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F L Kiechle

Publications and source records attributed to F L Kiechle.

At least 19 recordsLinked to original sources
PubMed1992

Fasting induced alterations in mitochondrial palmitoyl-CoA metabolism may inhibit adipocyte pyruvate dehydrogenase activity.

1. Adipocytes from fed and fasted (24 hr) groups of rats were fractionated into mitochondria, microsomes and plasma membranes. 2. Fasting significantly decreased the mitochondrial activity of palmitoyl-CoA synthetase, palmitoyl-CoA hydrolase, beta-oxidation and pyruvate dehydrogenase. 3. Fasting elevated intramitochondrial long-chain acyl-CoA. 4. Pyruvate dehydrogenase was inhibited 50% by addition of 30 microM palmitoyl-CoA. 5. Fasting-induced changes in palmitoyl-CoA metabolism may modulate pyruvate dehydrogenase activity in adipocyte mitochondria.

Adipose Tissue↗
PubMed1992

A flow cytometric method to detect anti-pyruvate dehydrogenase antibody in primary biliary cirrhosis.

Primary biliary cirrhosis (PBC) is an autoimmune disease characterized by the presence of anti-mitochondrial antibodies specifically directed against the M2 group of mitochondrial antigens. Recently, the E-1, the E-2, and protein X components of pyruvate dehydrogenase enzyme complex have been identified as the major antigens within the M2 group of autoantigens. An immunoassay using pyruvate dehydrogenase enzyme complex as a specific antigen for the diagnosis of PBC was developed. Pyruvate dehydrogenase enzyme complex was attached to polystyrene microbeads, incubated with sera from PBC patients (n = 18), normal controls (n = 50), or patients with other autoimmune diseases (n = 26), followed by incubation with a second fluorescein isothiocyanate conjugated goat anti-human immunoglobulin and then analyzed by flow cytometry. High numbers of fluorescence channels (mean, 1,693 +/- 846) were obtained for all PBC sera except for two patients. Compared to the conventional anti-mitochondrial antibody assay, the assay had a sensitivity rate of 94% and a specificity rate of 100%. The reactive antibodies are predominantly of the immunoglobulin G3 subclass. Their levels could be correlated with the histopathologic stages of PBC. These results were corroborated by immunoblotting. Sera from patients with later stages of PBC strongly reacted with pyruvate dehydrogenase enzyme complex components, E1 alpha, and protein X.

Antibodies↗
PubMed1992

Quality improvement in the laboratory assessment of in vitro fertilization.

There has been a trend to use fewer laboratory tests during ovarian hyperstimulation prior to oocyte recovery, in vitro fertilization, and embryo transfer. Estradiol is routinely monitored during ovulation induction protocols. Estradiol rises steadily to supraphysiologic concentration during ovarian hyperstimulation. Review of the College of American Pathologists proficiency testing data from 1990 reveal that the within-method interlaboratory coefficient of variation meets the proposed maximum allowable analytical error of 11.8%. The luteinizing hormone level increases during ovarian hyperstimulation with a skewed distribution. Molecular variants exist that may bind with different affinities to monoclonal antibodies used in two-site sandwich assays. Polyclonal radioimmunoassays usually overestimate luteinizing hormone concentration. The College of American Pathologists proficiency testing data reveal that only three of eight methods with greater than 20 participants had a within-method interlaboratory coefficient of variation that met the proposed maximum allowable analytical error of 10% or less. International standardization of reference standards, antibodies, and labeling methods may improve the performance of this assay. The low pregnancy rate experienced by in vitro fertilization protocols suggests that additional laboratory tests need to be developed to monitor the receptivity of the endometrium for implantation and the quality of the oocyte and embryo.

Fertilization in Vitro↗
PubMed1991

Fluorometric determination of phosphatidylcholine as a measure of phospholipid methylation.

The successive methylation of phosphatidylethanolamine to phosphatidylcholine (phospholipid methylation) has been measured by the incorporation of S-[methyl-3H]adenosylmethionine or colorimetric assay of phosphatidylcholine extracted from adipocyte plasma membranes. A fluorometric assay for phosphatidylcholine was developed to measure phospholipid methylation. This assay is 10 times more sensitive than the colorimetric assay and demonstrates no significant interference with other methylated phospholipids. The fluorometric assay was used to determine a biphasic insulin dose response in adipocyte plasma membranes. This fluorometric assay for phosphatidylcholine represents an alternative method for monitoring phospholipid methylation, especially when increased sensitivity is required.

Adipose Tissue↗
PubMed1990

The effect of amino acids, monoamines and polyamines on pyruvate dehydrogenase activity in mitochondria from rat adipocytes.

The ability of polyamines and other cationic compounds including monoamines, amino acids, poly-L-arginine, poly-D-lysine and poly-L-lysine, to alter pyruvate dehydrogenase (PDH) activity in mitochondria from rat epididymal adipocytes was determined. PDH was assayed with the substrate [1-14C] pyruvate in the presence of 0.05 mM Ca2+ and Mg2+. Nine of the fourteen compounds tested at 0.1 mM caused a significant increase (procaine, 3-(beta-morpholinopropionyl) benzo [b]thiophene [VII], spermine, spermidine, putrescine, lysine and tryptophan) or decrease (poly-L-arginine, 3-(beta-piperidinopropionyl) benzo[b]thiophene) in PDH activity. None of these compounds nonenzymatically decarboxylated [1-14C] pyruvate to release 14CO2. NaF, a PDH phosphatase inhibitor, suppressed the stimulatory effects of those compounds tested: procaine, tryptophan, VII, spermine and spermidine. These results imply that these five compounds activate PDH activity through stimulation of the PDH phosphatase. When the Mg2+ concentration was increased from 0.05 to 4.5 mM, the stimulatory effect of spermine was increased, consistent with the finding by others that spermine lowers the Km of the enzyme for Mg2+. However, at Mg2+ concentrations greater than 0.3 mM, the stimulatory effect of VII was unaltered, procaine failed to alter PDH activity, lysine inhibited PDH activity, and poly-L-lysine stimulated PDH activity. Therefore, polyamines and other positively charged small molecules may be physiologic regulators of PDH activity.

Adipose Tissue↗
PubMed1990

A comparison of chromogen test strip (Chemstrip bG) and serum glucose values in newborns.

Although glucose oxidase-peroxidase chromogen test strips are frequently used to estimate serum glucose values in newborns, previous studies have not evaluated multiobserver variability of test strip readings and have included few infants with hypoglycemia. We compared values of 272 samples of serum glucose with values simultaneously obtained by chromogen test strips (Chemstrip bG) in newborns. The diagnostic sensitivity of a chromogen test strip less than 2.2 mmol/L for predicting a serum glucose level less than 1.9 mmol/L was 86% (95% confidence interval [CI], 75% to 94%), with 78% specificity (95% CI, 73% to 84%). The positive predictive value in our specimens, with a 21% prevalence of serum glucose levels less than 1.9 mmol/L, was 52% (95% CI, 41% to 62%), with a negative predictive value of 95% (95% CI, 91% to 100%). Fifty-eight of our serum glucose values were less than 1.9 mmol/L and the levels obtained by chromogen test strip were greater than or equal to 2.2 mmol/L in 8 of these cases. Review of these 8 cases showed that a delay in performing the laboratory glucose oxidase serum glucose could account for the discrepancy in 2 cases. Chromogen test strips are readily available and easy to use, but more sensitive, specific, accurate, and precise methods of serum glucose screening in newborns are needed.

Blood Glucose↗
PubMed1989

Phosphatidylglycerol in amniotic fluid. Comparison of an "ultrasensitive" immunologic assay with TLC and enzymatic assay.

Phosphatidylglycerol (PG) in amniotic fluid is recognized as a good indicator of fetal lung maturity and is unaffected by moderate amounts of blood or meconium contamination. A rapid immunologic agglutination assay, Ultrasensitive AmnioStat-FLM (FLM), was compared with two-dimensional thin-layer chromatography (TLC) and an enzymic, colorimetric procedure (E-PG). Eighty amniotic fluid specimens were analyzed. FLM results were reported as high (H), intermediate (I), or low positive (L). TLC was compared with FLM:H (n = 27), mean 0.14 (fraction of total phospholipids); I (n = 7), mean 0.11; L (n = 9), mean 0.03; negative results had no detectable PG by TLC. In 33 cases E-PG was compared with FLM:H (n = 9), mean 7.0 mumol/L; I (n = 5), mean 8.1 mumol/L; L (n = 3), mean 3.0 mumol/L; negative (n = 16), mean 3.2 mumol/L. Records were reviewed in 70 cases. Thirty cases were excluded: sample to delivery time was greater than 72 hours; steroids were given or sepsis was documented. Fetal lung immaturity was clinically present in six cases: respiratory distress syndrome in three cases and transient tachypnea of the newborn (TTN) in three cases. One false positive result was identified (TTN, FLM:H). FLM sensitivity for fetal lung maturity was 85.3%, specificity was 83.3%, and the positive predictive value for fetal lung maturity was 96.7%. FLM is a fast, reliable indicator of fetal lung maturity.

Agglutination Tests↗
PubMed1987

Effect of S-adenosylhomocysteine on insulin-independent release of pyruvate dehydrogenase activator from rat adipocyte plasma membranes.

The addition of insulin to adipocyte plasma membranes has been shown to release a low molecular weight, acid stable mediator which activates mitochondrial pyruvate dehydrogenase. The insulin-dependent release of this activator is dependent on the method used to prepare the plasma membranes. Adipocyte plasma membranes prepared in 0.25 M sucrose, 10 mM MOPS, pH 7.4 released an activator of pyruvate dehydrogenase in an insulin-independent manner. Insulin is required to stimulate phospholipid methylation in these membranes. The inhibition of insulin-stimulated phospholipid methylation with 1 mM S-adenosylhomocysteine resulted in a significant increase in amount and/or activity of the pyruvate dehydrogenase activator. The insulin-dependent dependent release of mediators of insulin action from adipocyte plasma may be regulated by phospholipid methylation.

Adipose Tissue↗
PubMed1987

Association of high-molecular-mass and electrophoretically atypical alkaline phosphatases.

Polyacrylamide gel electrophoresis of alkaline phosphatase may yield abnormally migrating fractions; these include high-molecular-mass alkaline phosphatase, which remains at the gel origin, and immunoglobulin-alkaline phosphatase complexes, which have a mobility approximately 1/3 that of liver isoenzyme. We performed a retrospective study of 19 patients whose sera exhibited atypical alkaline phosphatase fractions, defined as bands whose mobility was slower than bone, liver, or intestinal alkaline phosphatase; 17 had a mobility approximately 1/3 that of liver isoenzyme and 16 also exhibited gel origin enzyme activity or high-molecular-mass bands. The strong association of the atypical and high-molecular-mass alkaline phosphatases suggests that they may be structurally related, both consisting of either immunoglobulin-enzyme complexes or membrane-alkaline phosphatase complexes. This hypothesis is supported by (1) one serum available for investigation containing alkaline phosphatase-immunoglobulin complexes in both abnormally migrating fractions, but on detergent treatment showing no evidence of membrane-bound enzyme; (2) detergent treatment of serum from patients with only high-molecular-mass alkaline phosphatase creating bands with a mobility of approximately 1/3 that of the liver isoenzyme.

Adolescent↗
PubMed1987

Electrophoresis and the isomune-LD and LD-1 immuno methods compared for measurement of lactate dehydrogenase isoenzyme-1.

We evaluated three different methods for measuring lactate dehydrogenase (EC 1.1.1.27) isoenzyme LD-1 activity--agarose gel electrophoresis and two immunoassays, Isomune-LD (Roche) and LD-1 Immuno (Seragen)--in patients' samples for which measurement of creatine kinase-MB was ordered. Regression analyses of the comparisons gave the following results: LD-1 (%) from Isomune-LD (y) vs electrophoresis (x) (n = 51), y = 1.05x + 1.99, r = 0.92; LD-1 (%) from LD-1 Immuno (y) vs electrophoresis (n = 27), y = 1.05x + 3.94, r = 0.88; LD-1 (%) from LD-1 Immuno (y) vs Isomune-LD (x) (n = 41), y = 1.06x + 0.48, r = 0.95. Comparison by Student's paired t-test yielded significant differences between the mean values by electrophoresis and both Isomune-LD (P less than 0.005) and LD-1 Immuno (P less than 0.001), but no significant difference between the two immunoassays (P greater than 0.2). Analyzing these results by the overlap index, we conclude that electrophoresis shows the best clinical correlation followed, in order, by the Isomune-LD and the LD-1 Immuno methods. Both immunoassays are simpler and more rapid than electrophoresis, but in our hands the Isomune-LD method demonstrated greater precision and better correlation with electrophoretic values.

Electrophoresis↗
PubMed1986

Stimulation of phosphatidylcholine synthesis by insulin and ATP in isolated rat adipocyte plasma membranes.

Added individually or together, insulin and/or ATP significantly and rapidly increased the concentration of phosphatidylcholine in an enriched plasma membrane preparation from rat adipocytes. The increase in phosphatidylcholine synthesis mediated by insulin or ATP was suppressed by the phospholipid methyltransferase inhibitor, S-adenosylhomocysteine. These results suggest that the activity of phospholipid methyltransferase from adipocyte plasma membranes may be increased by phosphorylation and that insulin may further increase the activity of the phosphorylated phospholipid methyltransferase by an alternative pathway.

Adenosine Triphosphate↗
PubMed1986

Potential problems in serum protein electrophoresis.

Potential problems are described that one could encounter in carrying out an electrophoretic procedure including its ancillary phases of visualization (staining) and quantification (densitometry). Endpoint-like measurements for separated isoenzymes may provide artifactual kinetic values as well, because stain measurement is fixed at a single time whereas reagent blanking in the electrophoretic medium is substituted for the conventional serum initial absorbance readings of test-tube determinations. Truncation of separated electrophoretic zones or opacity of an electrophoretic anti-convection medium such as uncleared cellulose acetate may also interfere with absolute quantification procedures.

Albumins↗
PubMed1986

The clinical use of alkaline phosphatase enzymes.

The enzyme alkaline phosphatase is an important serum analyte and its elevation in serum is correlated with the pressure of bone, liver, and other diseases. The analysis of the isoenzymes of alkaline phosphatase is an aid in diagnosing liver and/or bone disease, especially the high molecular weight isoenzymes that appear in cholestatic liver disease.

Alkaline Phosphatase↗