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OBJECTIVE: Intercellular communication within pancreatic ductal adenocarcinoma (PDAC) dramatically contributes to metastatic processes. The underlying mechanisms are poorly understood, resulting in a lack of targeted therapy to counteract stromal-induced cancer cell aggressiveness. Here, we investigated whether ion channels, which remain understudied in cancer biology, contribute to intercellular communication in PDAC. DESIGN: We evaluated the effects of conditioned media from patient-derived cancer-associated fibroblasts (CAFs) on electrical features of pancreatic cancer cells (PCC). The molecular mechanisms were deciphered using a combination of electrophysiology, bioinformatics, molecular and biochemistry techniques in cell lines and human samples. An orthotropic mouse model where CAF and PCC were co-injected was used to evaluate tumour growth and metastasis dissemination. Pharmacological studies were carried out in the Pdx1-Cre, Ink4afl/fl LSL-KrasG12D (KICpdx1) mouse model. RESULTS: We report that the K+ channel SK2 expressed in PCC is stimulated by CAF-secreted cues (8.84 vs 2.49 pA/pF) promoting the phosphorylation of the channel through an integrin-epidermal growth factor receptor (EGFR)-AKT (Protein kinase B) axis. SK2 stimulation sets a positive feedback on the signalling pathway, increasing invasiveness in vitro (threefold) and metastasis formation in vivo. The CAF-dependent formation of the signalling hub associating SK2 and AKT requires the sigma-1 receptor chaperone. The pharmacological targeting of Sig-1R abolished CAF-induced activation of SK2, reduced tumour progression and extended the overall survival in mice (11.7 weeks vs 9.5 weeks). CONCLUSION: We establish a new paradigm in which an ion channel shifts the activation level of a signalling pathway in response to stromal cues, opening a new therapeutic window targeting the formation of ion channel-dependent signalling hubs.
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Carcinoma Ductal Pancreático , Neoplasias Pancreáticas , Humanos , Animais , Camundongos , Proteínas Proto-Oncogênicas c-akt , Carcinogênese , Transformação Celular Neoplásica , Transdução de Sinais , Neoplasias PancreáticasRESUMO
Inflammatory alterations of the extracellular matrix shape the tumor microenvironment and promote all stages of carcinogenesis. This study aims to determine the impact of cellular fibronectin on inflammatory facets of tumor-associated macrophages (TAMs) in breast cancer. Cellular fibronectin (FN) harboring the alternatively spliced extra domain A (FN-EDA) was determined to be a matrix component produced by the triple-negative breast cancer (TNBC) cells. High levels of FN-EDA correlated with poor survival in breast cancer patients. The proinflammatory cytokine IL-1ß enhanced the expression of cellular fibronectin including FN-EDA. TAMs were frequently observed in the tumor areas rich in FN-EDA. Conditioned media from TNBC cells induced the differentiation of CD206+CD163+ macrophages and stimulated the STAT3 pathway, ex vivo. In the macrophages, the STAT3 pathway enhanced FN-EDA-induced IL-1ß secretion and NF-κB signaling. In conclusion, our data indicate a self-reinforcing mechanism sustained by FN-EDA and IL-1ß through NF-κB and STAT3 signaling in TAMs which fosters an inflammatory environment in TNBC.
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NF-kappa B , Neoplasias de Mama Triplo Negativas , Humanos , NF-kappa B/genética , NF-kappa B/metabolismo , Fibronectinas/genética , Fibronectinas/metabolismo , Fibronectinas/farmacologia , Retroalimentação , Neoplasias de Mama Triplo Negativas/genética , Transdução de Sinais , Microambiente Tumoral/genéticaRESUMO
Cellular fibronectin (FN; also known as FN1) variants harboring one or two alternatively spliced so-called extra domains (EDB and EDA) play a central bioregulatory role during development, repair processes and fibrosis. Yet, how the extra domains impact fibrillar assembly and function of the molecule remains unclear. Leveraging a unique biological toolset and image analysis pipeline for direct comparison of the variants, we demonstrate that the presence of one or both extra domains impacts FN assembly, function and physical properties of the matrix. When presented to FN-null fibroblasts, extra domain-containing variants differentially regulate pH homeostasis, survival and TGF-ß signaling by tuning the magnitude of cellular responses, rather than triggering independent molecular switches. Numerical analyses of fiber topologies highlight significant differences in variant-specific structural features and provide a first step for the development of a generative model of FN networks to unravel assembly mechanisms and investigate the physical and functional versatility of extracellular matrix landscapes.This article has an associated First Person interview with the first author of the paper.
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Processamento Alternativo , Fibronectinas , Células Cultivadas , Matriz Extracelular/metabolismo , Fibroblastos/metabolismo , Fibronectinas/genética , Fibronectinas/metabolismo , HumanosRESUMO
Anti-human cytomegalovirus (HCMV) antibodies are considered triggers of systemic sclerosis (SSc), but such a hypothesis has been assessed in limited sub-dominant epitopes. Our aim was to systematically assess the potential association of HCMV antibodies targeting most immunodominant and subdominant viral antigens, as this would reveal immunopathogenic associations. Our study included 110 SSc patients, 60 multiple sclerosis (MS) patients, and 51 healthy controls (HC). Anti-HCMV abs were tested by immunoblotting. IgG anti-HCMV was broader in SSc and MS compared to HC. Anti- UL57 and UL55 were more frequent in SSc versus MS forms. Reactivity to multiple viral antigens was more frequent in SSc than MS forms. Anti-viral antibodies levels were higher in specific autoantibody-positive SSc patients compared to seronegative cases. In conclusion, more prevalent and/or stronger antigen-specific HCMV responses are noted in SSc compared to controls, implying a role of these viral responses in SSc development.
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Anticorpos Antivirais/sangue , Antígenos Virais/imunologia , Citomegalovirus/imunologia , Epitopos Imunodominantes/imunologia , Escleroderma Sistêmico/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Especificidade de Anticorpos , Estudos de Casos e Controles , Feminino , Humanos , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/imunologiaRESUMO
OBJECTIVES: To study immunoreactivity against human cytomegalovirus (HCMV) in systemic sclerosis (SSc), since HCMV has been put forward as a candidate infectious cause. METHODS: Eighty four patients with SSc (67 females; median age 60 years, range 25-81), 30 patients with multiple sclerosis (MS) (23 females; median age 44, range 20-69 years) and 28 healthy controls (NCs), all pre-tested positive for IgG anti-HCMV antibodies, were studied. IgG anti-UL83 HCMV antibodies were tested by western immunoblotting and expressed in arbitrary units (AUs). Reactivity to UL83 HCMV was assessed in relation to clinical manifestations and SSc-related autoantibodies (autoAbs), tested by an IgG SSc autoantibody profile line immunoassay (Euroimmun) that detects autoAbs against Scl-70, CENPA, CENPB, RNA polymerase III subunit 11 (RP11), RP155, fibrillarin, NOR90, Th/To, PM-Scl100, PM-Scl75, Ku, PDGFR and Ro-52. RESULTS: Fifty patients (59.5%) were anti-UL83 clear positive (UL83+), including 21/40 (52.5%) lcSSc and 29/44 (65.6%) dcSSc, compared to 15/30 (50%) patients with MS (SSc vs MS, p=ns and 11/28 (39.29%) of NCs (SSc vs NC, p=ns MS vs NC, p=ns). Anti-UL83 antibody AU levels (mean±SD) were higher in SSc (64.3 ± 26) compared to MS (49.1±21.6, p=0.05) or NCs (40.4±13.7, p<0.001; MS vs NCs, p=ns) and were associated with pulmonary fibrosis. CONCLUSIONS: Immunoreactivity to UL83 HCMV is frequent and strong in patients with SSc, implying a possible pathogenic role for this disease.
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Autoanticorpos/sangue , Fosfoproteínas/imunologia , Escleroderma Sistêmico/imunologia , Proteínas da Matriz Viral/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/imunologia , Escleroderma Sistêmico/etiologiaRESUMO
Due to the complex architectural diversity of biological networks, there is an increasing need to complement statistical analyses with a qualitative and local description of their spatial properties. One such network is the extracellular matrix (ECM), a biological scaffold for which changes in its spatial organization significantly impact tissue functions in health and disease. Quantifying variations in the fibrillar architecture of major ECM proteins should considerably advance our understanding of the link between tissue structure and function. Inspired by the analysis of functional magnetic resonance imaging (fMRI) images, we propose a novel statistical analysis approach embedded into a machine learning paradigm, to measure and detect local variations of meaningful ECM parameters. We show that parametric maps representing fiber length and pore directionality can be analyzed within the proposed framework to differentiate among various tissue states. The parametric maps are derived from graph-based representations that reflect the network architecture of fibronectin (FN) fibers in a normal, or disease-mimicking in vitro setting. Such tools can potentially lead to a better characterization of dynamic matrix networks within fibrotic tumor microenvironments and contribute to the development of better imaging modalities for monitoring their remodeling and normalization following therapeutic intervention.
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BACKGROUND: Our aims were to investigate the time-course effects of a futsal match on performance, oxidative stress, and muscle damage markers, as well as inflammatory and antioxidant responses during a 6-day post-match period. METHODS: Thirty-four female high-level futsal players were assessed on several oxidative stress, inflammation, subjective muscle soreness, subjective rate perceived exertion, and performance tests before a futsal match, immediately after, and 24 h to 144 h after. RESULTS: Counter movement jump, 20 m, and 10 m sprints performance significantly decreased immediately after the match (p < 0.05) and returned to baseline 72 h post-match (p > 0.05). Delayed onset muscle soreness peaked 24 h post-match and rate perceived exertion peaked post-match (p < 0.05) and returned to baseline 96 h post-match (p > 0.05). Inflammatory biomarkers peaked at 24 h (p < 0.05) and remained significantly elevated for 72 h after the match (p < 0.05). Muscle damage biomarkers peaked at 24 h (p < 0.05) and remained significantly (p < 0.05) elevated for at least 72 h after the match. Oxidative stress markers peaked at 24 h-48 h (p < 0.05) and returned to baseline 120 h post-match (p > 0.05). In respect to antioxidant responses, these peaked at 24 h-48 h post-match (p < 0.05) and returned to baseline 120 h after the match (p > 0.05). CONCLUSIONS: A single futsal match induces short/mid-term changes in performance, inflammation, oxidative stress, and muscle damage markers for about 72 h-96 h post-match.
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Cachexia is an early result of rheumatoid arthritis (RA) (rheumatoid cachexia, RC), characterised mainly by involuntary loss of fat-free mass. RC is apparent in 1-67% of patients with RA, depending on the diagnostic criteria applied and the method used for the assessment of body composition. RC is associated with increased inflammation and disability, lower health perception, and greater mortality risk. These changes in body composition are driven by the inflammation process, the low levels of physical activity, the underlying testosterone deficiency and hypogonadism, and the pharmacotherapy required for RA. Chronic inflammation enhances resting energy expenditure as a response to stress, inducing an energy deficit, further propelling protein turnover. The use of corticosteroids and tumour necrosis factor α (TNF-α) inhibitors tend to increase fat accumulation, whereas other disease-modifying antirheumatic drugs (DMARDs) appear to induce increments in fat-free mass. The present review presents all information regarding the prevalence of RC, diagnostic criteria, and comorbidities, as well as the effects of pharmacotherapy and medical nutrition therapy on body composition of patients with RA.
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BACKGROUND: Ferredoxins are small iron-sulfur proteins belonging to all domains of life. A sub-group binds two [4Fe-4S] clusters with unequal and extremely low values of the reduction potentials. These unusual properties are associated with two specific fragments of sequence. The functional importance of the very low potential ferredoxins is unknown. RESULTS: A bioinformatic screening of the sequence features defining very low potential 2[4Fe-4S] ferredoxins has revealed the almost exclusive presence of the corresponding fdx gene in the Proteobacteria phylum, without occurrence in Archaea and Eukaryota. The transcript was found to be monocistronic in Pseudomonas aeruginosa, and not part of an operon in most bacteria. Only fdx genes of bacteria which anaerobically degrade aromatic compounds belong to operons. As this pathway is not present in all bacteria having very low potential 2[4Fe-4S] ferredoxins, these proteins cannot exclusively be reductants of benzoyl CoA reductases. Expression of the ferredoxin gene did not change in response to varying growth conditions, including upon macrophage infection or aerobic growth with 4-hydroxy benzoate as carbon source. However, it increased along the growth curve in Pseudomonas aeruginosa and in Escherichia coli. The sequence immediately 5' upstream of the coding sequence contributed to the promotor activity. Deleting the fdx gene in Pseudomonas aeruginosa abolished growth, unless a plasmid copy of the gene was provided to the deleted strain. CONCLUSIONS: The gene of the very low potential 2[4Fe-4S] ferredoxin displays characteristics of a housekeeping gene, and it belongs to the minority of genes that are essential in Pseudomonas aeruginosa. These data identify a new potential antimicrobial target in this and other pathogenic Proteobacteria.
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Proteínas de Bactérias/metabolismo , Ferredoxinas/metabolismo , Pseudomonas aeruginosa/metabolismo , Animais , Bactérias/química , Bactérias/genética , Bactérias/crescimento & desenvolvimento , Bactérias/metabolismo , Infecções Bacterianas/microbiologia , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Linhagem Celular , Humanos , Camundongos , Dados de Sequência Molecular , Pseudomonas aeruginosa/química , Pseudomonas aeruginosa/genética , Alinhamento de SequênciaRESUMO
Normal tissue homeostasis and architecture restrain tumor growth. Thus, for a tumor to develop and spread, malignant cells must overcome growth-repressive inputs from surrounding tissue and escape immune surveillance mechanisms that curb cancer progression. This is achieved by promoting the conversion of a physiological microenvironment to a pro-tumoral state and it requires a constant dialog between malignant cells and ostensibly normal cells of adjacent tissue. Pro-tumoral reprogramming of the stroma is accompanied by an upregulation of certain extracellular matrix (ECM) proteins and their cognate receptors. Fibronectin (FN) is one such component of the tumor matrisome. This large multidomain glycoprotein dimer expressed over a wide range of human cancers is assembled by cell-driven forces into a fibrillar array that provides an obligate scaffold for the deposition of other matrix proteins and binding sites for functionalization by soluble factors in the tumor microenvironment. Encoded by a single gene, FN regulates the proliferation, motile behavior and fate of multiple cell types, largely through mechanisms that involve integrin-mediated signaling. These processes are coordinated by distinct isoforms of FN, collectively known as cellular FN (as opposed to circulating plasma FN) that arise through alternative splicing of the FN1 gene. Cellular FN isoforms differ in their solubility, receptor binding ability and spatiotemporal expression, and functions that have yet to be fully defined. FN induction at tumor sites constitutes an important step in the acquisition of biological capabilities required for several cancer hallmarks such as sustaining proliferative signaling, promoting angiogenesis, facilitating invasion and metastasis, modulating growth suppressor activity and regulating anti-tumoral immunity. In this review, we will first provide an overview of ECM reprogramming through tumor-stroma crosstalk, then focus on the role of cellular FN in tumor progression with respect to these hallmarks. Last, we will discuss the impact of dysregulated ECM on clinical efficacy of classical (radio-/chemo-) therapies and emerging treatments that target immune checkpoints and explore how our expanding knowledge of the tumor ECM and the central role of FN can be leveraged for therapeutic benefit.
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Helicobacter pylori (Hp) is a likely trigger of systemic sclerosis (SSc), but systemic antigen-specific antibody (Ab) responses in a well-defined cohort of SSc patients have not been thoroughly assessed. Line immunoassay and immunoblotting testing Abs against 15 Hp antigens were performed in 91 SSc patients and 59 demographically matched healthy controls (HCs). Results were validated in an independent cohort of 35 SSc patients. Anti-Hp positivity was detected in 67% SSc patients vs 76.3% HCs. Among anti-Hp (+) individuals, anti-p67-FSH was less frequent in SSc than HCs (p = 0.016), whereas reactivity to the remaining 14 Hp antigens did not differ between patients and HCs. Anti-p67 Abs were less frequent in diffuse cutaneous SSc (dcSSc) compared with HCs (p = 0.018). Anti-p57 and anti-p33 Ab levels were lower in SSc vs HCs (p = 0.007 and p = 0.035, respectively). Anti-p57 and anti-p33 Ab levels were lower in limited cutaneous SSc (lcSSc) (p = 0.010) and dcSSc (p = 0.024), respectively, compared with HCs. Anti-p50 and anti-p17 Ab titers tended to be higher in dcSSc than in lcSSc. Sera from the independent SSc cohort showed comparable results. Anti-VacA Abs were more frequent in pulmonary arterial hypertension (p = 0.042), and anti-p30 Abs were more frequent in calcinosis (p = 0.007), whereas anti-VacA Ab levels were higher in lung fibrosis (p = 0.02). In conclusion, anti-Hp Abs are neither more frequent nor elevated in SSc compared with healthy population, the only exception being the higher frequency and levels of anti-VacA Abs in pulmonary hypertension and lung fibrosis, respectively. These results suggest that Hp is unlikely to be involved in the development of SSc.
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Infecções por Helicobacter/imunologia , Helicobacter pylori/fisiologia , Escleroderma Sistêmico/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antibacterianos/metabolismo , Antígenos de Bactérias/imunologia , Estudos de Coortes , Epitopos/imunologia , Feminino , Humanos , Imunidade Humoral , Masculino , Pessoa de Meia-IdadeRESUMO
The crystal structures of the C57A and V13G molecular variants of Allochromatium vinosum 2[4Fe-4S] ferredoxin (AlvinFd) and that of the homologous ferredoxin from Escherichia coli (EcFd) have been determined at 1.05-, 1.48-, and 1.65-A resolution, respectively. The present structures combined with cyclic voltammetry studies establish clear effects of the degree of exposure of the cluster with the lowest reduction potential (cluster I) towards less negative reduction potentials (E degrees ). This is better illustrated by V13G AlvinFd (high exposure, E degrees = -594 mV) and EcFd (low exposure, E degrees = -675 mV). In C57A AlvinFd, the movement of the protein backbone, as a result of replacing the noncoordinating Cys57 by Ala, leads to a +50-mV upshift of the potential of the nearby cluster I, by removal of polar interactions involving the thiolate group and adjustment of the hydrogen-bond network involving the cluster atoms. In addition, the present structures and other previously reported accurate structures of this family of ferredoxins indicate that polar interactions of side chains and water molecules with cluster II sulfur atoms, which are absent in the environment of cluster I, are correlated to the approximately 180-250 mV difference between the reduction potentials of clusters I and II. These findings provide insight into the significant effects of subtle structural differences of the protein and solvent environment around the clusters of [4Fe-4S] ferredoxins on their electrochemical properties.
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Proteínas de Bactérias/química , Chromatiaceae/química , Cristalografia por Raios X , Escherichia coli/química , Ferredoxinas/química , Sequência de Aminoácidos , Eletroquímica , Ferredoxinas/genética , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Conformação Proteica , Alinhamento de SequênciaRESUMO
BACKGROUND: Autoantibodies (autoAbs) help in diagnosis and predicting clinical phenotypes in systemic sclerosis (SSc). AIM OF THE STUDY: To determine the clinical utility of 13 SSc-related autoAbs in SSc patients. MATERIAL AND METHODS: A total of 131 consecutive patients with SSc (111 female, mean age 58.1 ± 14 years; 49 with diffused cutaneous SSc [dcSSc] and 82 with limited cutaneous SSc [lcSSc]) were analysed by a multiplex line immunoassay (Euroimmun) for autoantibodies (autoAbs) against 13 SSc-related antigens. A total of 22 patients with primary Raynaud phenomenon (RP), and 22 healthy controls were also analysed. RESULTS: ANA by indirect immunofluorescence was present in 128 (97.7%) patients with SSc. Excluding anti-Ro52, 113 (89.3%) SSc patients were positive for at least one autoAb: anti-Topoisomerase I (anti-Topo) I abs in 54 (41.2%), anti-centromere proteins (anti-CENP) in 37 (28.2%, all reactive with centromere protein-A (CENPA) and centromere protein B (CENPB)), anti-RNA polymerase III(RP11) in 19 (14.5%), anti-RNA polymerase III(RP155) in 13 (9.9%), anti-fibrillarin in 4 (3.1%), anti-Ku in 6 (4.6%), anti-nucleolus-organizing region (anti-NOR90) in 8 (6.1%), anti-PM-Scl100 in 2 (1.5%), and anti-PM-Scl75 in 4 (3.1%). There was no immunoreactivity for Th/To or platelet-derived growth factor receptor (PDGFR). Overall, 102 (77.9%) SSc patients had autoAbs against Topo I, CENPA or CENPB, RP11 or RP155. Anti-Topo I abs were strongly associated with dcSSc, interstitial lung disease (ILD) (p < .001), pulmonary hypertension (PH) (p = .019) and ILD-PH (p = .003). Anti-CENPB abs were associated with lcSSc, and negatively associated with ILD. Anti-RP11 and anti-NOR90 abs were associated with male gender, and anti-NOR90 associated with ILD. CONCLUSIONS: Anti-Topo I, anti-CENP, and anti-RNA pol III are the most prevalent autoAbs in SSc. Anti-Topo I and anti-NOR90 abs are associated with ILD and/or PAH.
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Autoanticorpos/imunologia , Autoimunidade , Escleroderma Sistêmico/imunologia , Adulto , Idoso , Autoantígenos/imunologia , Epitopos/imunologia , Feminino , Humanos , Imunoensaio , Imunoglobulina G/imunologia , Masculino , Pessoa de Meia-Idade , PrevalênciaRESUMO
To assess whether Helicobacter pylori (Hp) antibody (ab) reactivity against individual Hp antigens is pathogenetically relevant to multiple sclerosis (MS), we systematically investigated prevalence and clinical significance of abs against 14 immunodominant and subdominant Hp antigens by ELISA and immunoblotting in 139 consecutive MS patients with relapsing-remitting (RRMS, n = 102) or secondary progressive (SPMS, n = 37). Sera from 39 patients with Parkinson's disease (PD), 21 with Alzheimer's disease (ALZ) and 68 healthy controls (HCs), were also tested. Anti-flagellin (18.3%) and anti-p41 (25.0%) abs in MS were less frequent than in HCs (39.4%, 48.5%, respectively). Abs against 5 of the 14 antigens were less frequent in RRMS than HCs, including p41, p54-flagellin, p29-UreA, p67-FSH, and p120-CagA. Anti-VacA abs were more frequent in SPMS than in HCs (42.1 vs 12.1%, p = 0.019). Anti-p54, anti-p29-UreA and anti-p26 correlated with extended disability status scale (EDSS) (p = 0.017, p = 0.005, p = 0.002, respectively). Anti-p26 and anti-p17 correlated with the number of relapses (p = 0.037 and p = 0.047, respectively). This is the first comprehensive analysis of ab reactivities against most Hp antigens in MS patients. Ab responses differ between MS and HCs and between RRMS and SPMS, being more prevalent in SPMS than RRMS, thus suggesting an association between anti-Hp and the former type of MS.
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Anticorpos Antibacterianos/sangue , Antígenos de Bactérias/imunologia , Infecções por Helicobacter/patologia , Helicobacter pylori/imunologia , Esclerose Múltipla Crônica Progressiva/epidemiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Ensaio de Imunoadsorção Enzimática , Feminino , Infecções por Helicobacter/complicações , Humanos , Immunoblotting , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Crônica Progressiva/etiologia , Prevalência , RecidivaRESUMO
The role of human cytomegalovirus (HCMV) has been postulated as a trigger of systemic sclerosis (SSc). The aim of the study was to assess the prevalence of antibodies against HCMV UL44 and UL57 antigens not tested in the past. Sixty SSc patients, 40 multiple sclerosis and 17 normal controls (NCs), all anti-HCMV positive, were tested by immunoblotting. Reactivity to HCMV antigens, expressed as arbitrary units (AUs), was assessed for correlation with clinical and immunological parameters, including types of SSc-related autoantibodies. Anti-UL44 and anti-UL57 HCMV antibodies were present in 3/60 (5%) and 58/60 (96.7%) SSc patients, respectively (p < 0.001). Anti-UL57 antibodies were present in 35/40 (87.5%) MS patients and 16/17 (94.1%) NCs (SSc vs MS, MS vs NC, p = ns). Strong (50-75 AU) and very strong (75-100 AU) anti-UL57 immunoreactivity was found in 24 (41.4%) and 22 (37.9%) SSc patients, respectively (p = ns). Dilution experiments showed anti-UL57 antibody persistence in up to 1/5000. Overall, there was no difference in the frequency or the magnitude of anti-UL57 immunoreactivity between diffuse cutaneous systemic sclerosis and limited cutaneous systemic sclerosis patients (96.67 vs 96.67%; 65.45 ± 20.19 vs 64.31 ± 21.11 AU, p > 0.05) but strong anti-UL57 reactivity were more frequent in SSc compared to NCs (p = 0.007). Anti-UL57 reactivity was not inhibited by SSc-specific autoantigens. Anti-UL57 seropositivity did not correlate with demographic, clinical or immunological features of SSc. Anti-HCMV UL57 antibodies are universally present in anti-HCMV-positive patients with SSc, while those against UL44 are rarely seen. Because anti-UL57 lack disease specificity and are not involved in cross-reactive responses, their immunopathogenetic potential is to be questioned.
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Anticorpos Antivirais/sangue , Infecções por Citomegalovirus/imunologia , Proteínas de Ligação a DNA/imunologia , Escleroderma Sistêmico/imunologia , Proteínas Virais/imunologia , Adulto , Idoso , Formação de Anticorpos , Autoanticorpos/sangue , Autoantígenos/imunologia , Estudos de Casos e Controles , Citomegalovirus , Feminino , Grécia , Humanos , Masculino , Pessoa de Meia-Idade , Escleroderma Sistêmico/virologiaRESUMO
In view of published data suggesting that Helicobacter pylori (Hp) is a trigger of multiple sclerosis (MS), we assessed anti-heat shock protein 60 (hsp60)Hp antibody reactivity in 129 MS patients and 48 demograpically-matched healthy controls (HCs). Anti-Hp antibodies by ELISA were more elevated in MS than HCs but did not differ between different MS phenotypes. All anti-Hp-positive MS sera, irrespectively of their clinical phenotype, were anti-anti-hsp60 positive. Anti-hsp60 Hp seropositivity correlated with age at disease onset. In conclusion, anti-hsp60 Hp antibodies are present in all anti-Hp positive MS patients, and their relevance to disease pathogenesis is questionable.
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Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/imunologia , Chaperonina 60/imunologia , Infecções por Helicobacter/complicações , Helicobacter pylori/patogenicidade , Esclerose Múltipla , Adulto , Fatores Etários , Feminino , Helicobacter pylori/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla/sangue , Esclerose Múltipla/etiologia , Esclerose Múltipla/imunologiaRESUMO
The structure of the 2[4Fe-4S] ferredoxin (PaFd) from Pseudomonas aeruginosa, which belongs to the Allochromatium vinosum (Alvin) subfamily, has been determined by X-ray crystallography at 1.32-A resolution, which is the highest up to now for a member of this subfamily of Fds. The main structural features of PaFd are similar to those of AlvinFd. However, the significantly higher resolution of the PaFd structure makes possible a reliable comparison with available high-resolution structures of [4Fe-4S]-containing Fds, in an effort to rationalize the unusual electrochemical properties of Alvin-like Fds. Three major factors contributing to the reduction potential values of [4Fe-4S]2+/+ clusters of Fds, namely, the surface accessibility of the clusters, the N-H...S hydrogen-bonding network, and the volume of the cavities hosting the clusters, are extensively discussed. The volume of the cavities is introduced in the present work for the first time, and can in part explain the very negative potential of cluster I of Alvin-like Fds.
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Proteínas de Bactérias/química , Ferredoxinas/química , Pseudomonas aeruginosa/metabolismo , Sequência de Aminoácidos , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cristalografia por Raios X , Eletroquímica , Ferredoxinas/genética , Ferredoxinas/metabolismo , Ligação de Hidrogênio , Proteínas Ferro-Enxofre/química , Proteínas Ferro-Enxofre/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Oxirredução , Estrutura Terciária de Proteína , Pseudomonas aeruginosa/genética , Alinhamento de Sequência , Difração de Raios XRESUMO
Aerobic reactions of Co(O(2)CMe)(2).4H(2)O with di-2-pyridyl ketone oxime (Hpko) in the presence of counterions (ClO(4)(-), PF(6-)) give the tetranuclear, mixed-valence cobalt(II/III) clusters [Co(II)(2)Co(III)(2)(OR)(2)(O(2)CMe)(2)(pko)(4)S(2)]X(2) [R = H, S = MeOH, X = ClO(4) (1); R = Me, S = EtOH, X = PF(6) (2)] depending on the solvent mixture. Complexes 1 and 2 are the first Co members in the family of metallacrowns adopting the extremely rare inverse 12-metallacrown-4 motif.