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1.
Curr Issues Mol Biol ; 44(1): 105-116, 2021 Dec 25.
Artigo em Inglês | MEDLINE | ID: mdl-35723387

RESUMO

Cancer metastasis is a stage of the disease where therapy is mostly ineffective; hence, the need to find reliable markers of its onset. The metalloproteinase-9 (MMP-9, gelatinase B) in its 82 kDa active form, is a good candidate, but here we show that the correspondent little known 65 kDa active MMP-9 isoform, often misrepresented with the other gelatinase MMP-2, is a more suitable marker. Sera from patients with lung and breast cancer were analyzed by bidimensional zymography to detect the activity of MMP-9 and MMP-2. Enzyme identity was confirmed by comparison with MMP-9 standards and by western blotting. The 65 kDa isoform of MMP-9 is a suitable biomarker to monitor tumor progression from tissue neoplasms to metastatic stage, as its activity begins to appear when disease severity increases and becomes very high in metastasis. Moreover, the 65 kDa MMP-9, which derives from the 82 kDa MMP-9, no longer responds to natural MMP-9 inhibitors. As its activity cannot be controlled, its appearance may warn that the pathological process is becoming irreversible. Identification and inhibition of the enzymes converting the inhibitor-sensitive 82 kDa MMP-9 into the corresponding "wild" 65 kDa MMP-9 may allow to develop therapies capable of blocking metastases.

2.
J Cell Mol Med ; 18(2): 242-52, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24616914

RESUMO

The matrix metalloproteinases (MMPs) gelatinase A (MMP-2) and gelatinase B (MMP-9) are mediators of brain injury in multiple sclerosis (MS) and valuable biomarkers of disease activity. We applied bidimensional zymography (2-DZ) as an extension of classic monodimensional zymography (1-DZ) to analyse the complete pattern of isoforms and post-translational modifications of both MMP-9 and MMP-2 present in the sera of MS patients. The enzymes were separated on the basis of their isoelectric points (pI) and apparent molecular weights (Mw) and identified both by comparison with standard enzyme preparations and by Western blot analysis. Two MMP-2 isoforms, and at least three different isoforms and two different states of organization of MMP-9 (the multimeric MMP-9 and the N-GAL-MMP-9 complex) were observed. In addition, 2-DZ revealed for the first time that all MMP-9 and MMP-2 isoforms actually exist in the form of charge variants: four or five variants in the NGAL complex, more charge variants in the case of MMP-9; and five to seven charge variants for MMP-2. Charge variants were also observed in recombinant enzymes and, after concentration, also in sera from healthy individuals. Sialylation (MMP-9) and phosphorylation (MMP-2) contributed to molecular heterogeneity. The detection of charge variants of MMP-9 and MMP-2 in MS serum samples illustrates the power of 2-DZ and demonstrates that in previous studies MMP mixtures, rather than single molecules, were analysed. These observations open perspectives for better diagnosis and prognosis of many diseases and need to be critically interpreted when applying other methods for MS and other diseases.


Assuntos
Metaloproteinase 2 da Matriz/sangue , Metaloproteinase 9 da Matriz/sangue , Esclerose Múltipla/enzimologia , Processamento de Proteína Pós-Traducional , Western Blotting , Estudos de Casos e Controles , Eletroforese em Gel de Poliacrilamida , Ensaios Enzimáticos , Feminino , Humanos , Isoenzimas/sangue , Isoenzimas/genética , Masculino , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/genética , Esclerose Múltipla/sangue , Esclerose Múltipla/diagnóstico , Esclerose Múltipla/genética , Fosforilação , Ácidos Siálicos/metabolismo , Eletricidade Estática
3.
Nutrients ; 16(13)2024 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-38999763

RESUMO

Vitamin D3 deficiency and insufficiency are becoming a common global issue for us, especially in the most industrially developed countries. The only acknowledged activity of vitamin D3 in vertebrates is to promote the absorption of calcium and, therefore, allow for the mineralization of bones. Accordingly, its deficiency is associated with diseases such as rickets. Other numerous vital functions associated with vitamin D3 are yet to be considered, and the function of vitamin D2 in plants is unknown. Thus, 100 years after its discovery, the importance of vitamin D still seems to be unacknowledged (except for rickets), with little attention given to its decrease throughout the world. In this review, I suggest that vitamin D deficiency and insufficiency may be linked to the westernized lifestyle in more developed countries. Furthermore, I suggest that, rather than the calcemic activity, the main function of vitamin D is, in general, that of strengthening living organisms. I conclude with the hypothesis that vitamin D deficiency may represent a marker for a greater risk of chronic inflammatory diseases and a shorter life expectancy.


Assuntos
Deficiência de Vitamina D , Vitamina D , Deficiência de Vitamina D/epidemiologia , Humanos , Vitamina D/sangue , Luz Solar , Raquitismo/etiologia , Raquitismo/epidemiologia , Raquitismo/prevenção & controle , Estilo de Vida , Saúde Global
4.
Neurochem Res ; 36(3): 518-27, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21207142

RESUMO

We investigated whether polyphenols modulate the expression and activity of the enzymes gelatinases A (MMP-2) and B (MMP-9), involved in the pathogenesis of multiple sclerosis (MS). LPS-activated primary rat astrocytes were treated with the flavonoids quercetin (QRC) and cathechins [green tea extract (GTE)] and the non-flavonoids resveratrol (RSV) and tyrosol/hydroxytyrosol (Oliplus). As assessed by zymography and RT-PCR, RSV and Oliplus, but not QRC and GTE, dose-dependently inhibited the LPS-induced levels and mRNA expression of MMP-2 and MMP-9. By contrast, in cell-free systems direct inhibition of gelatinase activity in MS sera was determined by QRC and GTE, but not by RSV. Oliplus was only partially effective. Our results indicate that the flavonoids and non-flavonoids tested exert their inhibitory effect on MMPs, displaying different mechanisms of action, possibly related to their structure. Therefore, their combined use may represent a powerful tool for the down-regulation of MMPs in the course of MS.


Assuntos
Antioxidantes , Astrócitos/efeitos dos fármacos , Astrócitos/enzimologia , Flavonoides , Inibidores de Metaloproteinases de Matriz , Esclerose Múltipla , Fenóis , Animais , Antioxidantes/química , Antioxidantes/farmacologia , Antioxidantes/uso terapêutico , Astrócitos/citologia , Células Cultivadas , Suplementos Nutricionais , Relação Dose-Resposta a Droga , Flavonoides/química , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Humanos , Lipopolissacarídeos/farmacologia , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Estrutura Molecular , Esclerose Múltipla/sangue , Esclerose Múltipla/tratamento farmacológico , Esclerose Múltipla/enzimologia , Fenóis/química , Fenóis/farmacologia , Fenóis/uso terapêutico , Polifenóis , Ratos , Relação Estrutura-Atividade
5.
Mar Drugs ; 9(7): 1220-1231, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21822412

RESUMO

Crustaceans Munida (fam. Galatheideae, ord. Decapodi) were fished in the Southern Adriatic Sea and their proteolytic activities were characterized and tested for potential application in cheese manufacturing. Enzymes extracted from whole crustaceans, mainly serine proteases, showed high caseinolytic and moderate clotting activities. Analysis by 2D zymography of the digestive enzymes extracted from Munida hepatopancreas, showed the presence of several isotrypsin- and isochymotrypsin-like enzymes in the range of 20-34 kDa and 4.1-5.8 pI. Moreover, specific enzymatic assays showed the presence of aminopeptidases and carboxypeptidases A and B. Overall, optimum activity was achieved at pH 7.5 and 40-45 °C. Caseinolytic activity, determined both spectrophotometrically and by SDS gel electrophoresis, indicated higher activity on ß-casein than on α-casein. Miniature cheddar-type cheeses and Pecorino-type cheeses were manufactured by adding starter, rennet and Munida extracts to milk. Reverse-phase HPLC and MALDI-ToF mass spectrometry showed a more complex pattern of proteolytic products in cheeses made using Munida instead of chymosin. Munida extracts were found to degrade the chymosin-derived ß-casein fragment f193-209, one of the peptides associated with bitterness in cheese. In conclusion, Munida digestive enzymes represent a promising tool for development of new cheese products and shorten cheese ripening when used either alone or in addition to calf rennet.


Assuntos
Produtos Biológicos/farmacologia , Queijo/análise , Decápodes/enzimologia , Manipulação de Alimentos/métodos , Manufaturas , Peptídeo Hidrolases/farmacologia , Animais , Produtos Biológicos/análise , Produtos Biológicos/isolamento & purificação , Produtos Biológicos/metabolismo , Caseínas/metabolismo , Indústria Alimentícia , Humanos , Peptídeo Hidrolases/análise , Peptídeo Hidrolases/metabolismo
6.
J Sci Food Agric ; 90(14): 2411-8, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-20672335

RESUMO

BACKGROUND: Proteinases present in kiwi fruits are potentially allergenic enzymes belonging to the papain family of cysteine proteinases. Actinidin is a prominent kiwi enzyme. The study of kiwi proteinases is important for the follow-up of fruit maturation, a deeper insight in the allergenic properties of individual proteins, and the application of kiwi proteinases for meat tenderisation and other industrial purposes. RESULTS: Kiwi crude extracts were analysed by two-dimensional zymography on gelatin-containing gels. The digestion by the reactivated proteolytic enzymes after electrophoresis resulted in insights into kiwi proteinases. A mixture of several enzyme isotypes with the same pI but different molecular mass was observed. Clear spots, corresponding to the proteolytic activities, were excised, digested with trypsin, and submitted to MALDI-ToF mass spectrometry for protein identification. The most representative enzyme was actinidin. CONCLUSIONS: The innovative achievements of the present study are the: (1) two-dimensional zymographic map of kiwi gelatinases without the need for extensive purification; and (2) direct identification of proteinase isotypes by means of direct MALDI-ToF MS analysis of the zymographic spots.


Assuntos
Actinidia/enzimologia , Cisteína Endopeptidases/análise , Eletroforese em Gel Bidimensional/métodos , Endopeptidases/análise , Frutas/enzimologia , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/métodos , Hidrólise , Extratos Vegetais/química
7.
FEBS Lett ; 594(20): 3262-3271, 2020 10.
Artigo em Inglês | MEDLINE | ID: mdl-33011965

RESUMO

The recent explosive increase in the number of works on gut microbiota has been accompanied by the spread of rather vague or improper definitions, chosen more for common use than for experimental evidence. Among them are those defining the human gut microbiota as an organ of our body or as a commensal. But, is the human gut microbiota an organ or a commensal? Here, we address this issue to spearhead a reflection on the real roles of the human gut microbiota in our life. Actually, the misuse of the vocabulary used to describe the properties and functions of the gut microbiota may generate confusion and cause misunderstandings both in the scientific community and among the general public.


Assuntos
Microbioma Gastrointestinal , Simbiose , Biotransformação , Trato Gastrointestinal/microbiologia , Humanos
8.
Nutrients ; 11(11)2019 Nov 09.
Artigo em Inglês | MEDLINE | ID: mdl-31717475

RESUMO

As food is an active subject and may have anti-inflammatory or pro-inflammatory effects, dietary habits may modulate the low-grade neuroinflammation associated with chronic neurodegenerative diseases. Food is living matter different from us, but made of our own nature. Therefore, it is at the same time foreign to us (non-self), if not yet digested, and like us (self), after its complete digestion. To avoid the efflux of undigested food from the lumen, the intestinal barrier must remain intact. What and how much we eat shape the composition of gut microbiota. Gut dysbiosis, as a consequence of Western diets, leads to intestinal inflammation and a leaky intestinal barrier. The efflux of undigested food, microbes, endotoxins, as well as immune-competent cells and molecules, causes chronic systemic inflammation. Opening of the blood-brain barrier may trigger microglia and astrocytes and set up neuroinflammation. We suggest that what determines the organ specificity of the autoimmune-inflammatory process may depend on food antigens resembling proteins of the organ being attacked. This applies to the brain and neuroinflammatory diseases, as to other organs and other diseases, including cancer. Understanding the cooperation between microbiota and undigested food in inflammatory diseases may clarify organ specificity, allow the setting up of adequate experimental models of disease and develop targeted dietary interventions.


Assuntos
Dieta , Disbiose , Microbioma Gastrointestinal , Inflamação , Doenças Neurodegenerativas , Especificidade de Órgãos/imunologia , Animais , Transtorno Autístico/imunologia , Transtorno Autístico/microbiologia , Transtorno Autístico/fisiopatologia , Disbiose/imunologia , Disbiose/microbiologia , Disbiose/fisiopatologia , Humanos , Inflamação/imunologia , Inflamação/microbiologia , Inflamação/fisiopatologia , Camundongos , Doenças Neurodegenerativas/imunologia , Doenças Neurodegenerativas/microbiologia , Doenças Neurodegenerativas/fisiopatologia
9.
Oxid Med Cell Longev ; 2019: 8056904, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31485299

RESUMO

Neuroinflammation, a hallmark of chronic neurodegenerative disorders, is characterized by sustained glial activation and the generation of an inflammatory loop, through the release of cytokines and other neurotoxic mediators that cause oxidative stress and limit functional repair of brain parenchyma. Dietary antioxidants may protect against neurodegenerative diseases by counteracting chronic neuroinflammation and reducing oxidative stress. Here, we describe the effects of a number of natural antioxidants (polyphenols, carotenoids, and thiolic molecules) in rescuing astrocytic function and neuronal viability following glial activation by reducing astrocyte proliferation and restoring astrocytic and neuronal survival and basal levels of reactive oxygen species (ROS). All antioxidant molecules are also effective under conditions of oxidative stress and glutamate toxicity, two maladaptive components of neuroinflammatory processes. Moreover, it is remarkable that their antioxidant and anti-inflammatory activity occurs through differential modulation of NF-κB binding activity in neurons and astrocytes. In fact, we show that inflammatory stimuli promote a significant induction of NF-κB binding activity in astrocytes and its concomitant reduction in neurons. These changes are prevented in astrocytes and neurons pretreated with the antioxidant molecules, suggesting that NF-κB plays a key role in the modulation of survival and anti-inflammatory responses. Finally, we newly demonstrate that effective antigliosis and neuroprotective activity is achieved with a defined cocktail of four natural antioxidants at very low concentrations, suggesting a promising strategy to reduce inflammatory and oxidative damage in neurodegenerative diseases with limited side effects.


Assuntos
Antioxidantes/metabolismo , Astrócitos/metabolismo , NF-kappa B/genética , Doenças Neurodegenerativas/genética , Neuroproteção/genética , Estresse Oxidativo/genética , Humanos
10.
Sci Rep ; 9(1): 642, 2019 01 24.
Artigo em Inglês | MEDLINE | ID: mdl-30679613

RESUMO

Compact myelin forms the basis of nerve insulation essential for higher vertebrates. Dozens of myelin membrane bilayers undergo tight stacking, and in the peripheral nervous system, this is partially enabled by myelin protein zero (P0). Consisting of an immunoglobulin (Ig)-like extracellular domain, a single transmembrane helix, and a cytoplasmic extension (P0ct), P0 harbours an important task in ensuring the integrity of compact myelin in the extracellular compartment, referred to as the intraperiod line. Several disease mutations resulting in peripheral neuropathies have been identified for P0, reflecting its physiological importance, but the arrangement of P0 within the myelin ultrastructure remains obscure. We performed a biophysical characterization of recombinant P0ct. P0ct contributes to the binding affinity between apposed cytoplasmic myelin membrane leaflets, which not only results in changes of the bilayer properties, but also potentially involves the arrangement of the Ig-like domains in a manner that stabilizes the intraperiod line. Transmission electron cryomicroscopy of native full-length P0 showed that P0 stacks lipid membranes by forming antiparallel dimers between the extracellular Ig-like domains. The zipper-like arrangement of the P0 extracellular domains between two membranes explains the double structure of the myelin intraperiod line. Our results contribute to the understanding of PNS myelin, the role of P0 therein, and the underlying molecular foundation of compact myelin stability in health and disease.


Assuntos
Membrana Celular/metabolismo , Proteína P0 da Mielina/química , Proteína P0 da Mielina/metabolismo , Humanos , Modelos Moleculares , Conformação Proteica
11.
Biochim Biophys Acta ; 1770(2): 178-86, 2007 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-17137715

RESUMO

Gelatinase B/matrix metalloproteinase-9 (MMP-9) is a multidomain enzyme functioning in acute and chronic inflammatory and neoplastic diseases. It belongs to a family of more than 20 related zinc proteinases. Therefore, the discovery and the definition of the action mechanism of selective MMP inhibitors form the basis for future therapeutics. The monoclonal antibody REGA-3G12 is a most selective inhibitor of human gelatinase B. REGA-3G12 was found to recognize the aminoterminal part and not the carboxyterminal O-glycosylated and hemopexin protein domains. A variant of gelatinase B, lacking the two carboxyterminal domains, was expressed in insect cells and fragmented with purified proteinases. The fragments were probed by one- and two-dimensional Western blot and immunoprecipitation experiments with REGA-3G12 to map the interactions between the antibody and the enzyme. The interaction unit was identified by Edman degradation analysis as the glycosylated segment from Trp(116) to Lys(214) of gelatinase B. The sequence of this segment was analysed by hydrophobicity/hydrophilicity, accessibility and flexibility profiling. Four hydrophilic peptides were chemically synthesized and used in binding and competition assays. The peptide Gly(171)-Leu(187) in molar excess inhibited partially the binding of MMP-9 to REGA-3G12 and thus refines the structure of the conformational binding site. These results define part of the catalytic domain of gelatinase B/MMP-9, and not the zinc-binding or fibronectin domains, as target for the development of selective inhibitors.


Assuntos
Anticorpos Monoclonais/farmacologia , Inibidores de Metaloproteinases de Matriz , Sequência de Aminoácidos , Animais , Sítios de Ligação , Domínio Catalítico , Clonagem Molecular , DNA Complementar/genética , Inibidores Enzimáticos/farmacologia , Fibronectinas/química , Fibronectinas/metabolismo , Variação Genética , Humanos , Insetos , Metaloproteinase 9 da Matriz/química , Metaloproteinase 9 da Matriz/genética , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Inibidores de Proteases/farmacologia , Proteínas Recombinantes/antagonistas & inibidores , Proteínas Recombinantes/química , Zinco/metabolismo
12.
J Neurochem ; 107(1): 291-301, 2008 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-18680558

RESUMO

It is now well established that plasma membranes, such as the myelin sheath, are made of different microdomains with different lipid and protein composition. Lipid rafts are made mainly of sphingolipids and cholesterol, whereas the non-raft regions are made mainly of phosphoglycerides. Most myelin proteins may distribute themselves in raft and non-raft microdomains but the driving force that gives rise to their different distribution is not known yet. In this paper, we have studied the distribution of protein zero (P0), the most representative protein of PNS myelin, in the membrane microdomains. To this end, we have purified P0 from both non-raft (soluble P0, P0-S) and raft (P0-R) regions of PNS. Purified proteins were analyzed by two-dimensional gel electrophoresis and identified and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. A detailed structural description of the two P0 forms is given in terms of amino acid sequence, post-translational modifications, and composition of associated lipids. Our findings suggest that structural differences between the two proteins, mainly related to the glycogroups, might be responsible for their different localization.


Assuntos
Microdomínios da Membrana/metabolismo , Proteína P0 da Mielina/metabolismo , Bainha de Mielina/metabolismo , Fibras Nervosas Mielinizadas/metabolismo , Nervos Periféricos/metabolismo , Sequência de Aminoácidos/fisiologia , Animais , Bovinos , Feminino , Glicosilação , Proteína P0 da Mielina/análise , Proteína P0 da Mielina/química , Bainha de Mielina/ultraestrutura , Fibras Nervosas Mielinizadas/ultraestrutura , Nervos Periféricos/ultraestrutura , Isoformas de Proteínas/análise , Isoformas de Proteínas/química , Isoformas de Proteínas/metabolismo , Processamento de Proteína Pós-Traducional/fisiologia
13.
Neurotherapeutics ; 15(1): 75-91, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-29067566

RESUMO

Central to the understanding of the relationships between diet, gut microbiota, and vitamins D and A in multiple sclerosis is low-grade inflammation, which is involved in all chronic inflammatory diseases and is influenced by each of the above effectors. We show that food components have either proinflammatory or anti-inflammatory effects and influence both the human metabolism (the "metabolome") and the composition of gut microbiota. Hypercaloric, high-animal-fat Western diets favor anabolism and change gut microbiota composition towards dysbiosis. Subsequent intestinal inflammation leads to leakage of the gut barrier, disruption of the blood-brain barrier, and neuroinflammation. Conversely, a vegetarian diet, rich in fiber, is coherent with gut eubiosis and a healthy condition. Vitamin D levels, mainly insufficient in a persistent low-grade inflammatory status, can be restored to optimal values only by administration of high amounts of cholecalciferol. At its optimal values (>30 ng/ml), vitamin D requires vitamin A for the binding to the vitamin D receptor and exert its anti-inflammatory action. Both vitamins must be supplied to the subjects lacking vitamin D. We conclude that nutrients, including the nondigestible dietary fibers, have a leading role in tackling the low-grade inflammation associated with chronic inflammatory diseases. Their action is mediated by gut microbiota and any microbial change induced by diet modifies host-microbe interactions in a consequent way, to improve the disease or worsen it.


Assuntos
Dieta , Microbioma Gastrointestinal , Esclerose Múltipla/dietoterapia , Esclerose Múltipla/microbiologia , Vitamina A/metabolismo , Vitamina D/metabolismo , Animais , Disbiose/complicações , Encefalite/microbiologia , Humanos , Metaboloma , Esclerose Múltipla/metabolismo
14.
AIDS ; 21(6): 677-84, 2007 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-17413688

RESUMO

OBJECTIVE: To investigate whether antiretroviral therapy (ART) influences the release of matrix metalloproteinase (MMP)-9 from peripheral blood mononuclear cells (PBMC) of HIV-infected individuals. DESIGN: Culture supernatants were collected from PBMC isolated from 46 HIV-infected subjects and 19 healthy donors (HD). Among the HIV-infected subjects, 30 were receiving ART therapy, while 16 were naive for any ART treatment. METHODS: Zymography of culture supernatants was followed by determination of MMP-9 using computered scanning densitometry. MMP-9 net enzymatic activity was determined by the sensitive fluorescent-activated substrate conversion assay (FASC) to complement the zymography data. RESULTS: MMP-9 was significantly elevated in culture supernatants from PBMC of ART-naive subjects in comparison with HD. In the supernatants from 30 HIV-infected subjects receiving ART therapy, MMP-9 was significantly lower than that in those from ART-naive subjects. Analysis by the reverse transcriptase polymerase chain reaction indicated that MMP-9 expression was increased in ART-naive subjects in comparison with HD but ART induced a decrease of MMP-9 expression to levels comparable with those of HD. FASC used as a functional assay showed conversion of fluorescent gelatine in ART-naive subjects, indicating the presence of active MMP-9. By contrast, in both HD and ART-treated subjects, there was no MMP-9 activity, indicating that MMP-9 was completely blocked by binding to its natural tissue inhibitor TIMP-1. CONCLUSIONS: The present findings show for the first time that ART can reduce the capacity of PBMC from HIV-infected patients to secrete increased amounts of MMP-9.


Assuntos
Antirretrovirais/uso terapêutico , Infecções por HIV/imunologia , Leucócitos Mononucleares/enzimologia , Inibidores de Metaloproteinases de Matriz , Adulto , Idoso , Feminino , Citometria de Fluxo/métodos , Expressão Gênica , Infecções por HIV/tratamento farmacológico , Infecções por HIV/enzimologia , Inibidores da Protease de HIV/uso terapêutico , Humanos , Leucócitos Mononucleares/imunologia , Masculino , Metaloproteinase 9 da Matriz/genética , Pessoa de Meia-Idade , RNA Mensageiro/análise , RNA Viral/análise , Inibidores da Transcriptase Reversa/uso terapêutico , Carga Viral
15.
Ann Ital Chir ; 78(4): 283-8; discussion 288-9, 2007.
Artigo em Inglês | MEDLINE | ID: mdl-17990602

RESUMO

BACKGROUND: With term GIST is now defined a group of mesenchimal tumours of the gastrointestinal tract expressing immunopositivity for kit protein kinase (CD117). Surgical therapy remains the gold standard for these rare tumours. Imatinib Mesylate (STI-571) is a potent inhibitor of Kit Kinase activity and different reports demonstrated its efficacy in unresectable or metastatic Gists. AIM OF THE PAPER: To value the incidence of GISTs among gastric mesenchimal neoplasms and analyzed their clinical presentation, prognostic parameters and surgical treatment. The response to Imatinib Mesylate in a case of metastatic GIST is then valued. METHODS: Twelve cases of gastric mesenchimal neoplasms are retrospectively reviewed and tested by CD117 immmunopositivity identifyng 8 GISTs. The median follow-up was 37 (range7-120) months. We describe in details the case of a metastatic Gist treated for 15 months with Imatinib Mesylate. RESULTS: The 67 per cent of mesenchimal gastric tumours were CD117+. Gastrointestinal bleeding was the most common presenting symptom. The 50% of patiens with malignant GISTS had a palpable abdominal mass at diagnosis. All tumors < 5 cm in diameter had a mitotic count (MC) <5/50 high-power fields (HPFs) except a case of high grade leiomyosarcoma. Surgical therapy was complete tumour resection with free margins. No recurrence was observed in lesions <5cm and < 5 mitosis/50 High Power Fields (HPFs). A good response to Imatinib Mesylate was reported in a metastatic GIST. CONCLUSION: The surgeon's role in gastric Gist's treatment is to achieve a complete cancer resection with free margins. In advanced lesions, even in presence of hepatic metastases, surgical resection of the mass is indicated because is possible to obtain a stabilization or a partial remission with Imatinib Mesylate palliative treatment in some patients.


Assuntos
Tumores do Estroma Gastrointestinal , Idoso , Antineoplásicos/uso terapêutico , Benzamidas , Biomarcadores Tumorais/análise , Tumores do Estroma Gastrointestinal/diagnóstico , Tumores do Estroma Gastrointestinal/epidemiologia , Tumores do Estroma Gastrointestinal/terapia , Humanos , Mesilato de Imatinib , Incidência , Masculino , Piperazinas/uso terapêutico , Inibidores de Proteínas Quinases/uso terapêutico , Proteínas Proto-Oncogênicas c-kit/análise , Pirimidinas/uso terapêutico , Estudos Retrospectivos
16.
Ann Ital Chir ; 78(5): 413-7; discussion 417-8, 2007.
Artigo em Italiano | MEDLINE | ID: mdl-18338550

RESUMO

BACKGROUND: Sentinel node biopsy is a minimally invasive technique alternative to routine axillary dissection in breast cancer staging. This technique selects women with positive nodes who may benefit from axillary dissection, avoiding unnecessary operations in negative node biopsies. AIMS: In this article we report a 5 year multi disciplinary experience in sentinel node biopsy involving the General Surgery Unit of Imola Hospital in collaboration with Radiologist, Pathologists and Specialists in Nuclear Medicine. METHODS: From 2000 to 2004 the Authors treated 209 women performing 214 sentinel node biopsies (in 5 cases the tumor was bilateral). Sentinel node identification was undertaken by lymphoscintigraphy; in 15 cases we associated intradermal injection of blue dye. Lymph nodes were examined by at least 60 hematoxylin and eosin stained sections and when nodes found negative were further studied with immunohistochemical stains for cytokeratins. RESULTS: Sentinel node identification rate was 99.1%. In 62 patients sentinel node was metastatic and in 17 such nodes micrometastases were detected. In 6 cases with single metastatic cells, axillary dissection was not perfomed, in accordance to current opinions. In 50 of 62 women with metastatic axillary nodes (80.6%) the sentinel node was the only metastatic one. Number of axillary dissections decreased of more than 70% in four years, from theoretical 214 to 62. CONCLUSION: Sentinel node biopsy is currently a validated technique and many breast cancer patients are spared a regional lymph node dissection without compromising local control and the accuracy of staging.


Assuntos
Neoplasias da Mama/patologia , Neoplasias da Mama/cirurgia , Biópsia de Linfonodo Sentinela , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade
17.
Artigo em Inglês | MEDLINE | ID: mdl-16260192

RESUMO

Histamine content in fish may increase by decarboxylation of free histidine to values that can be toxic, if storage conditions are not well controlled. We have studied the influence of storage temperature and time of freezing on histamine formation in the anchovy, Engraulis encrasicholus (L., 1758), for which little information is available. Analysis, carried out by capillary zone electrophoresis (CZE) without sample pre-treatment, was very simple, fast and reproducible. Results indicate that temperatures above 20 degrees C notably increase histamine production, whereas freezing can clearly prevent or slow down the process.


Assuntos
Eletroforese Capilar/métodos , Congelamento , Histamina/análise , Animais , Peixes , Temperatura
18.
Exp Biol Med (Maywood) ; 241(6): 620-35, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26785711

RESUMO

The aim of this work was to assess the influence of nutritional intervention on inflammatory status and wellness in people with multiple sclerosis. To this end, in a seven-month pilot study we investigated the effects of a calorie-restricted, semi-vegetarian diet and administration of vitamin D and other dietary supplements (fish oil, lipoic acid, omega-3 polyunsaturated fatty acids, resveratrol and multivitamin complex) in 33 patients with relapsing-remitting multiple sclerosis and 10 patients with primary-progressive multiple sclerosis. At 0/3/6 months, patients had neurological examination, filled questionnaires and underwent anthropometric measurements and biochemical analyses. Serum fatty acids and vitamin D levels were measured as markers of dietary compliance and nutritional efficacy of treatment, whereas serum gelatinase levels were analyzed as markers of inflammatory status. All patients had insufficient levels of vitamin D at baseline, but their values did not ameliorate following a weekly administration of 5000 IU, and rather decreased over time. Conversely, omega-3 polyunsaturated fatty acids increased already after three months, even under dietary restriction only. Co-treatment with interferon-beta in relapsing-remitting multiple sclerosis was irrelevant to vitamin D levels. After six months nutritional treatment, no significant changes in neurological signs were observed in any group. However, serum levels of the activated isoforms of gelatinase matrix metalloproteinase-9 decreased by 59% in primary-progressive multiple sclerosis and by 51% in relapsing-remitting multiple sclerosis patients under nutritional intervention, including dietary supplements. This study indicates that a healthy nutritional intervention is well accepted by people with multiple sclerosis and may ameliorate their physical and inflammatory status.


Assuntos
Anti-Inflamatórios/uso terapêutico , Dieta/métodos , Esclerose Múltipla Crônica Progressiva/terapia , Adulto , Antropometria , Biomarcadores/análise , Ácidos Graxos/sangue , Feminino , Gelatinases/sangue , Humanos , Masculino , Pessoa de Meia-Idade , Esclerose Múltipla Crônica Progressiva/patologia , Exame Neurológico , Projetos Piloto , Recidiva , Soro/química , Inquéritos e Questionários , Resultado do Tratamento , Vitamina D/sangue , Adulto Jovem
19.
J Neuroimmunol ; 168(1-2): 111-7, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16120465

RESUMO

Pixantrone is an immunesuppressor similar to mitoxantrone but with lower cardiotoxicity. We evaluated the effect of pixantrone on B cells and lymphomononuclear cells in the course of acute EAE. Pixantrone reduced the number of B cells and suppressed myelin basic protein (MBP) specific IgG production. In vitro, pixantrone induced apoptosis of rat B lymphocytes in a way similar to mitoxantrone. In addition, pixantrone inhibited antigen specific and mitogen induced lymphomononuclear cell proliferation, as well as IFN-gamma production, during EAE. These findings suggest a similar mechanism of action for pixantrone and mitoxantrone on the effector function of lymphomonocyte B and T cells.


Assuntos
Linfócitos B/efeitos dos fármacos , Encefalomielite Autoimune Experimental/tratamento farmacológico , Imunossupressores/uso terapêutico , Isoquinolinas/uso terapêutico , Análise de Variância , Animais , Anticorpos/sangue , Antígenos CD/imunologia , Antígenos CD/metabolismo , Apoptose/efeitos dos fármacos , Linfócitos B/fisiologia , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Citocinese , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Citometria de Fluxo/métodos , Proteína Básica da Mielina/administração & dosagem , Proteína Básica da Mielina/imunologia , Ratos , Ratos Endogâmicos Lew , Linfócitos T/efeitos dos fármacos , Fatores de Tempo
20.
Mol Nutr Food Res ; 49(10): 926-31, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16189794

RESUMO

Deep-water fish are becoming an interesting object of studies and research due to the development of deep fishery activities. This paper analyses the chemical composition and nutritional value of the fish species Mora moro (Risso, 1810) inhabiting deep Mediterranean waters. The fatty acid profile and the principal water-soluble proteins present in the white muscle of this fish species have also been determined. The major fatty acids were 22 : 6n-3, 16 : 0, 18 : 1n-9, 20 : 4n-6 and 20 : 5n-3. The polyunsaturated fatty acid (PUFA) content was higher than that of saturated (SFA) and monounsaturated fatty acids, but the ratio PUFA/SFA was lower than the value reported in other studies. Both the atherogenic index and thrombogenic index were very low. Water-soluble proteins were characterised by monodimensional native PAGE and 2-D SDS-gel electrophoresis. Protein patterns showed the presence of parvalbumins and of the principal myofibrillar proteins. Therefore, the deep-water fish M. moro could represent an interesting target for deep-sea fishery and commercial exploitation.


Assuntos
Ácidos Graxos/análise , Peixes , Proteínas Musculares/análise , Músculos/química , Valor Nutritivo , Animais , Cromatografia Gasosa , Eletroforese em Gel de Poliacrilamida , Ácidos Graxos Insaturados , Carne/análise , Solubilidade , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Água
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