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1.
J Cancer Res Clin Oncol ; 149(18): 16391-16406, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37707574

RESUMO

BACKGROUND: Ovarian cancer (OC) is a prevalent gynecological malignancy with the highest mortality rate, which generally diagnosed at late stages due to the lack of effective early screening methods and the nonspecific symptoms. Hence, here we aim to identify new metastasis markers and develop a novel detection method with the characteristics of high sensitivity, rapid detection, high specificity, and low cost when compared with other conventional detection technologies. METHODS: Blood from OC patients with or without metastasis were collected and analyzed by 4D Label free LC - MS/MS. Surgically resect samples from OC patients were collected for Single cell RNA sequencing (sc-RNA seq). Short hairpin RNA (shRNA) was used to silence SAA1 expression in SKOV3 and ID8 to verify the relationship between endogenous SAA1 and tumor invasion or metastasis. The functional graphene chips prepared by covalent binding were used for SAA1 detection. RESULTS: In our study, we identified Serum Amyloid A1 (SAA1) as a hematological marker of OC metastasis by comprehensive analysis of proteins in plasma from OC patients with or without metastasis using 4D Label free LC - MS/MS and gene expression patterns from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) databases. Further validation using tumor tissues and plasma from human OC and mouse OC model confirmed the correlation between SAA1 and tumor metastasis. Importantly, sc-RNA seq of human OC samples revealed that SAA1 was specifically expressed in tumor cells and upregulated in the metastasis group. The functional role of SAA1 in metastasis was demonstrated through experiments in vitro and in vivo. Based on these findings, we designed and investigated a graphene-based platform for SAA1 detection to predict the risk of metastasis of OC patients. CONCLUSION: Our study suggests that SAA1 is a biomarker of OC metastasis, and we have developed a rapid and highly sensitive platform using graphene chips to detection of plasma SAA1 for the early assessment of metastasis in OC patients.


Assuntos
Grafite , Neoplasias Ovarianas , Animais , Camundongos , Humanos , Feminino , Espectrometria de Massas em Tandem , Neoplasias Ovarianas/diagnóstico , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , RNA Interferente Pequeno/metabolismo , Proteínas , Proteína Amiloide A Sérica/genética , Proteína Amiloide A Sérica/metabolismo
2.
PLoS One ; 14(6): e0217633, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31211768

RESUMO

Acute pancreatitis (AP) is acute inflammation of the pancreas, mainly caused by gallstones and alcohol, driven by changes in communication between cells. Heparin-binding proteins (HBPs) play a central role in health and diseases. Therefore, we used heparin affinity proteomics to identify extracellular HBPs in pancreas and plasma of normal mice and in a caerulein mouse model of AP. Many new extracellular HBPs (360) were discovered in the pancreas, taking the total number of HBPs known to 786. Extracellular pancreas HBPs form highly interconnected protein-protein interaction networks in both normal pancreas (NP) and AP. Thus, HBPs represent an important set of extracellular proteins with significant regulatory potential in the pancreas. HBPs in NP are associated with biological functions such as molecular transport and cellular movement that underlie pancreatic homeostasis. However, in AP HBPs are associated with additional inflammatory processes such as acute phase response signalling, complement activation and mitochondrial dysfunction, which has a central role in the development of AP. Plasma HBPs in AP included known AP biomarkers such as serum amyloid A, as well as emerging targets such as histone H2A. Other HBPs such as alpha 2-HS glycoprotein (AHSG) and histidine-rich glycoprotein (HRG) need further investigation for potential applications in the management of AP. Pancreas HBPs are extracellular and so easily accessible and are potential drug targets in AP, whereas plasma HBPs represent potential biomarkers for AP. Thus, their identification paves the way to determine which HBPs may have potential applications in the management of AP.


Assuntos
Biomarcadores/sangue , Pancreatite/genética , Proteoma/genética , alfa-2-Glicoproteína-HS/genética , Animais , Modelos Animais de Doenças , Heparina/genética , Homeostase , Humanos , Camundongos , Pâncreas/metabolismo , Pâncreas/patologia , Pancreatite/sangue , Pancreatite/patologia , Ligação Proteica/genética , Proteínas/genética , Proteômica/métodos , Proteína Amiloide A Sérica/metabolismo
3.
Int J Mol Sci ; 19(8)2018 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-30065196

RESUMO

Despite many years of studies, ovarian cancer remains one of the top ten cancers worldwide. Its high mortality rate is mainly due to lack of sufficient diagnostic methods. For this reason, our research focused on the identification of blood markers whose appearance would precede the clinical manifestation of the disease. ITRAQ-tagging (isobaric Tags for Relative and Absolute Quantification) coupled with mass spectrometry technology was applied. Three groups of samples derived from patients with: ovarian cancer, benign ovarian tumor, and healthy controls, were examined. Mass spectrometry analysis allowed for highlighting the dysregulation of several proteins associated with ovarian cancer. Further validation of the obtained results indicated that five proteins (Serotransferrin, Amyloid A1, Hemopexin, C-reactive protein, Albumin) were differentially expressed in ovarian cancer group. Interestingly, the addition of Albumin, Serotransferrin, and Amyloid A1 to CA125 (cancer antigen 125) and HE4 (human epididymis protein4) improved the diagnostic performance of the model discriminating between benign and malignant tumors. Identified proteins shed light on the molecular signaling pathways that are associated with ovarian cancer development and should be further investigated in future studies. Our findings indicate five proteins with a strong potential to use in a multimarker test for screening and detection of ovarian cancer.


Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias Ovarianas/metabolismo , Proteômica/métodos , Antígeno Ca-125/metabolismo , Feminino , Humanos , Proteínas/metabolismo , Proteína Amiloide A Sérica/metabolismo , Transferrina/metabolismo , Proteína 2 do Domínio Central WAP de Quatro Dissulfetos
4.
J Chem Theory Comput ; 12(11): 5656-5666, 2016 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-27767301

RESUMO

Recent experiments suggest that an amino acid sequence encodes not only the native fold of a protein but also other forms that are essential for its function or are important during folding or association. These various forms populate a multifunnel folding and association landscape where mutations, changes in environment, or interaction with other molecules switch between the encoded folds. We introduce replica exchange with tunneling as a way to efficiently simulate switching between distinct folds of proteins and protein aggregates. The correctness and efficiency of our approach are demonstrated in a series of simulations covering a wide range of proteins, from a small 11-residue large designed peptide to two 56-residue large mutants of the A and B domains of protein G.


Assuntos
Proteínas/química , Sequência de Aminoácidos , Simulação de Dinâmica Molecular , Dobramento de Proteína , Proteínas/metabolismo , Proteína Amiloide A Sérica/química , Proteína Amiloide A Sérica/metabolismo , Temperatura , Termodinâmica
5.
Acta Pharmacol Sin ; 37(11): 1458-1466, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27546006

RESUMO

AIM: Metrnl is a novel secreted protein, but its physiological roles remain elusive. In this study, we investigated the tissue expression pattern of Metrnl in humans and explored its possible physiological role in the tissues with most highly expressed levels. METHODS: A human tissue microarray containing 19 types of tissues from 69 donors was used to examine the tissue expression pattern of Metrnl, and the expression pattern was further verified in fresh human and mouse tissues. Intestinal epithelial cell-specific Metrnl knockout mice were generated, which were used to analyze the physiological roles of Metrnl. RESULTS: Metrnl was the most highly expressed in the human gastrointestinal tract, and was specifically expressed in the intestinal epithelium. Consistent with this, Metrnl mRNA was also most highly expressed in the mouse gastrointestinal tract among the 14 types of tissues tested. In the intestinal epithelial cell-specific Metrnl knockout mice, the Metrnl levels in the gut fluid were significantly reduced, whereas the Metrnl serum levels showed a trend towards a reduction, but this change was not statistically significant. This cell-specific deletion of Metrnl did not affect body weight, food intake, blood glucose, colon length and histology, intestinal permeability, mucus content or mucin 2 expression under physiological conditions, but statistically decreased the expression of antimicrobial peptides, such as regenerating islet-derived 3 gamma (Reg3g) and lactotransferrin. CONCLUSION: Metrnl is highly expressed in the intestinal epithelial cells of humans and mice, which mainly contributes to the local gut Metrnl levels and affects the serum Metrnl level to a lesser extent. Metrnl plays a role in maintaining gut antimicrobial peptides.


Assuntos
Adipocinas/metabolismo , Peptídeos Catiônicos Antimicrobianos/metabolismo , Mucosa Intestinal/metabolismo , Fatores de Crescimento Neural/metabolismo , Adulto , Idoso , Animais , Colo/metabolismo , Células Epiteliais/metabolismo , Feminino , Humanos , Lactoferrina/metabolismo , Masculino , Camundongos Knockout , Pessoa de Meia-Idade , Fatores de Crescimento Neural/genética , Especificidade de Órgãos , Proteínas Associadas a Pancreatite , Proteínas/metabolismo , Proteína Amiloide A Sérica/metabolismo , Análise Serial de Tecidos
6.
J Alzheimers Dis ; 46(4): 947-61, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25881911

RESUMO

BACKGROUND: Four previously reported studies have tested for association of blood proteins with neocortical amyloid-ß burden (NAB). If shown to be robust, these proteins could have utility as a blood test for enrichment in clinical trials of Alzheimer's disease (AD) therapeutics. OBJECTIVE: This study aimed to investigate whether previously identified blood proteins also show evidence for association with NAB in serum samples from the Australian Imaging, Biomarker and Lifestyle Flagship Study of Ageing (AIBL). The study considers candidate proteins seen in cohorts other than AIBL and candidates previously discovered in the AIBL cohort. METHODS: Our study used the SOMAscan platform for protein quantification in blood serum. Linear and logistic regressions were used to model continuous NAB and dichotomized NAB respectively using single proteins as a predictor. Multiple protein models were built using stepwise regression techniques and support vectors machines. Age and APOEɛ4 carriage were used as covariates for all analysis. RESULTS: Of the 41 proteins previously reported, 15 AIBL candidates and 20 non-AIBL candidates were available for testing. Of these candidates, pancreatic polypeptide (PPY) and IgM showed a significant association with NAB. Notably, IgM was found to associate with continuous NAB across cognitively normal control subjects. CONCLUSIONS: We have further demonstrated the association of PPY and IgM with NAB, despite technical differences between studies. There are several reasons for a lack of significance for the other candidates including platform differences and the use of serum rather than plasma samples. To investigate the possibility of technical differences causing lack of replication, further studies are required.


Assuntos
Doença de Alzheimer/sangue , Doença de Alzheimer/patologia , Peptídeos beta-Amiloides/metabolismo , Proteínas Sanguíneas/metabolismo , Neocórtex/metabolismo , Idoso , Idoso de 80 Anos ou mais , Envelhecimento/sangue , Envelhecimento/patologia , Compostos de Anilina/metabolismo , Apolipoproteínas E/genética , Austrália , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tomografia por Emissão de Pósitrons , Proteínas , Proteômica , Tiazóis/metabolismo
7.
Tumour Biol ; 34(5): 2645-50, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23640061

RESUMO

The aim of this study was to evaluate the prognostic and predictive efficacy of the human epididymis secretory protein 4 (HE4) and serum amyloid-A (S-AA) together with the other tumor markers (CA 125, CA 15-3, CEA, and CA 19-9) in endometrial cancer patients. The study group consisted of 64 patients with defined stage and grade of endometrial cancer and 60 women with benign uterine diseases. Thirty-four healthy women were defined as the control group. Fasting blood samples were collected prior to surgery and tumor marker levels were determined in blood samples by E170 autoanalyzer. S-AA concentrations were measured by particle-enhanced immunonephelometry. Preoperative serum HE4 and S-AA levels were significantly higher in endometrial cancer patients than in controls, whereas the other measured parameters were not significantly different. Serum levels of HE4 were related to both the stage and grade of tumor. The best cutoff point for HE4 was determined to be 59.7 pmol/L; with 75 % sensitivity and 65.5 % specificity. For S-AA, the cutoff point was 8.8 U/mL, with 68.7 % sensitivity and 58.6 % specificity. The combination of HE4, CA 125, CEA, and S-AA raised the sensitivity to 84 %. Preoperative measurement of serum HE4 and S-AA may be of help in early detection of endometrial cancer. Preoperative screening with these markers may provide important information about the patient's outcome and prognosis.


Assuntos
Adenocarcinoma/metabolismo , Biomarcadores Tumorais/sangue , Neoplasias do Endométrio/metabolismo , Proteínas/metabolismo , Proteína Amiloide A Sérica/metabolismo , Adenocarcinoma/patologia , Adulto , Antígeno Ca-125/sangue , Estudos de Casos e Controles , Diagnóstico Diferencial , Neoplasias do Endométrio/patologia , Feminino , Humanos , Proteínas de Membrana/sangue , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Curva ROC , Proteína 2 do Domínio Central WAP de Quatro Dissulfetos
8.
Anal Biochem ; 434(1): 178-80, 2013 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-23201266

RESUMO

The Bradford assay is a simple method for protein quantitation, but variation in the results between proteins is a matter of concern. In this study, we compared and normalized quantitative values from two models for protein quantitation, where the residues in the protein that bind to anionic Coomassie Brilliant Blue G-250 comprise either Arg and Lys (Method 1, M1) or Arg, Lys, and His (Method 2, M2). Use of the M2 model yielded much more consistent quantitation values compared with use of the M1 model, which exhibited marked overestimations against protein standards.


Assuntos
Modelos Teóricos , Proteínas/análise , Corantes de Rosanilina/química , Espectrofotometria , Peptídeos beta-Amiloides/química , Peptídeos beta-Amiloides/metabolismo , Animais , Arginina/química , Bovinos , Histidina/química , Humanos , Insulinas/química , Insulinas/metabolismo , Lisina/química , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , Ligação Proteica , Proteínas/metabolismo , Ratos , Corantes de Rosanilina/metabolismo , Albumina Sérica/química , Albumina Sérica/metabolismo
9.
J Med Chem ; 55(24): 10823-43, 2012 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-23075044

RESUMO

Protein misfolding is a process in which proteins are unable to attain or maintain their biologically active conformation. Factors contributing to protein misfolding include missense mutations and intracellular factors such as pH changes, oxidative stress, or metal ions. Protein misfolding is linked to a large number of diseases such as cystic fibrosis, Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and less familiar diseases such as Gaucher's disease, nephrogenic diabetes insipidus, and Creutzfeldt-Jakob disease. In this Perspective, we report on small molecules that bind to and stabilize the aberrant protein, thereby helping it to attain a native or near-native conformation and restoring its function. The following targets will be specifically discussed: transthyretin, p53, superoxide dismutase 1, lysozyme, serum amyloid A, prions, vasopressin receptor 2, and α-1-antitrypsin.


Assuntos
Doenças Neurodegenerativas/tratamento farmacológico , Dobramento de Proteína , Proteínas/química , Proteínas/fisiologia , Deficiências na Proteostase/tratamento farmacológico , Bibliotecas de Moléculas Pequenas/química , Amiloide/metabolismo , Animais , Humanos , Modelos Moleculares , Muramidase/química , Muramidase/fisiologia , Mutação , Doenças Neurodegenerativas/metabolismo , Pré-Albumina/química , Pré-Albumina/fisiologia , Príons/química , Príons/fisiologia , Ligação Proteica , Conformação Proteica , Deficiências na Proteostase/metabolismo , Receptores de Vasopressinas/química , Receptores de Vasopressinas/fisiologia , Proteína Amiloide A Sérica/química , Proteína Amiloide A Sérica/fisiologia , Bibliotecas de Moléculas Pequenas/farmacologia , Superóxido Dismutase/química , Superóxido Dismutase/fisiologia , Superóxido Dismutase-1 , Proteína Supressora de Tumor p53/química , Proteína Supressora de Tumor p53/fisiologia , Resposta a Proteínas não Dobradas , alfa 1-Antitripsina/química , alfa 1-Antitripsina/fisiologia
10.
Equine Vet J Suppl ; (43): 12-6, 2012 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23447871

RESUMO

REASONS FOR PERFORMING STUDY: Serum amyloid A (SAA) in synovial fluid has recently been used as a marker for septic arthritis in horses but the effects of repeated intra-articular (IA) administration of amikacin on synovial SAA concentrations are unknown. OBJECTIVES: To report the effect of repeated IA administration of amikacin on SAA, total protein (TP), nucleated cell count (NCC) and differential NCC in synovial fluid of healthy equine joints. METHODS: A controlled, 2 period crossover study was performed on 5 clinically healthy horses. Each intercarpal joint received one of 2 treatments every 48 h for 5 consecutive times: arthrocentesis alone (control group) or arthrocentesis combined with IA administration of 500 mg of amikacin (treatment group). Clinical and lameness examinations were performed daily. Serum SAA and synovial SAA, TP, NCC and differential NCC were measured and statistically compared. Significance level was set at P < 0.05. RESULTS: Horses remained healthy and nonlame throughout the study. Baseline values for all variables were not significantly different between groups. Values for TP in the treatment group were significantly higher than in the control group after the first sample (P < 0.05). In both groups NCC increased significantly (P < 0.05) after the first sample. No significant changes were identified in differential NCC. In both groups, all synovial and most serum SAA concentrations remained below the lower limit of quantification. CONCLUSIONS: Repeated IA administration of amikacin caused increased values of TP and NCC in synovial fluid, with some TP concentrations falling within the range reported for septic arthritis. In contrast, synovial SAA concentrations did not increase in either group. POTENTIAL RELEVANCE: Synovial SAA could serve as a more reliable marker than TP and NCC when evaluating a joint previously sampled or treated with amikacin.


Assuntos
Amicacina/efeitos adversos , Artropatias/veterinária , Proteínas/metabolismo , Proteína Amiloide A Sérica/metabolismo , Líquido Sinovial/química , Líquido Sinovial/citologia , Amicacina/administração & dosagem , Animais , Estudos Cross-Over , Feminino , Doenças dos Cavalos/induzido quimicamente , Cavalos , Injeções Intra-Articulares , Artropatias/induzido quimicamente , Proteínas/química , Proteína Amiloide A Sérica/química
11.
J Nephrol ; 16(3): 431-4, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12832747

RESUMO

Familial Mediterranean fever (FMF) is an autosomal recessive disease, which primarily affects the population surrounding the Mediterranean basin. It is characterized by recurrent attacks of fever and peritonitis, pleuritis, arthritis or erysipelas-like erythema. Amyloidosis, causing renal failure, is one of the most severe complications of the disease. The gene associated with FMF (MEFV) has been recently isolated. Phenotype-genotype correlation studies revealed that amyloidosis was more common in FMF patients originating from North-Africa who were homozygous for the M694V mutation. Such a correlation was not found in Turkish patients. The risk of amyloidosis is increased in male FMF patients and in patients bearing polymorphism a/a in the SAA1 gene. Colchicine is the chosen drug for the treatment of FMF and can prevent amyloidosis.


Assuntos
Amiloidose/genética , Amiloidose/metabolismo , Rim/metabolismo , Proteína Amiloide A Sérica/metabolismo , Amiloidose/terapia , Proteínas do Citoesqueleto , Febre Familiar do Mediterrâneo , Genótipo , Humanos , Mutação , Fenótipo , Prognóstico , Proteínas/genética , Pirina
12.
Immunology ; 109(1): 127-36, 2003 May.
Artigo em Inglês | MEDLINE | ID: mdl-12709026

RESUMO

We have previously reported that the expressions of TLR2 and TLR4 mRNA are differentially regulated in mouse liver and in the parenchymal cells. In the present study, we investigated the mechanism of the up-regulatory effects of interleukin-1alpha (IL-1alpha), tumour necrosis factor-alpha (TNF-alpha), lipopolysaccharide (LPS), or bacterial lipoprotein (BLP) on TLR2 mRNA expression in primary cultured murine hepatocytes. Although TLR2 mRNA stability was not affected, these treatments enhanced NF-kappaB activity and TLR2 gene transcription simultaneously. The up-regulation of TLR2 transcription in response to these reagents was completely inhibited by blocking the NF-kappaB activation pathway, demonstrating a pivotal role of NF-kappaB activation in the regulation of hepatocyte TLR2 transcription. The expression of TLR2 protein by hepatocytes was also remarkably up-regulated by IL-1alpha and, to a lesser extent, by TNF-alpha as well, but not by LPS or BLP. In addition, pretreatment of mice with IL-1alpha markedly increased the BLP (a ligand for TLR2)-induced serum level of serum amyloid A (SAA), an acute-phase protein predominantly produced by hepatocytes, indicating that IL-1alpha may also up-regulate functional TLR2 in vivo. These results demonstrate that IL-1alpha, through activating the TRAF6-NF-kappaB pathway, serves as the most potent inducer for TLR2 up-regulation, and plays an important role in the regulation of hepatocyte functions by augmenting the hepatocyte response to bacteria or bacterial products.


Assuntos
Hepatócitos/imunologia , Interleucina-1/imunologia , Glicoproteínas de Membrana/metabolismo , NF-kappa B/imunologia , Proteínas/imunologia , Receptores de Superfície Celular/metabolismo , Animais , Proteínas de Bactérias/imunologia , Células Cultivadas , Feminino , Ligantes , Lipopolissacarídeos/imunologia , Glicoproteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos ICR , RNA Mensageiro/genética , Receptores de Superfície Celular/genética , Proteína Amiloide A Sérica/biossíntese , Fator 6 Associado a Receptor de TNF , Receptor 2 Toll-Like , Receptor 4 Toll-Like , Receptores Toll-Like , Fator de Necrose Tumoral alfa/imunologia , Regulação para Cima/imunologia
13.
Arthritis Rheum ; 48(4): 1149-55, 2003 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-12687559

RESUMO

OBJECTIVE: The clinical profile in familial Mediterranean fever (FMF), including its major manifestation, amyloidosis, is influenced by MEFV allelic heterogeneity and other genetic and/or environmental factors. In this study, we analyzed the contribution of genotypes at the MEFV and SAA1 loci to disease severity and to the development of amyloidosis, and further defined the factors affecting the clinical profile of FMF. METHODS: We investigated a sample of 277 FMF patients (154 men and 123 women), including 62 patients with nephropathic amyloidosis, in whom both FMF alleles had been identified. A detailed chart review, interview, and physical examination were undertaken to determine the patients' demographic characteristics, medical history, clinical manifestations, and treatment. The disease severity score was calculated from the Tel-Hashomer key. Genotypes at the SAA1 locus (isoforms alpha, beta, and gamma) were determined in all patients. The SAA1 13C/T polymorphism of the SAA1 promotor was analyzed in a subset of cases. RESULTS: The male:female ratio (154:123, or 1.3) was higher among patients with amyloidosis (40:22, or 1.8) compared with patients without amyloidosis (114:101, or 1.1). Logistic regression analysis showed that homozygosity for the M694V allele (odds ratio [OR] 4.27, 95% confidence interval [95% CI] 2.01-9.07), the presence of the SAAalpha/alpha genotype (OR 2.99, 95% CI 1.47-6.09), the occurrence of arthritis attacks (OR 2.43, 95% CI 1.17-5.06), and male sex (OR 1.73, 95% CI 0.90-3.33) were significantly and independently associated with renal amyloidosis. Disease severity was mainly influenced by MEFV mutations and was not associated with genotypes at the SAA1 locus. The SAA1 13T allele was rare, being associated mainly with the SAA gamma isoform, and not related to renal amyloidosis. CONCLUSION: Overall, disease severity and the development of amyloidosis in FMF are differentially affected by genetic variations within and outside the MEFV gene.


Assuntos
Amiloidose/genética , Febre Familiar do Mediterrâneo/genética , Predisposição Genética para Doença , Proteínas/genética , Proteína Amiloide A Sérica/metabolismo , Amiloidose/complicações , Amiloidose/patologia , Proteínas do Citoesqueleto , Análise Mutacional de DNA , Febre Familiar do Mediterrâneo/complicações , Febre Familiar do Mediterrâneo/patologia , Feminino , Genótipo , Humanos , Masculino , Reação em Cadeia da Polimerase , Pirina
15.
Ann Rheum Dis ; 60(8): 777-80, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11454642

RESUMO

BACKGROUND: Familial Mediterranean fever (FMF) is a periodic febrile disorder, characterised by fever and serositis. The acute phase response during attacks of FMF results from the release of cytokines, which in turn induce increased expression and changed glycosylation of acute phase proteins. A recent study indicated that attacks in FMF are accompanied by a rise of plasma concentrations of serum amyloid A (SAA) and C reactive protein (CRP), which remain significantly raised during remission relative to healthy controls. Another study suggested that obligatory heterozygotes also display an inflammatory acute phase response. OBJECTIVE: To determine the state of inflammation in homozygotic and heterozygotic MEFV genotypes. METHODS: CRP and SAA were studied by enzyme linked immunosorbent assay (ELISA). The glycosylation of the acute phase protein, alpha(1)-acid glycoprotein (AGP), was visualised with crossed affinoimmunoelectrophoresis with concanavalin A as diantennary glycan-specific component and Aleuria aurantia lectin as fucose-specific affinity component. RESULTS: FMF attacks were associated with an increase (p<0.05) in the serum inflammation parameters CRP, SAA, and AGP. The glycosylation of AGP showed an increase (p<0.05) in fucosylated AGP glycoforms, whereas the branching of the glycans remained unaffected. The glycosylation of AGP in the MEFV carrier group, compared with that in a healthy control group, was characterised by a significant increase (p<0.05) in branching of the glycans, whereas the fucosylation remained unaffected. CONCLUSION: The findings suggest an FMF-specific release of cytokines, resulting in a different glycosylation of AGP between a homozygotic and heterozygotic MEFV genotype.


Assuntos
Febre Familiar do Mediterrâneo/metabolismo , Heterozigoto , Orosomucoide/metabolismo , Proteínas/genética , Adolescente , Adulto , Proteína C-Reativa/análise , Estudos de Casos e Controles , Proteínas do Citoesqueleto , Eletroforese em Gel Bidimensional , Ensaio de Imunoadsorção Enzimática/métodos , Febre Familiar do Mediterrâneo/genética , Feminino , Glicosilação , Homozigoto , Humanos , Masculino , Pessoa de Meia-Idade , Distribuição Normal , Pirina , Proteína Amiloide A Sérica/análise , Estatísticas não Paramétricas
16.
Biochem Biophys Res Commun ; 264(2): 395-403, 1999 Oct 22.
Artigo em Inglês | MEDLINE | ID: mdl-10529375

RESUMO

Serum amyloid A (SAA) is a major acute-phase protein whose expression can be dramatically induced in response to tissue damage, infection, and inflammation. Its expression is highly tissue-specific, restricted almost exclusively to liver hepatocytes. We have shown that a 320-bp fragment of the rat SAA1 promoter could confer liver-cell-specific expression on a reporter gene when transfected into cultured cells. Here we report the identification of a 29-bp regulatory element that possesses inhibitory activities on SAA1 promoter in HeLa cells but has no such effects in liver cells. Moreover, this regulatory element has properties of a transcriptional repressor; in that, it can function with a heterologous promoter and is independent of orientation and distance from the transcription initiation site. Protein binding studies showed that this regulatory element can form specific protein-DNA complexes with nuclear proteins from several nonliver cell lines (HeLa, 10T(1/2), and C2) and placenta. However, the same DNA-protein complex was not detected in extracts from liver or liver-derived cell lines (HepG2 and Hep3B). Taken together, our results demonstrate the presence of a DNA-binding protein, termed tissue-specific repressor, found only in nonhepatocytes which may function to repress SAA1 gene expression by interacting with a repressor element. Thus, liver-specific expression of the SAA1 gene is apparently regulated by both positive and negative regulatory elements and their interacting transcription factors to ensure that it is expressed only in suitable cell types.


Assuntos
Fígado/metabolismo , Proteínas/genética , Proteína Amiloide A Sérica/genética , Animais , Subunidade RIalfa da Proteína Quinase Dependente de AMP Cíclico , Proteínas Quinases Dependentes de AMP Cíclico , Regulação da Expressão Gênica , Células HeLa , Humanos , Plasmídeos , Regiões Promotoras Genéticas , Ratos , Proteína Amiloide A Sérica/biossíntese , Transcrição Gênica , Transfecção , Células Tumorais Cultivadas
17.
Biochem Biophys Res Commun ; 252(2): 492-6, 1998 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-9826558

RESUMO

Biosensor technology was employed to study the specific interactions of different lipopolysaccharide (LPS)-binding proteins and peptides with LPS, using an LPS-coated surface. Two methods to immobilize biotinylated LPS to streptavidin-coated sensor chips (SA-chips) were evaluated. Biotinylated LPS in PBS or biotinylated LPS, pretreated with EDTA and sodium-desoxycholate, were injected across an SA-chip, resulting in a 'high-' and 'low- mass' LPS chip, respectively. While the 'high mass' LPS chip appeared to be unstable, the 'low mass' LPS chip resulted in reproducible binding curves for bactericidal/permeability-increasing protein (rBPI21) with a binding affinity corresponding to the literature (Kd: 3.75 nM). New Kd values were obtained for serum amyloid P component (SAP, Kd: 3.9 nM), a recently discovered new LPS-binding protein, and cationic protein 18 (CAP18, Kd: 0.58 nM). Moreover, binding affinities of bioactive BPI- and SAP-derived peptides could be determined. This study shows for the first time the applicability of biosensor technology to study interactions of proteins and peptides with LPS, using an LPS-coated sensor chip.


Assuntos
Proteínas de Fase Aguda , Técnicas Biossensoriais , Lipopolissacarídeos/metabolismo , Glicoproteínas de Membrana , Proteínas de Membrana , Peptídeos/metabolismo , Proteínas/metabolismo , Sequência de Aminoácidos , Peptídeos Catiônicos Antimicrobianos , Proteínas Sanguíneas/genética , Proteínas Sanguíneas/metabolismo , Proteínas de Transporte/química , Proteínas de Transporte/metabolismo , Humanos , Técnicas In Vitro , Cinética , Dados de Sequência Molecular , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Peptídeos/genética , Ligação Proteica , Proteínas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Componente Amiloide P Sérico/genética , Componente Amiloide P Sérico/metabolismo , Estreptavidina
18.
Biochem Biophys Res Commun ; 242(3): 480-3, 1998 Jan 26.
Artigo em Inglês | MEDLINE | ID: mdl-9464241

RESUMO

Angiogenin (Ang), a potent mediator of neovascularization, is secreted by and is critical for the growth of human tumor cells in experimental animals. However, control mechanisms that regulate its expression under normal physiological conditions have not been described. We have determined previously that Ang is present in normal human serum and that its concentration, normally falling within a narrow range, can vary widely in hospitalized patients. This observation, plus a report that Ang is synthesized in the adult liver, led us to investigate whether it can be regulated as an acute phase protein (APP). Ang concentration in the serum of mice placed into the acute phase by injection with 3% thioglycollate do indeed increase transiently as is typical for APPs. Moreover, a liver-specific rise and subsequent fall in Ang mRNA transcripts also follows entrance into acute inflammation. We conclude that Ang can be regulated in vivo in a manner that is characteristic of an APP and, therefore, may contribute to the angiogenic component of tissue repair that accompanies host response to inflammation and trauma. To our knowledge, this is the first demonstration that a well-characterized angiogenic mediator can be regulated as an APP.


Assuntos
Proteínas de Fase Aguda/metabolismo , Fígado/metabolismo , Proteínas/metabolismo , Ribonuclease Pancreático , Animais , Northern Blotting , Regulação da Expressão Gênica/genética , Inflamação/induzido quimicamente , Camundongos , Biossíntese de Proteínas , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Radioimunoensaio , Proteína Amiloide A Sérica/metabolismo , Tioglicolatos/farmacologia , Fatores de Tempo
20.
Am J Kidney Dis ; 30(6): 923-7, 1997 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9398142

RESUMO

Hypoalbuminemia predicts mortality in dialysis patients. It has been postulated that hypoalbuminemia in the dialysis population is a consequence of poor protein intake resulting from inadequate dialysis. To establish the cause of hypoalbuminemia in a group of 27 patients on peritoneal dialysis (PD), we determined the relationship between serum albumin concentration and a group of parameters including dialysis dose delivered (Kt/V), normalized protein catabolic rate (PCRn), transperitoneal and urinary albumin losses, and the serum concentration of two acute-phase proteins, C-reactive protein (CRP), and serum amyloid A (SAA). Serum albumin concentration could be predicted by a combination of transperitoneal albumin loss and either the serum concentration of CRP or of SAA. There was no relationship between weekly Kt/V or PCRn and serum albumin concentration. CRP and SAA significantly correlated with one another, but neither correlated with transperitoneal albumin losses. Hypoalbuminemia in PD patients is a consequence of transperitoneal albumin losses and of the acute phase response.


Assuntos
Albuminas/análise , Proteína C-Reativa/análise , Soluções para Diálise/análise , Diálise Peritoneal , Albumina Sérica/análise , Proteína Amiloide A Sérica/análise , Reação de Fase Aguda/sangue , Reação de Fase Aguda/urina , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Albuminúria/urina , Soluções para Diálise/administração & dosagem , Proteínas na Dieta/administração & dosagem , Proteínas na Dieta/metabolismo , Feminino , Previsões , Humanos , Masculino , Pessoa de Meia-Idade , Peritônio/metabolismo , Proteínas/metabolismo , Taxa de Sobrevida
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