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1.
J Proteomics ; 285: 104941, 2023 08 15.
Artigo em Inglês | MEDLINE | ID: mdl-37285906

RESUMO

There has been little information about the proteome of bovine faeces or about the contribution to the faecal proteome of proteins from the host, the feed or the intestinal microbiome. Here, the bovine faecal proteome and the origin of its component proteins was assessed, while also determining the effect of treating barley, the major carbohydrate in the feed, with either ammonia (ATB) or sodium propionate (PTB) preservative. Healthy continental crossbreed steers were allocated to two groups and fed on either of the barley-based diets. Five faecal samples from each group were collected on Day 81 of the trial and analysed by quantitative proteomics using nLC-ESI-MS/MS after tandem mass tag labelling. In total, 281 bovine proteins, 199 barley proteins, 176 bacterial proteins and 190 archaeal proteins were identified in the faeces. Mucosal pentraxin, albumin and digestive enzymes were among bovine proteins identified. Serpin Z4 a protease inhibitor was the most abundant barley protein identified which is also found in barley-based beer, while numerous microbial proteins were identified, many originating bacteria from Clostridium, while Methanobrevibacter was the dominant archaeal genus. Thirty-nine proteins were differentially abundant between groups, the majority being more abundant in the PTB group compared to the ATB group. SIGNIFICANCE: Proteomic examination of faeces is becoming a valuable means to assess the health of the gastro-intestinal tract in several species, but knowledge on the proteins present in bovine faeces is limited. This investigation aimed to characterise the proteome of bovine faecal extracts in order to evaluate the potential for investigations of the proteome as a means to assess the health, disease and welfare of cattle in the future. The investigation was able to identify proteins in bovine faeces that had been (i) produced by the individual cattle, (ii) present in the barley-based feed eaten by the cattle or (iii) produced by bacteria and other microbes in the rumen or intestines. Bovine proteins identified included mucosal pentraxin, serum albumin and a variety of digestive enzymes. Barley proteins found in the faeces included serpin Z4, a protease inhibitor that is also found in beer having survived the brewing process. Bacterial and archaeal proteins in the faecal extracts were related to several pathways related to the metabolism of carbohydrates. The recognition of the range of proteins that can be identified in bovine faeces raises the possibility that non-invasive sample collection of this material could provide a novel diagnostic approach to cattle health and welfare.


Assuntos
Proteínas Arqueais , Hordeum , Serpinas , Bovinos , Animais , Serpinas/análise , Proteoma/análise , Cerveja/análise , Proteômica , Espectrometria de Massas em Tandem , Dieta/veterinária , Fezes/microbiologia , Bactérias , Extratos Vegetais , Ração Animal/análise
2.
Mikrochim Acta ; 190(6): 223, 2023 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-37184586

RESUMO

This study aimed to design a sandwich electrochemiluminescence (ECL) immunosensor with double co-reaction accelerators for sensitively detecting squamous cell carcinoma antigen (SCCA). First, silver orthophosphate (Ag3PO4) nanoparticles were modified on the surface of EuPO4 nanowires to improve their poor dispersibility/solubility. At the same time, EuPO4 was used as a co-reaction accelerator to catalyze S2O82- to produce more intermediates (SO4•-), significantly enhancing the ECL signal of Ag3PO4. Ag nanoparticles (AgNP) modified on Ag3PO4@EuPO4 composite nanomaterials were used not only as linkers of luminescence groups and biomarkers but also as a co-reaction accelerator to effectively enhance ECL signal. The designed ECL immunosensor displayed several advantages, including good stability and reproducibility. Under the optimal conditions, its linear range in detecting SCCA was 0.0001-50 ng·mL-1, the detection limit was 25 fg·mL-1 (S/N = 3), the recovery was 96.6-100.4%, and the relative standard deviation was less than 4.8%. It was successfully applied to detect SCCA in human serum.


Assuntos
Antígenos de Neoplasias , Técnicas Biossensoriais , Nanopartículas Metálicas , Serpinas , Humanos , Técnicas Eletroquímicas , Imunoensaio , Medições Luminescentes , Reprodutibilidade dos Testes , Prata , Antígenos de Neoplasias/análise , Serpinas/análise
3.
Biomed Res Int ; 2022: 8592729, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35535041

RESUMO

Objective: The objective is to investigate the relationship and correlation between PEDF and TGF-ß in aqueous humor and serum and high myopia CNV lesions. Methods: For each group of patients (namely, group A: patients with high myopia CNV (mCNV); group B: patients with high myopia without CNV; group C: patients with CNV caused by other eye diseases; and group D (control group): patients with simple cataract (without CNV and high myopia)), 20 patients were collected. A total of 40 patients have been collected since the beginning of the study in December 2020, including 7 patients in group A, 13 patients in group B, 10 patients in group C, and 10 patients in group D. Serum and aqueous humor samples were collected, and PEDF and TGF-ß levels in serum and aqueous humor were detected by enzyme-linked immunosorbent assay (ELISA). SPSS 26.0 statistical software was used to process the data. Independent sample t-test was used to compare the data of the same factor in the same group between serum and aqueous humor. Comparisons of the same factors between different groups were performed using a one-way analysis of variance (ANOVA). Correlation analysis was conducted by the Pearson correlation coefficient test. P < 0.05 indicated that the difference was statistically significant. Results: There were no significant differences in age, gender, and course of disease among all groups (P > 0.05). The concentration of PEDF in aqueous humor in group A and group C was higher than that in group B and group D. There was no significant correlation between serum PEDF content and the above-mentioned diseases. The concentration of TGF-ß in aqueous humor in groups A, B, and C was significantly higher than that in group D. However, there was no significant correlation between TGF-ß content in serum and the above-mentioned diseases. There was no significant correlation between aqueous humor and serum PEDF. There was no significant correlation between the content of TGF-ß in aqueous humor and serum. Conclusion: TGF-ß in aqueous humor may be involved in the development of high myopia and intraocular CNV disease. However, PEDF in aqueous humor may be involved in the development of intraocular CNV disease and has no significant correlation with high myopia. At the same time, TGF-ß and PEDF in serum had no significant correlation with high myopia and intraocular CNV disease. There was no significant correlation between the concentrations of TGF-ß and PEDF in aqueous humor and serum.


Assuntos
Catarata , Miopia , Serpinas , Humor Aquoso/química , Ensaio de Imunoadsorção Enzimática , Humanos , Serpinas/análise , Fator de Crescimento Transformador beta
4.
Mikrochim Acta ; 189(3): 101, 2022 02 12.
Artigo em Inglês | MEDLINE | ID: mdl-35152310

RESUMO

Three disposable stochastic sensors using nanolayer deposition of a graphene nanocomposite comprising graphene nanoparticles and gold nanoparticles, on different supports: silk, plastic, and paper, and modified with chitosan, were characterized and validated for molecular recognition and quantification of maspin in biological samples. Very low limits of determination (of pg mL-1 magnitude order) were recorded (5.12 pg mL-1 for the sensor based on silk at pH 7.40 and on copy paper at pHs 3.00 and 7.40; 16 ng mL-1 at pH 7.40 for the sensor based on plastic, 41.00 pg mL-1 for the sensor based on silk at pH 3.00, and 204.00 pg mL-1 for the sensor based on plastic, at pH 3.00), with wide linear concentration ranges (5.12 × 10-12-2.00 × 10-6 g mL-1 for the sensors based on silk at pH 7.40, and on copy paper at pH 3.00; 5.12 × 10-12-8.00 × 10-7 g mL-1 for the sensor based on copy paper at pH 7.40; 1.60 × 10-8-2.00 × 10-6 g mL-1 for the sensor based on plastic at pH 7.40; 4.10 × 10-14-2.00 × 10-6 g mL-1 for the sensor based on silk, at pH 3.00; and 2.04 × 10-13-8.00 × 10-7 g mL-1 for the sensor based on plastic at pH 3.00) allowing the molecular recognition and quantification of maspin in healthy people and patients with gastric cancer, when a potential of 125 mV vs. Ag/AgCl was applied. The recoveries of maspin in whole blood, saliva, urine, and tissue samples were higher than 95.00%, with a relative standard deviation lower than 1.0%.


Assuntos
Técnicas Biossensoriais , Líquidos Corporais/química , Técnicas Eletroquímicas , Serpinas/análise , Ouro/química , Grafite/química , Humanos , Nanopartículas/química , Processos Estocásticos
5.
Asian Pac J Cancer Prev ; 22(10): 3299-3307, 2021 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-34711007

RESUMO

BACKGROUND AND STUDY AIM: One of the problems in diagnosing pancreatic ductal adenocarcinoma (PDAC) is differentiation between PDAC cells and benign pancreatic tissue cells in cytologic samples. This study aimed to evaluate the usefulness of Maspin, CK17 and Ki-67 immunocytochemistry (ICC) in differentiation between these two groups of cells. MATERIALS AND METHODS: This retrospective study was carried on 80 cases of PDAC and 25 cell blocks of benign pancreatic tissue cells as a control group for evaluation of Maspin, CK17 and Ki-67 ICC. PDAC cases were sampled by endoscopic ultrasound guided fine needle aspiration cytology (EUS-FNAC), while cell blocks of control group were aspirated from benign pancreatic tissues that were obtained from the pancreatic surgically resected specimens. Immunostaining patterns, sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy of each antibody as well as possible antibody combined panels of these markers in differentiation between the two groups were evaluated. RESULTS: Positive immunoreactivity for Maspin, CK17 and Ki-67 were 92.5%, 80% and 72.5% in PDAC cases, respectively. In contrast to PDAC cases, all the cell blocks of benign pancreatic tissue cells were negative for these markers. Regarding different panels, combined use of Maspin, CK17 and Ki-67 together as a triple test (at least one of them is positive) achieved the highest sensitivity of 98.8%, specificity of 100%, PPV of 100%, NPV of 96.2% and accuracy of 99% in the differentiation between PDAC and benign pancreatic tissue. CONCLUSION: Employing this short panel [Maspin, CK17 and Ki-67] is helpful for better differentiation between PDAC and benign pancreatic tissue.


Assuntos
Carcinoma Ductal Pancreático/química , Iminas/análise , Antígeno Ki-67/análise , Pâncreas/química , Neoplasias Pancreáticas/química , Serpinas/análise , Tiazinas/análise , Adulto , Idoso , Biomarcadores Tumorais/análise , Antígeno CA-19-9/sangue , Antígeno Carcinoembrionário/sangue , Carcinoma Ductal Pancreático/patologia , Diagnóstico Diferencial , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico , Feminino , Humanos , Imuno-Histoquímica , Imunofenotipagem , Masculino , Pessoa de Meia-Idade , Pâncreas/citologia , Neoplasias Pancreáticas/patologia , Valor Preditivo dos Testes , Estudos Retrospectivos
6.
Parasitol Int ; 83: 102346, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-33857597

RESUMO

Schistosomiasis remains a worldwide public health problem, especially in sub-Saharan Africa. The World Health Organization targets the goal for its elimination as a public health problem in the 2030 Neglected Tropical Diseases (NTDs) Roadmap. Concerted action and agile responses to challenges will be necessary to achieve the targets. Better diagnostic tests can accelerate progress towards the elimination by monitoring disease trends and evaluating the effectiveness of interventions; however, current examinations such as Kato-Katz technique are of limited power to detect light-intensity infections. The point-of-care circulating cathodic antigen (POC-CCA) test shows a higher sensitivity compared to the reference standard, Kato-Katz technique, but it still lacks sufficient sensitivity with low infection intensity. In this study, we examined antibody reactions against recombinant protein antigens; Schistosoma mansoni serine protease-inhibitor (SmSerpin) and RP26, by enzyme-linked immunosorbent assay (ELISA) in plasma samples with light-intensity infection. The sensitivity using the cocktail antigen of recombinant SmSerpin and RP26 showed 83.7%. The sensitivity using S. mansoni soluble egg antigen (SmSEA) was 90.8%, but it showed poor specificity (29.7%), while the cocktail antigen presented improved specificity (61.4%). We conclude that antibody detection to the SmSerpin and RP26 protein antigens is effective to detect S. mansoni light-intensity infections. Our study indicates the potential of detecting antibody against recombinant protein antigens to monitor the transmission of schistosomiasis in low endemicity contexts.


Assuntos
Testes Diagnósticos de Rotina/estatística & dados numéricos , Ensaio de Imunoadsorção Enzimática , Schistosoma mansoni/isolamento & purificação , Esquistossomose mansoni/diagnóstico , Adolescente , Animais , Antígenos de Helmintos/análise , Criança , Pré-Escolar , Feminino , Proteínas de Helminto/análise , Humanos , Quênia/epidemiologia , Masculino , Prevalência , Esquistossomose mansoni/epidemiologia , Esquistossomose mansoni/parasitologia , Serpinas/análise
7.
Anticancer Res ; 41(4): 2053-2058, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33813413

RESUMO

AIM: To investigate potential associations between selected oncomarkers [carcinoembryonic antigen (CEA), C-terminus of cytokeratin 19 (CYFRA 21-1, CYFRA), and squamous cell carcinoma antigen (SCC)] and outcomes in patients with NSCLC treated with bevacizumab plus chemotherapy. PATIENTS AND METHODS: We retrospectively analysed 105 patients with NSCLC from the Czech TULUNG registry treated at University Hospital in Pilsen with bevacizumab plus chemotherapy. Response to therapy was tested by Fisher's exact test. Survival statistics were evaluated using the Kaplan-Meier method and Cox analysis. RESULTS: Only normal values of CYFRA (not CEA or SCC) were associated with significantly better overall and progression-free survival in univariate analysis. We also observed a trend for a better disease control rate in patients with normal levels of CYFRA. In a multivariate Cox model, only CYFRA was associated with significantly better overall but not progression-free survival. CONCLUSION: In our retrospective study, we point out the possibility of using CYFRA as a prognostic marker in patients with NSCLC treated with chemotherapy plus bevacizumab.


Assuntos
Antígenos de Neoplasias/fisiologia , Antineoplásicos/uso terapêutico , Bevacizumab/administração & dosagem , Carcinoma Pulmonar de Células não Pequenas/tratamento farmacológico , Queratina-19/fisiologia , Neoplasias Pulmonares/tratamento farmacológico , Idoso , Antígenos de Neoplasias/análise , Antígenos de Neoplasias/sangue , Bevacizumab/efeitos adversos , Biomarcadores Farmacológicos/análise , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/fisiologia , Antígeno Carcinoembrionário/análise , Antígeno Carcinoembrionário/sangue , Carcinoma Pulmonar de Células não Pequenas/diagnóstico , Carcinoma Pulmonar de Células não Pequenas/mortalidade , Carcinoma Pulmonar de Células não Pequenas/patologia , Progressão da Doença , Feminino , Humanos , Queratina-19/análise , Queratina-19/sangue , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/mortalidade , Neoplasias Pulmonares/patologia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Valor Preditivo dos Testes , Prognóstico , Estudos Retrospectivos , Serpinas/análise , Serpinas/sangue , Resultado do Tratamento
8.
Anticancer Res ; 41(3): 1203-1212, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33788711

RESUMO

BACKGROUND/AIM: Hepatocellular carcinoma (HCC) remains one of the biggest medical issues. Pigment epithelial-derived factor (PEDF) is a glycoprotein that belongs to the superfamily of serine protease inhibitors. PEDF interacts with its two receptors, adipose triglyceride lipase (ATGL) and laminin receptor (LR). MATERIALS AND METHODS: We conducted immunohistochemical staining for PEDF, LR and ATGL in 151 resected HCCs and their background liver tissues. RESULTS: High expression of LR in HCC was associated with high histological grade and portal vein invasion, while high expression of PEDF in HCC was associated with absence of portal vein invasion. High LR expression in background liver was statistically associated with low serum albumin levels and was an independent prognostic factor of worse outcomes. No cases with more than 5% fatty degeneration in the background liver tissue showed high PEDF expression. CONCLUSION: PEDF/LR/ATGL could be potential biomarkers in HCC and various chronic hepatic disorders.


Assuntos
Carcinoma Hepatocelular/química , Proteínas do Olho/análise , Lipase/análise , Neoplasias Hepáticas/química , Fígado/química , Fatores de Crescimento Neural/análise , Receptores de Laminina/análise , Receptores de Neuropeptídeos/análise , Serpinas/análise , Carcinoma Hepatocelular/patologia , Feminino , Humanos , Imuno-Histoquímica , Neoplasias Hepáticas/patologia , Masculino , Invasividade Neoplásica , Prognóstico , Albumina Sérica/análise
9.
J Clin Pathol ; 74(11): 712-717, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33046564

RESUMO

OBJECTIVE: Sessile serrated lesions without dysplasia (SSL-ND) are epitomised by dilated crypts with epithelial serrations and architectural distortions portraying boot-shapes, L-shapes or inverted-T shapes. Recently, crypts in asymmetric fission were detected in SSL-ND. The purpose was to assess the frequency of crypts in asymmetric fission in a cohort of SSL-ND. METHODS: The frequency of crypts in fission was assessed in 60 SSL-ND, the distribution of cell proliferation in 48 SSL-ND and the expression of maspin, a tumour-suppressor protein, in 29 SSL-ND. RESULTS: Out of the 60 SSL-ND, 40 (66.7%) showed crypts in fission: 39 (65%) SSL-ND had crypts in asymmetric fission and one SSL-ND (1.7%), in symmetric fission (p<0.05). Of 1495 crypts recorded in the 60 SSL-ND, 73 (4.9%) were in asymmetric fission but only one (0.06%), in symmetric fission (p<0.05). Out of the 48 Ki67-immunostained SSL-ND,15 (31%) showed randomly distributed proliferating cell-domains. All 29 SSL-ND revealed maspin-upregulation (including crypts in asymmetric and symmetric fission). In contrast, the normal colon mucosa showed occasional single crypts in symmetric fission, proliferating cell-domains limited to the lower thirds of the crypts, absence of crypts in asymmetric fission and remained maspin negative. CONCLUSIONS: SSL-ND thrive with crypts in asymmetric fission displaying randomly distributed proliferating cell-domains and maspin-upregulation. These histo-biological aberrations disclose pathological cryptogenesis and suggest possibly unfolding somatic mutations in SSL-ND. The present findings may open new vistas on the parameters pertinent to the susceptibility of SSL-ND to develop dysplasia and carcinoma.


Assuntos
Proliferação de Células , Colo/patologia , Mucosa Intestinal/patologia , Estudos de Coortes , Neoplasias do Colo/etiologia , Humanos , Antígeno Ki-67 , Serpinas/análise
10.
J Comp Neurol ; 529(1): 141-158, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32427349

RESUMO

Pigment epithelium-derived factor (PEDF) is a multifunctional protein which was initially described in the retina, although it is also present in other tissues. It functions as an antioxidant agent promoting neuronal survival. Recently, a PEDF receptor has shown an elevated binding affinity for PEDF. There are no relevant data regarding the distribution of both proteins in the brain, therefore the main goal of this work was to investigate the spatiotemporal presence of PEDF and PEDFR in the adult mouse brain, and to determine the PEDF blood level in mouse and human. The localization of both proteins was analyzed by different experimental methods such as immunohistochemistry, western-blotting, and also by enzyme-linked immunosorbent assay. Differential expression was found in some telencephalic structures and positive signals for both proteins were detected in the cerebellum. The magnitude of the PEDFR labeling pattern was higher than PEDF and included some cortical and subventricular areas. Age-dependent changes in intensity of both protein immunoreactions were found in the cortical and hippocampal areas with greater reactivity between 4 and 8 months of age, whilst others, like the subventricular zones, these differences were more evident for PEDFR. Although ubiquitous presence was not found in the brain for these two proteins, their relevant functions must not be underestimated. It has been described that PEDF plays an important role in neuroprotection and data provided in the present work represents the first extensive study to understand the relevance of these two proteins in specific brain areas.


Assuntos
Química Encefálica/fisiologia , Encéfalo/metabolismo , Proteínas do Olho/análise , Proteínas do Olho/biossíntese , Fatores de Crescimento Neural/análise , Fatores de Crescimento Neural/biossíntese , Receptores de Neuropeptídeos/análise , Receptores de Neuropeptídeos/biossíntese , Serpinas/análise , Serpinas/biossíntese , Adolescente , Adulto , Fatores Etários , Animais , Criança , Pré-Escolar , Feminino , Humanos , Masculino , Camundongos , Camundongos da Linhagem 129 , Camundongos Endogâmicos C57BL , Adulto Jovem
11.
Sci Rep ; 10(1): 9033, 2020 06 03.
Artigo em Inglês | MEDLINE | ID: mdl-32493989

RESUMO

Recently, we have been seeing emerging applications of non-invasive approaches using serum biomarkers including miRNA and proteins in detection of multiple cancers. Currently, majority of these methods only use solitary type of biomarkers, which often lead to non-satisfactory sensitivity and specificity in clinical applications. To this end, we established a unique biomarker panel in this study, which determined both squamous cell carcinoma antigen (SCC Ag) degree and miRNA-29a, miRNA-25, miRNA-486-5p levels in blood for detection of early-stage cervical cancer. We designed our study with two phases: a biomarker discovery phase, followed by an independent validation phase. In total of 140 early-stage cervical cancer patients (i.e., AJCC stage I and II) and 140 healthy controls recruited in the biomarker discovery phase, we achieved sensitivity of 88.6% and specificity of 92.9%. To further assess the predictive power of our panel, we used it to an independent patient cohort that consisted of 60 early-stage cervical cancer individuals as well as 60 healthy controls, and successfully achieved both high sensitivity (80.0%) and high specificity (96.7%). Our study indicated combining analyses of multiple serum biomarkers could improve the accuracy of non-invasive detection of early-stage cervical cancer, and potentially serve as a new liquid biopsy approach for detecting early-stage cervical cancer.


Assuntos
Neoplasias do Colo do Útero/diagnóstico , Neoplasias do Colo do Útero/genética , Adulto , Antígenos de Neoplasias/análise , Antígenos de Neoplasias/sangue , Biomarcadores Tumorais/genética , Proteínas Sanguíneas/genética , Detecção Precoce de Câncer/métodos , Feminino , Perfilação da Expressão Gênica/métodos , Testes Genéticos/métodos , Humanos , Biópsia Líquida/métodos , MicroRNAs/genética , Curva ROC , Sensibilidade e Especificidade , Serpinas/análise , Serpinas/sangue , Neoplasias do Colo do Útero/sangue
12.
Pathol Res Pract ; 216(5): 152925, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32273198

RESUMO

Morphology plays an important role in the distinction of autoimmune pancreatitis (AIP) from pancreatic ductal adenocarcinoma (PDAC). However, we aimed to determine the utility of immunohistochemical tumor markers to contribute in the distinction of these entities. In surgical specimens with AIP (n = 20), PDAC (n = 20) and normal pancreas (n = 20), the expression of pVHL, maspin, IMP3, S100P and Ki67 was examined. We evaluated intralobular reactive ducts / acinoductal metaplasia (ILDs) and extralobular ducts (ELDs) in AIP, neoplastic glands in PDAC, and ductal epithelium in the normal pancreas, using a five-tiered scoring system. The Ki67 hot spot index (Ki67-HSPI) was determined manually and using automated digital imaging analysis of virtual double stains of Ki67 and CK8. Besides, sequential dual-immunohistochemical staining of maspin/pVHL, maspin/IMP3 and Ki67/maspin was performed in a subset of the specimens. Strong overexpression of IMP3, maspin, S100P and Ki67 and loss of pVHL was observed in PDAC compared to AIP and normal pancreas. In AIP however, focal and weak aberrant expression was observed with the following proportions in ILDs/ELDs: pVHL in 45 %/85 %, maspin in 30 %/70 %, IMP3 in 55 %/5%, S100P in 10 %/35 % and Ki67-HSPI >20 % in 15 %/70 %. At least two markers were aberrantly expressed in ILDs/ELDs in 45 %/60 %. The aberrant expression was more pronounced in type 2 AIP compared to type 1. In conclusion, our data indicate that pVHL, maspin, IMP3, S100P and Ki67 can be focal and weak aberrantly expressed in AIP. However, when used as a panel, these markers seem to be useful for the differentiation of AIP from PC.


Assuntos
Pancreatite Autoimune/diagnóstico , Biomarcadores Tumorais/análise , Carcinoma Ductal Pancreático/diagnóstico , Neoplasias Pancreáticas/diagnóstico , Adulto , Idoso , Proteínas de Ligação ao Cálcio/análise , Proteínas de Ligação ao Cálcio/biossíntese , Diagnóstico Diferencial , Feminino , Humanos , Antígeno Ki-67/análise , Antígeno Ki-67/biossíntese , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/análise , Proteínas de Neoplasias/biossíntese , Ribonucleoproteínas Nucleolares Pequenas/análise , Ribonucleoproteínas Nucleolares Pequenas/biossíntese , Serpinas/análise , Serpinas/biossíntese , Proteína Supressora de Tumor Von Hippel-Lindau/análise , Proteína Supressora de Tumor Von Hippel-Lindau/biossíntese
13.
Mikrochim Acta ; 187(2): 128, 2020 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-31938893

RESUMO

A method is described for the electrochemical determination of squamous cell carcinoma (SCC) antigen, and by testing the effect of 30 nm gold nanoparticles (GNPs). Three comparative studies were performed in the presence and absence of GNPs, and with agglomerated GNPs. The divalent ion Ca(II) was used to induce a strong agglomeration of GNPs, as confirmed by colorimetry and voltammetry. Herein, colorimetry was used to test the best amount of salt needed to aggregate the GNPs. Despite, voltammetry was used to determine the status of biomolecules on the sensor. The topography of the surface of ZnO-coated interdigitated electrodes was analyzed by using 3D-nano profilometry, scanning electron microscopy, atomic force microscopy and high-power microscopy. The interaction between SCC antigen and antibody trigger vibrations on the sensor and cause dipole moment, which was measured using a picoammeter with a linear sweep from 0 to 2 V at 0.01 V step voltage. The sensitivity level was 10 fM by 3σ calculation for the dispersed GNP-conjugated antigen. This indicates a 100-fold enhancement compared to the condition without GNP conjugation. However, the sensitivity level for agglomerated GNPs conjugated antibody was not significant with 100 fM sensitivity. Specificity was tested for other proteins in serum, namely blood clotting factor IX, C-reactive protein, and serum albumin. The SCC antigen was quantified in spiked serum and gave recoveries that ranged between 80 and 90%. Graphical abstractSchematic representation of SCC (squamous cell carcinoma) antigen determination using divalent ion induced agglomerated GNPs. Sensitivity increment depends on the occurrence of more SCC antigen and antibody binding event via GNPs integration. Notably, lower detection limit was achieved at femto molar with proper orientation of biological molecules.


Assuntos
Antígenos de Neoplasias/análise , Técnicas Biossensoriais/métodos , Ouro , Nanopartículas Metálicas/química , Serpinas/análise , Anticorpos/imunologia , Reações Antígeno-Anticorpo , Antígenos de Neoplasias/imunologia , Cálcio/farmacologia , Cátions Bivalentes/farmacologia , Técnicas Eletroquímicas , Eletrodos , Humanos , Limite de Detecção , Serpinas/imunologia
14.
Spectrochim Acta A Mol Biomol Spectrosc ; 226: 117610, 2020 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-31606675

RESUMO

Vaspin is a protein present in human serum that can cause type-2 diabetes, obesity, and other cardiovascular diseases. We report fluorescent upconverting nanoparticles (UCNPs)-based lateral flow biosensor for ultrasensitive detection of Vaspin. A pair (primary and secondary) of cognate aptamers was used that has duo binding with Vaspin. UCNPs with a diameter of around 100 nm were used as a tag to label a detection probe (secondary aptamer). A primary aptamer (capture probe) was immobilized on the test zone. Sandwich type hybridization reactions among the conjugate probe, target Vaspin, and primary aptamer were performed on the lateral flow biosensor. In the presence of target Vaspin, UCNPs were captured on the test zone of the biosensor and the fluorescent intensity of the captured UCNPs was measured through a colorimetric app under NIR. Fluorescence intensity indicates the quantity of Vaspin present in the sample. A range of Vaspin concentration across 0.1-55 ng ml-1 with a Limit of detection (LOD) 39 pg ml-1 was tested through this UCNPs based LFSA with high sensitivity, reproducibility and repeatability, whereas it's actual range in human blood is from 0.1 to 7 ng ml-1. Therefore, this research provides a well-suited lateral flow strip with an ultrasensitive and low-cost approach for the early diagnosis of type-2 diabetes and this could be applied to any targets with a duo of aptamers generated.


Assuntos
Técnicas Biossensoriais/métodos , Análise Química do Sangue/métodos , Corantes Fluorescentes , Nanopartículas/química , Serpinas/análise , Aptâmeros de Nucleotídeos/análise , Diabetes Mellitus Tipo 2/sangue , Diabetes Mellitus Tipo 2/diagnóstico , Diagnóstico Precoce , Fluorescência , Corantes Fluorescentes/síntese química , Corantes Fluorescentes/química , Corantes Fluorescentes/farmacocinética , Humanos , Limite de Detecção , Ligação Proteica , Reprodutibilidade dos Testes , Serpinas/metabolismo
15.
Biosens Bioelectron ; 150: 111951, 2020 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-31818758

RESUMO

Catalytic reactions contribute a lot to electrochemical sensing by amplifying electrochemical signals to elevating sensitivity, allowing ultrasensitive sensing of bioindicators. However, the unsatisfactory catalytical performance of catalysts results in low efficiency, limiting its practice in rapid immunosensing. Herein, we demonstrated the potential of photo-induced microscale hyperthermia in accelerating catalysis to enhance sensitivity in the short time. Under near-infrared (NIR) laser irradiation, the period of Fenton-like reaction was significantly reduced, further allowing rapid change of electrical signal on electrode in situ. By constructing a novel immunosensor, efficacious sensing of Squamous Cell Carcinoma Antigen (SCCA) was achieved with improved sensitivity for two times, high timeliness within several minutes and advanced performance of electrochemical immunosensor (linear detection range: 0.1 pg mL-1-1 µg mL-1; limit of detection: 120.2 fg mL-1). To the best of our knowledge, this research is the first work that typifies the photothermal-enhanced catalysis in the electrochemical immunoassays, which illuminates a great direction of developing advanced electrochemical sensing protocol with both favorable capacities and accelerated process.


Assuntos
Anticorpos Imobilizados/química , Antígenos de Neoplasias/sangue , Técnicas Biossensoriais/métodos , Indóis/química , Nanopartículas de Magnetita/química , Polímeros/química , Serpinas/sangue , Antígenos de Neoplasias/análise , Catálise , Técnicas Eletroquímicas/métodos , Temperatura Alta , Humanos , Imunoensaio/métodos , Nanopartículas de Magnetita/ultraestrutura , Processos Fotoquímicos , Serpinas/análise
16.
Biomed Res Int ; 2019: 5320902, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31886225

RESUMO

Pregnancy is a period in a woman's life in which changes can occur that affect different physiological processes. Common conditions during this period include vascular changes, such as lower extremity venous insufficiency (VI). This is an observational, analytical, and prospective cohort study in which 114 pregnant women were analyzed, of which 62 were clinically diagnosed with VI. In parallel, 52 control patients without VI (HC) were studied. The aim of this study was to observe changes in angiogenesis and inflammation markers as well as the presence of calcium deposits. The expression of vascular endothelial growth factor (VEGF), transforming growth factor-ß (TGF-ß), and pigment epithelium-derived factor (PEDF) was analyzed by immunohistochemistry and RT-qPCR. The presence of calcium deposits was revealed using the von Kossa method. In the placentas of mothers with VI, gene expression of VEGF (34.575 [32.380-36.720] VI vs 32.965 [30.580-36.320] HC) and PEDF (25.417 [24.459-27.675] VI vs 24.400 [23.102-30.223] HC) significantly increased, as was protein expression in the placental villi. An increase in calcium deposits was observed in the placentas of women with VI (72.58% VI/53.84% HC). This study revealed the existence of cellular damage in the placental villi of mothers with VI with tissue implications such as increased calcification.


Assuntos
Calcinose/metabolismo , Proteínas do Olho/análise , Fatores de Crescimento Neural/análise , Placenta , Complicações Cardiovasculares na Gravidez/metabolismo , Serpinas/análise , Fator A de Crescimento do Endotélio Vascular/análise , Insuficiência Venosa/metabolismo , Adolescente , Adulto , Calcinose/fisiopatologia , Proteínas do Olho/química , Proteínas do Olho/metabolismo , Feminino , Humanos , Extremidade Inferior/fisiopatologia , Fatores de Crescimento Neural/química , Fatores de Crescimento Neural/metabolismo , Placenta/irrigação sanguínea , Placenta/química , Placenta/patologia , Gravidez , Complicações Cardiovasculares na Gravidez/fisiopatologia , Estudos Prospectivos , Serpinas/química , Serpinas/metabolismo , Regulação para Cima , Fator A de Crescimento do Endotélio Vascular/química , Fator A de Crescimento do Endotélio Vascular/metabolismo , Insuficiência Venosa/fisiopatologia , Adulto Jovem
17.
J Colloid Interface Sci ; 555: 647-654, 2019 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-31408763

RESUMO

Squamous cell carcinoma antigen (SCCA) has great diagnostic values for its high specificity (90-96%) in diagnosis of squamous cell carcinoma especially cancers, where electrochemical immunoassay is powerful for its accurate and reliable detection of specific tumor biomarkers at very low levels. In this work, well-defined three-dimensional dendritic core-shell rhodium@platinum-cobalt nanocrystals (Rh@PtCo NCs) were facilely prepared via a simple one-pot solvothermal strategy. Taking advantage of dramatically enhanced catalytic activity of the Rh@PtCo NCs for catechol oxidation, a label-free electrochemical immunosensor was developed for highly sensitive assay of SCCA. Under optimal conditions, the electrochemical signals were linearly correlated with the logarithm of the SCCA concentration, showing a broad linear range from 0.0001 to 10.0 ng mL-1 and a ultralow detection limit of 0.04 pg mL-1 (S/N = 3). The resultant immunosensor was further exploited for actual analysis of serum samples with convinced results. This immunosensor has good expectation in actual samples analysis and provides a universal approach for detection of other tumor markers and disease surveillance.


Assuntos
Antígenos de Neoplasias/análise , Cobalto/química , Técnicas Eletroquímicas , Imunoensaio , Nanopartículas Metálicas/química , Platina/química , Ródio/química , Serpinas/análise , Dendritos/química , Humanos , Tamanho da Partícula , Propriedades de Superfície
18.
Biosens Bioelectron ; 142: 111540, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31376714

RESUMO

Metal-organic framework nanocrystal (Zn-MOF) was synthesized by using 3,3'-{(propane-1,3-diyl)bis[1-(4-carboxybenzyl)-1H-imidazol-3-ium]} hexafluorophosphate ionic liquid as the functional monomer and Zn2+ as the central metal ion under hydrothermal conditions. Spatially confined gold nanoparticles (Au-NP) were prepared by in-situ reduction of chloroauric acid in the nanopores of Zn-MOF using acetic acid as the reducing agent to fabricate Au-NP@Zn-MOF nanocomposites. Au-NPs@Zn-MOF was further functionalized with 1H-imidazolium-1,3-bis(2-aminoethyl)bromide ionic liquid (IBABr) to prepare IBABr-Au@Zn-MOF nanocomposites. All abovementioned nanomaterials were thoroughly characterized by TEM, SEM, XPS, FTIR, and nitrogen-adsorption surface area analysis. IBABr-Au@Zn-MOFnanocomposites were then deposited onto a glassy carbon electrode and used as the photoactive element to fabricate a label-free photoelectrochemical (PEC) immunosensor by immobilizing anti-squamous cell carcinoma antigen (anti-SCCA). The PEC sensing principle is based on the photocurrent decline due to the blocking effect of SCCA on the electron and mass transfer after binding SCCA to anti-SCCA. The photocurrent variation related to the specific recognition of SCCA shows a linear relationship to the logarithm of SCCA concentration in the range of 5.0 pg mL-1 to 15.0 ng mL-1. The detection limit is as low as 2.34 pg mL-1. Such a signal-off PEC immunosensor is highly selective, sensitive, stable, and reproducible towards SCCA detection. Its performance is comparable to enzyme-linked immunosorbent assay from the studies on clinical samples. This immunosensor is promising for the label-free determining SCCA in clinical human serum samples.


Assuntos
Antígenos de Neoplasias/análise , Técnicas Biossensoriais/métodos , Líquidos Iônicos/química , Estruturas Metalorgânicas/química , Serpinas/análise , Zinco/química , Anticorpos Imobilizados/química , Antígenos de Neoplasias/sangue , Técnicas Eletroquímicas/métodos , Ouro/química , Humanos , Imunoensaio/métodos , Limite de Detecção , Nanopartículas Metálicas/química , Processos Fotoquímicos , Serpinas/sangue
19.
Biotechnol Appl Biochem ; 66(6): 939-944, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31468573

RESUMO

Ovarian cancer starts in the ovaries in its earlier stages and then spreads to the pelvis, uterus, and abdominal region. The success of an ovarian cancer treatment depends on the stage of the cancer and the diagnostic system. Squamous cell carcinoma antigen (SCC-Ag) is one of the most efficient cancer biomarkers, and elevated levels of SCC-Ag in ovarian cancer cells have been used to identify ovarian cancer. Carbon is a potential material for biosensing applications due to its thermal, electrical, and physical properties. Multiwalled carbon nanotubes (MWCNTs) are carbon-based materials that can be used here to detect SCC-Ag. Anti-SCC-Ag antibody was immobilized on the amine-modified MWCNT dielectric sensing surface to detect SCC-Ag. The uniformity of the surface structure was measured with a 3D nanoprofiler, and the results confirmed the detection of SCC-Ag at ∼80 pM. The specific detection of SCC-Ag was confirmed with two control proteins (factor IX and human serum albumin), and the system did not show biofouling. This experimental set-up with MWCNTs a dielectric sensing surface can lead to the detection of ovarian cancer in its initial stages.


Assuntos
Antígenos de Neoplasias/análise , Biomarcadores Tumorais/análise , Técnicas Biossensoriais , Técnicas Eletroquímicas , Nanotubos de Carbono/química , Neoplasias Ovarianas/diagnóstico , Serpinas/análise , Eletrodos , Feminino , Humanos , Propriedades de Superfície
20.
Arch Dermatol Res ; 311(8): 637-641, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31292731

RESUMO

Vaspin is a serine protease inhibitor of the serpin family which has an anti-inflammatory effect. It has an important role in the pathogenesis of some inflammatory diseases such as psoriasis. There are no previous studies comparing the effect of narrowband ultraviolet B (NB-UVB) radiation on tissue vaspin levels in psoriasis. So we aimed in this case-control study to estimate the possible role of vaspin in the pathogenesis of psoriasis, and to evaluate the effect of NB-UVB radiation on tissue vaspin in psoriasis. This study included 21 non-obese patients with moderate psoriasis and 20 non-obese clinically healthy age and sex matched controls. Patients underwent 24 sessions of NB-UVB radiation. A 4 mm punch skin biopsy was taken from all patients before and after treatment and from the controls for estimation of tissue vaspin level by enzyme-linked immunosorbent assay. Vaspin levels was significantly lower in patients before NB-UVB (99.72 pg/mg ± 12.11 pg/mg) compared to controls (257.34 pg/mg ± 28.11 pg/mg) with (P < 0.001). In addition, high significant difference was detected between vaspin levels in patients before (99.72 pg/mg ± 12.39 pg/mg) and after NB-UVB (190.92 pg/mg ± 27.61 pg/mg) with (P < 0.001). In conclusion, improvement of psoriatic plaques by NB-UVB is associated with an upregulation of tissue vaspin levels. Therefore, we suggest that vaspin has an important role in psoriasis pathogenesis.


Assuntos
Psoríase/patologia , Serpinas/metabolismo , Pele/patologia , Terapia Ultravioleta/métodos , Adulto , Biópsia , Estudos de Casos e Controles , Regulação para Baixo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Psoríase/radioterapia , Serpinas/análise , Pele/efeitos da radiação , Adulto Jovem
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