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Biomedical subjects

K D Gerbitz

Publications and source records attributed to K D Gerbitz.

At least 55 records · Page 3Linked to original sources
PubMed1995

Immunological determination of advanced glycosylation end-products in human blood and urine.

Advanced glycosylation end-products (AGE) in human blood and urine were investigated with the aid of an AGE-specific enzyme-linked immunosorbent assay. Evidence is presented that AGE are natural constituents of human serum and urine. In diabetics with normal renal function only a small increase in serum AGE levels was found as compared to normal controls, while no difference in urinary excretion rate was discernible. Urinary excretion rate of AGE from diabetic and non-diabetic patients with end-stage renal disease was reduced, while high serum AGE levels were observed. AGE in serum occur in a low-molecular-weight fraction and in a possibly protein-bound high-molecular-weight fraction. In urine from normal controls three immunologically reactive fractions were detected whose apparent molecular mass ranged from 100 to 1000 Daltons, while in urine from patients with end-stage renal failure additional high-molecular-weight fractions appeared.

Adolescent↗
PubMed1994

Atherogenic levels of low density lipoprotein alter the permeability and composition of the endothelial barrier.

In the present study we investigated the influence of elevated low density lipoprotein (LDL) concentration on endothelial permeability. Endothelial cells were cultured on microporous membranes until confluence and albumin, dextran and LDL transfer across endothelial monolayers was determined to assess macromolecular permeability. Exposure of proliferating aortic endothelial cells to LDL levels of more than 1 mg/ml LDL-cholesterol induced a concentration-dependent exponential increase in the permeability of confluent endothelial monolayers. Acute addition of high LDL concentration did not alter macromolecular permeability. Once elevated permeability was induced, it persisted. It was not readily reversible after addition of normal LDL levels. Change in permeability was accompanied by a selective decrease in basement membrane associated heparan sulfate proteoglycan (HSPG) content. The apparent parallel between the loss in endothelial barrier function and HSPG decrease implicates a connection between the two events. Prolonged, but not acute, incubation with antiserum directed against the core-protein of HSPG also led to increased permeability, suggesting a causal role of HSPG for the proper function of endothelium. The fact that non-atherogenic LDL-cholesterol levels had no effect indicates that a 'threshold' concentration for LDL-cholesterol may exist, leading to nondenuding injury in the endothelial barrier as an early event in development of atherosclerosis.

Animals↗
PubMed1994

Heparin induces endothelial extracellular matrix alterations and barrier dysfunction.

We investigated the influence of heparin on the composition of the subendothelial matrix and on endothelial permeability to elucidate the structure-function relationship of matrix composition and permeability. Albumin flux across the confluent endothelial monolayers was used to assess the macromolecular permeability. Lowest values were obtained when 100% serum was used as medium for permeability studies. The endothelial matrix components, fibronectin and basement membrane-associated heparan sulfate proteoglycan (HSPG), were measured by enzyme immunoassay. Treatment of proliferating endothelial cells with heparin (0-900 micrograms/ml) induced a dose-dependent decrease in endothelial HSPG content, whereas the fibronectin content was unaltered. This structural change was accompanied by an increase in albumin permeability. Both heparin effects exhibited similar dose-response curves with half-maximal effects at approximately 5 micrograms/ml heparin. Acute addition of 300 micrograms/ml heparin had no effect on permeability or HSPG content. When endothelial cells were preincubated with an HSPG antiserum, the endothelial permeability increased nearly threefold. Our results indicate that heparin-induced loss of HSPG may cause the increase in endothelial permeability. The data underline the importance of HSPG for the integrity of the endothelial barrier.

Animals↗
PubMed1994

Impaired immunoglobulin G Fc fragment function in diabetics is caused by a mechanism different from glycation.

Glycation and fluorescence at 440 nm (excitation at 370 nm) were found to be increased in immunoglobulin G (IgG) from diabetics, strongly indicating the presence of IgG-linked advanced glycosylation end products. In contrast, levels of IgG-linked advanced glycosylation end products were low or undetectable in both normal and diabetic subjects when an advanced glycosylation end products specific antibody was employed for immunological determination of advanced glycosylation end products. Furthermore, no correlation exists between IgG glycation, fluorescence and immunoreactivity. In diabetics, the Fc fragment of IgG showed decreased protein A binding and decreased fixation of complement. This impairment of biological activity was not correlated with the immunologically determined level of advanced glycosylation end products, whereas IgG-linked fluorescence was inversely related to complement fixing activity. These results indicate that mechanisms different from glycation or browning are responsible for changes in the functional properties of IgG.

Adolescent↗
PubMed1993

In-vitro carboxymethylation of low density lipoprotein alters its metabolism via the high-affinity receptor.

Carboxymethylation of lysine residues has been shown to result from oxidation of glycated proteins in vivo and in vitro leading to an augmentation of proteins' net negative charge. The metabolism of carboxymethylated low density lipoprotein (LDL) was studied in cultured human fibroblasts and mouse peritoneal macrophages. In vitro carboxymethylation was achieved by incubation of LDL with glyoxylic acid in the presence of Na(CN)BH3. Carboxymethylation inhibited metabolism of LDL via the high affinity receptor in fibroblasts as did methylation. The uptake of LDL into mouse peritoneal macrophages via the scavenger receptor, which was stimulated by acetylation, was not affected.

Acetylation↗
PubMed1993

Upregulation of myo-inositol transport compensates for competitive inhibition by glucose. An explanation for the inositol paradox?

High glucose concentrations inhibit the uptake of myo-inositol into cells. However, whether this leads to a depletion of intracellular myo-inositol levels has been debated, because unchanged, decreased, and increased cellular myo-inositol levels all have been reported for diabetic tissues. To evaluate whether cells are capable of counterregulating impaired uptake, we have investigated myo-inositol uptake in porcine aortic endothelial cells under short- and long-term hyperglycemic conditions. Although increasing glucose concentrations inhibited acute myo-inositol uptake competitively, the uptake was increased markedly, when cells were already preincubated in a high glucose medium for > 6 h. The stimulation was maximal at 20 mM of glucose with no further increase at 40 mM glucose. The same stimulation of uptake could be induced by 5 mM of glucose plus 35 mM of raffinose, whereas 35 mM of sorbitol or mannitol, which do not compete for myo-inositol uptake, were ineffective. Lineweaver-Burk analysis revealed an increased Vmax for the induced myo-inositol transport activity, whereas the Km for myo-inositol remained constant (18 microM). The upregulated inositol transporter was still Na+ and ATP dependent, indicating that the same carrier system was operating. Uptake returned to control values when cells were again exposed to normoglycemic medium conditions for an additional 24 h. When endothelial cells were incubated with D-[U-14C]glucose and 10 microM myo-[2-3H]inositol of equal specific radioactivity for 24 h, no 14C radioactivity was found in intracellular myo-inositol, indicating that conversion of glucose to myo-inositol was rather low.

Animals↗
PubMed1993

Specific glycation of albumin depends on its half-life.

Some suggest that measurements of plasma fructosamine concentration should be corrected for the plasma protein (or albumin) concentration because the extent of glycation per volume depends on both protein and glucose concentrations. Several reports, however, demonstrate a poor correlation between plasma fructosamine and albumin concentrations in diabetic patients. In vitro kinetic and in vivo studies have shown that glycation is also dependent on the half-lives of plasma proteins. Because a decrease in plasma albumin diminishes its catabolism, we speculated that low albumin concentrations are associated with a greater extent of glycation on a molar basis (specific glycation) and vice versa. To test this hypothesis, we studied plasma albumin, total protein, and fructosamine in 63 nondiabetic subjects with normal plasma fasting glucose concentrations and hemoglobin A1c between 5.1% and 5.9%. Plasma fructosamine was poorly correlated with albumin concentration (r = 0.348) but a logarithmic plot of the specific glycation of albumin vs albumin concentration showed a better correlation (r = -0.842), suggesting that the kinetic considerations were operating in vivo. Therefore, because lower specific glycation of plasma albumin "compensates" for higher concentration, correction of fructosamine for albumin content in patients will overestimate mean blood glucose when albumin is low and underestimate it when albumin is high.

Blood Glucose↗
PubMed1992

No genetic differences between affected and unaffected members of a German family with Leber's hereditary optic neuropathy (LHON) with respect to ten mtDNA point mutations associated with LHON.

In order to investigate possible synergistic influences of different mtDNA mutations on penetrance and severity of Leber's hereditary optic neuropathy (LHON), a large German LHON pedigree is characterized with respect to 10 different mutations associated with LHON. All members of the family carry three different mtDNA mutations (at nucleotide 4,216, 11,778 and 13,708) in a homoplasmic form, regardless of whether or not they are clinically affected. Testing for another 7 mutations reveals negative results in all family members. Hence, the variable disease expression of the family members cannot be explained by varying combinations of LHON-associated mtDNA mutations.

Base Sequence↗
PubMed1992

Elevated glucose decreases the content of a basement membrane associated heparan sulphate proteoglycan in proliferating cultured porcine mesangial cells.

The effect of elevated glucose concentrations on the synthesis of basement membrane components was investigated in proliferating cultured porcine mesangial cells. Basement membrane associated heparan sulphate proteoglycan was determined by enzyme immunoassay with a specific antiserum recognizing the core protein of the heparan sulphate proteoglycan. When cells were exposed to increasing glucose concentrations up to 40 mmol/l, the heparan sulphate proteoglycan content was concomitantly decreased to 53% when compared to cells cultured under normal glucose concentrations or in the presence of 40 mmol/l sorbitol. The fibronectin content was essentially unchanged under these conditions. No significant effect of insulin on these basement membrane components was found. The results indicate that hyperglycaemia may be responsible for a decrease of mesangial heparan sulphate proteoglycan content in diabetes mellitus. This supports the view that loss of heparan sulphate proteoglycan may be an important step or even an initial event of mesangial alterations in diabetic glomerulopathy.

Animals↗
PubMed1992

In situ hybridization of mitochondrial DNA in the heart of a patient with Kearns-Sayre syndrome and dilatative cardiomyopathy.

Previous studies have revealed cytochrome-c-oxidase-deficient cardiomyocytes and the 4,977 base pair deletion ("common deletion") of mitochondrial DNA (position 8,482-13,459) in the heart of a patient with dilatative cardiomyopathy and Kearns-Sayre syndrome. In the present investigation the co-localization of the enzymatic and genomic defects was studied. In situ hybridization of mitochondrial DNA (mtDNA) revealed different hybridization patterns in the cytochrome-c-oxidase-deficient cells: (1) a selective reduction of the hybridization signal with an mtDNA probe recognizing the common deletion, indicating predominance of the deleted over the nondeleted mtDNA molecules in the cytochrome-c-oxidase-deficient cells; (2) a reduced hybridization signal with different mtDNA probes, indicating depletion of mtDNA; and (3) normal hybridization signals with different probes in single cytochrome-c-oxidase-deficient cardiomyocytes. These results indicate that different mechanisms may co-exist in Kearns-Sayre syndrome and may lead to defective respiratory chain function. The question of the pathogenetic interrelationship is discussed.

Adult↗
PubMed1991

Respiratory chain activity in tissues from patients (MELAS) with a point mutation of the mitochondrial genome [tRNA(Leu(UUR))].

A heteroplasmic point mutation (transition A to G at position 3243 in the mitochondrial tRNA(Leu(UUR)) gene is indicative for myo-encephalopathy with lactic acidosis and stroke-like episodes (MELAS). Decreased respiratory chain complex activities measured in different tissues from four patients with MELAS syndrome do not correlate with the proportion of mutated mitochondrial genome.

Acidosis, Lactic↗
PubMed1991

A specific point mutation in the mitochondrial genome of Caucasians with MELAS.

The mitochondrial DNA (mtDNA) of Japanese patients suffering from the syndrome of mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes (MELAS) exhibits a specific heteroplasmic A----G transition in the tRNA(Leu) at position 3243. In this study, we investigated mtDNA from skeletal muscle, cardiac muscle, brain, liver, diaphragm, fibroblasts and blood cells of four Caucasians with MELAS, one younger healthy sister of two MELAS patients, and eleven controls. We found that 1) the mutation was present in all investigated tissues of Caucasians with MELAS but not in controls, 2) within a single patient, the tissue-specific variation of the copy number of mutated mtDNA covered the same range as in the skeletal muscle of different patients, 3) the mutation was also present in the blood cells of the healthy sister of two MELAS siblings.

Acidosis, Lactic↗
PubMed1991

Biochemistry and molecular genetics of muscle diseases.

The inherited skeletal muscle diseases form a highly heterogeneous group of disorders covering single enzyme defects, complex metabolic disorders, storage diseases, dystrophies and malignant hyperthermia. Whereas these myopathies may be caused by a large number of different biochemical and genetic defects their clinical presentation by contrast is relatively monotonous. with only a few specific findings pointing to a particular molecular defect. This review of the biochemical and molecular genetic basis of these diseases concentrates on 1) disorders in fuel utilization and energy production. 2) disorders in structural integrity or mechanical function, and 3) disorders in contractility and electrophysiological properties of the muscle cell. The authors address the questions of organ-specificity and of a possible relationship between clinical, biochemical, and genetic heterogeneity of metabolic defects, and also try to present the current state of chromosome mapping for such disorders.

Biochemical Phenomena↗
PubMed1990

Different copy numbers of apparently identically deleted mitochondrial DNA in tissues from a patient with Kearns-Sayre syndrome detected by PCR.

An apparently identical deletion of 4.977 bp in length (position 8,483-13,459) was detectable in the mitochondrial DNA from skeletal muscle, heart muscle, kidney, and liver of a patient with Kearns-Sayre syndrome. The proportion of deleted genome varied from 60% for the skeletal muscle to 15% for heart muscle and kidney, and was below 5% in the liver. The mtDNA heteroplasmy of the liver was only detectable after amplification by PCR. In skeletal and heart muscle histochemical and immunocytochemical findings concerning cytochrome c oxidase were in good correlation with the proportion of deleted mitochondrial DNA.

Base Sequence↗
PubMed1990

High concentrations of low density lipoprotein decrease basement membrane-associated heparan sulfate proteoglycan in cultured endothelial cells.

The effect of increasing low density lipoprotein (LDL) concentrations on the synthesis of basement membrane components was investigated in proliferating porcine aortic endothelial cells (PAEC) in culture. Basement membrane-associated heparan sulfate proteoglycan (HSPG) and fibronectin were determined by enzyme immunoassay. Low extracellular LDL-levels increase, high extracellular LDL-levels decrease the HSPG content of PAEC. Fibronectin synthesis was only slightly affected while proliferation and metabolic activity as assessed by lactate production were constant. Insulin or high extracellular glucose did not influence the effect of LDL on basement membrane components.

Animals↗