™Real-Time PCR Detection System (Bio-Rad Laboratories, Inc., USA). Polymerase chain reaction (PCR) for TaqMan genotyping was carried out according to the kit instructions (Applied Biosystems, USA). Our research identified that that genotype variants of VDR rs2228570 are not risk factors for reduced serum 25-OH vitamin D or vitamin D deficiency in patients with various forms of thyroid pathology patients in the West-Ukrainian population. Vitamin D levels were significantly lower in the carriers of AA and AG genotypes with hypothyroidism caused by autoimmune thyroiditis. In AA genotype carriers with postoperative hypothyroidism, 25-OH vitamin D levels were significantly lower compared to AA genotype carriers with autoimmune thyroiditis.


Assuntos
Receptores de Calcitriol , Vitamina D , Estudos de Casos e Controles , Predisposição Genética para Doença , Genótipo , Humanos , Polimorfismo Genético/genética , Polimorfismo de Nucleotídeo Único/genética , Receptores de Calcitriol/genética , Soro , Glândula Tireoide
11.
Int J Mol Sci ; 22(19)2021 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-34638936

RESUMO

Nicotinamide adenine dinucleotide (NAD+) and its reduced form (NADH) are coenzymes employed in hundreds of metabolic reactions. NAD+ also serves as a substrate for enzymes such as sirtuins, poly(ADP-ribose) polymerases (PARPs) and ADP-ribosyl cyclases. Given the pivotal role of NAD(H) in health and disease, studying NAD+ metabolism has become essential to monitor genetic- and/or drug-induced perturbations related to metabolic status and diseases (such as ageing, cancer or obesity), and its possible therapies. Here, we present a strategy based on liquid chromatography-tandem mass spectrometry (LC-MS/MS), for the analysis of the NAD+ metabolome in biological samples. In this method, hydrophilic interaction chromatography (HILIC) was used to separate a total of 18 metabolites belonging to pathways leading to NAD+ biosynthesis, including precursors, intermediates and catabolites. As redox cofactors are known for their instability, a sample preparation procedure was developed to handle a variety of biological matrices: cell models, rodent tissues and biofluids, as well as human biofluids (urine, plasma, serum, whole blood). For clinical applications, quantitative LC-MS/MS for a subset of metabolites was demonstrated for the analysis of the human whole blood of nine volunteers. Using this developed workflow, our methodology allows studying NAD+ biology from mechanistic to clinical applications.


Assuntos
Metaboloma , NAD/biossíntese , Plasma/metabolismo , Soro/metabolismo , Espectrometria de Massas em Tandem/métodos , Urina/fisiologia , Animais , Doadores de Sangue , Cromatografia Líquida/métodos , Células Hep G2 , Humanos , Interações Hidrofóbicas e Hidrofílicas , Metabolômica/métodos , Camundongos , Camundongos Endogâmicos C57BL , Monitorização Fisiológica/métodos , Oxirredução , Projetos Piloto , Plasma/química , Soro/química , Urina/química
12.
Anal Chem ; 93(43): 14527-14536, 2021 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-34668374

RESUMO

Highly abundant serum proteins tend to mask the low- and ultralow-abundance proteins, making low-abundance species detection extremely challenging. While traditional highly abundant protein depletion techniques are effective, they suffer from nonspecific binding problems and laborious sample manipulation procedures, and the kinetics of release of current separation systems is inadequately long, causing dilution of the eluted low-abundance protein samples. Here, we introduce an on-chip light-controlled reusable platform for the direct and fast depletion of highly abundant proteins from serum biosamples. Our nanoarrays display fast and highly selective depletion capabilities, up to 99% depletion of highly abundant protein species, with no undesired depletion effects on the concentration of low-abundance protein biomarkers. Displaying an ultrahigh surface area, ∼3400 m2 g-1, alongside a light-triggerable ultrafast release, this platform allows for a high depletion performance, together with high-yield reusability capabilities. Furthermore, this nanostructured light-controlled separation device could easily be integrated with downstream analytical technologies in a single lab-on-a-chip platform.


Assuntos
Nanoestruturas , Silício , Proteínas Sanguíneas , Dispositivos Lab-On-A-Chip , Soro
13.
Anal Chem ; 93(41): 13774-13782, 2021 10 19.
Artigo em Inglês | MEDLINE | ID: mdl-34622651

RESUMO

N-linked protein glycosylation is a key regulator in various biological functions. Previous studies have shown that aberrant glycosylation is associated with many diseases. Therefore, it is essential to elucidate protein modifications of glycosylation by quantitatively profiling intact N-linked glycopeptides. Data-independent acquisition (DIA) mass spectrometry (MS) is a cost-effective, flexible, and high-throughput method for global proteomics. However, substantial challenges are still present in the quantitative analysis of intact glycopeptides with high accuracy at high throughput. In this study, we have established a novel integrated platform for the DIA analysis of intact glycopeptides isolated from complex samples. The established analysis platform utilizes a well-designed DIA-MS method for raw data collection, a spectral library constructed specifically for intact glycopeptide quantification providing accurate results by the inclusion of Y ions for quantification and filtering of quantified intact glycopeptides with low-quality MS2 spectra automatically using a set of criteria. Intact glycopeptides isolated from human serum were used to evaluate the performance of the integrated platform. By utilizing 100 isolation windows for DIA data acquisition, a well-constructed human serum spectral library containing 1123 nonredundant intact glycopeptides with Y ions, and automated data inspection, 620 intact glycopeptides were quantified with high confidence from DIA-MS. In summary, our integrated platform can serve as a reliable quantitative tool for characterizing intact glycopeptides isolated from complex biological samples to assist our understanding of biological functions of N-linked glycosylation.


Assuntos
Glicopeptídeos , Proteômica , Glicopeptídeos/metabolismo , Glicosilação , Humanos , Espectrometria de Massas , Soro/metabolismo
14.
Clin Lab ; 67(10)2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34655201

RESUMO

BACKGROUND: Primary nephrotic syndrome (PNS), a clinically prevalent glomerular disease, mostly results in a large loss of plasma albumin, and its predominant clinical manifestations are proteinuria, hypoalbuminemia, edema, and hyperlipidemia. Research has uncovered [9] that sPD-L1 and sPD-1 modulate the PD and PD-L1 pathway in the development of various autoimmune diseases. METHODS: We randomly selected 80 PNS patients treated for PNS in our institution from October 2017 to October 2018 as the case group and 78 healthy volunteers examined in our hospital during the same period as the control group. Not only sPD-1 but also sPD-L1 level in serum and urine was assayed via ELISA. We compared the distribution of T lymphocyte subsets in peripheral blood and mALB and NAG levels in urine. Pearson's correlation analysis was adopted for assessing the relationship of serum and urine sPD-1, sPD-L1 with T lymphocyte subsets and mALB. RESULTS: (1) In contrast to the control group, the case group harbored higher pretreatment serum and urine sPD-1 and sPD-L1 contents (p < 0.05). (2) Before treatment, sPD-1 in the serum and urine held a positive relationship with sPD-L1 level (r was 0.683 and 0.235, respectively, p < 0.05); serum sPD-1 harbored a positive link with urine sPD-1 (r = 0.287, p < 0.01), whereas no relationship was discovered in serum sPD-L1 and urine sPD-L1. (3) In contrast to the control group, the CD4+ level in the case group abated, CD8+ increased, the CD4+/CD8+ ratio assuaged, and mALB level in urine increased (all p < 0.05), whereas NAG harbored no statistical difference (p > 0.05). (4) In the case group, the CD4+/CD8+ ratio possessed positive association with serum sPD-1 (r = 0.384, p < 0.001), and the mALB had a positive relationship with urine sPD-1 (r = 0.704, p < 0.001). (5) After treatment, in comparison with the remission group, the serum sPD-1 level of the non-remission group was increased (p < 0.05), whereas sPD-L1 value was not statistically different (p > 0.05); the sPD-1 level in urine was not statistically significant (p > 0.05), while sPD-L1 content was elevated (p < 0.05). CONCLUSIONS: Serum and urine sPD-1/sPD-L1 levels of PNS patients change dynamically. Detecting sPD-L1 and sPD-1 has certain clinical value for the prognosis of PNS.


Assuntos
Síndrome Nefrótica , Estudos de Casos e Controles , Humanos , Síndrome Nefrótica/diagnóstico , Prognóstico , Soro , Subpopulações de Linfócitos T
15.
Anal Chem ; 93(38): 12848-12853, 2021 09 28.
Artigo em Inglês | MEDLINE | ID: mdl-34520178

RESUMO

Quantum dot (QD)-based digital immunoassays play an important role in ultrasensitive biomarker detection. However, the requirement of an objective with a high numerical aperture (NA) limits the application of this immunoassay. Here, high-quality imaging of massive single-QDs was achieved by the combination of an air objective (20×/0.4 NA) and liquid-immersed microspheres (150 µm, n = 2.2). The signal-to-noise ratio was comparable to that of a 100×/1.4 NA oil objective. Digital analysis of prostate-specific antigen (PSA) was performed within the dynamic range of 0-50 ng/mL and a limit of detection of 0.17 ng/mL. The measured serum data from the PSA were close to the values provided by a hospital. Using a low-magnification and low-NA objective may reduce the barrier of microscopy miniaturization and is beneficial to popularize biomolecular digital analysis.


Assuntos
Pontos Quânticos , Humanos , Imunoensaio , Masculino , Microesferas , Antígeno Prostático Específico , Soro
16.
Wiad Lek ; 74(8): 1925-1930, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34537745

RESUMO

OBJECTIVE: The aim is to study the level of insulin-like growth factor-1 (IGF-1) and insulin-like growth factor-2 (IGF-2) in the blood serum of patients with papillary thyroid cancer, depending on the main clinical and morphological features of the disease. PATIENTS AND METHODS: Materials and methods: The material was the information about 60 patients with papillary thyroid cancer (group 1). In group 2 there were 10 patients without oncopathology. All patients underwent clinical examination after total thyroidectomy before special treatment (radioiodine therapy): ultrasound diagnosis of the neck, confirmed diagnosis of papillary thyroid cancer by morphological examination of operative material. All patients underwent anthropometric studies (height, weight), on the basis of which the body mass index (BMI) was calculated. The study program also included determination of the level of thyroid-stimulating hormone of the pituitary gland (TSH), thyroglobulin (TG), antibodies to thyroglobulin (AB-TG). It was also determined the serum glucose level. In order to assess insulin resistance, the HOMA-IR index was calculated. All patients were tested for serum IGF-1 and IGF-2. RESULTS: Results: In the blood serum of patients with papillary thyroid cancer in 63% of patients the level of IGF-1 and in 85% - IGF-2 was probably higher than in the control group. There is a relationship between the level of IGF-1, IGF-2 and elevated level of proliferating factor - insulin in the serum of patients with papillary thyroid cancer. This may indicate an aggressive potential of the disease (i.e. clinical data on the prevalence of papillary thyroid cancer coincide with laboratory data). There was found a relationship between the expression of IGF-1, IGF-2 and insulin: at elevated levels of insulin> 24.9 µIU/ml, IGF-1 increases 4.2 times, and IGF-2 - 2.5 times. Evaluation of the relationship between the level of IGF-1 and IGF-2 and cervical lymph node involvement shows that in the absence of lesion (N0) there is an increase in these indicators by 2.2 and 1.8 times, respectively. CONCLUSION: Conclusions: The signaling system of insulin-like growth factors (IGF-1 and IGF-2) plays an important role in the occurrence and progression of malignant tumors. It is especially true for papillary thyroid cancer, so its components can be considered as potential diagnostic and prognostic markers of the disease and targets for anticancer therapy.


Assuntos
Somatomedinas/análise , Câncer Papilífero da Tireoide , Neoplasias da Glândula Tireoide , Humanos , Radioisótopos do Iodo , Soro , Tireoglobulina , Câncer Papilífero da Tireoide/diagnóstico , Neoplasias da Glândula Tireoide/diagnóstico
17.
Biosensors (Basel) ; 11(9)2021 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-34562937

RESUMO

Extracellular vesicles (EVs) have emerged as novel biomarkers and therapeutic material. However, the small size (~200 nm) of EVs makes efficient separation challenging. Here, a physical/chemical stress-free separation of EVs based on diffusion through a nanoporous membrane chip is presented. A polycarbonate membrane with 200 nm pores, positioned between two chambers, functions as the size-selective filter. Using the chip, EVs from cell culture media and human serum were separated. The separated EVs were analyzed by nanoparticle tracking analysis (NTA), scanning electron microscopy, and immunoblotting. The experimental results proved the selective separation of EVs in cell culture media and human serum. Moreover, the diffusion-based separation showed a high yield of EVs in human serum compared to ultracentrifuge-based separation. The EV recovery rate analyzed from NTA data was 42% for cell culture media samples. We expect the developed method to be a potential tool for EV separation for diagnosis and therapy because it does not require complicated processes such as immune, chemical reaction, and external force and is scalable by increasing the nanoporous membrane size.


Assuntos
Vesículas Extracelulares , Nanoporos , Meios de Cultura , Humanos , Dispositivos Lab-On-A-Chip , Nanopartículas , Soro
18.
Environ Pollut ; 291: 118140, 2021 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-34555793

RESUMO

In this biomonitoring study, we evaluated the concentrations of 8 polychlorinated biphenyls (PCBs), 11 organochlorinated pesticides (OCPs), 33 brominated flame retardants (BFRs), 7 novel brominated and chlorinated flame retardants (novel FRs) and 30 per- and polyfluoroalkylated substances (PFAS) in human serum samples (n = 274). A total of 89 persistent organic pollutants (POPs) were measured in blood serum samples of city policemen living in three large cities and their adjacent areas (Ostrava, Prague, and Ceske Budejovice) in the Czech Republic. All samples were collected during the year 2019 in two sampling periods (spring and autumn). The identification/quantification of PCBs, OCPs, BFRs, novel FRs and PFAS was performed by means of gas chromatography coupled to (tandem) mass spectrometry (GC-MS/(MS)) and ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry (UHPLC-MS/MS). The most frequently detected pollutants were perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorooctanesulfonate (PFOS), perfluorohexanesulfonate (PFHxS), 2,2',3,4,4',5'-hexachlorobiphenyl (CB 138), 2,2',4,4',5,5'-hexachlorobiphenyl (CB 153), 2,2',3,3',4,4',5-heptachlorobiphenyl (CB 170), 2,2',3,4,4',5,5'-heptachlorobiphenyl (CB 180), hexachlorobenzene (HCB), and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) quantified in 100% of serum samples. In the serum samples, the concentrations of determined POPs were in the range of 0.108-900 ng g-1 lipid weight (lw) for PCBs, 0.106-1016 ng g-1 lw for OCPs, <0.1-618 ng g-1 lw for FRs and <0.01-18.3 ng mL-1 for PFAS, respectively. Locality, sampling season, and age were significantly associated with several POP concentrations. One of the important conclusions was that within the spring sampling period, statistically significant higher concentrations of CB 170 and CB 180 were observed in the samples from Ostrava (industrial area) compared to Prague and Ceske Budejovice. Older policemen had higher concentrations of five PCBs and two OCPs in blood serum.


Assuntos
Poluentes Ambientais , Hidrocarbonetos Clorados , Praguicidas , Bifenilos Policlorados , Monitoramento Biológico , Cidades , República Tcheca , Monitoramento Ambiental , Poluentes Ambientais/análise , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Hidrocarbonetos Clorados/análise , Praguicidas/análise , Bifenilos Policlorados/análise , Soro/química , Espectrometria de Massas em Tandem
19.
ACS Chem Biol ; 16(9): 1671-1679, 2021 09 17.
Artigo em Inglês | MEDLINE | ID: mdl-34469105

RESUMO

Vaccination represents the most effective way to prevent invasive pneumococcal diseases. The glycoconjugate vaccines licensed so far are obtained from capsular polysaccharides (CPSs) of the most virulent serotypes. Protection is largely limited to the specific vaccine serotypes, and the continuous need for broader coverage to control the outbreak of emerging serotypes is pushing the development of new vaccine candidates. Indeed, the development of efficacious vaccine formulation is complicated by the high number of bacterial serotypes with different CPSs. In this context, to simplify vaccine composition, we propose the design of new saccharide fragments containing chemical structures shared by different serotypes as cross-reactive and potentially cross-protective common antigens. In particular, we focused on Streptococcus pneumoniae (Sp) 19A and 19F. The CPS repeating units of Sp 19F and 19A are very similar and share a common structure, the disaccharide ManNAc-ß-(1→4)-Glc (A-B). Herein, we describe the synthesis of a small library of compounds containing different combinations of the common 19F/19A disaccharide. The six new compounds were tested with a glycan array to evaluate their recognition by antibodies in reference group 19 antisera and factor reference antisera (reacting against 19F or 19A). The disaccharide A-B, phosphorylated at the upstream end, emerged as a hit from the glycan array screening because it is strongly recognized by the group 19 antisera and by the 19F and 19A factor antisera, with similar intensity compared with the CPSs used as controls. Our data give a strong indication that the phosphorylated disaccharide A-B can be considered a common epitope among different Sp 19 serotypes.


Assuntos
Epitopos/química , Glicoconjugados/análise , Proteínas Imobilizadas/química , Polissacarídeos Bacterianos/análise , Anticorpos/química , Técnicas Biossensoriais , Reações Cruzadas , Glicoconjugados/metabolismo , Hexosaminas/química , Polissacarídeos Bacterianos/metabolismo , Sorogrupo , Soro/química , Espectrometria de Fluorescência , Streptococcus pneumoniae/metabolismo , Propriedades de Superfície
20.
Talanta ; 234: 122631, 2021 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-34364440

RESUMO

Some metabolites have been found to play key roles in inflammation and immunity events that are associated with diseases such as cancer, diabetes and cytokine release syndrome. Characterization upon the inflammation and immunity-related metabolites (IIMs) will be helpful to the assessment of related pathological states. Although these metabolites have been partially reported in previous studies, the methods for specific measurement of them remain lacking. In the present study, a liquid chromatography - mass spectrometry based method was developed for the targeted analyses of 45 IIMs including amino acids, organic acids, phosphatidylcholines (PCs), polyunsaturated fatty acids and hormones selected based on the literature knowledge. Direct extraction with dansyl-chloride in acetonitrile was proved to be the most efficient and time-saving strategy, in which precipitation, extraction and derivatization were integrated. IIMs derivatized for 4 min and quenched for 2 min revealed the most comprehensive abundance. Based on the defined conditions, all the IIMs had a low limit of detection smaller than 1 ng/mL with the linear range greater than three orders of magnitude. The relative standard derivations of intra-day and inter-day precisions were ranged from 2.2% to 13.4% and 1.7% to 19.5%, respectively. The recovery rates and accuracy in low concentration were 98.9% ± 5.6% and 106.7% ± 11.6%, in medium concentration were 97.1% ± 6.8% and 106.9% ± 9.5%, and in high concentration were 98.4% ± 8.9% and 98.1% ± 8.1%, respectively. Matrix effect and stability were ranged from -37.8% to 35.6% and 2.9% to 14.2%, respectively. To show the usefulness of the method, serum IIMs in hepatitis B virus (HBV) infected patients and healthy subjects were determined and compared. Bile acids, lipoxygenase-mediated lipid mediators and non-enzymatic products showed global increases, whereas most of LysoPCs and cyclooxygenase-mediated prostaglandin D2 decreased in HBV serum samples. This study provided a robust approach for the characterization of IIMs.


Assuntos
Soro , Espectrometria de Massas em Tandem , Cromatografia Líquida de Alta Pressão , Cromatografia Líquida , Humanos , Inflamação
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