Search results for "Epsin"
showing 10 items of 218 documents
Aspartic Proteinase from Barley Seeds is Related to Animal Cathepsin D
1991
In contrast to the well-characterized mammalian aspartic proteinases, plant aspartic proteinases have received little attention so far. Aspartic proteinase activity has been detected, for example, in resting seeds of scots pine (Salmia et al., 1978), soybean (Bond & Bowles, 1983), barley and wheat (Morris et al., 1985) as well as in leaves of orange (Garcia-Martinez & Moreno, 1986) and barley (Kervinen et al., 1990). Aspartic proteinases have been purified from the seeds of rice (Doi et al., 1980), cucumber, squash (Polanowski et al 1985) and wheat (Dunaevsky et al., 1989) as well as from the leaves of tomato (Rodrigo et al., 1989). The plant aspartic proteinases have been reported to enhan…
Identification of Casein Phosphopeptides in β-casein and Commercial Hydrolysed Casein by Mass Spectrometry
2006
Casein phosphopeptides (CPPs) in commercial hydrolysed casein (CE90CPP) and in β-CN (β-CN) after simulated gastrointestinal digestion (gastric stage pepsin, pH =2, 37°C 2h) and intestinal stage (pancreatic-bile extract, pH =5.2, 37°C 2h) were sequenced by on-line reversed-phase high performance liquid chromatography coupled to electrospray ionisation tandem mass spectrometry (RP-HPLC-ESIMS/MS). In β-CN digest five peptides that contained four to five phosphate groups and the cluster sequence SpSpSpEE (residues 17-21) were identified. All CPPs with one exception β-CN(1-24)4P, had the protein fragment β-CN(1-25)4P, which is one of the main CPPs produced in vivo digestion of casein and the re…
Effect of Enzyme Amounts Used in Gastrointestinal Digestion Upon Solubility and Caco-2 Cell Uptake Assays of Minerals from Infant Formulas
2005
The effect of enzyme amounts used in gastrointestinal in vitro digestion upon the solubility and Caco-2 cell uptake of calcium, iron and zinc from infant formulas (IFs) was studied. Different amounts of enzymes (g enzyme/g IF), pepsin (0.002 and 0.048), pancreatin (0.0005, 0.002 and 0.01) and bile extract (0.003, 0.125 and 0.0625) were assayed. Mineral soluble contents and mineral uptakes by Caco-2 cells were affected by the enzyme amounts used in digestion. Although the highest mineral solubility (Ca 98.6 vs 46.2%; Fe 98.1 vs 83.9%; Zn 98.4 vs 83%) was obtained when the lowest enzyme (pepsin 0.002 vs 0.048; pancreatin 0.0005 vs 0.01g/g IF) and bile extract (0.003 vs 0.0625g/g IF) amounts …
Improvement of Analytical Conditions of Mineral Caco-2 Cell Uptake Assays
2004
Caco-2 cell mineral uptake assays are used to estimate the bioavailability of minerals from foods. The uptake of minerals by Caco-2 cells can be affected by several factors – particularly the conditions of the in vitro digestion process and the growth conditions of the cell culture. Therefore, a standardisation of the assays conditions is required to obtain reproducible results. This work determined the effect of enzyme demineralisation, the inactivation of the proteolytic activity of the digest and the replacement of distilled-deionised water by cell culture degree water. Treatment with Chelex-100 resin was applied to remove calcium, iron and zinc from pepsin and pancreatin-bile salts. Th…
Aspartic proteinase from barley grains is related to mammalian lysosomal cathepsin D
1991
Resting barley (Hordeum vulgare L.) grains contain acid-proteinase activity. The corresponding enzyme was purified from grain extracts by affinity chromatography on a pepstatin-Sepharose column. The pH optimum of the affinity-purified enzyme was between 3.5 and 3.9 as measured by hemoglobin hydrolysis and the enzymatic activity was completely inhibited by pepstatin a specific inhibitor of aspartic proteinases (EC 3.4.23). Further purification on a Mono S column followed by activity measurements and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the affinity-purified enzyme preparation contained two active heterodimeric aspartic proteinases: a larger 48k Da enzyme, c…
Neuropeptide Y (NPY) in cerebrospinal fluid from patients with Huntington's Disease: increased NPY levels and differential degradation of the NPY1-30…
2016
Huntington's disease (HD) is an inherited and fatal polyglutamine neurodegenerative disorder caused by an expansion of the CAG triplet repeat coding region within the HD gene. Progressive dysfunction and loss of striatal GABAergic medium spiny neurons (MSNs) may account for some of the characteristic symptoms in HD patients. Interestingly, in HD, MSNs expressing neuropeptide Y (NPY) are spared and their numbers is even up-regulated in HD patients. Consistent with this, we report here on increased immuno-linked NPY (IL-NPY) levels in human cerebrospinal fluid (hCSF) from HD patients (Control n = 10; early HD n = 9; mid HD n = 11). As this antibody-based detection of NPY may provide false pos…
Amorphous, Smart, and Bioinspired Polyphosphate Nano/Microparticles: A Biomaterial for Regeneration and Repair of Osteo-Articular Impairments In-Situ
2018
Using femur explants from mice as an in vitro model, we investigated the effect of the physiological polymer, inorganic polyphosphate (polyP), on differentiation of the cells of the bone marrow in their natural microenvironment into the osteogenic and chondrogenic lineages. In the form of amorphous Ca-polyP nano/microparticles, polyP retains its function to act as both an intra- and extracellular metabolic fuel and a stimulus eliciting morphogenetic signals. The method for synthesis of the nano/microparticles with the polyanionic polyP also allowed the fabrication of hybrid particles with the bisphosphonate zoledronic acid, a drug used in therapy of bone metastases in cancer patients. The r…
The potential of cystatin C as a predictive biomarker in breast cancer
2020
Breast cancer (BCa) is the leading cause of cancer-related deaths among women. Numerous efforts are being directed toward identifying novel tissue and/or circulating molecular markers that may help clinicians in detecting early-stage BCa patients and in providing an accurate estimation of the prognosis and prediction of response to clinical treatments. In this setting, emerging evidence has indicated Cystatin C (Cyst C), as the most potent endogenous inhibitor of cysteine cathepsins, as a possible useful marker in the clinical management of BCa patients.This review analyzes the results of emerging studies underpinning a potential clinical role of Cyst C, as additional marker in BCa.Cyst C e…
Clinical Impact of Cystatin C/Cathepsin L and Follistatin/Activin A Systems in Breast Cancer Progression: A Preliminary Report.
2016
This study was directed to assess the clinical impact of the circulating cathepsin L, cystatin C, activin A, and follistatin in breast cancer patients. The serum concentrations of these molecules were determined by immunoenzymatic assays, and their association with some clinico-pathological parameters of breast cancer progression was evaluated. Our results identified cystatin C and activin A as predictive markers for the presence of breast cancer and bone metastasis, respectively. Therefore, these proteins may have a clinical role as circulating biomarkers in the diagnosis and therapeutic monitoring of breast cancer patients.
Development of Novel Peptide-Based Michael Acceptors Targeting Rhodesain and Falcipain-2 for the Treatment of Neglected Tropical Diseases (NTDs)
2017
This paper describes the development of a class of peptide-based inhibitors as novel antitrypanosomal and antimalarial agents. The inhibitors are based on a characteristic peptide sequence for the inhibition of the cysteine proteases rhodesain of Trypanosoma brucei rhodesiense and falcipain-2 of Plasmodium falciparum. We exploited the reactivity of novel unsaturated electrophilic functions such as vinyl-sulfones, -ketones, -esters, and -nitriles. The Michael acceptors inhibited both rhodesain and falcipain-2, at nanomolar and micromolar levels, respectively. In particular, the vinyl ketone 3b has emerged as a potent rhodesain inhibitor (k2nd = 67 × 106 M-1 min-1), endowed with a picomolar b…