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1.
BMC Med ; 21(1): 90, 2023 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-36894970

RESUMO

BACKGROUND: Pre-eclampsia (PE) is one of the leading causes of maternal and fetal morbidity/mortality during pregnancy, and alpha-2-macroglobulin (A2M) is associated with inflammatory signaling; however, the pathophysiological mechanism by which A2M is involved in PE development is not yet understood. METHODS: Human placenta samples, serum, and corresponding clinical data of the participants were collected to study the pathophysiologic mechanism underlying PE. Pregnant Sprague-Dawley rats were intravenously injected with an adenovirus vector carrying A2M via the tail vein on gestational day (GD) 8.5. Human umbilical artery smooth muscle cells (HUASMCs), human umbilical vein endothelial cells (HUVECs), and HTR-8/SVneo cells were transfected with A2M-expressing adenovirus vectors. RESULTS: In this study, we demonstrated that A2M levels were significantly increased in PE patient serum, uterine spiral arteries, and feto-placental vasculature. The A2M-overexpression rat model closely mimicked the characteristics of PE (i.e., hypertension in mid-to-late gestation, histological and ultrastructural signs of renal damage, proteinuria, and fetal growth restriction). Compared to the normal group, A2M overexpression significantly enhanced uterine artery vascular resistance and impaired uterine spiral artery remodeling in both pregnant women with early-onset PE and in pregnant rats. We found that A2M overexpression was positively associated with HUASMC proliferation and negatively correlated with cell apoptosis. In addition, the results demonstrated that transforming growth factor beta 1 (TGFß1) signaling regulated the effects of A2M on vascular muscle cell proliferation described above. Meanwhile, A2M overexpression regressed rat placental vascularization and reduced the expression of angiogenesis-related genes. In addition, A2M overexpression reduced HUVEC migration, filopodia number/length, and tube formation. Furthermore, HIF-1α expression was positively related to A2M, and the secretion of sFLT-1 and PIGF of placental origin was closely related to PE during pregnancy or A2M overexpression in rats. CONCLUSIONS: Our data showed that gestational A2M overexpression can be considered a contributing factor leading to PE, causing detective uterine spiral artery remodeling and aberrant placental vascularization.


Assuntos
Placenta , Pré-Eclâmpsia , Animais , Feminino , Humanos , Gravidez , Ratos , Células Endoteliais/metabolismo , Macroglobulinas/metabolismo , Placenta/metabolismo , Fator de Crescimento Placentário/metabolismo , Ratos Sprague-Dawley , Artéria Uterina/metabolismo
2.
Sci Rep ; 13(1): 4579, 2023 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-36941303

RESUMO

Human α2-macroglobulin (hα2M) is a large homotetrameric protein involved in the broad inhibition of endopeptidases. Following cleavage within a bait region, hα2M undergoes stepwise transitions from its native, expanded, highly flexible, active conformation to an induced, compact, triggered conformation. As a consequence, the peptidase is entrapped by an irreversible Venus flytrap mechanism. Given the importance of hα2M, biochemical studies galore over more than seven decades have attempted to ascertain its role, typically using authentic hα2M purified from frozen and non-frozen fresh blood plasma, and even outdated plasma. However, hα2M is sensitive once isolated and purified, and becomes heterogeneous during storage and/or freezing, raising concerns about the functional competence of frozen plasma-derived hα2M. We therefore used a combination of native and sodium dodecylsulfate polyacrylamide gel electrophoresis, affinity and ion-exchange chromatography, multi-angle laser light scattering after size-exclusion chromatography, free cysteine quantification, and peptidase inhibition assays with endopeptidases of two catalytic classes and three protein substrates, to characterize the biochemical and biophysical properties of hα2M purified ad hoc either from fresh plasma or frozen fresh plasma after thawing. We found no differences in the molecular or functional properties of the preparations, indicating that protective components in plasma maintain native hα2M in a functionally competent state despite freezing.


Assuntos
Endopeptidases , Peptídeo Hidrolases , Plasma , Humanos , Congelamento , Macroglobulinas
3.
Mol Reprod Dev ; 89(10): 459-470, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35901249

RESUMO

The present study evaluated the effects of in vitro maturation (IVM) on the proteome of cumulus-oocyte complexes (COCs) from ewes. Extracted COC proteins were analyzed by LC-MS/MS. Differences in protein abundances (p < 0.05) and functional enrichments in immature versus in vitro-matured COCs were evaluated using bioinformatics tools. There were 2550 proteins identified in the COCs, with 89 and 87 proteins exclusive to immature and mature COCs, respectively. IVM caused downregulation of 84 and upregulation of 34 proteins. Major upregulated proteins in mature COCs were dopey_N domain-containing protein, structural maintenance of chromosomes protein, ubiquitin-like modifier-activating enzyme 2. Main downregulated proteins in mature COCs were immunoglobulin heavy constant mu, inter-alpha-trypsin inhibitor heavy chain 2, alpha-2-macroglobulin. Proteins exclusive to mature COCs and upregulated after IVM related to immune response, complement cascade, vesicle-mediated transport, cell cycle, and extracellular matrix organization. Proteins of immature COCs and downregulated after IVM were linked to metabolic processes, immune response, and complement cascade. KEGG pathways and miRNA-regulated genes attributed to downregulated and mature COC proteins related to complement and coagulation cascades, metabolism, humoral response, and B cell-mediated immunity. Thus, IVM influenced the ovine COC proteome. This knowledge supports the future development of efficient IVM protocols for Ovis aries.


Assuntos
Células do Cúmulo , MicroRNAs , Ovinos , Animais , Feminino , Células do Cúmulo/metabolismo , Proteoma/metabolismo , Carneiro Doméstico , Cromatografia Líquida , Espectrometria de Massas em Tandem , Oócitos/metabolismo , Ubiquitinas/metabolismo , Ubiquitinas/farmacologia , Imunoglobulinas/metabolismo , Macroglobulinas/metabolismo , Macroglobulinas/farmacologia , MicroRNAs/metabolismo , Técnicas de Maturação in Vitro de Oócitos/métodos
4.
Matrix Biol ; 114: 108-137, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-35618217

RESUMO

Extracellular matrices (ECMs) in the intervertebral disc (IVD), lung and artery are thought to undergo age-dependant accumulation of damage by chronic exposure to mechanisms such as reactive oxygen species, proteases and glycation. It is unknown whether this damage accumulation is species-dependant (via differing lifespans and hence cumulative exposures) or whether it can influence the progression of age-related diseases such as atherosclerosis. Peptide location fingerprinting (PLF) is a new proteomic analysis method, capable of the non-targeted identification of structure-associated changes within proteins. Here we applied PLF to publicly available ageing human IVD (outer annulus fibrosus), ageing mouse lung and human arterial atherosclerosis datasets and bioinformatically identified novel target proteins alongside common age-associated differences within protein structures which were conserved between three ECM-rich organs, two species, three IVD tissue regions, sexes and in an age-related disease. We identify peptide yield differences across protein structures which coincide with biological regions, potentially reflecting the functional consequences of ageing or atherosclerosis for macromolecular assemblies (collagen VI), enzyme/inhibitor activity (alpha-2 macroglobulin), activation states (complement C3) and interaction states (laminins, perlecan, fibronectin, filamin-A, collagen XIV and apolipoprotein-B). Furthermore, we show that alpha-2 macroglobulin and collagen XIV exhibit possible shared structural consequences in IVD ageing and arterial atherosclerosis, providing novel links between an age-related disease and intrinsic ageing. Crucially, we also demonstrate that fibronectin, laminin beta chains and filamin-A all exhibit conserved age-associated structural differences between mouse lung and human IVD, providing evidence that ECM, and their associating proteins, may be subjected to potentially similar mechanisms or consequences of ageing across both species, irrespective of differences in lifespan and tissue function.


Assuntos
Aterosclerose , Degeneração do Disco Intervertebral , Disco Intervertebral , Camundongos , Animais , Humanos , Fibronectinas/metabolismo , Filaminas/análise , Filaminas/metabolismo , Proteômica/métodos , Disco Intervertebral/metabolismo , Degeneração do Disco Intervertebral/metabolismo , Colágeno/metabolismo , Envelhecimento/metabolismo , Laminina/metabolismo , Peptídeos/metabolismo , Aterosclerose/genética , Aterosclerose/metabolismo , Macroglobulinas/análise , Macroglobulinas/metabolismo
5.
Am J Physiol Heart Circ Physiol ; 322(4): H537-H548, 2022 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-35089808

RESUMO

Both skin wound healing and the cardiac response to myocardial infarction (MI) progress through similar pathways involving inflammation, resolution, tissue repair, and scar formation. Due to the similarities, we hypothesized that the healing response to skin wounding would predict future response to MI. Mice were given a 3-mm skin wound using a disposable biopsy punch and the skin wound was imaged daily until closure. The same set of animals was given MI by permanent coronary artery ligation 28 days later and followed for 7 days. Cardiac physiology was measured by echocardiography at baseline and MI days 3 and 7. Animals that survived until day 7 were grouped as survivors, and animals that died from MI were grouped as nonsurvivors. Survivors had faster skin wound healing than nonsurvivors. Faster skin wound healing predicted MI survival better than commonly used cardiac functional variables (e.g., infarct size, fractional shortening, and end diastolic dimension). N-glycoproteome profiling of MI day 3 plasma revealed α2-macroglobulin and ELL-associated factor 1 as strong predictors of future MI death and progression to heart failure. A second cohort of MI mice validated these findings. To investigate the clinical relevance of α2-macroglobulin, we mapped the plasma glycoproteome in patients with MI 48 h after admission and in healthy controls. In patients, α2-macroglobulin was increased 48 h after MI. Apolipoprotein D, another plasma glycoprotein, detrimentally regulated both skin and cardiac wound healing in male but not female mice by promoting inflammation. Our results reveal that the skin is a mirror to the heart and common pathways link wound healing across organs.NEW & NOTEWORTHY Faster skin wound healers had more efficient cardiac healing after myocardial infarction (MI). Two plasma proteins at D3 MI, EAF1 and A2M, predicted MI death in 66% of cases. ApoD regulated both skin and cardiac wound healing in male mice by promoting inflammation. The skin was a mirror to the heart and common pathways linked wound healing across organs.


Assuntos
Infarto do Miocárdio , Remodelação Ventricular , Animais , Humanos , Inflamação/metabolismo , Macroglobulinas/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Miocárdio/metabolismo , Fatores de Transcrição/metabolismo , Cicatrização/fisiologia
6.
Int J Mol Sci ; 22(13)2021 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-34203120

RESUMO

Activated alpha-2 Macroglobulin (α2M*) is specifically recognized by the cluster I/II of LRP1 (Low-density lipoprotein Receptor-related Protein-1). LRP1 is a scaffold protein for insulin receptor involved in the insulin-induced glucose transporter type 4 (GLUT4) translocation to plasma membrane and glucose uptake in different types of cells. Moreover, the cluster II of LRP1 plays a critical role in the internalization of atherogenic lipoproteins, such as aggregated Low-density Lipoproteins (aggLDL), promoting intracellular cholesteryl ester (CE) accumulation mainly in arterial intima and myocardium. The aggLDL uptake by LRP1 impairs GLUT4 traffic and the insulin response in cardiomyocytes. However, the link between CE accumulation, insulin action, and cardiac dysfunction are largely unknown. Here, we found that α2M* increased GLUT4 expression on cell surface by Rab4, Rab8A, and Rab10-mediated recycling through PI3K/Akt and MAPK/ERK signaling activation. Moreover, α2M* enhanced the insulin response increasing insulin-induced glucose uptake rate in the myocardium under normal conditions. On the other hand, α2M* blocked the intracellular CE accumulation, improved the insulin response and reduced cardiac damage in HL-1 cardiomyocytes exposed to aggLDL. In conclusion, α2M* by its agonist action on LRP1, counteracts the deleterious effects of aggLDL in cardiomyocytes, which may have therapeutic implications in cardiovascular diseases associated with hypercholesterolemia.


Assuntos
Membrana Celular/metabolismo , Insulina/metabolismo , Proteína-1 Relacionada a Receptor de Lipoproteína de Baixa Densidade/metabolismo , Macroglobulinas/metabolismo , Miócitos Cardíacos/metabolismo , Animais , Western Blotting , Linhagem Celular , Glucose/metabolismo , Insulina/farmacologia , Lipoproteínas LDL/genética , Lipoproteínas LDL/metabolismo , Proteína-1 Relacionada a Receptor de Lipoproteína de Baixa Densidade/genética , Camundongos , Microscopia Confocal , Reação em Cadeia da Polimerase em Tempo Real , Transdução de Sinais/fisiologia
7.
Nagoya J Med Sci ; 81(1): 55-64, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30962655

RESUMO

The major hallmarks of Alzheimer's disease (AD) are the extracellular accumulation of pathological amyloid beta (Aß) in the brain parenchyma and Aß deposition in cerebral blood walls (cerebral amyloid angiopathy; CAA). Although CAA occurs in more than 80% of AD patients, the mechanisms of Aß deposition and clearance around the vessel walls are unknown. We found Aß-degrading activity in human serum during analysis of the regulatory mechanism of Aß production in human endothelial cells. To elucidate the metabolic dynamics of Aß surrounding the brain microvessels, we identified Aß-degrading activity in human serum (blood Aß-degrading activity: BADA) by column chromatography and LC/MS. BADA exhibited characteristics of an acidic protein, pI 4.3, which had two different protein surface charges (low and high affinity cations). Both BADA fractions had a relative molecular mass of greater than 400 kDa. Furthermore, BADA in the low affinity cation fraction was inhibited by the serine protease inhibitor 4-(2-Aminoethyl) benzenesulfonyl fluoride hydrochloride (AEBSF). We clarified alpha-2-macroglobulin (a2M) and several serine proteases from this BADA by LC-MS. Moreover, we demonstrated that BADA is increased by approximately 5000-fold in human serum by column chromatography. Therefore, BADA may play an important role in the circulation and metabolism of Aß in human brain microvessels.


Assuntos
Doença de Alzheimer/metabolismo , Peptídeos beta-Amiloides/metabolismo , Encéfalo/irrigação sanguínea , Encéfalo/patologia , Encéfalo/fisiologia , Angiopatia Amiloide Cerebral/patologia , Cromatografia Líquida , Células Endoteliais da Veia Umbilical Humana/metabolismo , Humanos , Macroglobulinas/metabolismo , Espectrometria de Massas , Microvasos/patologia , Microvasos/fisiologia , Serina Proteases/metabolismo
11.
Biochemistry (Mosc) ; 82(10): 1200-1206, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29037141

RESUMO

The primary role of insulin-like growth factor binding proteins (IGFBPs) is to regulate availability of IGFs for interacting with receptors, but IGFBPs perform IGF-independent actions as well. The availability and activity of IGFBPs in the circulation is influenced primarily by their concentration and structural modifications, but possibly also by interaction with major plasma proteins such as transferrin, alpha-2-macroglobulin (α2M), and fibrinogen. Four types of circulating IGFBP complexes were examined in this study by immuno- and ligand-binding assays in adults of different age. The amounts of IGFBP-3/transferrin and IGFBP-1/fibrinogen complexes were similar in middle- and old-aged persons, whereas the amounts of IGFBP-1 (or -2)/α2M monomer complexes were lower in the old-aged group and negatively correlated with total IGFBP-1 (or -2) amounts in blood. In contrast to IGFBP-1, IGFBP-2 was present in significantly greater quantities in complexes with α2M dimer than α2M monomer in older individuals. IGFBP complexes did not bind 125I-labeled IGF-I in amounts detectable by ligand blotting. According to the results of this study, the quantities of IGFBP-1 and IGFBP-2, which interact with α2M, are age-dependent and, in the case of complexes with α2M monomer, they are negatively correlated with the total circulating levels of these two IGFBPs.


Assuntos
Envelhecimento , Proteínas Sanguíneas/metabolismo , Western Blotting , Eletroforese , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/sangue , Adulto , Idoso , Idoso de 80 Anos ou mais , Proteínas Sanguíneas/química , Feminino , Fibrinogênio/química , Fibrinogênio/metabolismo , Humanos , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/química , Proteínas de Ligação a Fator de Crescimento Semelhante a Insulina/metabolismo , Radioisótopos do Iodo/química , Ligantes , Macroglobulinas/química , Macroglobulinas/metabolismo , Masculino , Pessoa de Meia-Idade , Ligação Proteica , Carbonilação Proteica , Transferrina/química , Transferrina/metabolismo
13.
Biochem J ; 473(10): 1315-27, 2016 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-26936971

RESUMO

Insulin increases glucose uptake by increasing the rate of exocytosis of the facilitative glucose transporter isoform 4 (Glut4) relative to its endocytosis. Insulin also releases Glut4 from highly insulin-regulated secretory compartments (GSVs or Glut4 storage vesicles) into constitutively cycling endosomes. Previously it was shown that both overexpression and knockdown of the small GTP-binding protein Rab14 decreased Glut4 translocation to the plasma membrane (PM). To determine the mechanism of this perturbation, we measured the effects of Rab14 knockdown on the trafficking kinetics of Glut4 relative to two proteins that partially co-localize with Glut4, the transferrin (Tf) receptor and low-density-lipoprotein-receptor-related protein 1 (LRP1). Our data support the hypothesis that Rab14 limits sorting of proteins from sorting (or 'early') endosomes into the specialized GSV pathway, possibly through regulation of endosomal maturation. This hypothesis is consistent with known Rab14 effectors. Interestingly, the insulin-sensitive Rab GTPase-activating protein Akt substrate of 160 kDa (AS160) affects both sorting into and exocytosis from GSVs. It has previously been shown that exocytosis of GSVs is rate-limited by Rab10, and both Rab10 and Rab14 are in vitro substrates of AS160. Regulation of both entry into and exit from GSVs by AS160 through sequential Rab substrates would provide a mechanism for the finely tuned 'quantal' increases in cycling Glut4 observed in response to increasing concentrations of insulin.


Assuntos
Adipócitos/metabolismo , Endossomos/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Células 3T3-L1 , Adipócitos/efeitos dos fármacos , Animais , Endocitose/genética , Endocitose/fisiologia , Citometria de Fluxo , Insulina/farmacologia , Macroglobulinas/genética , Macroglobulinas/metabolismo , Camundongos , Transporte Proteico/fisiologia , Transferrina/metabolismo , Proteínas rab de Ligação ao GTP/genética
14.
Ann Hepatol ; 15(2): 200-6, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26845597

RESUMO

UNLABELLED:  Background and rationale. Acoustic radiation force impulse (ARFI) is a non-invasive tool used in the evaluation of liver fibrosis in HCV positive immune-competent patients. This study aimed to assess the accuracy of ARFI in discriminating liver transplanted patients with different graft fibrosis severity and to verify whether ARFI, eventually combined with non-invasive biochemical tests, could spare liver biopsies. This prospective study included 51 HCV positive liver transplanted patients who consecutively underwent to annual liver biopsy concomitantly with ARFI and blood chemistry tests measurements needed to calculate several non-invasive liver fibrosis tests. RESULTS: Overall ARFI showed an AUC of 0.885 in discriminating between patients without or with significant fibrosis (Ishak score 0-2vs. 3-6). Using a cut-off of 1.365 m/s, ARFI possesses a negative predictive value of 100% in identifying patients without significant fibrosis. AUC for Fibrotest was 0.848 in discriminating patients with Ishak fibrosis score 0-2 vs. 3-6. The combined assessment of ARFI and Fibro-test did not improve the results obtained by ARFI alone. CONCLUSION: ARFI measurement in HCV positive liver transplanted patients can be considered an easy and accurate non-invasive tool in identify patients with a benign course of HCV recurrence.


Assuntos
Hepatite C Crônica/diagnóstico por imagem , Cirrose Hepática/diagnóstico por imagem , Transplante de Fígado , Fígado/diagnóstico por imagem , Idoso , Alanina Transaminase/sangue , Apolipoproteína A-I/sangue , Área Sob a Curva , Aspartato Aminotransferases/sangue , Bilirrubina/sangue , Biópsia , Técnicas de Imagem por Elasticidade , Feminino , Haptoglobinas/metabolismo , Hepatite C Crônica/metabolismo , Hepatite C Crônica/patologia , Hepatite C Crônica/cirurgia , Humanos , Fígado/patologia , Cirrose Hepática/metabolismo , Cirrose Hepática/patologia , Cirrose Hepática/cirurgia , Macroglobulinas/metabolismo , Masculino , Pessoa de Meia-Idade , Valor Preditivo dos Testes , Estudos Prospectivos , Recidiva
15.
J Biol Chem ; 290(4): 2334-50, 2015 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-25488663

RESUMO

The solution structure of complement C3b is crucial for the understanding of complement activation and regulation. C3b is generated by the removal of C3a from C3. Hydrolysis of the C3 thioester produces C3u, an analog of C3b. C3b cleavage results in C3c and C3d (thioester-containing domain; TED). To resolve functional questions in relation to C3b and C3u, analytical ultracentrifugation and x-ray and neutron scattering studies were used with C3, C3b, C3u, C3c, and C3d, using the wild-type allotype with Arg(102). In 50 mm NaCl buffer, atomistic scattering modeling showed that both C3b and C3u adopted a compact structure, similar to the C3b crystal structure in which its TED and macroglobulin 1 (MG1) domains were connected through the Arg(102)-Glu(1032) salt bridge. In physiological 137 mm NaCl, scattering modeling showed that C3b and C3u were both extended in structure, with the TED and MG1 domains now separated by up to 6 nm. The importance of the Arg(102)-Glu(1032) salt bridge was determined using surface plasmon resonance to monitor the binding of wild-type C3d(E1032) and mutant C3d(A1032) to immobilized C3c. The mutant did not bind, whereas the wild-type form did. The high conformational variability of TED in C3b in physiological buffer showed that C3b is more reactive than previously thought. Because the Arg(102)-Glu(1032) salt bridge is essential for the C3b-Factor H complex during the regulatory control of C3b, the known clinical associations of the major C3S (Arg(102)) and disease-linked C3F (Gly(102)) allotypes of C3b were experimentally explained for the first time.


Assuntos
Ativação do Complemento , Complemento C3/metabolismo , Complemento C3b/metabolismo , Complemento C3c/metabolismo , Complemento C3d/metabolismo , Arginina/química , Cristalografia por Raios X , Humanos , Macroglobulinas/metabolismo , Mutagênese , Mutação , Conformação Proteica , Multimerização Proteica , Espalhamento de Radiação , Ressonância de Plasmônio de Superfície , Ultracentrifugação
16.
Ukr Biochem J ; 86(2): 79-88, 2014.
Artigo em Ucraniano | MEDLINE | ID: mdl-24868914

RESUMO

The main goal of the study was to determine the ability of histones to induce production of the proteolytically active IgG-antibodies in BALB/c mice. In order to perform this study 8 mice were immunized with the fraction of total calf thymus histones. IgGs were isolated from the serum of the immunized and not immunized animals by means of precipitation with 33% ammonium sulfate, followed by affinity chromatography on protein G-Sepharose column. Histones, myelin basic protein (MBP), lysozyme, BSA, ovalbumin, macroglobulin, casein and cytochrome c served as substrates for determining the proteolytic activity. It was found that IgGs from the blood serum of immunized mice are capable of hydrolyzing histone H1, core histone and MBP. On the contrary, the proteolytic activity of IgGs from the blood serum of not immunized mice was not detected. The absence of proteolytical enzymes in the fraction of IgGs was proven by HPLC chromatography. High levels of proteolytic activity toward histones have been also detected in affinity purified IgGs from blood serum of patients with rheumatoid arthritis, but not in healthy donors. These data indicate that eukaryotic histones may induce production of protabzymes in mammals. The possible origin of these protabzymes and their potential biological role in mammalians is discussed.


Assuntos
Anticorpos Catalíticos/química , Artrite Reumatoide/sangue , Histonas/administração & dosagem , Soros Imunes/química , Imunoglobulina G/química , Animais , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Caseínas/química , Bovinos , Cromatografia de Afinidade , Cromatografia Líquida de Alta Pressão , Citocromos c/química , Histonas/imunologia , Histonas/isolamento & purificação , Humanos , Imunização , Macroglobulinas/química , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Muramidase/química , Proteína Básica da Mielina/química , Ovalbumina/química , Proteólise , Especificidade por Substrato , Timo/química
18.
Toxicol Appl Pharmacol ; 260(2): 105-14, 2012 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-22366155

RESUMO

Identification of biomarkers assists in the diagnosis of disease and the assessment of health risks from environmental exposures. We hypothesized that rats exposed to Libby amphibole (LA) would present with a unique serum proteomic profile which could help elucidate epidemiologically-relevant biomarkers. In four experiments spanning varied protocols and temporality, healthy (Wistar Kyoto, WKY; and F344) and cardiovascular compromised (CVD) rat models (spontaneously hypertensive, SH; and SH heart failure, SHHF) were intratracheally instilled with saline (control) or LA. Serum biomarkers of cancer, inflammation, metabolic syndrome (MetS), and the acute phase response (APR) were analyzed. All rat strains exhibited acute increases in α-2-macroglobulin, and α1-acid glycoprotein. Among markers of inflammation, lipocalin-2 was induced in WKY, SH and SHHF and osteopontin only in WKY after LA exposure. While rat strain- and age-related changes were apparent in MetS biomarkers, no LA effects were evident. The cancer marker mesothelin was increased only slightly at 1 month in WKY in one of the studies. Quantitative Intact Proteomic profiling of WKY serum at 1 day or 4 weeks after 4 weekly LA instillations indicated no oxidative protein modifications, however APR proteins were significantly increased. Those included serine protease inhibitor, apolipoprotein E, α-2-HS-glycoprotein, t-kininogen 1 and 2, ceruloplasmin, vitamin D binding protein, serum amyloid P, and more 1 day after last LA exposure. All changes were reversible after a short recovery regardless of the acute or long-term exposures. Thus, LA exposure induces an APR and systemic inflammatory biomarkers that could have implications in systemic and pulmonary disease in individuals exposed to LA.


Assuntos
Reação de Fase Aguda/induzido quimicamente , Amiantos Anfibólicos/toxicidade , Inflamação/induzido quimicamente , Síndrome Metabólica/induzido quimicamente , Reação de Fase Aguda/imunologia , Adiponectina/sangue , Animais , Biomarcadores/sangue , Inflamação/imunologia , Leptina/sangue , Lipocalina-2 , Lipocalinas/sangue , Macroglobulinas/metabolismo , Masculino , Síndrome Metabólica/imunologia , Orosomucoide/metabolismo , Osteopontina/sangue , Proteômica , Ratos , Ratos Endogâmicos F344 , Ratos Endogâmicos SHR , Ratos Endogâmicos WKY , Estudos Retrospectivos
19.
Cell Biochem Biophys ; 62(1): 257-65, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21913005

RESUMO

To evaluate the effectiveness and safety of autologous cytokine-induced killer (CIK) cells in elderly patients with diffuse large B-cell lymphoma. Peripheral blood mononuclear cells (PBMC) were isolated from nine elderly patients with diffuse large B-cell lymphoma. PBMCs were augmented by priming with interferon gamma (IFN-γ) followed by IL-2 and monoclonal antibody (mAb) against CD3. Autologous CIK cells (range 5 × 10(9)-1 × 10(10)) were then infused back to individual patients; infusion was repeated every 4 weeks for 32 weeks (eight cycles). Patients were assessed for changes in lymphocyte subgroup, tumor-related biological parameters, imaging characteristics, the condition of remission, quality of life (QOL), and survival. Prior to CIK infusion, two patients were in complete remission and seven patients were in partial remission. After autologous CIK cell transfusions, the proportion of CD3+, CD3+CD8+, and CD3+CD56+ cells were significantly increased compared with baseline (P < 0.05); whereas serum levels of ß2-microglobulin and LDH were significantly decreased (P < 0.05). The lymphoma symptoms were reduced and QOL was improved (P < 0.05) in all patients. All patients achieved complete remission at study endpoint. No adverse reactions were reported. Autologous CIK cell immunotherapy is safe and efficacious for the treatment of elderly patients with diffuse large B-cell lymphoma.


Assuntos
Células Matadoras Induzidas por Citocinas/transplante , Imunoterapia , Linfoma Difuso de Grandes Células B/terapia , Idoso , Idoso de 80 Anos ou mais , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/farmacologia , Complexo CD3/imunologia , Complexo CD3/metabolismo , Antígeno CD56/metabolismo , Antígenos CD8/metabolismo , Células Cultivadas , Células Matadoras Induzidas por Citocinas/imunologia , Feminino , Humanos , Interferon gama/farmacologia , Interleucina-2/farmacologia , L-Lactato Desidrogenase/sangue , Linfoma Difuso de Grandes Células B/diagnóstico por imagem , Linfoma Difuso de Grandes Células B/metabolismo , Linfoma Difuso de Grandes Células B/mortalidade , Macroglobulinas/análise , Masculino , Pessoa de Meia-Idade , Imagem Multimodal , Tomografia por Emissão de Pósitrons , Análise de Sobrevida , Tomografia Computadorizada por Raios X , Transplante Autólogo , Resultado do Tratamento
20.
Biomed Khim ; 58(4): 400-10, 2012.
Artigo em Russo | MEDLINE | ID: mdl-23413684

RESUMO

This review will focus on the systematization of knowledge about structure of macroglobulin signaling system, which includes macroglobulin family proteins (alpha-2-macroglobulin, alpha-2-glycoprotein, pregnancy associated plasma protein A), their receptors (LRP, grp78), ligands (proteinases, cytokines, hormones, lipids, et al.) transforming and transcriptional factors for regulation of macroglobulins synthesis. After reviewing the functions of macroglobulin signaling system, and mechanisms of their realization, we discuss the complex and significant role of this system in different physiological and pathological processes.


Assuntos
Macroglobulinas/metabolismo , Transdução de Sinais/fisiologia , Animais , Transporte Biológico/fisiologia , Citocinas/metabolismo , Chaperona BiP do Retículo Endoplasmático , Feminino , Proteínas de Choque Térmico/metabolismo , Hormônios/metabolismo , Humanos , Ligantes , Masculino , Peptídeo Hidrolases/metabolismo
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