ABSTRACT
Novel assemblies of DNA-functionalized gold nanoparticles (DNA-GNPs) have received considerable interest due to their fascinating properties which are desired for various detection applications. In this study, we present innovative GNP assemblies which have a cage-shaped protein ferritin in the center, and discrete GNPs sterically surrounding the central ferritin. These assemblies were constructed by hybridizing DNA-GNP to chemically DNA-modified ferritin, which has a hollow cavity or an iron NP core. Subsequent gel electrophoresis purification and transmission electron microscopy observation showed that ferritin/DNA/GNP assemblies were successfully constructed and can be isolated as independent functional units, which can be used to investigate not only the interaction between the GNPs of complicated GNP clusters but also the interaction between the GNPs and the internalized NP.
Subject(s)
DNA/chemistry , Ferritins/chemistry , Gold/chemistry , Metal Nanoparticles/chemistry , Nucleic Acid Hybridization/methods , Animals , Electrophoresis, Agar Gel , Electrophoresis, Polyacrylamide Gel , Horses , Iron/chemistry , Metal Nanoparticles/ultrastructure , Surface PropertiesABSTRACT
Ferritin has a mono-dispersed structure and biomineralization properties that allow it to form various kinds of nanoparticles and play an important role in modern nanotechnology. Independent nanoparticles synthesized in ferritin are valuable, but moreover a pair of nanoparticles can bring new properties different from those of the independent nanoparticles. In this study, by breaking ferritin's symmetry, we successfully produced ferritin dimers which provide real protein frameworks for nanoparticle dimer formation. Identical nickel hydro-oxide nanoparticle dimers were produced by simply biomineralizing ferritin dimers. The method presented here can produce multi-functional ferritin dimers with different kinds of nanoparticles.
Subject(s)
Ferritins/chemistry , Protein Multimerization , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , Ferritins/ultrastructure , Hydrogen-Ion Concentration , Light , Mutant Proteins/chemistry , Protein Structure, Secondary , Scattering, Radiation , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
Newly designed porter proteins, which catch gold nanoparticles and deliver the nanoparticles selectively to a silicon dioxide (SiO(2)) surface under the specific conditions were reported. Recombinant apoferritin subunits, each of which has gold-binding peptide and titanium-binding peptide at the C- and N-terminus, respectively, can efficiently encapsulate a gold nanoparticle. The bio-conjugate, a nanogold and surrounding mutant protein subunits, had a property which can deliver itself to the SiO(2) surface through the interaction. In theory, our genetically manipulated apoferritin subunits can encapsulate gold nanoparticles of various sizes, which is a promising property for applications involving surface plasmon resonance.
Subject(s)
Drug Delivery Systems/methods , Ferritins/metabolism , Gold/metabolism , Metal Nanoparticles/chemistry , Recombinant Proteins/metabolism , Adsorption , Ferritins/chemistry , Ferritins/genetics , Gold/chemistry , Metal Nanoparticles/ultrastructure , Models, Molecular , Mutation , Peptides/chemistry , Peptides/metabolism , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Silicon Dioxide/chemistry , Surface Plasmon ResonanceSubject(s)
Adenocarcinoma/complications , Amyloidosis/complications , Intestinal Diseases/complications , Intestinal Neoplasms/complications , Adenocarcinoma/pathology , Adenocarcinoma/surgery , Aged , Amyloidosis/pathology , Humans , Intestinal Diseases/pathology , Intestinal Neoplasms/pathology , Intestinal Neoplasms/surgery , Intestine, Small/pathology , MaleABSTRACT
An enzyme-linked-immunosorbent assay (ELISA) with HCl heat-extracted antigen of Fusobacterium necrophorum was conducted to detect specific immunoglobulins G and M in infected cattle. The ELISA revealed an increase (> 0.40) in specific IgG in most of the animals with hepatic abscesses but not that in specific IgM. All the lesions were positive for F. necrophorum. These findings indicated that the ELISA for immunoglobulin G detection may prove to be a useful tool for predictive serodiagnosis of F. necrophorum infection in cattle.
Subject(s)
Antibodies, Bacterial/blood , Fusobacterium necrophorum/immunology , Immunoglobulin G/blood , Immunoglobulin M/blood , Animals , Cattle , Enzyme-Linked Immunosorbent Assay , Fusobacterium Infections/diagnosis , Serologic TestsSubject(s)
Glomerulonephritis, IGA/immunology , IgA Vasculitis/immunology , Immunoglobulins/analysis , Nephritis/immunology , Adolescent , Child , Child, Preschool , Complement C3/analysis , Female , Fluorescent Antibody Technique , Glomerular Mesangium/analysis , Humans , Immunoglobulin A/analysis , MaleSubject(s)
Nephritis, Hereditary/epidemiology , Adolescent , Adult , Age Factors , Child , Child, Preschool , Female , Humans , Japan , Kidney Diseases/genetics , Male , Sex FactorsABSTRACT
Fibrinogen fragments (X, Y, D and E) and fibrin fragments (D-dimer and E) were examined in the urine of 52 patients with various types of renal diseases. One or more of the fibrinogen fragments were detectable in all the urine specimens. D-Dimer together with fibrinogen fragments was found in 38 of the 52 patients. The clearance ratio of D-dimer to IgG, which indicates D-dimer generated in the kidney, was lower than 1 in all the patients with the minimal changes nephrotic syndrome, and was greater than 1 in the majority of patients with acute glomerulonephritis, rapidly progressive glomerulonephritis, mesangial proliferative glomerulonephritis and membranoproliferative glomerulonephritis. Our results suggest that urinary fibrin/fibrinogen degradation products (FDP) in renal diseases are derived primarily from increased filtration of FDP from the plasma through a damaged glomerular basement membrane, and that the mechanism of lysis of cross-linked fibrin deposited in the glomeruli occurs simultaneously in some types of glomerulonephritis. It seems that the determination of the clearance ratio of D-dimer to IgG may be useful in assessing the activation of the coagulation and fibrinolysis systems in the kidney in patients with renal diseases.
Subject(s)
Fibrin Fibrinogen Degradation Products/urine , Kidney Diseases/urine , Adolescent , Child , Child, Preschool , Female , Fibrin Fibrinogen Degradation Products/blood , Humans , Immunoglobulin G/analysis , Kidney Diseases/blood , Kidney Diseases/immunology , Male , Nephrotic Syndrome/urine , ProteinuriaABSTRACT
Plasma levels of coagulation factor XIII determined quantitatively were found to have a tendency to be decreased in more severely affected cases of acute poststreptococcal glomerulonephritis and Henoch-Schönlein nephritis, and found to be decreased in patients with rapidly progressive glomerulonephritis and chronic renal failure in comparison with normal controls. It seems that the degree of decrease in factor XIII reflects the intensity of glomerular damage. In contrast, the patients with the nephrotic syndrome showed significant elevation of plasma factor XIII levels and it was closely correlated with the increase in serum triglycerides. The elevation of factor XIII levels in the nephrotic syndrome is suggested to be mainly attributed to enhanced protein synthesis in connection with urinary loss of proteins.
