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Biomedical subjects

K Arnold

Publications and source records attributed to K Arnold.

At least 109 records · Page 6Linked to original sources
PubMed1995

NMR studies of the action of hypochlorous acid on native pig articular cartilage.

The action of sodium hypochlorite on pig articular cartilage was studied by 1H-NMR spectroscopy to model some aspects of degradation processes of cartilage during rheumatoid arthritis. Two effects of NaOCl on cartilage polysaccharides have been observed. Hypochlorous acid causes an enhanced release of oligomeric polysaccharides from cartilage. The second effect concerns the degradation of N-acetyl side chains of carbohydrates to acetate via a chlorinated transient product. Signal intensities for N-acetyl groups (approximately 2.0 ppm) increase during the first 2 h of incubation of cartilage with NaOCl. Then they decrease again. However, acetate (1.90 ppm) as the final product of degradation of N-acetyl side chains increases continuously over the period of incubation with NaOCl. In addition to polysaccharides, effects of NaOCl were only observed in cartilage samples on amino acids like alanine. The alanine resonance disappeared already at NaOCl concentrations where only small effects on cartilage polysaccharides have been observed.

Animals↗
PubMed1995

Identification and preliminary characterization of a chitinase gene in the Autographa californica nuclear polyhedrosis virus genome.

A functional chitinase gene (chiA) has been identified in the genome of the Autographa californica nuclear polyhedrosis virus (AcMNPV). It is expressed in the late phase of virus replication in insect cells. High levels of both endo- and exochitinase activity were detected by 12 hr p.i. and remained stable throughout infection. An AcMNPV chiA protein-specific antibody was prepared using recombinant material prepared in bacteria. This was used to demonstrate that a product of approximately 58 kDa was synthesised in virus-infected cells. Immunofluorescence analysis of virus-infected cells showed that most chitinase was located in the cytoplasm. Primer extension analysis of mRNA from AcMNPV-infected cells confirmed that transcription initiated from a baculovirus late start site (TAAG), 14 nucleotides upstream from the putative translation initiation codon. The predicted protein sequence of the AcMNPV chiA shares extensive sequence similarity with chitinases from bacteria and, in particular, the Serratia marcescens chitinase A (60.5% identical residues). Phylogenetic analyses indicate that AcMNPV, or an ancestral baculovirus, acquired the chitinase gene from a bacterium via horizontal gene transfer.

Amino Acid Sequence↗
PubMed1995

Identification of the major site of apolipoprotein B modification by advanced glycosylation end products blocking uptake by the low density lipoprotein receptor.

Advanced glycosylation end products (AGEs) arise from glucose-derived Amadori products and have been implicated in the pathogenesis of diabetic vascular disease. We recently reported the presence of an AGE-modified form of low density lipoprotein (LDL) that circulates in high amounts in patients with diabetes or renal insufficiency and that exhibits impaired plasma clearance kinetics. We utilized AGE-specific antibodies to identify the major sites of AGE modification within protease-digested preparations of apolipoprotein B that impair the binding of the AGE-modified form of LDL by human fibroblast LDL receptors. The predominant site of AGE immunoreactivity was found to lie within a single, 67-amino acid region located 1791 residues NH2-terminal of the putative LDL receptor binding domain. These data point to the high reactivity and specificity of this site for AGE formation and provide further evidence for important structural interactions between the LDL receptor binding domain and remote regions of the apolipoprotein B polypeptide.

Amino Acid Sequence↗
PubMed1995

Action of compression and cations on the proton and deuterium relaxation in cartilage.

In this paper, investigations are described on the influence of osmotic pressures and of varying cation concentrations on water relaxation times in cartilage (pig articular cartilage and bovine nasal cartilage). Both water content and relaxation times decrease strongly with increasing osmotic pressure. This relaxation behavior can be explained in terms of a fast chemical exchange between unbound and bound water. Na+ does not influence water content or relaxation times, whereas Ca2+ causes a small reduction in these parameters.

Animals↗
PubMed1995

Immunoglobulin A class fibroblast antibodies in patients with Graves' disease and pretibial myxedema.

The involvement of autoantibodies in the extrathyroidal manifestations of Graves' disease has been the subject of extensive investigation, with fairly inconclusive results to date. We investigated the presence of immunoglobulin A (IgA) and IgG antibodies in patients with Graves' disease and pretibial myxedema (PTM; n = 21) as well as those with Graves' disease with thyroid-associated ophthalmopathy (TAO; n = 10), Graves' disease with no clinical evidence of extrathyroidal manifestations (n = 11), Hashimoto's thyroiditis (n = 9), type 1 diabetes mellitus (n = 10), systemic lupus erythematosus (n = 9) and normal individuals (n = 17). We looked for antibodies to both retroocular muscle and dermal fibroblasts as well as to thyroid peroxidase, thyroid microsomal antigen, thyroglobulin, and human eye muscle membranes. IgA class antibodies to microsomal antigen (30-50% of patients), thyroid peroxidase (5-20%), and human eye muscle membrane (0-26%) antigens were found in the various groups of patients with Graves' disease. With each of these antigens, serum from patients with PTM showed the greatest binding. Highly significant IgA binding was shown by PTM serum to both dermal (P < 0.001) and retroocular muscle (P < 0.001) fibroblasts from 12 different donors. Serum from Graves' patients with and without TAO and that from Hashimoto's thyroiditis patients reacted significantly with 4 of the 12 fibroblasts lines. In contrast, IgG binding was only found for 3 of the 12 fibroblast lines using patient serum. The IgA binding to fibroblasts shown by PTM patients was predominantly of the IgA2 subclass. The activity was absorbed out by both fibroblasts and thyroid cells. In immunoblotting studies, PTM patient serum reacted with a 54-kilodalton dermal fibroblast antigen and a 66-kilodalton retroocular fibroblast antigen. No binding to these antigens was seen with serum from normal controls or patients without PTM. Further elucidation of the nature of this fibroblast antigen will help to determine the role of IgA autoantibodies in the extrathyroidal manifestations of Graves' disease.

Adult↗
PubMed1995

[Comparison of the effectiveness of sodium azide and merthiolate as bacteriocidal agents--a NMR study].

The pseudohalogenide sodium azide (NaN3) and the organometallic mercury compound merthiolate (sodium salt of ethyl mercurithiosalicylate) are commonly used as preserving solutions for biomaterials. In the present paper, these preserving solutions were compared for their efficacy in preserving porcine articular cartilage. For this purpose, porcine articular cartilage was incubated with phosphate buffer containing the corresponding amounts of NaN3 or merthiolate. Supernatants were assayed using NMR spectroscopy, and the content of soluble saccharides was determined using the dye Alcian blue. The results obtained clearly indicate that merthiolate is more effective than sodium azide. This makes the former a more suitable preserving agent for the storage of cartilage. A significant decrease in the lactate concentration, and enhancement of the acetate and the N-acetyl resonances indicate degenerative changes in the cartilage. However, when a large excess of sodium azide is used, its efficacy approaches that of merthiolate.

Animals↗
PubMed1994

Peroxidation of egg yolk phosphatidylcholine liposomes by hypochlorous acid.

The powerful neutrophil-derived oxidant hypochlorous acid HOCl/OCl- is assumed to contribute to tissue injury in a number of pathological states accompanied by massive accumulation of neutrophils. The production of malondialdehyde to indicate lipid peroxidation was studied in egg yolk phosphatidylcholine liposomes upon treatment with NaOCl as a source for hypochlorous acid. Its accumulation was inhibited by alpha-tocopherol and butylated hydroxytoluene. Singlet oxygen, hydroxyl radicals or superoxide anion radicals derived from direct reactions of hypochlorous acid seem not to be involved in initiation of lipid peroxidation because the malondialdehyde accumulation was unaffected by hydrogen peroxide, catalase, superoxide dismutase, ferrous sulphate or ferric chloride. Double bonds of fatty acid residues seem to be the primary target for NaOCl. Their number is continuously diminished in liposomes (2 mg lipids/ml) after incubation with increasing amounts of NaOCl at 37 degrees C for 40 min as detected by two independent methods (iodine bromide reduction and 1H-NMR spectroscopy). A 1:1 molar ratio between the loss of double bonds and NaOCl added was found only at low NaOCl concentrations. Then double bonds are decreased with a lower efficiency. A continuous increase of lipid peroxidation products was only observed up to 0.5-0.7 mmol/l NaOCl. The yield of lipid hydroperoxides kept constant at higher NaOCl concentrations. However, diene conjugates and malondialdehyde exhibit a maximum at 0.7-1 mmol/l or 0.5 mmol/l NaOCl, respectively, while the concentration of these products decreases at higher doses of NaOCl. The decrease of malondialdehyde was more pronounced than for diene conjugates. These results were discussed from the background that at minimum two (diene conjugates) or three (malondialdehyde) double bonds in a fatty acid residue are necessary for formation of lipid peroxidation products.

Egg Yolk↗
PubMed1994

Partitioning of high-density lipoproteins in charge-sensitive two-phase systems.

Aqueous polymer two-phase systems characterized by a difference in the electrical potential between the upper and the lower phase (charged systems) are useful tools for the detection of changes in the surface charge and hydrophobicity of high-density lipoproteins (HDL). While the large particle size of low-density lipoproteins (LDL) leads to accumulation at the interface, the smaller diameter and the higher surface charge density of the native HDL particles allows partitioning without aggregation at the interface. Charged two-phase systems can be used to check the native state of HDL samples. Moreover, these systems would be suitable for investigating the hydrophobicity and surface charge of modified HDL.

Dextrans↗
PubMed1994

Interaction of melittin with lipid membranes.

Interaction of melittin with lipid membranes was studied systematically with respect to its adsorption onto membranes, its effect on membrane leakage and fusion, and micellization at various melittin/lipid ratios. It was found that melittin has a strong affinity for adsorption onto lipid membranes. The analysis of the measured electrophoretic mobilities by use of a Gouy-Chapman double layer theory, shows that melittin is adsorbed onto the phosphatidylserine membrane several times more than the phosphatidylcholine membrane. However, it was observed that the phosphatidylcholine membrane is more susceptible to membrane leakage, vesicle fusion and micellization at a lower level of melittin adsorbed than the phosphatidylserine membrane. For small unilamellar phosphatidylcholine vesicles in 0.1 M NaCl, membrane leakage started at melittin to lipid ratio of 1:2000, a large increase in the rate of membrane leakage occurred at a ratio of about 1:500 or higher, membrane fusion occurred at a ratio of 1:200, and membrane micellization at a ratio of 1:10. On the other hand, for small unilamellar phosphatidylserine vesicles, the respective concentrations of melittin to result in membrane leakage, vesicle fusion, and membrane micellization were several times higher. Surface pressure measurements of lipid monolayers showed that the increase in surface pressure of the phosphatidylcholine monolayer due to the presence of melittin in the subphase solution was greater than that for the phosphatidylserine monolayer at any melittin concentration in the subphase solution. These experimental results indicate that melittin tends to be adsorbed on the surface of the negatively charged phosphatidylserine membrane due to the electrostatic binding so that the melittin molecule can stay out more on the surface of the membrane, while melittin appears to be adsorbed more into the hydrophobic membrane core for the electrically neutral phosphatidylcholine membrane.

Adsorption↗
PubMed1994

Quenching and dequenching of octadecyl Rhodamine B chloride fluorescence in Ca(2+)-induced fusion of phosphatidylserine vesicles: effects of poly(ethylene glycol).

Ca(2+)-induced fusion of SUV and LUV composed of ox brain phosphatidylserine (PS) was studied as a function of temperature and concentration of Ca2+ using octadecyl Rhodamine B chloride (R-18). Ca2+ was added to a 1:1 mixture of labelled (8 mol%) and unlabelled vesicles (assay conditions) or to samples containing only labelled liposomes (control conditions). Both, in SUV and LUV the dependence of differences in fluorescence between assay and control samples on temperature can be divided into three regions. At temperatures lower than 20 degrees C the differences in fluorescence increase only slightly in SUV or remain unchanged in LUV after the addition of Ca2+. At 28 degrees C and higher temperatures the differences of fluorescence intensities increase much more drastically, whereby SUV exhibit higher fusion rates than LUV. Between 20 degrees C and 28 degrees C exists an intermediate region for both SUV and LUV. Here the fluorescence changes continuously from one behaviour to the other independent of the concentration of Ca2+. A drastic quenching of R-18 fluorescence occurs in LUV composed of PS below 10 degrees C, where the lipids are in the gel state. In SUV the fluorescence is only weakly changed in this temperature region. It is assumed that a demixing between dye and phospholipid molecules occurs below phase transition. During fusion the phase transition of PS is shifted from 8-10 degrees C to about 24-28 degrees C as revealed by polarization measurements using diphenylhexatriene. Because the differences in R-18 fluorescence between assay and control samples depend strongly on temperature we assume that the shift in phase transition temperature of PS occurs immediately after the addition of Ca2+ to SUV or LUV. Poly(ethylene glycol) 6000 accelerates fusion in both SUV and LUV under all conditions where a fusion takes place. Further, the threshold concentration of Ca2+ to induce fusion is diminished from about 1 mmol/l without polymers to about 0.5 mmol/l in the presence of 10% (w/v) PEG 6000. The intermediate region of changes in fluorescence properties of R-18 in the Ca(2+)-induced fusion of PS is not changed by PEG.

Calcium↗
PubMed1994

Partitioning of chemically modified low-density lipoprotein in aqueous polymer two-phase systems.

Aqueous polyethylene glycol (PEG)-dextran two-phase systems containing 10 mM Tris.HCl (pH 7.4) were used for the partitioning of chemically modified low-density lipoprotein (LDL). Anionic modification connected with an increase in the negative surface charge of lipoproteins favours the accumulation of modified LDL in the top phase. The partition coefficient increases depending on the extent of modification. Cationic modification yields lower values for the partition coefficient. Positively charged LDL favours a bottom-phase accumulation. With weakly charged and nearly neutral particles, the Van der Waals interaction between polymer and particle preponderates over electrostatic interactions, leading to a favoured accumulation of LDL in the PEG-rich top phase. Results of measurements of the relative electrophoretic mobility and the determination of free amino groups are in agreement with the calculated values of the partition coefficient. Because the partitioning of LDL is accompanied by aggregation at the interface, experimental techniques have to be carefully standardized. Subtle differences in the surface properties of modified LDL can be detected.

Acetylation↗
PubMed1994

Early treatment of malaria in the community using artemisinin--hope or hazard?

Most malaria morbidity and mortality occurs in the community. Treatment aimed at managing patients at this level of the health care system should bring health and economic benefits to the population. Artemisinin suppositories have been successfully used in adults and children to treat, uncomplicated and complicated malaria. Their use early in the course of the disease may reduce complications and patient referral to tertiary care hospitals. A single dose of artemisinin followed by a single dose of mefloquine, given at the household or hamlet level, may be a therapeutic approach that has advantages for developing countries. However, widespread and uncontrolled use of artemisinin could result in unexpected and undesirable side-effects, improper dose schedules and poor compliance which, in turn, might cause primary treatment failures, recrudescences, and possibly resistance to the drug. Such problems can be avoided or managed by regulating and monitoring its use.

Adult↗
PubMed1994

T cell responses to orbital antigens in thyroid-associated ophthalmopathy.

Thyroid-associated ophthalmopathy (TAO) is most likely to be a T cell-mediated disease, in which cytokines released in the extraocular muscles activate fibroblasts, increasing glycosaminoglycan production. The nature of the orbital antigen recognized by the infiltrating T cells is unclear, although it is possible that there is cross-reactivity between this and a thyroid autoantigen to explain the close association with thyroid autoimmunity. We have tested the ability of human and porcine eye muscle antigen preparations to stimulate proliferation of circulating T cells from healthy subjects and patients with TAO or Graves' disease without clinical TAO. Occasional responses were seen, particularly after depletion of CD8+ T cells, and two out of 10 TAO patients responded to eye muscle proteins of 25-50 kD after fractionation of antigens on gels and subsequent elution. There was no disease-specific response of T cells to R1, R14, D1 and 1D3, recombinant proteins identified from screening an eye muscle cDNA library with sera from patients with autoimmune thyroid disease. We have also found that interferon-gamma (IFN-gamma) production by T cells from TAO patients was not stimulated by eye muscle membrane antigens or by 1D3. These results suggest that the frequency of circulating T cells responding to eye muscle antigens in TAO is low, and that several candidate orbital antigens, including the 64-kD protein 1D3, are unlikely to be important T cell autoantigens in this condition.

Animals↗
PubMed1994

Immunoglobulin class and subclass distribution of eye muscle and fibroblast antibodies in patients with thyroid-associated ophthalmopathy.

OBJECTIVE: The investigation of the antibody response in thyroid-associated ophthalmopathy (TAO) using different antigens and assays has given inconsistent results. We have analysed antibodies against eye muscle and control antigens in a large group of TAO patients to assess whether specific eye muscle antibodies exist in TAO. We have also evaluated the presence of IgA and IgM class antibodies and examined IgG subclass distribution. DESIGN: Sera were obtained from all patients (TAO, Graves' disease without ophthalmopathy and Hashimoto's thyroiditis) within one year of diagnosis. Sera were also collected from healthy controls, with no family history of autoimmune thyroid disease. PATIENTS: Thirty-eight patients had Graves' disease with Grade III or greater TAO; 15 patients had Graves' disease without ophthalmopathy and nine had Hashimoto's thyroiditis without any eye signs. The control group consisted of 14 subjects. MEASUREMENTS: Antibodies against porcine eye and skeletal muscle, human eye (membrane and soluble antigen) and skeletal muscle, human thyroid microsomal and thyroglobulin antigens and dermal and orbital fibroblast antigens were assessed using ELISA. Antibody isotypes and IgG subclasses were studied for porcine and human eye muscle antibodies. Eye muscle (porcine and human) and orbital fibroblast antibodies were further analysed by immunoblotting. RESULTS: There were no significant differences in the ability of either IgG or IgA in sera from the different groups to bind porcine and human eye muscle antigens. There was a significant correlation (P < 0.0001) between the binding to porcine eye muscle and skeletal muscle antigens (for both IgG and IgA). There was no difference between sera from TAO patients and control subjects in their binding to eye muscle fibroblasts for both IgG and IgA antibodies. However, IgA antibody activity against dermal fibroblasts differed significantly between TAO patients and controls (P < 0.05). By immunoblotting, the frequency of IgA antibodies recognizing 21 kDa (40% of patients) and 62 kDa (52%) bands in porcine eye muscle blots and 20, 24 and 38 kDa bands in blots of human eye muscle (soluble) antigen differed significantly between patients with TAO and controls (P < 0.05 in all cases). IgG antibodies recognizing 80 and 92 kDa bands in blots of the subcellular membrane antigen prepared from orbital fibroblasts were found more frequently in patients with TAO compared with controls (P < 0.05 in both cases). CONCLUSIONS: We found no evidence that eye muscle membrane or fibroblast antibodies are present in a significant proportion of TAO patients, using ELISAs based on antigens prepared from several sources. We have also failed to demonstrate the presence of previously described specific, TAO-associated antibodies, including those directed against a 64 kDa protein in eye muscle and a 23 kDa protein in fibroblasts. IgA class antibodies reactive with orbital components appeared to be more strongly associated with TAO than those of the IgG class, though even this relationship is weak. These results suggest that antibodies are of secondary importance in the pathogenesis of TAO, which is most likely a T cell-mediated disorder.

Animals↗
PubMed1994

The action of hypochlorous acid on polymeric components of cartilage.

The action of sodium hypochlorite on N-acetylglucosamine, N-acetylgalactosamine, chondroitinsulfate and hyaluronic acid was studied by 1H-nuclear magnetic resonance (1H-NMR) in order to model some aspects of degradation processes caused by neutrophils on carbohydrate polymers of cartilage in rheumatoid arthritis. N-Acetyl side groups of carbohydrate monomers and chondroitinsulfate yield a resonance at 2.01-2.04 ppm in proton NMR-spectra. This resonance is observed in hyaluronic acid solutions only after a prolonged incubation to yield shorter polymeric chains. Sodium hypochlorite causes a continuous decrease of the line for N-acetyl groups. Two new resonances appear in the 1H-NMR spectra. An intermediate product, assumed as a chlorinated product of N-acetyl side chains, shows a chemical shift of about 2.35 ppm. This intermediate is hydrolyzed to a carbohydrate ring and acetate (1.90 ppm). Sodium hypochlorite acts in all systems investigated mainly on N-acetyl groups. Only small effects on the carbohydrate ring were found under our experimental conditions.

Acetylgalactosamine↗
PubMed1993

Effect of glycosaminoglycans and PEG on fusion of Sendai virus with phosphatidylserine vesicles.

The fusion of Sendai virus with phosphatidylserine vesicles was monitored by a pyrene-phosphatidylcholine fluorescence assay. A strong influence of pH and ionic strength on the extent of fusion was observed. The negatively-charged polymers (dextran sulfate, heparin and chondroitin sulfate) inhibited the ability of the viruses to fuse with the liposomes. The extent of inhibition, for a given amount (w/v) of the polymers, was the greatest for dextran sulfate followed by heparin and chondroitin sulfate. The extent of inhibition depended on the pH and ionic strength of the solution; the lower the pH of the solution, the more effective the fusion inhibition by the polymers. The molecular weight of dextran sulfate (DS) influenced the inhibition effect, i.e., DS with higher molecular weight exhibited a stronger inhibition effect. The presence of sodium sulfate, even in excess concentration, had no inhibitory effect on fusion. On the other hand, PEG had an opposite effect on fusion compared to the negatively-charged polymers, and it decreased their inhibition effect when both were present in the same media. It is concluded that the inhibition of the fusion activity of Sendai virus results from the adherence of negatively-charged polymers to the virus surface preventing close contacts between the virus and liposome surface.

Dextran Sulfate↗
PubMed1993

Randomized comparative study of fleroxacin and chloramphenicol in typhoid fever.

This study compared fleroxacin, 400 mg daily for 7 or 14 days, with chloramphenicol, 50 mg/kg per day for 14 days, for the treatment of patients with typhoid fever in a multicenter study. A total of 184 patients were randomly assigned to the three treatment groups. Efficacy was determined by culture of blood and stool, overall clinical response, and time to defervescence. Safety was assessed by occurrence of adverse events and laboratory abnormalities. A total of 97 patients were evaluable for efficacy, 28 in the fleroxacin 7-day group, 35 in the fleroxacin 14-day group, and 34 in the chloramphenicol group. All showed rapid defervescence with high cure rates. Bacteriologic cure rates were 96% in the fleroxacin 7-day group, 97% in the fleroxacin 14-day group, and 85% in the chloramphenicol group. Clinical cure rates were 83-100% with fleroxacin and 82% with chloramphenicol. The time to defervescence was shorter for patients treated with fleroxacin than for those treated with chloramphenicol. All three treatment regimens were well tolerated. Fleroxacin, 400 mg daily for 7 days, appears to be satisfactory for the treatment of typhoid fever and compares favorably with the standard 14-day therapy with chloramphenicol.

Adolescent↗