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1.
Artigo em Inglês | MEDLINE | ID: mdl-36674183

RESUMO

There is evidence in previous studies that high levels of heavy metals may play a key role in the development of COPD due to the induction of chronic inflammation and oxidative stress. In this preliminary study, we used atomic absorption spectrophotometry to measure the levels of four heavy metals (Cu, Zn, Cd, and Pb) in blood serum of COPD patients and controls over 2 years. Clinical data on disease progression or absence were collected in patients living in the industrial region of Stara Zagora, Bulgaria. The mean values of Cu in the serum of patients with COPD and the control group were 374.29 ± 15.03 µg/L and 238.55 ± 175.31 µg/L, Zn-2010.435 ± 670.006 µg/L and 1672.78 ± 934.27 µg/L, Cd-0.334 ± 0.0216 µg/L and 0.395 ± 0.110 µg/L and Pb-0.0732 ± 0.009 µg/L and 0.075 ± 0.0153 µg/L. This is probably because these elements are biogenic and are used in the body for its anti-oxidant protection. In fact, it cannot be stated with certainty that elevated levels of Cu and Zn in the environment have a negative impact in COPD patients. There was a trend towards higher levels of the toxicants lead and cadmium in COPD patients compared to the control group of patients. There is a statistically unproven trend toward higher levels of lead and cadmium in COPD patients compared to controls, which to some extent supports our hypothesis that there is a relationship between environmental lead and cadmium levels and the COPD manifested. In COPD patients, a positive correlation was found between BMI and serum Cu levels (r = 0.413, p = 0.005). A higher concentration of serum Cu was found in men with BMI ≥ 30, compared to those with BMI < 30. There is also a positive correlation to a lesser extent between CRP and cadmium (r = 0.380; p = 0.019) and lead (r = 0.452; p = 0.004). The correlation of lead and cadmium with PSA also shows that these elements may also be associated with the presence of inflammatory processes. A significant negative correlation exists between Pb in the serum of patients with COPD and their blood hemoglobin (r = -356; p = 0.028). The results of our study suggest that higher doses of the trace elements Cu and Zn do not always have a negative effect in patients with COPD, while the toxicants Pb and Cd may be involved in COPD exacerbation and can be used as prognostic biomarkers for progression. Further studies are warranted to confirm these preliminary results.


Assuntos
Cádmio , Metais Pesados , Masculino , Humanos , Chumbo , Soro/química , Metais Pesados/análise , Zinco , Monitoramento Ambiental/métodos , Cobre
2.
J Clin Lab Anal ; 37(1): e24818, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36550640

RESUMO

BACKGROUND: Even though the serum anion gap (AG) is frequently measured in clinical practice, there is not much research that has examined long-term mortality in unselected adult patients. Our study's objective was to investigate how serum anion gap levels could be used to predict death in unselected participants. METHODS: The relationship between baseline serum AG levels and short-, intermediate-, and long-term all-cause mortality in unselected adult patients is examined using the Cox proportional risk analysis, smoothed curve fitting, subgroup analysis, and Kaplan-Meier survival curves. RESULTS: After screening the database using the appropriate method, a total of 26,270 patients were enrolled in our study for the final data analysis. Our study used smoothed curve fit plots and COX proportional risk regression models incorporating cubic spline functions to evaluate the association between AG levels and all-cause mortality in a non-selected population, and the results indicated a non-linear relationship. In the fully adjusted model, we found that AG levels were positively associated with 30-day, 90-day, 365-day, and 4-year all-cause mortality in unselected adult patients with HRs of 1.08 95% CIs (1.06, 1.09); 1.08 95% CIs (1.06, 1.09); 1.08 95% CIs (1.07, 1.08); 1.07 95% CIs (1.06, 1.07). CONCLUSION: Serum anion gap levels were positively correlated with all-cause mortality in unselected adult patients, with increasing levels of serum anion gap increasing patient mortality.


Assuntos
Equilíbrio Ácido-Base , Soro , Humanos , Adulto , Estudos Retrospectivos , Medição de Risco
3.
Anal Biochem ; 663: 115031, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36580994

RESUMO

The rapid development of proteomics technology in the past decades has led to further human understanding of tumor research, and in some ways, the technology plays a very important supporting role in the early detection of tumors. Human serum has been shown to contain a variety of proteins closely related to life activities, and the dynamic change in proteins can often reflect the physiological and pathological conditions of the body. Serum has the advantage of easy extraction, so the application of proteomics technology in serum has become a hot spot and frontier area for the study of malignant tumors. However, there are still many difficulties in the standardized use of proteomic technologies, which inevitably limit the clinical application of proteomic technologies due to the heterogeneity of human proteins leading to incomplete whole proteome populations, in addition to most serum protein markers being now not highly specific in aiding the early detection of tumors. Nevertheless, further development of proteomics technologies will greatly increase our understanding of tumor biology and help discover more new tumor biomarkers with specificity that will enable medical technology.


Assuntos
Neoplasias , Proteômica , Humanos , Biomarcadores Tumorais , Neoplasias/diagnóstico , Soro , Tecnologia , Proteoma
4.
Talanta ; 253: 123922, 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36122435

RESUMO

This work highlights the efficient approach to highly sensitive determination of dipeptides that can present in biological liquids at very low and trace quantities. The approach involves preliminary derivatization of peptides with tris(2,4,6-trimethoxyphenyl)-methyl carbenium hexafluoroborate followed by ESI and MALDI high-resolution mass spectrometry. Using model dipeptides with various amino acid compositions and sequences, it was shown that the derivatization reaction proceeded smoothly in mild conditions and gave rise to pink-red colored salt derivatives. Ready cations of interest for the analysis are easily desorbed from the salt-derivatives providing strong signals in ESI and MALDI mass spectra and this ensures high sensitivity of the analysis. Another positive aspect is the removal of the target signal from the region of a matrix noise, since the introduced fragment possesses a large mass increment (359 Da). High resolution mass spectrometry, which provides the determination of accurate weights and elemental compositions of ions, was used to reliably detect model dipeptides added to artificial urine and blood serum. A number of these dipeptides was shown to be present in real blood serum collected from volunteers. Collision induced dissociation of precursor cations composed of derivatizing reagent and dipeptide moieties gives rise to characteristic and simple fragmentation mass spectra. A comparison of limits of detection (LOD) measured for non-modified and derivatized dipeptides showed that the latter derivatives provide the highest sensitivity when LOD is determined by using multiple reaction monitoring (MRM) transitions. The suggested derivatization approach was shown to be useful for unambiguous identification of special dipeptides in artificial media and dietary supplements.


Assuntos
Projetos de Pesquisa , Soro , Humanos , Espectrometria de Massas , Peptídeos
5.
J Microbiol Methods ; 204: 106652, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36503053

RESUMO

Haemorrhagic septicaemia (HS) is an acute infection of cattle and buffaloes caused by the B:2 serotype of Pasteurella multocida. This disease is highly endemic in South Asia. In some peracute cases, there is 100% mortality in infected animals within a few hours of infection. Therefore, timely diagnosis of infection may contribute to its treatment and control to minimize economic losses. The current work reported the development of ELISA-based assays for the detection of anti-P. multocida antibodies and pathogen i.e. P. multocida. Owing to high immunogenicity, membrane proteins (MPs) extracted from local isolates of P. multocida serotype B:2 (PM1, PM2, and PM3) were employed as a potential diagnostic antigen for the development of indirect ELISA (i-ELISA) to detect HS antibodies in animals. MPs extracted from PM1, PM2 and PM3 isolates showed very low heterogeneity; hence MPs from the PM3 isolate were selected for the development of i-ELISA. The concentration of MPs (as coating antigen) of 3.13 µg/well and test sera dilution 1:100 was found to be optimal to perform i-ELISA. The developed method was validated through the detection of anti-P. multocida antibodies in sera of mice, immunized with MPs and formalin killed cells from the three local isolates (PM1, PM2 and PM3) of P. multocida. The significantly higher antibody titer in immunized mice was determined compared to unimmunized mice with the cut off value of 0.139. To detect P. multocida directly from the blood of infected animals, whole cell-based ELISA (cb-ELISA) assay was developed. A better detection signal was observed in the assay where bacterial cells were directly adsorbed on plate wells as compared to poly L-lysine (PLL) assisted attachment at a cell concentration of 106 CFU and 107 CFU respectively. The developed assays can be scaled up and potentially be used for the rapid detection of HS antibodies to gauge the immune status of the animal as well as vaccination efficacy and pathogen detection.


Assuntos
Septicemia Hemorrágica , Infecções por Pasteurella , Pasteurella multocida , Camundongos , Animais , Bovinos , Septicemia Hemorrágica/diagnóstico , Septicemia Hemorrágica/veterinária , Soro , Ensaio de Imunoadsorção Enzimática/veterinária , Búfalos , Infecções por Pasteurella/diagnóstico , Infecções por Pasteurella/veterinária , Infecções por Pasteurella/microbiologia
6.
Wiad Lek ; 75(10): 2497-2500, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36472287

RESUMO

OBJECTIVE: The aim: To study the features of changes in the level of prostaglandins (I2 and F2α) in blood serum of patients GERD on the background of OH of the cervical and thoracic spine and obesity. PATIENTS AND METHODS: Materials and methods: The examined patients included 56 patients with GERD and OH of the cervical and thoracic spine. All patients had their blood serum prostaglandin (Pg) F2α and 6-keto prostaglandin F1α (blood prostacyclin - Pg I2) levels examined using the method of immunoassay analysis. RESULTS: Results: In all patients with GERD and OH an excessive body weight or obesity of varying degrees was found while analyzing anthropometric study results. The determination of prostaglandin F2α and prostacyclin (Pg I2) levels in blood serum in patients with GERD and OH and healthy individuals was performed. A more pronounced increase of Pg I2 and Pg F2α in blood serum in patients with GERD and OH with III degree obese was found and the smallest concentration of prostaglandines in blood serum was diagnosed in patients with excessive weight (p<0.05). CONCLUSION: Conclusions: 1. In patients with GERD and OH, an increase in levels of prostaglandins F2α and I2 in blood serum has been established. 2. The relationship between the duration of excess body weigh, obesity and the dynamics of the level of prostaglandin Pg I2 and F2α in blood serum in patients with GERD on the background of OH has been established.


Assuntos
Refluxo Gastroesofágico , Osteocondrose , Doenças da Coluna Vertebral , Humanos , Epoprostenol , Refluxo Gastroesofágico/diagnóstico , Obesidade/complicações , Prostaglandinas , Soro , Doenças da Coluna Vertebral/complicações
7.
Sci Rep ; 12(1): 21379, 2022 12 09.
Artigo em Inglês | MEDLINE | ID: mdl-36494437

RESUMO

Twenty-four blood serum samples from patients with acute methanol poisoning (M) from the mass methanol poisoning outbreak in the Czech Republic in 2012 were compared with 46 patient samples taken four years after poisoning (S) (overlap of 10 people with group M) and with a control group (C) of 24 samples of patients with a similar proportion of chronic alcohol abuse. When comparing any two groups, tens to hundreds of proteins with a significant change in concentration were identified. Fifteen proteins showed significant changes when compared between any two groups. The group with acute methanol poisoning showed significant changes in protein concentrations for at least 64 proteins compared to the other groups. Among the most important identified proteins closely related to intoxication are mainly those involved in blood coagulation, metabolism of vitamin A (increased retinol-binding protein), immune response (e.g., increased complement factor I, complement factors C3 and C5), and lipid transport (increased apolipoprotein A I, apolipoprotein A II, adiponectin). For blood coagulation, the most affected proteins with significant changes in the methanol poisoning group were von Willebrand factor, carboxypeptidase N, alpha-2-antiplasmin (all increased), inter-alpha-trypsin inhibitor heavy chain H4, kininogen-1, plasma serine protease inhibitor, plasminogen (all decreased). However, heparin administration used for the methanol poisoning group could have interfered with some of the changes in their concentrations. Data are available via ProteomeXchange with the identifier PXD035726.


Assuntos
Alcoolismo , Intoxicação , Humanos , Metanol , Soro , Proteoma , Coagulação Sanguínea , Intoxicação/epidemiologia
8.
Lasers Med Sci ; 38(1): 24, 2022 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-36571665

RESUMO

A proof-of-concept of colloidal surface-enhanced Raman scattering (SERS) substrate for rapid selective detection of overexpressed CA 15-3 biomarker in breast cancer serum (BCS) is suggested using PEGylated gold nanourchins (GNUs) conjugated with anti-CA 15-3 monoclonal antibody (mAb). UV-vis spectroscopy provided conformational information about mAb where the initial aromatic amino acid peak was red-shifted from 271 to 291 nm. The fluorescence peak of tyrosine in mAb was reduced by ≈ 77%, and red-shifted by ≈ 3 nm after incubation in BCS. Fourier transform near-infrared spectroscopy and SERS were used to study the composition and the molecular structure of the mAb and BCS. Some of the most dominant Raman shifts after GNU-PEG-mAb interaction with BCS are 498, 736, 818, 1397, 1484, 2028, 2271, and 3227 cm-1 mainly corresponding to C-N-C in amines, vibrational modes of amino acids, C-H out-of-plane bend, C-O stretching carboxylic acid, the vibrational mode in phospholipids, NH3+ amine salt, C≡N stretching in nitriles, and O-H stretching. The intensity of SERS signals varied per trial due to the statistical behavior of GNU in BCS, agglomeration, laser power, and the heating effect. Despite very small amount of plasmonic heating, the result of the ANOVA test demonstrated that under our experimental conditions, the heating effect on signal variation is negligible and that the differences in the laser power are insignificant for all SERS observations (p > 0.6); thus, other parameters are responsible. The absorbance of mAb-conjugated GNU was decreased after five minutes of irradiation at 8 mW in the BCS.


Assuntos
Nanopartículas Metálicas , Neoplasias , Análise Espectral Raman/métodos , Ouro/química , Biomarcadores Tumorais , Soro , Anticorpos Monoclonais , Nanopartículas Metálicas/química
9.
Lasers Med Sci ; 38(1): 22, 2022 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-36564570

RESUMO

This study aimed to identify the differences presented in the Raman spectrum of blood serum from normal subjects compared to leukemic and non-leukemic subjects and the differences between the leukemics and non-leukemics, correlating the spectral differences with the biomolecules. Serum samples from children and adolescents were subjected to Raman spectroscopy (830 nm, laser power 350 mW; n = 566 spectra, being 72 controls, 269 leukemics, and 225 non-leukemics). Exploratory analysis based on principal component analysis (PCA) of the serum sample's spectra was performed. Classification models based on partial least squares discriminant analysis (PLS-DA) were developed to classify the spectra into normal, leukemic, and non-leukemic, as well as to discriminate spectra of leukemic from non-leukemic. The exploratory analysis showed principal components with peaks related to amino acids, proteins, lipids, and carotenoids. The spectral differences between normal, leukemic, and non-leukemic showed features assigned to proteins (serum features), amino acids, and carotenoids. The PLS-DA model classified the spectra of the normal group versus leukemic and non-leukemic groups with accuracy of 66%, sensitivity of 99%, and specificity of 57%. The PLS-DA discriminated the spectra of the leukemic and non-leukemic groups with accuracy of 67%, sensitivity of 72%, and specificity of 60%. The study showed that Raman spectroscopy is a technique that may be used for the biochemical differentiation of leukemias and other types of cancer in serum samples of children and adolescents. Nevertheless, building an extensive data library of Raman spectra from serum samples of controls, leukemics, and non-leukemics of different age groups is necessary to understand the findings better.


Assuntos
Leucemia , Neoplasias , Humanos , Adolescente , Criança , Soro , Leucemia/diagnóstico , Análise Discriminante , Análise Espectral Raman/métodos , Análise de Componente Principal , Carotenoides , Aminoácidos
10.
Medicine (Baltimore) ; 101(50): e31966, 2022 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-36550793

RESUMO

The present study aimed to explore gut microbiota alterations and host cytokine responses in a population with elevated serum diamine oxidase (DAO) disorder. A total of 53 study participants were included in this study, segregated into 2 groups: subjects with high-level DAO (DAO-H, n = 22) subjects with normal DAO level (DAO-N, n = 31). We investigated the clinical and demographic parameters of study participants. The fecal bacterial communities and serum cytokines in 2 groups were assessed by 16S ribosomal RNA gene sequencing and immunoassay. High-pressure liquid chromatography was used to determine hemoglobin Alc. Flow cytometry was used to find the cytokine level in the blood serum. There is no difference in age, total cholesterol (TCHO), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), hemoglobin Alc, fasting plasma glucose (FPG) and homocysteine between the 2 groups. No significant difference were found in α-diversity between the 2 groups, however, the gut microbiota of subjects in DAO-H were characterized by marked interindividual differences, decreased abundance of Phocaeicola, Lachnospira, Bacteroides, Alistipes, Agathobacter, Lachnospira and Bactetoides and increased abundances of Mediterraneibacter, Blautia, Faecallibacterium, Agathobacter, and Parasutterella. Furthermore, the cytokines were no related to the DAO level in both groups and exhibited no significant differences between DAO-H and DAO-N. This study adds a new dimension to our understanding of the DAO and gut microbiota, and revealed that an increase in the DAO level in the intestinal mucosa could alter the gut microbiota composition, which can cause gut-related complications. Research is needed to extensively evaluate downstream pathways and provide possible protective or treatment measures pertaining to relevant disorders.


Assuntos
Amina Oxidase (contendo Cobre) , Microbioma Gastrointestinal , Humanos , Citocinas , Soro , Microbioma Gastrointestinal/fisiologia , Mucosa Intestinal/metabolismo , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo
11.
Biosensors (Basel) ; 12(12)2022 Nov 25.
Artigo em Inglês | MEDLINE | ID: mdl-36551046

RESUMO

The blood ammonia (NH3) level is one of the most important hepatic biomarkers for the diagnosis and monitoring of liver pathologies and infections. In this work, we developed an optimized optical biosensing method to extract and quantify the ammonia contained in complex-matrix samples emulating the blood serum. First, the approach was tested with solutions of phosphate-buffered saline (PBS) and ammonia chloride. Then, further trials were carried out with solutions of fetal bovine serum (FBS). The ammonia was extracted from the tested samples through a customized cell, and it was optically quantified by exploiting the indophenol reaction. The extraction cell included a cation-exchange membrane in Nafion, which was chemically pre-treated through cleaning procedures of sulfuric acid and hydrogen peroxide to keep a basic pH in the ammonia solution and to avoid contaminants in the membrane. From the NH3 solution, the indophenol reaction produced light-reactive indophenol dye molecules, which were used as colorimetric indicators. Through absorbance measurements of the indophenol dye solution at 670 nm wavelength, we were able to detect and quantify the ammonia level in the samples both with a spectrophotometer and a customized miniaturized read-out system, obtaining a detection limit of 0.029 µmol/mL.


Assuntos
Amônia , Dispositivos Ópticos , Soro , Indofenol/química
12.
Biosensors (Basel) ; 12(12)2022 Dec 14.
Artigo em Inglês | MEDLINE | ID: mdl-36551133

RESUMO

In this work, immobilizing anti-GFAP antibodies via covalent attachment onto L-cysteine/gold nanoparticles that were modified with screen-printed carbon electrodes (Anti-GFAP/L-cys/AuNps/SPCE) resulted in the development of a sensitive label-free impedance immunosensor for the detection of Glial Fibrillary Acidic Protein (GFAP). The immunosensor's stepwise construction was studied using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). L-cysteine was chosen as the linker between GFAP antibodies and Au NPs/SPCE because it enables the guided and stable immobilization of GFAP antibodies, thus resulting in increased immunosensor sensitivity. As a redox probe, 5 mM of [Fe(CN)6]3-/4- was used to measure the electron-transfer resistance (Ret), which was raised by the binding of antigens to the immobilized anti-GFAP on the surface of the modified electrode. A linear correlation between Rct and GFAP concentration was achieved under optimum conditions in the range of 1.0-1000.0 pg/mL, with an extraordinarily low detection limit of 51.0 fg/mL. The suggested immunosensor was successfully used to detect the presence of GFAP in human blood serum samples, yielding good findings. As a result, the proposed platform may be utilized to monitor central nervous system injuries.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Humanos , Ouro/química , Imunoensaio/métodos , Soro , Proteína Glial Fibrilar Ácida , Técnicas Biossensoriais/métodos , Cisteína , Nanopartículas Metálicas/química , Eletrodos , Limite de Detecção , Técnicas Eletroquímicas/métodos
13.
Int J Mol Sci ; 23(24)2022 Dec 13.
Artigo em Inglês | MEDLINE | ID: mdl-36555448

RESUMO

Like in many other pathologies, oxidative stress is involved in the development of neurodegenerative disorders. Human serum albumin (HSA) is the main protein in different body fluids including cerebrospinal fluid (CSF). By its redox state in terms of cysteine-34, albumin serves as marker for oxidative burden. We aimed to evaluate the redox state of HSA in patients with multiple sclerosis in serum and CSF in comparison to controls to identify possible correlations with disease activity and severity. Samples were stored at -70 °C until analysis by HPLC for the determination of albumin redox state in terms of the fractions of human mercaptalbumin (HMA), human nonmercaptalbumin1 (HNA1), and human nonmercaptalbumin2 (HNA2). Albumin in CSF showed significantly higher fractions of the reduced form HMA and decreased HNA1 and HNA2. There was no difference between albumin redox states in serum of patients and controls. In CSF of patients HNA2 showed a trend to higher fractions compared to controls. Albumin redox state in serum was associated with physical disability in remission while albumin redox state in CSF was related to disease activity. Thus, albumin redox state in serum and CSF of patients in relation to disease condition merits further investigation.


Assuntos
Esclerose Múltipla , Albumina Sérica Humana , Humanos , Albumina Sérica Humana/metabolismo , Projetos Piloto , Soro/metabolismo , Oxirredução
14.
BMC Vet Res ; 18(1): 451, 2022 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-36564781

RESUMO

BACKGROUND: In human and veterinary medicine calprotectin is most widely used in diagnosing different gastro-intestinal diseases. The aim of this study was to assess the stability of canine calprotectin (cCP) in serum after storage at low temperatures and imprecision of the method. METHODS: Blood samples were collected from dogs with different clinical diagnoses. Twenty-two dogs were included in this study. Calprotectin concentration was measured 4 hours after serum separation (T0), and after being frozen at - 80 °C for 8 (T1) and 16 weeks (T2). The maximum permissible difference (MPD) was derived from the equation for calculating total error (TE) TE = %Bias + (1.96 x %CV), where bias and coefficient of variation (CV) were defined by the manufacturer. The dogs enrolled in this study were patients admitted during the morning (9-12 a.m.), on the day the first measurement was performed. All sample analysis for determination of stability were done in duplicates. For determination of within-run precision, the two patients' serum samples were analyzed in 20 replicates. Imprecision was assessed by analyzing 20 replicates on one plate on two samples where high and low concentrations were anticipated. RESULTS: The calculated value of MPD was 32.52%. Median calprotectin concentrations were higher at T1 114.08 µg/L (IQR = 55.05-254.56) and T2 133.6 µg/L (IQR = 100.57-332.98) than at T0 83.60 µg/L (IQR = 50.38-176.07). Relative and absolute bias at T1 (49.3%; 45.98 µg/L) and T2 (109.93%; 94.09 µg /L) have shown that calprotectin concentrations increase after long term storage at - 80 °C. CONCLUSION: The results of the present study indicate that c-CP was not stable for 16 weeks at low storage temperature (- 80 °C). Considering the observed change in the concentration of c-CP at T1, a storage time of 8 weeks should be safely applied. The method imprecision was not satisfactory, especially in the lower concentration range.


Assuntos
Complexo Antígeno L1 Leucocitário , Soro , Humanos , Cães , Animais , Temperatura , Complexo Antígeno L1 Leucocitário/análise , Congelamento , Soro/química
15.
Clin Nutr ; 41(12): 2729-2739, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36368258

RESUMO

BACKGROUND & AIMS: Nutrient status may affect the risk of microbial infections and play a role in modulating the immune response against such infections. The aim of this study was to determine whether serum 25-hydroxyvitamin D [25(OH)D] and serum fatty acids in infancy are associated with microbial infections by the age of 18 months. METHODS: Altogether 576 newborn infants from Trial to Reduce IDDM in the Genetically at Risk (TRIGR) born between 2002 and 2007 were included. The concentration of 25(OH)D vitamin and proportions of 26 fatty acids (presented as % of total fatty acids) were analyzed in cord blood serum and in sera taken at 6, 12, and 18 months of age. The cord blood samples and mean of 6-18-month values were used as exposures. Infections were detected by screening IgG antibodies against 10 microbes using enzyme immunoassay and antibodies against 6 coxsackievirus B serotypes by plaque neutralization assay in serum samples taken at 18 months of age. RESULTS: A higher proportion of n-3 polyunsaturated fatty acids (PUFAs) and especially long-chain n-3 PUFAs at birth and at the age of 6-18 months was associated with decreased risk of coxsackievirus B2 infection unadjusted and adjusted for region, case-control status, and maternal type 1 diabetes. Higher proportion of docosapentaenoic acid (DPA, 22:5 n-3) at birth was associated with a decreased risk of respiratory syncytial virus infection. 25(OH)D vitamin concentration was not consistently associated with the risk of infections. When only infected children were included docosahexaenoic acid (DHA, 22:6 n-3) and arachidonic acid (20:4 n-6) proportions were positively associated with IgG antibody levels against influenza A virus. 25(OH)D vitamin concentration showed an inverse association with rotavirus IgG levels among children with rotavirus seropositivity. CONCLUSIONS: In young children with increased susceptibility to type 1 diabetes, long-chain n-3 PUFAs may influence the risk of viral infections and immune response against the infections. However, this association may depend on the type of virus suggesting virus-specific effects.


Assuntos
Diabetes Mellitus Tipo 1 , Ácidos Graxos Ômega-3 , Lactente , Recém-Nascido , Criança , Humanos , Pré-Escolar , Ácidos Graxos , Soro , Ácidos Docosa-Hexaenoicos , Calcifediol , Vitaminas , Imunoglobulina G
16.
Bioanalysis ; 14(18): 1241-1249, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-36378607

RESUMO

Background: HMBD-001 is an IgG1 humanized monoclonal antibody specifically targeting HER3, a receptor highly expressed on cancer cells in certain tumors. A bioanalytical method was required to quantify HMBD-001 in human serum, with high selectivity and without interference from HER3. Methods and results: A bridging ELISA using an anti-idiotypic monoclonal capture and detection was developed and validated for quantitative measurement of HMBD-001 in human serum. The assay is sensitive, with a lower limit of quantification of 250 ng/ml, has a broad dynamic range of 250-7000 ng/ml HMBD-001, and exhibits excellent precision and overall accuracy. Conclusion: We have developed and validated a sensitive and selective method for measuring HMBD-001 in human serum. This assay is now being used in a clinical trial setting.


Assuntos
Anticorpos Monoclonais , Soro , Humanos , Ensaio de Imunoadsorção Enzimática/métodos , Anticorpos Monoclonais Humanizados , Imunoglobulina G
17.
Folia Biol (Praha) ; 68(2): 45-49, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36384261

RESUMO

Autologous serum eye drops (ASEDs) are used as a treatment for severe dry eye disease. The concentration and stability of various growth factors in ASEDs is determinative for their efficiency. We therefore assessed the concentrations of transforming growth factor beta 1 (TGF-ß1), epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF-1) in ASEDs following storage at 4-8, -20, -80 and -156 °C. Twenty % and 100% sera from eight healthy volunteers were analysed by the sandwich enzyme immunoassay at different time intervals up to seven months. The mean levels of TGF-ß1 and EGF in undiluted and 20% serum did not differ significantly from the baseline levels in fresh serum for any storage conditions after 7 days at 4-8 °C, as well as after 4- and 7-month preservation at sub-zero temperatures. In 20% serum, no IGF-1 concentration decrease was found following 7 days of preservation at 4-8 °C. However, a decrease to 78 % and 81 % (P < 0.01) of baseline values was found in 20% serum after 4-month storage at -20 °C and 7-month storage at -156 °C, respectively. A more pronounced decrease in IGF-1 was observed in undiluted serum. All assessed growth factors present in 20% frozen serum remained stable for up to 7 months. The highest stability was achieved at -80 °C. At -20 and -156 °C, some decrease in IGF-1 occurred. Our results indicate that 20% ASEDs can be stored frozen up to 7 months under proper conditions.


Assuntos
Fator de Crescimento Epidérmico , Fator de Crescimento Insulin-Like I , Humanos , Fator de Crescimento Insulin-Like I/metabolismo , Fator de Crescimento Transformador beta1 , Temperatura , Soro/metabolismo , Soluções Oftálmicas
18.
Chempluschem ; 87(11): e202200325, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36410784

RESUMO

The electrochemical aptamer-based (E-AB) biosensor usually has a long reaction time when detecting thrombin. This work reports the design of an E-AB biosensor with dual recognition sites to quickly detect thrombin. Specifically, two specific recognition sites of thrombin were used to design three aptamer sequences (TBA-15, TBA-29 and TBA-U), followed by fabrication of corresponding sensors. First, we tested these three types of biosensors in tris buffer solution, and found that the response time of the TBA-U sensor to the same concentration of thrombin was about 2 hours, which is shorter than TBA-15 and TBA-29 sensors. Then, we also did the same test in 50 % diluted serum with 500 nM thrombin. The response time of the TBA-U sensor was about 2 hours, which is still faster than the 3 hours of TBA-15 sensor and the 5.5 hours for TBA-29 sensor. In addition, in terms of dynamic range and specificity, TBA-U has good performance.


Assuntos
Técnicas Biossensoriais , Trombina , Soro , Oligonucleotídeos , Trometamina
19.
Biomolecules ; 12(11)2022 11 08.
Artigo em Inglês | MEDLINE | ID: mdl-36359007

RESUMO

Mild cognitive impairment (MCI) is an early stage of memory loss that affects cognitive abilities, such as language or virtual/spatial comprehension. This cognitive decline is mostly observed with the aging of individuals. Recently, MCI has been considered as a prodromal phase of Alzheimer's disease (AD), with a 10-15% conversion rate. However, the existing diagnostic methods fail to provide precise and well-timed diagnoses, and the pathophysiology of MCI is not fully understood. Alterations of serum N-glycan expression could represent essential contributors to the overall pathophysiology of neurodegenerative diseases and be used as a potential marker to assess MCI diagnosis using non-invasive procedures. Herein, we undertook an LC-MS/MS glycomics approach to determine and characterize potential N-glycan markers in depleted blood serum samples from MCI patients. For the first time, we profiled the isomeric glycome of the low abundant serum glycoproteins extracted from serum samples of control and MCI patients using an LC-MS/MS analytical strategy. Additionally, the MRM validation of the identified data showed five isomeric N-glycans with the ability to discriminate between healthy and MCI patients: the sialylated N-glycans GlcNAc5,Hex6,Neu5Ac3 and GlcNAc6,Hex7,Neu5Ac4 with single AUCs of 0.92 and 0.87, respectively, and a combined AUC of 0.96; and the sialylated-fucosylated N-glycans GlcNAc4,Hex5,Fuc,Neu5Ac, GlcNAc5,Hex6,Fuc,Neu5Ac2, and GlcNAc6,Hex7,Fuc,Neu5Ac3 with single AUCs of 0.94, 0.67, and 0.88, respectively, and a combined AUC of 0.98. According to the ingenuity pathway analysis (IPA) and in line with recent publications, the identified N-glycans may play an important role in neuroinflammation. It is a process that plays a fundamental role in neuroinflammation, an important process in the progression of neurodegenerative diseases.


Assuntos
Disfunção Cognitiva , Soro , Humanos , Cromatografia Líquida/métodos , Soro/química , Espectrometria de Massas em Tandem/métodos , Polissacarídeos/análise , Glicoproteínas , Biomarcadores , Disfunção Cognitiva/diagnóstico
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