Sera Of Cystic Fibrosis Patients Research Articles

Selenium speciation in human serum of cystic fibrosis patients compared to serum from healthy persons

A formerly developed Se speciation method was applied to human serum. Chomatographic performance was checked regularly by measuring control standards after four sample run, each, to ensure sufficient separation of the species and sensitivity of the method even after a considerable number of serum samples. Detection limits of investigated species were similar to those reported and showed values below 1 microg/L related to Se for each Se compound. Sera of cystic fibrosis (CF) patients were investigated in comparison to sera from healthy volunteers with respect to total selenium and Se species. Generally, CF sera showed lower values of total Se, Selenocystine (SeC), and cationic/neutral Se compounds compared to serum of healthy persons. No significant gender-specific differences were found. Total Se in sera of healthy persons was determined at 102+/-12 microg/L (n = 12 individuals, mean value from male and female, age 4-38 years), whereas CF patients showed 58+/-10 microg/L (n = 31 individuals, mean value from male and female, age 3-35 years). Se-cystine showed significant differences between the CF and healthy group with a lowered SeC value in sera of CF patients by -75% (mean ca. 26 microg/L in healthy sera compared to about 6.5 microg/L (mean) in CF sera). A similar situation is seen for neutral/cationic Se compounds, which partly may comprise of Se proteins. The lowered SeC values together with lowered cationic/neutral Se compounds (probably Se enzymes) point to a Se-depleted regulated pathway combined with a reduced capability of protective functions such as protection from peroxides.

Selenium speciation in human serum of cystic fibrosis patients compared to serum from healthy persons

A formerly developed Se speciation method was applied to human serum. Chomatographic performance was checked regularly by measuring control standards after four sample run, each, to ensure sufficient separation of the species and sensitivity of the method even after a considerable number of serum samples. Detection limits of investigated species were similar to those reported and showed values below 1 μg/L related to Se for each Se compound. Sera of cystic fibrosis (CF) patients were investigated in comparison to sera from healthy volunteers with respect to total selenium and Se species. Generally, CF sera showed lower values of total Se, Selenocystine (SeC), and cationic/neutral Se compounds compared to serum of healthy persons. No significant gender-specific differences were found. Total Se in sera of healthy persons was determined at 102 ± 12 μg/L ( n = 12 individuals, mean value from male and female, age 4–38 years), whereas CF patients showed 58 ± 10 μg/L ( n = 31 individuals, mean value from male and female, age 3–35 years). Se-cystine showed significant differences between the CF and healthy group with a lowered SeC value in sera of CF patients by −75% (mean ca. 26 μg/L in healthy sera compared to about 6.5 μg/L (mean) in CF sera). A similar situation is seen for neutral/cationic Se compounds, which partly may comprise of Se proteins. The lowered SeC values together with lowered cationic/neutral Se compounds (probably Se enzymes) point to a Se-depleted regulated pathway combined with a reduced capability of protective functions such as protection from peroxides.

Differential IgE recognition of recombinant Aspergillus fumigatus allergens by cystic fibrosis patients with allergic bronchopulmonary aspergillosis or Aspergillus allergy.

Allergic bronchopulmonary aspergillosis (ABPA), an intense inflammatory reaction to Aspergillus in the lung, is recognized as a severe complication in patients with cystic fibrosis (CF). The diagnosis of ABPA in CF patients sensitized to Aspergillus fumigatus is complicated by interfering laboratory and clinical findings shared by the diseases. We have used cDNA encoding A. fumigatus allergens which were cloned from a cDNA library displayed on phage surface to produce recombinant proteins in Escherichia coli. Differential IgE responses to the allergens in A. fumigatus-sensitized CF patients with or without ABPA and CF controls without sensitization to A. fumigatus were demonstrated. A secreted ribotoxin (rAsp f 1) and a peroxisomal protein (rAsp f 3) were recognized by sera from A. fumigatus-sensitized CF-patients with or without ABPA. An intracellular manganese superoxide dismutase (rAsp f 6) and rAsp f 4, a protein with unknown function, were recognized exclusively by IgE from sera of CF patients with ABPA. Therefore, Asp f 4 and Asp f 6 represent specific markers for ABPA and allow a sensitive, fully specific diagnosis of the disease. The data suggest distinct IgE responses to colonization of the bronchial tree in CF patients with ABPA or A. fumigatus allergy and therefore a differential recognition of the pathogen in the two IgE-related inflammatory diseases.

Serum immunoglobulin G directed against porcine trypsin in the serum of cystic fibrosis children receiving porcine pancreatic enzyme supplements

Cystic fibrosis (CF) patients require pancreatic enzyme replacement therapy to correct pancreatic insufficiency. These enzymes are derived from porcine pancreas and are known to be antigenic. To determine the possible clinical consequences, a specific ELISA was developed to detect IgG antibody directed against porcine trypsin (PTAb) in the sera of CF patients. The assay was used to evaluate the occurrence of PTAb in a cross sectional study of 103 CF patients in relation to the introduction of porcine enzyme therapy, clinical status and genotype. Antibodies against porcine trypsin were detected in the sera of 63% of patients unrelated to the age of commencement or the duration of enzyme therapy. No differences were observed in the clinical status of CF patients who had developed PTAb (n = 65) and those who had no detectable PTAb (n = 38) as determined from: the current prescribed dose of porcine pancreatic enzyme capsules; Z scores for height and weight; and respiratory function tests. It is suggested that the PTAb commonly found in the sera of CF patients are of doubtful clinical significance but the prospect of PTAb contributing to immune complex disease should be examined further.

Cystic fibrosis proteins detected by electrophoretic techniques.

For the detection of the cystic fibrosis protein (CFP) in serum of cystic fibrosis (CF) carriers, thin-layer polyacrylamide gel isoelectric focusing proved inappropriate as a diagnostic test, but was useful for screening fractions on purification of CFP by chromatofocusing on a Mono P column. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis an Mr 12,000 protein (P12) was found in most CFP-positive sera, indicating good correlation between these two CF-associated proteins. Detection of the P12 protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was well reproducible and less delicate than IEF. The technique was also used to purify P12 from serum by two successive preparative electrophoresis steps in a 7.5-15% gradient and 15% homogeneous gel. The use of silver staining revealed that P12, which was present in all sera of CF patients and carriers with variable intensities, was also present in trace amounts in normal sera.

Activity levels and properties of acid α-glucosidase from liver and neutral α-glucosidase from sera of cystic fibrosis patients and controls

The average activity levels of acid α-glucosidase are comparable in liver supernatant fluids for 15 cystic fibrosis patients and 12 controls (401 ± 131 and 347 ± 109 nmol/h/mg protein, respectively) and no significant differences were found for the cystic fibrosis and control liver acid α-glucosidases in their (a) apparent K m values for the 4-methylumbelliferyl substrate (1.1 mmol/1), (b) pH optima (4.2) and thermostability curves and (c) isoelectric profiles (one form with an isoelectric point of 4.5 ± 0.2). In contrast, average neutral α-glucosidase activity levels were significantly increased ( p < 0.0002) in sera from 21 cystic fibrosis patients compared to 15 controls (10.7 and 2.7 nmol/h/ml). This increased activity is not due to (a) different stability upon storage at −20°C, (b) the presence of activators in cystic fibrosis sera or inhibitors in normal sera (as determined by mixing studies), (c) altered K m values or (d) altered pH optima curves. Cystic fibrosis serum neutral α-glucosidase appears to be more thermostable and has a consistently altered isoelectric profile (greater percentage of activity above pI 4.8) when compared to the normal serum enzyme. This altered isoelectric composition may reflect changes in neutral α-glucosidase which contribute to its increased activity in cystic fibrosis sera.

The levels of galactosyltransferase activity in sera from normal children and patients with cystic fibrosis

The levels of galactosyltransferase have been studied in the sera obtained from 59 normal individuals varying in age from 5 – 19 years, and from 29 cystic fibrosis patients varying in age from 5 – 33 years. The enzyme level found in children was lower than that in the adults. The levels in the sera of cystic fibrosis patients was found to be about half of that in the normal sera for the corresponding age group. In those cystic fibrosis patients over 20 years of age, normal levels were found.

Cystic fibrosis: fractionation of fibroblast media demonstrating ciliary inhibition.

A chromatographic fraction has been identified in media from fibroblast cultures derived from 15 of 18 cystic fibrosis patients and 8 of 10 heterozygotes that inhibits ciliary activity, as judged by the oyster gill assay. The analogous fraction of media from cultures of fibroblasts derived from 11 normal individuals did not inhibit ciliary activity under the experimental conditions used. The chromatographic fractions containing the ciliary inhibitory activity from both fibroblast media and sera of cystic fibrosis patients and heterozygotes eluted in the same position from diethylaminoethyl-cellulose at pH 8.5. Although the serum ciliary inhibitor had consistently been detected in a fraction that contained predominantly immunoglobulin G, results of the fractionation of fibroblast media demonstrate that the ciliary inhibitor is present in a protein fraction containing three species of different molecular weight and in which IgG cannot be detected by the techniques used.