Search results for "recombinant"

Acetylcysteine therapy for chronic hepatitis C: are its effects synergistic with interferon alpha? A pilot study.

2008

This trial reports the 6-month results of a pilot study using lymphoblastoid interferon alpha (IFNalpha) and acetylcysteine (N-acetylcysteine) separately and in combination in patients with chronic hepatitis C, genotype 1b, who were nonresponders to previous treatment with recombinant IFNalpha alone.21 patients were randomly divided into three groups of seven each. Group A was treated with lymphoblastoid IFNalpha 6MU three times a week for 6 months; group B received the same schedule of lymphoblastoid IFNalpha as group A plus acetylcysteine 1200 mg/day per os in two administrations, and group C received only acetylcysteine 1200 mg/day per os in two administrations.Mean serum alanine aminotr…

Erythropoietin: a new paradigm for neuroprotection.

2006

Alfavīrusu vektoru izstrāde makrofāgu aktvācijai vēža imunoterapijā

2016

Audzēju-iefiltrējošiem makrofāgiem piemīt gan spēja veicināt audzēja attīstību, gan inhibēt, attiecībā no to aktivācijas stāvokļa. In vitro pētījumos ir ticis parādīts, ka, iedarbojoties ar interferonu-g (IFN-γ) kopā ar Toll-like receptora (TLR) ligandu, tika ierosināts M1 makrofāgu fenotips, kas tiek asociēts ar audzēja iznīcināšanu. Audzēja imūnterapiju varētu nodrošināt M1 makrofāgu fenotipa inducēšana audzēju-iefiltrējošos makrofāgos. Šajā darbā mēs izvērtējām Semliki meža vīrusa (SFV) vektoru potenciālu makrofāgu aktivācijā pret vēzi. SFV tika izmantots, lai konstruētu replikācijas defektīvus konstruktus ar ieklonētiem peļu audzēja nekrozes faktora (mTNFα), peļu IFNγ un cilvēka IFNγ gē…

2007

We have recently demonstrated that the rheumatoid arthritis (RA) shared epitope (SE) acts as a ligand that triggers nitric oxide (NO) signaling in opposite cells. Given the known pro-oxidative effect of NO and the proposed role of oxidative stress in the pathogenesis of RA, this study explores whether SE-triggered signaling can increase cellular oxidative stress. cAMP levels, adenylyl cyclase activity, and protein kinase A activity were measured using commercial kits. Generation of reactive oxygen species (ROS) was quantified using the fluorochrome dichlorofluorescein diacetate. Oxidative DNA damage was quantified using the single-cell electrophoresis technique. Here, we report that cells e…

Interaction of a Recombinant Prion Protein with Organo-Mineral Complexes as Assessed by FT-IR and CPMAS 13C NMR Analysis

2010

Prion proteins are considered as the main agents for the Transmissible Spongiform Encephalopathies (TSE). The misfolded form, PrP Sc , which is also indicated as the etiological agent for TSE, exhibits high resistance to degradation in environmental processes. Soil contamination by prion proteins is a real environmental issue since contaminated soils can become potential reservoir and diffuser for TSE infectivity. In this work, the interaction of prion protein with organo- mineral complexes was studied by using a recombinant non pathogenic prion protein and a model soil system. This latter was represented by a soil manganese mineral coated with polymerized catechol. FT-IR spectra showed ami…

PCR-Based Site-Specific Mutagenesis

1998

The alteration of gene structure through the substitution of specific nucleotides by site-specific mutagenesis is an important tool in modern recombinant DNA technology. Nucleotide changes are necessary not only for the analysis of the structural basis of gene and corresponding protein function, but also for the generation of novel gene products. The availability of the polymerase chain reaction (PCR) in the last decade has enabled the modification of DNA for different needs to be made more rapidly and easily than was previously possible. In the course of mutagenesis the relevant sequence changes can be introduced more readily by chemically synthesized oligonucleotide primers than by manipu…

Effects of KL4-Type Peptides on the Surface Activity and Stability of Pulmonary Surfactant Films as Evaluated in the Captive Bubble Surfactometer

2012

Although SP-B is the most critical protein in lung surfactant, recombinant or synthetic forms of SP-B as a basis for the development of therapeutic surfactants are still not available. An alternative is the design and production of peptides mimicking the structure and general properties of essential motifs in SP-B.In the present study the surface activity of different KL4-derived peptides, as sequence variations of the original peptide designed to replicate a general amphipathic motif of SP-B [1], has been assessed in the captive bubble surfactometer. The peptides were reconstituted in a surfactant lipid matrix: DPPC/POPC/POPG (50:25:15, w/w/w). This mixture was selected because it offers a…

Human neuroglobin: crystals and preliminary X-ray diffraction analysis

2002

Neuroglobin, a recently discovered member of the haemoglobin superfamily, is primarily expressed in the brain of humans and other vertebrates, where it has been proposed to enhance O(2) supply in response to hypoxia or ischaemia, protecting the neuron from hypoxic injury. Neuroglobin is the first example of a vertebrate haemoglobin in which a hexacoordinate haem geometry has been detected. A triple mutant (replacing three Cys residues) of human neuroglobin (151 amino acids) has been expressed in Escherichia coli, purified and crystallized in two crystal forms, the best of which diffracts to 1.95 A resolution using synchrotron radiation. The crystals belong to space group P2(1), with unit-ce…

High yield recombinant production of a self-assembling polycationic peptide for silica biomineralization.

2015

We report the recombinant bacterial expression and purification at high yields of a polycationic oligopeptide, P5S3. The sequence of P5S3 was inspired by a diatom silaffin, a silica precipitating peptide. Like its native model, P5S3 exhibits silica biomineralizing activity, but furthermore has unusual self-assembling properties. P5S3 is efficiently expressed in Escherichia coli as fusion with ketosteroid isomerase (KSI), which causes deposition in inclusion bodies. After breaking the fusion by cyanogen bromide reaction, P5S3 was purified by cation exchange chromatography, taking advantage of the exceptionally high content of basic amino acids. The numerous cationic charges do not prevent, b…

Peptide–Membrane Interactions Monitored by Fluorescence Lifetime Imaging: A Study Case of Transportan 10

2021

The interest on detailed analysis of peptide-membrane interactions is of great interest in both fundamental and applied sciences as these may relate to both functional and pathogenic events. Such interactions are highly dynamic and spatially heterogeneous, making the investigation of the associated phenomena highly complex. The specific properties of membranes and peptide structural details, together with environmental conditions, may determine different events at the membrane interface, which will drive the fate of the peptide-membrane system. Here, we use an experimental approach based on the combination of spectroscopy and fluorescence microscopy methods to characterize the interactions …