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PubMed1994

LDL modification by activated polymorphonuclear leukocytes: a cellular model of mild oxidative stress.

The purpose of this study was to determine the most potent activator of the respiratory burst of polymorphonuclear leukocytes (PMN) with respect to the oxidative modification of low density lipoprotein (LDL). Phorbol-12-myristate-13-acetate (PMA), n-formyl-methionyl-leucylphenylalanine (nFMLP), lipopolysaccharide (LPS), and opsonized zymosan (OZ) were tested. The generation of reactive oxygen species by PMN was assayed as superoxide anion production. Oxidative modification of LDL was monitored by thiobarbituric acid reactive substance (TBARS) activity, by conjugated dienes formation at 234nm and by electrophoretic mobility on agarose gel. PMA was the most potent activator of PMN, inducing a 6-fold increase in the superoxide anion production, followed by OZ (3-fold increase). PMA activation also induced the greatest modification of LDL by PMN: 700% increase of conjugated dienes formation, 222% increase of TBARS, and 70% increase in the electrophoretic mobility. The indices of oxidative modification significantly correlated with the superoxide anion generated by different activators. Also, LDL oxidation by PMN was inhibited by superoxide dismutase but not by catalase, methionine, or hydroxyl radical scavengers. Our data indicate that PMNs activated by PMA produce a mildly oxidized form of LDL by a mechanism that appears to involve the superoxide anion.

Humans↗
PubMed1988

Development and evaluation of modifications to the Sickness Impact Profile for head injury.

Three modifications were made to the Sickness Impact Profile, a behavior-based measure of health status, to improve its sensitivity to the effects of head injury. (1) Additional items were included to capture head injury sequelae and behaviors typical of young adults, the age group to which head injury most frequently occurs. (2) Subjects individually excluded behaviors irrelevant to them, thus allowing the score to better reflect injury-related changes. (3) The different areas of functioning on the Sickness Impact Profile were reweighted to reflect global judgments of the construct's contribution to overall functioning rather than the sum of the item contributions. Only the first modification is head-injury specific. The others, are relevant to any disease or injury. The performance of the modifications was evaluated in a longitudinal study of 102 head injured and 102 comparison subjects tested at 1 and 12 months after injury. The evaluation of the modifications was based on their ability to distinguish head injury from comparison subjects and on the strength of their relationship with measures of brain dysfunction. Despite a few statistically significant improvements in discrimination, differences of a practical degree were not obtained. The standard Sickness Impact Profile performed well and is recommended for evaluation of day-to-day functioning in head injury studies.

Activities of Daily Living↗
PubMed1996

Mechanisms involved in the in vitro modification of low density lipoprotein by human umbilical vein endothelial cells and copper ions.

In this study we evaluated the time course and mechanism of low density lipoprotein (LDL) oxidation induced by human umbilical vein endothelial cells (HUVECs), cell-free medium (CFM) and Cu2+. After incubating LDL (200 micrograms/ml) with HUVECs, CFM and Cu2+ (concentration adjusted to obtain the same degree of LDL modification as with HUVECs), the extent of LDL lipid peroxidation and apoprotein B modification was monitored at different times from 0 to 24 h. This involved evaluating the time course of LDL conjugated diene, peroxide, malonyldialdehyde (MDA), fluorescence, relative electrophoretic mobility (REM), vitamin E and monounsaturated and polyunsaturated fatty acids. After incubation with HUVECs, the LDL REM was significantly higher than that obtained in CFM (p < 0.01). When balanced for the same degree of LDL modification as obtained with HUVECs, Cu2+ gave a REM similar to that obtained with HUVECs. At the different times of incubation there was no statistical difference between conjugated diene and peroxide values after incubation with HUVECs and with CFM. The values obtained with Cu2+ were significantly higher than those obtained with HUVECs and CFM (p < 0.01). MDA and LDL fluorescence were significantly higher after exposure to HUVECs than to CFM (p < 0.01), values being similar to those obtained with Cu2+. There was no statistical difference between the values of LDL oleic, linoleic, arachidonic and eicosapentaenoic acids after incubation with HUVECs and CFM. Eicosatetraynoic acid (ETYA), a lipoxygenase inhibitor, determined dose-dependent reduction of MDA formation induced by the incubation of LDL with HUVECs; it did not affect LDL conjugated diene. ETYA did not have any effect on the MDA derived from LDL after incubation with Cu2+ or CFM. The results of this study demonstrate that, unlike Cu2+, the contribution of HUVECs to LDL modification does not involve only lipid peroxidation of the lipoprotein; it also includes intracellular radical and non-radical processes.

5,8,11,14-Eicosatetraynoic Acid↗
PubMed1992

The mechanisms for the effects of dietary lipid modification on autoimmune diseases: the role of membrane lipid composition in antigen presenting cells.

Several mechanisms have been proposed to explain the effects of dietary lipid modification on autoimmune diseases. One of these being the modification of cell membranes which affects immune functions. In order to further define this mechanism, the author proposes that dietary lipid modification could affect antigen presentation, an immune function responsible for initiating cell-mediated immune responses in body defense or autoimmune diseases, through membrane lipid composition modification.

Antigen-Presenting Cells↗
PubMed2005

A surface modification strategy on silicon nitride for developing biosensors.

A surface modification strategy for the use of giant magnetoresistive materials in the detection of protein-protein interactions is developed. This modification strategy is based on silanization of semiconductive materials. A native silicon nitride surface was treated with concentrated hydrofluoric acid to improve surface homogeneity. Nano-strip was used to oxidize silicon nitride to form a hydrophilic layer. Aminopropyltriethoxysilane was subsequently used to functionalize the treated surfaces to form amine groups, which were further activated with glutaraldehyde to introduce a layer of aldehyde groups. The effectiveness of this modification strategy was validated by chemiluminescence immunoassays of purified 6x His-HrpW of Pseudomonas syringae pv. tomato DC3000 and human transferrin. Signals with intensities related to concentrations of these two immobilized model proteins were observed. The modified surface was also validated by a more complex system: intercellular proteins secreted by DC3000. HrpW in these protein mixtures was successfully recognized by anti-HrpW antibodies when mixed proteins were immobilized onto activated surfaces. This surface modification strategy provides a platform onto which proteins can be directly immobilized for biosensor and protein array applications.

Biosensing Techniques↗
PubMed2005

Heme-protein covalent bonds in peroxidases and resistance to heme modification during halide oxidation.

Plant peroxidases, as typified by horseradish peroxidase (HRP), primarily catalyze the one-electron oxidation of phenols and other low oxidation potential substrates. In contrast, the mammalian homologues such as lactoperoxidase (LPO) and myeloperoxidase primarily oxidize halides and pseudohalides to the corresponding hypohalides (e.g., Br(-) to HOBr, Cl(-) to HOCl). A further feature that distinguishes the mammalian from the plant and fungal enzymes is the presence of two or more covalent bonds between the heme and the protein only in the mammalian enzymes. The functional roles of these covalent links in mammalian peroxidases remain uncertain. We have previously reported that HRP can oxidize chloride and bromide ions, but during oxidation of these ions undergoes autocatalytic modification of its heme vinyl groups that virtually inactivates the enzyme. We report here that autocatalytic heme modification during halide oxidation is not unique to HRP but is a general feature of the oxidation of halide ions by fungal and plant peroxidases, as illustrated by studies with Arthromyces ramosus and soybean peroxidases. In contrast, LPO, a prototypical mammalian peroxidase, is protected from heme modification and its heme remains intact during the oxidation of halide ions. These results support the hypothesis that the covalent heme-protein links in the mammalian peroxidases protect the heme from modification during the oxidation of halide ions.

Bromides↗
PubMed2005

Home modification to prevent falls by older ED patients.

This trial was conducted at 11 EDs to test the effectiveness of distributing fall prevention information to patients 65 years or older. Intervention patients were given 2 brochures and received a reminder call 2 weeks later. All patients were called at 1 month and asked if they made home safety modifications. Three hundred ninety-seven patients were enrolled (118 control, 279 intervention). Seventy-six percent had complete follow up interviews. Nine percent of control and 8% of intervention patients reported a home modification (95% confidence interval on difference, -8.1% to 5.5%). Patients who fell in the prior year had a 2.0 increased odds (95% confidence interval, 0.8-4.6) of making a home modification. The similar home modification rates in the 2 study groups suggest that even minimum discussion (eg, the informed consent procedure) may increase patients' fall prevention activities. The stronger association in patients who fell suggests that a targeted program may have added benefit.

Accidental Falls↗
PubMed2005

Influence of lifestyle modification on arterial stiffness and wave reflections.

Arterial stiffness and wave reflections exert a number of adverse effects on cardiovascular function and disease risk and are associated with a greater rate of mortality in patients with end-stage renal failure and essential hypertension. Accordingly, the prevention and treatment of arterial stiffness are of paramount importance. Because arterial stiffening is being recognized as a critical precursor of cardiovascular disease (CVD), it is essential to understand the role of lifestyle modifications on preventing and reversing arterial stiffening. Available evidence indicates that lifestyle modifications, in particular aerobic exercise and sodium restriction, appear to be clinically efficacious therapeutic interventions for preventing and treating arterial stiffening. Thus, sufficient evidence is available to recommend lifestyle modifications as part of a first-line therapeutic approach for arterial stiffening. However, more information is needed for a full understanding and optimal use of lifestyle modifications in the management of arterial stiffening.

Arteries↗
PubMed2005

Thermal modification of partial tears of the anterior cruciate ligament.

PURPOSE: Partial tears of the anterior cruciate ligament (ACL) are common, representing 10% to 28% of all ACL tears. Untreated partial tears of the ACL may go on to complete rupture in up to 42% of cases, and as few as 30% of patients return to their preinjury activity level. The purpose of this study was to evaluate the effectiveness of thermal modification for the treatment of partial tears of the ACL. TYPE OF STUDY: A prospective, nonrandomized consecutive case series. METHODS: Thirteen patients with a partial tear of a native ACL were treated with thermal modification of the ACL. All patients had a preoperative office examination significant for an end-point to Lachman examination and pivot-glide. All patients had bilateral preoperative KT-1000 measurements. All tears were confirmed by arthroscopy to constitute a 50% or less loss of structural integrity of the ligament. The ACL underwent thermal modification using the Oratec thermal probe (Oratec Interventions, Menlo Park, CA), and was examined by intraoperative KT-1000 testing. Patients were evaluated postoperatively at 6 weeks, and at 3, 6, 12, and 24 months by office examination and functional outcome scoring. RESULTS: KT-1000 arthrometer testing revealed a mean side-to-side difference of 4.35 mm preoperatively (SD = 1.1 mm). At the most recent follow-up, averaging 23 months (range, 18 to 28 months), 10 patients had a negative Lachman examination and no pivot-shift. Two patients had persistent grade II Lachman and complaints of giving way 3 months postoperative. Both patients underwent ACL reconstruction. One patient was lost to follow-up. Of the remaining 10 patients, all patients achieved full extension, and the average flexion range of motion was 131 degrees (SD = 5.6). Compared with the preoperative KT-1000 arthrometer testing, the most recent evaluation revealed a decrease in mean side-to-side difference to 1.9 mm (SD = 1.5 mm). At most recent follow-up, the mean Lysholm score was 96.3 (SD = 4.4), the mean Tegner score was 6.1 (SD = 1.2), and the mean Cincinnati score was 94 (SD = 3.0). CONCLUSIONS: With cautious application, thermal modification may be a viable treatment option for partial tears of the ACL in a select subset of patients. Further investigation is necessary to determine the long-term effectiveness of this procedure. LEVEL OF EVIDENCE: Level IV, Therapeutic Case Series Study (no or historical control group).

Adult↗
PubMed2005

Structural modifications of HDL and functional consequences.

High density lipoproteins (HDL) are susceptible to structural modifications mediated by various mechanisms including oxidation, glycation, homocysteinylation or enzymatic degradation. Structural alterations of HDL may affect their functional and atheroprotective properties. Oxidants, such as hypochlorous acid, peroxyl radicals, metal ions, peroxynitrite, lipoxygenases and smoke extracts, can alter both surface and core components of HDL. The formation of lipid peroxidation derivatives, such as thiobarbituric acid reactive substances, conjugated dienes, lipid hydroperoxides and aldehydes, is associated with changes of physical properties (fluidity, molecular order) and of apoprotein conformation. Non-enzymatic glycation, generally associated with lipoxidation, leads to form irreversible complexes called advanced glycation end products. These HDL modifications are accompanied with altered biological activities of HDL and associated enzymes, including paraoxonase, CETP and LCAT. Homocysteine-induced modification of HDL is mediated by homocysteine-thiolactone, and can be prevented by a calcium-dependent thiolactonase/paraoxonase. Tyrosylation of HDL induces the formation of dimers and trimers of apo AI, and alters cholesterol efflux. Phospholipases and proteolytic enzymes can also modify HDL lipid and apoprotein structure. HDL modification induces generally the loss of their anti-inflammatory and cytoprotective properties. This could play a role in the pathogenesis of atherosclerosis and neurodegenerative diseases such as Alzheimer's disease.

Antioxidants↗
PubMed2006

Protection against glycation and similar post-translational modifications of proteins.

Glycation and other non-enzymic post-translational modifications of proteins have been implicated in the complications of diabetes and other conditions. In recent years there has been extensive progress in the search for ways to prevent the modifications and prevent the consequences of the modifications. These areas are covered in this review together with newer ideas on possibilities of reversing the chemical modifications.

Aspirin↗
PubMed2005

ISG15 modification of ubiquitin E2 Ubc13 disrupts its ability to form thioester bond with ubiquitin.

ISG15 was the first ubiquitin-like modifier to be identified. However, the function of ISG15 modification has been an enigma for many years. At present, no data are available about the function of ISGylation for any target. In this paper, we report the identification of Ubc13, which forms a unique ubiquitin-conjugating enzyme (Ubc) complex with ubiquitin enzyme variant Mms2 and generates atypical Lys63-linked ubiquitin conjugates, as one of the targets of ISG15 modification. Furthermore, we identify Lys92 as the only ISG15 modification site in Ubc13, which is the first report about the ISG15 modification site. Using the "covalent affinity" purification assay, we found that unmodified Ubc13 can bind to the ubiquitin-agarose, whereas ISGylated Ubc13 cannot. This result indicates that ISGylation of Ubc13 disrupts its ability to form thioester bond with ubiquitin.

Amino Acid Sequence↗
PubMed2006

Detection of redox-based modification in two-dimensional electrophoresis proteomic separations.

Oxidative stress arises when cellular defenses against molecular oxygen and its by-products (reactive oxygen species; ROS) are overcome leading to covalent modification of lipids, DNA, and protein. Redox-based modification of proteins can be conveniently studied by proteomic analysis using two-dimensional electrophoresis (2D SDS-PAGE). Despite some technical shortcomings, this technique allows rapid and quantitative analysis of paired samples, the visualization of discrete protein spots, and provides a robust platform for subsequent analysis and identification of specific proteins. Exposure to oxidative stress introduces a wide range of reversible or irreversible alterations to amino acid side chains. These include carbonylation, glutathionylation, formation of mixed disulphides, effects on disulphide bridge patterns, ubiquitinylation, and racemization. Identification of proteins targeted for specific modification adds a deeper dimension to the dissection of effects of oxidative stress on the proteome with potentially far-reaching implications. This article describes key methodologies now available for identification of redox-based modifications in proteins separated by 2D SDS-PAGE.

Animals↗
PubMed2006

The effect of RGD fluorosurfactant polymer modification of ePTFE on endothelial cell adhesion, growth, and function.

We have synthesized and characterized a novel peptide fluorosurfactant polymer (PFSP) modification that facilitates the adhesion and growth of endothelial cells on expanded polytetrafluoroetheylene (ePTFE) vascular graft material. This PFSP consists of a poly(vinyl amine) (PVAm) backbone with integrin binding Arg-Gly-Asp (RGD) peptides and perfluorocarbon pendant branches for adsorption and stable adhesion to underlying ePTFE. Aqueous PFSP solution was used to modify the surface of fluorocarbon substrates. Following subconfluent seeding, endothelial cell (EC) adhesion and growth on PFSP was assessed by determining cell population at different time points. Spectroscopic results indicated successful synthesis of PFSP. PFSP modification of ePTFE reduced the receding water contact angle measurement from 120 degrees to 6 degrees , indicating successful surface modification. Quantification of cell population demonstrated reduced EC attachment efficiency but increased growth rate on RGD PFSP compared with fibronectin (FN). Actin staining revealed a well-developed cytoskeleton for ECs on RGD PFSP indicative of stable adhesion. Uptake of acetylated low-density lipoprotein and positive staining for VE-Cadherin confirm EC phenotype for adherent cells. Production of prostacyclin, a potent antiplatelet agent, was equivalent between ECs on FN and RGD PFSP surfaces. Our results indicate successful synthesis and surface modification with PFSP; this is a simple, quantitative, and effective approach to modifying ePTFE to encourage endothelial cell attachment, growth, and function.

Amino Acid Sequence↗
PubMed2005

Post-translational modifications of naturally processed MHC-binding epitopes.

A variety of different post-translational modifications of peptides displayed by class I and II MHC molecules have now been described. Some modifications promote the binding of peptides to MHC molecules, and might also influence the ability of the peptide to be produced by antigen processing pathways. In some instances, the antigen processing components themselves are actually responsible for generating post-translational modifications. Finally, evidence is accumulating that modifications can be altered as a consequence of inflammation, transformation, apoptosis and aging. This leads to altered repertories of MHC-associated peptides, which may be important in immune responses associated with autoimmune diseases, infection and cancer.

Animals↗
PubMed2005

Life-style modification in peripheral arterial disease.

OBJECTIVE: To review the published evidence supporting the use of life-style modification in peripheral arterial disease (PAD). DESIGN: A systematic search of the medical literature was performed for relevant studies. MATERIALS: The publications obtained were then searched for randomised clinical trials which reported end-points of mortality or major cardiovascular event rates with various life-style modifications. RESULTS: Only one randomised controlled trial was found reporting relevant end-points. Other trials were of other end-points such as walking distance or biochemical markers. CONCLUSIONS: There is a lack of randomised controlled data proving the benefit of life-style modification in improving mortality and reducing cardiovascular events in patients with PAD. Despite this there is sufficient evidence to recommend some life-style modification as part of the overall approach to risk reduction in these patients. There is compelling evidence to support smoking cessation, increased exercise and improved diet.

Arterial Occlusive Diseases↗
PubMed2005

Wobble modification deficiency in mutant tRNAs in patients with mitochondrial diseases.

Point mutations in mitochondrial (mt) tRNA genes are associated with a variety of human mitochondrial diseases. We have shown previously that mt tRNA(Leu(UUR)) with a MELAS A3243G mutation and mt tRNA(Lys) with a MERRF A8344G mutation derived from HeLa background cybrid cells are deficient in normal taurine-containing modifications [taum(5)(s(2))U; 5-taurinomethyl-(2-thio)uridine] at the anticodon wobble position in both cases. The wobble modification deficiency results in defective translation. We report here wobble modification deficiencies of mutant mt tRNAs from cybrid cells with different nuclear backgrounds, as well as from patient tissues. These findings demonstrate the generality of the wobble modification deficiency in mutant tRNAs in MELAS and MERRF.

Base Sequence↗
PubMed2006

Unusual histone modifications in Trypanosoma brucei.

To start to understand the role of chromatin structure in regulating transcription in trypanosomes, we analyzed covalent modifications on the four core histones of Trypanosoma brucei. We found unusually few modifications in the N-terminal tails, which are abundantly modified in other organisms and whose sequences, but not composition, are highly divergent in trypanosomes. In contrast, the C-terminal region of H2A appears to be hyper-acetylated. Surprisingly, the N-terminal alanines of H2A, H2B, and H4, were mono-methylated, a modification that has not been described previously for histones. Possible functions and evolutionary explanations for these unusual histone modifications are discussed.

Acetylation↗