ABSTRACT
PURPOSE: Recently, there was an increasing interest on the use of ancient grains because of their better health-related composition. The aim of this study was to evaluate in healthy human subjects the antioxidative and diabetes-preventive properties of ancient KAMUT® khorasan wheat compared to modern wheat. METHODS: The study was a randomized, non-blind, parallel arm study where the biochemical parameters of volunteers with a diet based on organic whole grain KAMUT® khorasan products, as the only source of cereal products were compared to a similar replacement diet based on organic whole grain modern durum wheat products. A total of 30 healthy volunteers were recruited and the intervention period lasted 16 weeks. Blood analyses were performed before and after the diet intervention. The effect of KAMUT® khorasan products on biochemical parameters was analyzed by multiple quantile regression adjusted for age, sex, physical activity and BMI compared to data at baseline. RESULTS: Subjects receiving KAMUT® khorasan products showed a significantly greater decrease of fat mass (b = 3.7%; CI 1.6-5.5; p = 0.042), insulin (b = 2.4 µU/ml; CI 0.2-4.2; p = 0.036) and a significant increase of DHA (b = - 0.52%; CI - 1.1 to - 0.12; p = 0.021). CONCLUSIONS: Our study provides evidence that a substitution diet with KAMUT® khorasan wheat products can reduce some markers associated to the development of type-2 diabetes compared to a diet of modern wheat.
Subject(s)
Antioxidants/pharmacology , Diabetes Mellitus, Type 2/prevention & control , Oxidative Stress/drug effects , Triticum , Adult , Edible Grain , Female , Healthy Volunteers , Humans , Male , Pilot Projects , Reference ValuesABSTRACT
Lunasin is a peptide whose anticancer properties are widely reported. Originally isolated from soybean seeds, lunasin was also found in cereal (wheat, rye, barley and Triticale), Solanum and amaranthus seeds. However, it was recently reported that searches of transcript and DNA sequence databases for wheat and other cereals failed to identify sequences with similarity to those encoding the lunasin peptide in soy. In order to clarify the presence or absence of lunasin in wheat varieties, a broad investigation based on chemical (LC-ESI-MS) and molecular (PCR) analyses was conducted. Both approaches pointed out the absence of lunasin in the investigated wheat genotypes; in particular no compounds with a molecular weight similar to that of lunasin standard and no lunasin-related sequences were found in the analysed wheat samples. These findings confirm the hypothesis, reported in recent researches, that lunasin is not a wheat-derived peptide.
Subject(s)
Plant Proteins/chemistry , Triticum/chemistry , DNA, Plant/genetics , Genotype , Molecular Weight , Seeds/chemistry , Sequence Analysis, DNA , Soybean Proteins , Glycine max/chemistryABSTRACT
The hepatitis C virus (HCV) serine protease is necessary for viral replication and represents a valid target for developing new therapies for HCV infection. Potent and selective inhibitors of this enzyme have been identified and shown to inhibit HCV replication in tissue culture. The optimization of these inhibitors for clinical development would greatly benefit from in vitro systems for the identification and the study of resistant variants. We report the use HCV subgenomic replicons to isolate and characterize mutants resistant to a protease inhibitor. Taking advantage of the replicons' ability to transduce resistance to neomycin, we selected replicons with decreased sensitivity to the inhibitor by culturing the host cells in the presence of the inhibitor and neomycin. The selected replicons replicated to the same extent as those in parental cells. Sequence analysis followed by transfection of replicons containing isolated mutations revealed that resistance was mediated by amino acid substitutions in the protease. These results were confirmed by in vitro experiments with mutant enzymes and by modeling the inhibitor in the three-dimensional structure of the protease.
Subject(s)
Carrier Proteins/genetics , Drug Resistance, Viral/genetics , Genetic Variation , Hepacivirus/drug effects , Serine Proteinase Inhibitors/pharmacology , Viral Nonstructural Proteins/genetics , Viral Proteins/genetics , Binding Sites/drug effects , Carrier Proteins/antagonists & inhibitors , Carrier Proteins/drug effects , Hepacivirus/enzymology , Hepacivirus/genetics , Humans , Intracellular Signaling Peptides and Proteins , Microbial Sensitivity Tests , Models, Molecular , Mutation , Replicon , Selection, Genetic , Tumor Cells, Cultured , Viral Nonstructural Proteins/antagonists & inhibitors , Viral Nonstructural Proteins/drug effects , Viral Proteins/antagonists & inhibitors , Viral Proteins/drug effectsABSTRACT
For many years our knowledge on hepatitis C virus (HCV) replication has been based on in vitro experiments or transfection studies. Recently, the first reliable system for studying viral replication in tissue culture cells was developed. Taking advantage of this system, we examined in detail the localization of viral nonstructural (NS) proteins in cells containing functional replication complexes. By fractionation experiments and immunomicroscopy, we observed that all NS proteins were associated with the endoplasmic reticulum (ER) membranes, confirming the hypothesis that the ER is the site of membrane-associated HCV RNA replication. Interestingly, NS3 and NS4A were preferentially localized in endoplasmic reticulum cisternae surrounding mitochondria, suggesting additional subcellular compartment-related functions for these viral proteins. Furthermore, the immunoelectron microscopy revealed the loss of the organization and other morphological alterations of the ER (convoluted cisternae and paracrystalline structures), resembling alterations observed in liver biopsies of HCV-infected individuals and in flavivirus-infected cells.
