102 research outputs found
The role of proteases in fibrinonectin matrix remodeling in thyroid epithelial cell monolayer cultures
No full textFischer rat thyroid (FRT) cells organize a matrix of extracellular fibronectin (FN) fibrils, which undergoes extensive remodeling according to cell culture confluence. In non-confluent cells FN forms a fibrillar array associated with the ventral cell surface. However, basal FN is progressively removed in confluent cultures and substituted by non-fibrillar FN deposits at lateral cell domains in regions of cell-cell contacts. FRT cells secrete and expose on the plasma membrane the tissue-type plasminogen activator and, in serum-free cultures, plasminogen induces a rapid loss of FN fibrils. Incubation with plasmin inhibitors greatly reduces this effect. FRT cells also express annexin II, a plasminogen receptor, suggesting that plasmin activity is associated with the pericellular enviroment. This is in agreement with the observation that a great reduction in FN degradation is observed if the cells are pre-incubated with carboxypeptidase B, which prevents plasminogen binding to the cells. A gelatinolytic activity with a molecular weigth equivalent to MMP-2 has been demonstrated by zymography of culture media, and the presence of MMP-2 and MT1-MMP on the cell plasma membrane has been detected by immunofluorescence. These results indicate that in the FN remodeling process, occurring during FRT epithelium maturation, both plasmin-dependent (tPA activated) and plasmin-independent proteolytic activities are involved
A microfluidic platform to unravel immune cells cross-talk in rheumatoid arthritis
No full textRheumatoid arthritis (RA) is a chronic, inflammatory autoimmune disease that can cause irreversible joint damage and significant disability. Joint inflammation in RA is closely related to infiltration of immune cells, among which circulating T-cells and synovial resident macrophages play a critical role. Thus, including immune system in in vitro models is of crucial importance to better understand RA pathogenesis. Here, we developed a novel 3D microfluidic platform aimed at evaluating the active role of immune cells involved in RA. In particular, the platform was used to study spontaneous migration of Th1 T-cells towards pro-inflammatory M1 macrophages, thanks to the insertion of specific valves mechanism, allowing the trapping of immune cells inside the device and the stimulation of different cell types separately
Metabolomic and Proteomic Profile of Dried Hop Inflorescences (Humulus lupulus L. cv. Chinook and cv. Cascade) by SPME-GC-MS and UPLC-MS-MS
No full textHop (Humulus lupulus L.) is grown mainly for the production of beer. The flowers of the female plant give it the bitter taste and pungent aroma. There are a large number of hop varieties differing in their α-acid content, essential oil levels and odor profiles. Aside from their use in brewing, more recently, hops have been used for the pharmacological properties of its derivatives that are of great importance to the pharmaceutical industry. Hop is known to have a fairly complex chemistry characterized by the presence of a variety of sesquiterpenoids, diterpenoids and triterpenoids, phytoestrogens and flavonoids. Additionally, considering the countless applications in the pharmacological sector in recent years, a chemical characterization of the different cultivars is essential to better identify the source of specific secondary metabolites. For this purpose, the dried inflorescences of two hop cultivars, Chinook and Cascade, were investigated using Solid-Phase Microextraction-Gas Chromatography-Mass Spectrometry and Liquid Chromatography-Tandem Mass Spectrometry (SPME-GC-MS and LC-MS-MS) to describe their metabolomic and proteomic profile. Furthermore, thanks to an in-depth statistical survey, it was possible to carry out a comparative study highlighting interesting implications deriving from this investigative study
A MICROFLUIDIC PLATFORM TO INVESTIGATE THE CROSS-TALK BETWEEN IMMUNE CELLS IN RHEUMATOID ARTHRITIS
No full textProtein-tyrosine phosphatase PTPD1 regulates focal adhesion kinase autophosphorylation and cell migration.
No full textPTPD1 is a cytosolic nonreceptor tyrosine phosphatase and a positive regulator of the Src-epidermal growth factor transduction pathway. We show that PTPD1 localizes along actin filaments and at adhesion plaques. PTPD1 forms a stable complex via distinct molecular modules with actin, Src tyrosine kinase, and focal adhesion kinase (FAK), a scaffold protein kinase enriched at adhesion plaques. Overexpression of PTPD1 promoted cell scattering and migration, short hairpin RNA-mediated silencing of endogenous PTPD1, or expression of PTPD1 mutants lacking either catalytic activity (PTPD1(C1108S)) or the FERM domain (PTPD1(Delta1-325)) significantly reduced cell motility. PTPD1 and Src catalytic activities were both required for epidermal growth factor-induced FAK autophosphorylation at its active site and for downstream propagation of ERK1/2 signaling. Our findings demonstrate that PTPD1 is a component of a multivalent scaffold complex nucleated by FAK at specific intracellular sites. By modulating Src-FAK signaling at adhesion sites, PTPD1 promotes the cytoskeleton events that induce cell adhesion and migration
Protein-tyrosine phosphatase PTPD1 regulates focal adhesion kinase autophosphorylation and cell migration
No full textPTPD1 is a cytosolic nonreceptor tyrosine phosphatase and a positive regulator of the Src-epidermal growth factor transduction pathway. We show that PTPD1 localizes along actin filaments and at adhesion plaques. PTPD1 forms a stable complex via distinct molecular modules with actin, Src tyrosine kinase, and focal adhesion kinase (FAK), a scaffold protein kinase enriched at adhesion plaques. Overexpression of PTPD1 promoted cell scattering and migration, short hairpin RNA-mediated silencing of endogenous PTPD1, or expression of PTPD1 mutants lacking either catalytic activity (PTPD1(C1108S)) or the FERM domain (PTPD1(Delta 1-325)) significantly reduced cell motility. PTPD1 and Src catalytic activities were both required for epidermal growth factor-induced FAK autophosphorylation at its active site and for downstream propagation of ERK1/2 signaling. Our findings demonstrate that PTPD1 is a component of a multivalent scaffold complex nucleated by FAK at specific intracellular sites. By modulating Src-FAK signaling at adhesion sites, PTPD1 promotes the cytoskeleton events that induce cell adhesion and migration
PENGARUH PERENDAMAN NATRIUM METABISULFIT (Na2S2O5) TERHADAP MUTU TEPUNG TERUNG HIJAU (Solanum melongena L.)
No full textABSTRAK
Terung hijau (Solanum melongena L.) merupakan salah satu produk hortikultura dengan jenis sayuran yang memiliki kandungan air dan serat pangan cukup tinggi. Terung hijau memiliki sifat yang tidak tahan lama setelah panen. Salah satu cara yang dapat dilakukan untuk memperpanjang umur simpan terung adalah menjadikannya sebagai olahan berbentuk tepung. Selama pengolahan setelah terung hijau dirajang akan terjadi proses pembentukan pigmen bewarna coklat (browning enzimatis). Untuk mempertahankan mutu warna dari tepung terung hijau salah satu usaha yang dapat dilakukan untuk menghambat terjadinya browning enzimatis adalah perendaman menggunakan natrium metabisulfit (Na2S2O5). Penelitian ini bertujuan untuk menganalisis pengaruh dan menentukan konsentrasi terbaik perendaman menggunakan natrium metabisulfit (Na2S2O5) terhadap mutu tepung terung hijau (solanum melongena L.). Penelitian ini menggunakan metode eksperimen dan analisis datanya mengunakan RAL dengan SPSS 23. Pada penelitian ini terung direndam selama 20 menit dengan konsentrasi natrium metabisulfit (Na2S2O5) 0%, 0,25% dan 0,3%. Berdasarkan hasil penelitian konsentrasi natrium metabisulfit (Na2S2O5) berpengaruh nyata terhadap pemberian natrium metabisulfit tetapi tidak berpengaruh nyata terhadap konsentrasi natrium metabisulfit diatas 0% seperti pada kadar air setelah perendaman, rendemen, persentase kehalusan tepung, efisiensi total, uji organoleptik rasa dan uji organoleptik tekstur karena pengelompokan yang sama pada uji Duncan kecuali pada kadar air tepung dan uji organoleptik warna . Hasil penelitian perendaman dengan natrium metabisulfit (Na2S2O5) dengan konsentrasi 0,3% merupakan perlakuan terbaik untuk mutu tepung terung hijau pada penelitian ini.
Kata kunci : Terung Hijau, Natrium Metabisulfit (Na2S2O5), mutu
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