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PubMed1996

Relationship between alcohol intake and immunoglobulin a immunoreactivity with acetaldehyde-modified bovine serum albumin.

Acetaldehyde, the main metabolite of ethanol, is a highly reactive species that reacts with macromolecules to produce unstable and stable adducts. Acetaldehyde-modified proteins are immunogenic and have been detected in the liver and blood of alcoholics. Furthermore, antibodies reactive with acetaldehyde-modified proteins have been detected in the plasma of social drinkers and alcoholics. However, the class distribution of immunoglobulins reactive with modified proteins was different in the two groups, being predominantly immunoglobulin (Ig)M in social drinkers, but IgM and IgA in alcoholics. In this study, we demonstrate that heavy drinkers (alcohol intake > 130 g/week for females and 150 g/week for males) also exhibit IgA reactivity with acetaldehyde-modified proteins. The IgA adduct-specific reactivity (IgA reactivity with acetaldehyde-modified bovine serum albumin-reactivity with native bovine serum albumin) showed a moderate correlation with self-reported alcohol intake, but did not correlate with markers such as plasma transaminase, gamma-glutamyltransferase activity, or mean corpuscular volume. IgA adduct-specific reactivity had similar specificity to the conventional tests of alcohol abuse, but had higher sensitivity than the other tests, especially with heavy drinkers. Data presented herein demonstrate that elevated IgA reactivity with acetaldehyde-modified epitopes is associated with heavy drinking and is a potential marker for high alcohol intake.

Acetaldehyde↗
PubMed1997

Comparison of the adsorption ability between tryptophan and modified tryptophan columns.

Immunoadsorption therapy (IAT) using the tryptophan column (TR:TR-350, Asahi Medical Co., Tokyo, Japan) is used for patients with Guillain-Barré syndrome or Fisher's syndrome. Recently, a modified tryptophan column (modified TR:TR-350S, Asahi) was developed to reduce adsorption of fibrinogens. However, it is not clear whether the new column can effectively adsorb the antiganglioside antibodies. We treated 2 patients with Guillain-Barré syndrome, 1 patient with Fisher's syndrome, and 1 patient with Bickerstaff's brainstem encephalitis by IAT using the TR and the modified TR. Samples were taken from the inlet and outlet of the affinity column. We compared the adsorption ability of antiganglioside antibodies between the TR and modified TR. The modified TR adsorbed antiganglioside antibodies less effectively than the TR. No significant bleeding was observed in the patients during each session. Furthermore, results of the clinical study were confirmed by batchwise adsorption test. We propose that the modified TR should not be used for treating patients with Guillain-Barré syndrome and related disorders.

Adsorption↗
PubMed1995

Physiological responses of horses competing at a modified 1 star 3-day-event.

The impending 1996 summer Olympic 3-day-event in Atlanta has focused attention on the need to determine what modifications to the demanding Endurance Test will be required to ensure safety of the horses competing. Three groups of horses participated in a Field Trial held in August of 1994 in northern Georgia to determine the safety and feasibility of conducting a modified 3-day-event in hot, humid weather. One group (TD) completed a modified 1 Star 3-day-event test, a control group (HT) completed a Horse Trial identical to the modified 1 Star test except for the omission of Phases B and C and the third group (E), comprised of European horses, completed the modified 1 Star test with a longer, faster Phase C than was used for TD. During the Endurance Test, the ambient temperature and relative humidity ranged from 24.3 degrees C and 98.9% in the morning to 30.2 degrees C and 51.6% in the afternoon. No horse failed to complete the Trial because of heat stress or fatigue. There were no significant (P < 0.05) differences detected in heart rate, rectal temperature, respiratory rate or net weight loss between HT and TD horses at any observation time. The highest rectal temperature recorded at the end of Phase C was 39.6 degrees C. These findings suggest that the modified 1 Star Endurance Test was as well tolerated by American horses as the control Horse Trial test. Rectal temperature was significantly higher for E than for TD or HT at the finish of Phase C. European horses had significantly greater decreases in weight than HT and TD at the end of Phases C and D and the next day. These findings probably reflect the faster and longer work effort of E horses during Phase C. Modification of Phase C and the rest-pause to ensure that recovery and heat dissipation occurred before the start of Phase D resulted in a 3-day-event that was safe for horses. The Field Trial provides a model for designing a modified Olympic Endurance Test. If the 1996 Olympic 3-day-event is held in hotter and more humid weather than the Field Trial, additional modifications to the Endurance Test (decreased distances, speeds and numbers of jumping efforts) will probably be required to ensure safety of competing horses.

Animals↗
PubMed1998

A modified enzyme-linked immunosorbent assay for measurement of antibody responses to meningococcal C polysaccharide that correlate with bactericidal responses.

The standardized enzyme-linked immunosorbent assay (ELISA) for measurement of serum immunoglobulin G (IgG) antibody responses to meningococcal C polysaccharide has been modified to employ assay conditions that ensure specificity and favor detection primarily of high-avidity antibodies. The modified and standard assays were used to measure IgG antibody concentrations in sera of toddlers vaccinated with meningococcal polysaccharide vaccine or a meningococcal C conjugate vaccine. The results were compared to the respective complement-mediated bactericidal antibody titers. In sera obtained after one or two doses of vaccine, the correlation coefficients, r, for the results of the standard assay and bactericidal antibody titers were 0.45 and 0.29, compared to 0.85 and 0.87, respectively, for the modified assay. With the standard assay, there were no significant differences between the geometric mean antibody responses of the two vaccine groups. In contrast, with the modified assay, 5- to 20-fold higher postvaccination antibody concentrations were measured in the conjugate than in the polysaccharide group. Importantly, the results of the modified assay, but not the standard ELISA, paralleled the respective geometric mean bactericidal antibody titers. Thus, by employing conditions that favor detection of higher-avidity IgG antibody, the modified ELISA provides results that correlate closely with measurements of antibody functional activity that are thought to be important in protection against meningococcal disease.

Antibodies, Bacterial↗
PubMed1993

Characterization of a heat-modifiable outer membrane protein of Haemophilus somnus.

In immunoblot analysis, a murine monoclonal antibody (MAb), 27-1, which was produced to an outer membrane protein (OMP) of Haemophilus somnus, showed that a major OMP is heat modifiable, having a molecular mass of 28 kDa when the N-lauroylsarcosine-insoluble OMP preparation was solubilized at 60 degrees C and a mass of 37 kDa when the OMP preparation was solubilized at 100 degrees C. The heat-modifiable OMP reacted intensely with convalescent sera obtained from calves with experimental H. somnus pneumonia in immunoblot analysis. Immunoelectron microscopic and antibody absorption studies revealed that the MAb 27-1 epitope was not surface exposed on the intact bacterium. However, a decrease in antibody reactivity to the heat-modifiable OMP in immunoblot analysis after absorption of convalescent serum with intact bacterial cells of H. somnus suggests that a surface-exposed portion of the heat-modifiable OMP is expressed on the intact bacterium. MAb 27-1 reacted with 45 of 45 strains of H. somnus tested in immunoblot analysis. The apparent molecular mass of the antigen varied among strains, and five reactivity patterns demonstrated by MAb 27-1 were observed. MAb 27-1 also reacted with six species in the family Pasteurellaceae, Escherichia coli, and Salmonella dublin, but not with the other eight species of gram-negative bacteria. The heat-modifiable OMP of H. somnus showed immunological cross-reactivity with the OmpA protein of E. coli K-12 and significant N-terminal amino acid sequence homology with the OmpA proteins of gram-negative bacteria. We conclude that a major, 37-kDa heat-modifiable OMP of H. somnus, which elicits an antibody response in H. somnus-infected animals, is a common antigen among H. somnus strains tested and is structurally related to the OmpA protein of E. coli.

Amino Acid Sequence↗
PubMed1982

Immunological detection of the Kanagawa phenomenon of vibrio parahaemolyticus on modified selective media.

Selective media for Vibrio parahaemolyticus, BTB-Teepol agar and modified arabinose-ammonium sulfate-cholate agar, were modified for use in immunological detection of the thermostable direct hemolysin produced by this organism. The modified BTB-Teepol agar and modified arabinose-ammonium sulfate-cholate agar were both found to be useful for the modified Elek test and immunohalo test with antiserum (or immunoglobulin G) against the thermostable direct hemolysin. With these modified media it is possible to isolate V. parahaemolyticus and identify the Kanagawa phenomenon on a single plate and thus save time in obtaining results.

Culture Media↗
PubMed1996

Modified technique to recover microsporidian spores in sodium acetate-acetic acid-formalin-fixed fecal samples by light microscopy and correlation with transmission electron microscopy.

Microsporidia are an emerging cause of significant disease, particularly in the immunocompromised host. Until recently, the diagnosis of enteric infections has required invasive sampling, the use of expensive technology, and considerable technological expertise. The purpose of the present study was to examine three modifications to the processing of fecal specimens for light microscopy (LM) examination for microsporidian spores: the use of pretreatment with potassium hydroxide, modified centrifugation conditions, and a modified staining technique. A sodium acetate-acetic acid-formalin-fixed fecal sample containing numerous microsporidian spores confirmed to be positive by transmission electron microscopy (TEM) was used in all studies performed. A simulation of a heavy to lightly infected individual was used. The results of LM were correlated with those of TEM. Duplicate smears were stained with Weber's modified trichrome and Giemsa (GS) stains. The stained slides were randomized and examined blindly by LM at x 625 and x 1,250 magnifications. A portion of the dilutions after centrifugation were fixed for TEM. The Weber modified trichrome stain performance rating was higher than the Giemsa stain rating because of ease of interpretation, and material stained with Weber modified trichrome stain required less examination time at a lower magnification. The number of positive smears and the quantity of spores detected were significantly higher following pretreatment of the sample with KOH. TEM was positive only when numerous spores were present, but the quality of the photomicrographs was superior after pretreatment with KOH. Pretreatment of sodium acetate-acetic acid-formalin-fixed fecal samples with 10% KOH and then a 5-min centrifugation time and staining with Weber modified trichrome stain provide for the excellent recovery of microsporidia in the routine diagnostic parasitology laboratory.

AIDS-Related Opportunistic Infections↗
PubMed2001

Nonmyeloablative immunosuppressive regimen prolongs In vivo persistence of gene-modified autologous T cells in a nonhuman primate model.

The in vivo persistence of gene-modified cells can be limited by host immune responses to transgene-encoded proteins. In this study we evaluated in a nonhuman primate model whether the administration of a nonmyeloablative regimen consisting of low-dose total-body irradiation with 200 cGy followed by immunosuppression with mycophenolate mofetil and cyclosporin A for 28 and 35 days, respectively, could be used to facilitate persistence of autologous gene-modified T cells when a transgene-specific immune response had already been established or to induce long-lasting tolerance in unprimed recipients. Two macaques (Macaca nemestrina) received infusions of T cells transduced to express either the enhanced green fluorescent protein and neomycin phosphotransferase genes or the hygromycin phosphotransferase and herpes simplex virus thymidine kinase genes. In the absence of immunosuppression, both macaques developed potent class I major histocompatibility complex-restricted CD8(+) cytotoxic T-lymphocyte (CTL) responses that rapidly eliminated the gene-modified T cells and that persisted long term as memory CTL. Treatment with the nonmyeloablative regimen failed to abrogate preexisting memory CTL responses but interfered with the induction of transgene-specific CTL and facilitated in vivo persistence of gene-modified cells in an unprimed host. However, sustained tolerance to gene-modified T cells was not achieved with this regimen, indicating that further modifications will be required to permit sustained persistence of gene-modified T cells.

Animals↗
PubMed1999

Retrospective comparison of the Amplatz thrombectomy device with modified pulse-spray pharmacomechanical thrombolysis in the treatment of thrombosed hemodialysis access grafts.

PURPOSE: To retrospectively evaluate the Amplatz thrombectomy device (ATD) in the treatment of thrombosed hemodialysis grafts and compare it with modified pulse-spray pharmacomechanical thrombolysis (PPT). MATERIALS AND METHODS: During a 4-month period, 79 patients presented with 126 episodes of graft occlusion. Percutaneous recanalization was performed by using the ATD (n = 57) or the modified PPT technique (n = 69). Evaluation included the technical success, complications, and primary patency rates. RESULTS: Technical success was achieved in 93% (53 to 57) of the cases treated with the ATD and in 96% (66 of 69) of the cases treated with modified PPT (P = .70). Complications occurred in 6% (four of 69) of modified PPT procedures and 16% (nine of 57) of ATD procedures. This difference was not statistically significant (P = .08); however, there were significantly more local complications in the ATD group (P = .04). The primary patency rates at 30, 90, and 180 days were 65% and 65%, 36% and 50%, and 26% and 33% for modified PPT and ATD, respectively. Survival curves were found not to differ significantly (P = .49). CONCLUSION: The ATD and modified PPT were similarly successful in the recanalization of thrombosed hemodialysis access grafts and achieved comparable primary patency rates. The higher rate of local complications and technical difficulties encountered with use of the 8-F ATD limit its usefulness for this indication.

Adult↗
PubMed1983

Antibodies against polyethylene glycol produced in animals by immunization with monomethoxy polyethylene glycol modified proteins.

Antibodies to polyethylene glycol (PEG) were raised in rabbits by immunization with monomethoxy polyethylene glycol modified ovalbumin (OA), bovine superoxide dismutase (SOD), and ragweed pollen extract (Rag), given in Freund's complete adjuvant (FCA). Immunogenicity depended on the nature of the protein and the degree of modification. With modified OA, in the presence of FCA, the majority of animals showed an anti-PEG response. With modified SOD and Rag only a small proportion of animals responded. In the absence of FCA, modified OA, given s.c., did not elicit any anti-PEG antibody response in rabbits and only a weak response in mice. PEG of MW 10,000 and 100,000 given in FCA was found nonimmunogenic in rabbits, and PEG of MW 5.9 X 10(6), given s.c. to mice, showed no or very poor immunogenic properties. Gel diffusion, heterologous passive anaphylaxis and passive hemagglutination were used to demonstrate anti-PEG antibodies raised to PEG-modified proteins. Specificity was confirmed by hapten inhibition of precipitation, inhibition of passive hemagglutination and cross-reactivity tests. PEG of MW greater than or equal to 4,000 produced specific precipitates, smaller molecules acted as monovalent haptens. From hapten inhibition of precipitation by PEG of MW 300 it appears that the antigenic determinant of PEG may be a sequence of 6-7 -CH2CH2O-units. Anti-PEG antibodies can be used analytically. By gel diffusion, Peg was detected in minimal concentrations of 0.1-1 microgram/ml. The clinical relevance of these findings with regard to therapy with PEG-modified enzymes and allergens in humans remains to be established.

Animals↗
PubMed1998

Accuracy of ICD-9-CM coding for the identification of patients with acute ischemic stroke: effect of modifier codes.

BACKGROUND AND PURPOSE: Discharge ICD-9-CM (International Classification of Diseases, 9th Revision, Clinical Modification) codes have been used to identify patients with acute stroke for epidemiological, quality of care, and cost studies. The aim of this study was to determine if the accuracy of the primary ICD-9-CM codes for ischemic stroke is improved by modifier codes and how specific codes reflect stroke subtype diagnoses. METHODS: Available hospital charts for all patients discharged from a single hospital between May 1995 and June 1997 with ICD-9-CM codes 433 (occlusion and stenosis of precerebral arteries), 434 (occlusion of cerebral arteries), or 436 (acute but ill-defined cerebrovascular disease) listed in the first position were reviewed. The primary discharge diagnosis was verified, and a presumed stroke subtype was assigned on the basis of information provided in the medical record. RESULTS: Charts were available for 175 of the 198 identified patients (88%). Of these, 61% had an acute ischemic stroke (code 433, 4%; 434, 82%; 436, 79%) with the remaining patients having other conditions. Of the 130 patients with a modifier code indicating cerebral infarction, 79% had an acute stroke; of the 45 patients with a modifier code indicating an absence of cerebral infarction, 7% had acute stroke (sensitivity, 0.97; specificity, 0.60). The codes with the highest proportions of ischemic stroke cases were 434.11 (embolic occlusion of cerebral arteries with infarction, 85%), 434.91 (unspecified occlusion of precerebral arteries with infarction, 82%), and 436 (79%), with a combined sensitivity of 0.81 and specificity of 0.90. On review, 73% of patients with code 434.11 had embolic strokes, and 47% of those with code 436 had an identified stroke cause. Of patients with code 434.91, 39% had stroke of uncertain cause, 25% "lacunar," 17% atherothrombosis, and 15% embolism. CONCLUSIONS: Despite the use of modifier codes, 15% to 20% of patients with the indicated primary ICD-9-CM codes have conditions other than acute ischemic stroke. Although the proportion of patients with acute stroke increased from 61% to 79% with the use of modifier codes, the inclusion of modifier codes did not have an appreciable effect on the accuracy of the coding if patients with code 433 are excluded. Assignment of presumed ischemic stroke subtype is particularly inaccurate.

Acute Disease↗
PubMed2001

Rat osteoblast functions on the o-carboxymethyl chitosan-modified poly(D,L-lactic acid) surface.

In this study, the functions of rat osteoblasts on o-carboxymethyl chitosan-modified poly(D,L-lactic acid) (PDLLA) films were investigated in vitro. The surface characterization was measured by contact angle and electron spectroscopy for chemical analysis (ESCA). Cell adhesion and proliferation were used to assess cell behavior on the modified surface and control. The MTT assay was used to determined cell viability and alkaline phosphatase (ALP) activity was performed to evaluate differentiated cell function. Compared to the control films, cell adhesion of osteoblasts on o-carboxymethyl chitosan-modified PDLLA films was significantly higher (p < 0.05) after 6 and 8 h culture, and osteoblast proliferation was also significantly higher (p < 0.01) between 4 and 7 days. The MTT assay suggested cell viability of osteoblasts cultured on o-carboxymethyl chitosan modified PDLLA films was significantly greater (p < 0.05) than that seeded on control one, and the ALP activity of cells cultured on modified PDLLA films was significantly higher (p < 0.01) than that found on control. These results give the first evidence that o-carboxymethyl chitosan could be used to modify PDLLA surface for improving biocompatibility.

Alkaline Phosphatase↗
PubMed1997

Synthesis of silane coupling agents containing fluorocarbon chain and applications to dentistry: plaque-controlling surface modifiers.

Silane coupling agents containing a fluorocarbon chain were prepared in high yields. It was found that silanes can be useful modifiers of the surfaces of glass, metals, and resin composites for dental use. The silane coupling agent CF3(CF2)9CH2CH2Si(OCH3)3 was the best modifier of these surfaces in terms of water and oil repellency. Colorants and experimental bacterial plaque detached much more easily from, and adhered less well to, surfaces modified with this silane coupling agent compared with unmodified surfaces. The surfaces of four teeth of a denture were modified with this silane coupling agent by spreading the agent on the surfaces with a small brush followed by brief drying with a hair drier. The modified tooth surfaces of the denture, which was worn for four months in a heavy smoker's oral cavity, were more stain-resistant than the unmodified tooth surfaces. It is expected that silane coupling agents containing a fluorocarbon chain will be surface modifiers for enhancement of oral health.

Biocompatible Materials↗
PubMed2003

Electrocatalytic reduction of chloramphenicol at multiwall carbon nanotube-modified electrodes.

A multiwall carbon nanotube-modified glassy carbon (GC) electrode was employed for the investigation of chloramphenicol (CAP) reduction. Carbon nanotube coating can greatly lower the overpotential of the electrochemical reduction of CAP and promote the electrode reaction. CAP undergoes an irreversible reduction process in phosphate buffer by the modified electrode. The reduction peak current (Ip) was significantly increased. Effects of some important factors, including pH, scan rate, and amount of modifier, on the determination of CAP were investigated. In the range of 3 x 10(-7) to 1.2 x 10(-5) M, the reduction peak current (Ip) has a good linear relationship with the concentration of CAP. When the signal-to-noise ratio is 3, the detection limit is 4.5 x 10(-8) M. The relative standard deviation of 10 measurements for 3 x 10(-6) M CAP is 5.3%, suggesting an excellent reproducibility of the modified electrode. Interfering experiments show that the modified electrode has excellent selectivity for the detection of CAP. The modified electrode was used to determine CAP in eyedrops, and the recoveries were approximately 100%.

Biosensing Techniques↗
PubMed1994

Stimulation with a monoclonal antibody (mAb4E4) of scavenger receptor-mediated uptake of chemically modified low density lipoproteins by THP-1-derived macrophages enhances foam cell generation.

mAb4E4, a murine monoclonal antibody that is specific for acetylated LDL and malondialdehyde-treated LDL, binds specifically to modified LDL present in human atherosclerotic lesions. It is directed against an epitope that is poorly exposed in delipidated and solubilized apolipoprotein B-100 from modified LDL. mAb4E4, as well as its F(ab')2 and Fab fragments, enhanced the uptake of both acetylated LDL and malondialdehyde-treated LDL by THP-1-derived macrophages resulting in a sixfold increase of cytoplasmic cholesteryl ester levels. The increased uptake of modified LDL/mAb4E4 complexes did not occur via the Fc receptor and did not depend on aggregation of modified LDL particles. However, their uptake was inhibited by blocking the scavenger receptors with fucoidin or by downregulation of receptor expression with endotoxins or interferon-gamma, indicating that their uptake is mediated via these receptors. Thus, generation of autoimmune antibodies against modified LDL and subsequent endocytosis of soluble modified LDL/antibody complexes via scavenger receptors may enhance foam cell generation. This mechanism may contribute to the progression of atherosclerotic lesions.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1995

Malondialdehyde-modified low density lipoproteins in patients with atherosclerotic disease.

The murine monoclonal antibody mAb-1H11 raised against malondialdehyde (MDA)-modified LDL, was used to detect cross-reacting material in human atheromatous tissue and in plasma. MDA-modified LDL levels in plasma were 0.19 +/- 0.02 mg/dl (mean +/- SEM) in 44 control subjects, 0.24 +/- 0.02 mg/dl in 15 patients with chronic stable angina pectoris (P = NS vs LDL cholesterol matched controls), 1.4 +/- 0.1 mg/dl in 60 patients with acute myocardial infarction (P < 0.001 vs controls), and 0.86 +/- 0.11 mg/dl in 22 patients with carotid atherosclerosis (P < 0.001 vs controls). Modified LDL, isolated from pooled LDL of 10 patients, showed a higher electrophoretic mobility on agarose gels, a higher content of thiobarbituric acid reactive substances, and a higher cholesterol/protein ratio than native LDL and had a similar reactivity (antigen/protein ratio) in the assay as the in vitro MDA-modified LDL used for calibration. Its apo B-100 moiety was not fragmented. Uptake of this modified LDL by macrophages resulted in foam cell generation. In conclusion, elevated plasma levels of atherogenic MDA-modified LDL may be a marker for unstable atherosclerotic cardiovascular disease.

Aged↗
PubMedDate not supplied

Modified release tizanidine: a review.

The pharmacokinetic and clinical properties of a modified release formulation of the alpha 2-adrenergic agonist tizanidine are reviewed. Therapy with conventional tizanidine tablets is effective in the relief of spasticity, but it has a short half-life, resulting in it having to be administered three or four times daily. The modified release capsules have retard characteristics which allow the drug to be given as a once-daily dosage. Single- and multiple-dose pharmacokinetic studies in healthy volunteers have shown that the relative bioavailability of the modified release formulation is similar to that of conventional tablets and is unaffected by food. Clinical studies have shown that modified release tizanidine improved spasticity and disability in approximately 94% and 79%, respectively, of spastic patients. Adverse effects were observed in about 33% of patients. Mild, transient muscular weakness was the most common effect, but it did not require termination of treatment. Body weight, blood pressure, heart rate and haematological or biochemical measures were not adversely affected. The initial recommended dose of modified release tizanidine is one capsule per day of 6 mg, which may be increased gradually to 24 mg. A daily dose of 12 mg modified release tizanidine was used in 50% of patients studied.

Biological Availability↗
PubMed1997

Use of the Modified Mini-Mental State Examination with children.

The validity and reliability of the modified version of the Mini-Mental State Examination with children was examined. The Modified Mini-Mental State Examination was administered to 99 children between 4 and 12 years of age (45 males and 54 females) to assess expected scores for nonclinical children and with a clinical sample. Concurrent validity was assessed through correlations of Modified Mini-Mental State Examination scores with Wechsler Intelligence scores and Child Behavior Checklist scores. The Modified Mini-Mental State Examination was administered to 26 children on two occasions to determine test-retest reliability. Means and standard deviations of scores are reported by age and grade level. Test-retest reliability coefficients were positively significant. For the nonclinical sample, Modified Mini-Mental State Examination scores were significantly and positively correlated with Verbal IQ and Child Behavior Checklist scores. Modified Mini-Mental State Examination scores were significantly correlated with Verbal IQ scores in the total and clinical samples.

Child↗