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Biomedical subjects

R Lorenc

Publications and source records attributed to R Lorenc.

At least 37 records · Page 2Linked to original sources
PubMed1994

Pyridoxal 5'-phosphate related changes in retention of 1,25-dihydroxy vitamin D-receptor ligands in rat intestinal mucosa cell nuclei.

After feeding rats a vitamin B-6-deficient diet, we observed a decrease in pyridoxal 5'-phosphate concentrations in intestinal mucosa cells to 32 and 48% of control in cytoplasm and cell nuclei, respectively. Correlation analysis suggested that there were two pyridoxal 5'-phosphate pools in the nuclei: a "mobile" pool (equivalent to about 5% the concentration of the cytoplasmic pyridoxal 5'-phosphate), and a "stable" pool, which was independent of cytoplasmic fluctuations of pyridoxal 5'-phosphate (about 9 pmol pyridoxal 5'-phosphate/mg DNA). Reduction in pyridoxal 5'-phosphate content in the cells of vitamin B-6-deficient animals was accompanied by a substantial increase in 1,25-dihydroxyvitamin D-receptor ligand concentration in the cell nuclei (76.6 +/- 19.7 vs 762 +/- 291 fmol/mg DNA, mean +/- SEM). The degree of 1,25-dihydroxyvitamin D accumulation in the nuclei appeared to be an exponential function of the "mobile" nuclear pyridoxal 5'-phosphate concentration. Semilogarithmic transformation of the data yielded a straight line, representing an inverse correlation between the cytoplasm-related nuclear pool of pyridoxal 5'-phosphate and the logarithm of the 1,25-dihydroxyvitamin D concentration in the nuclei (r = -0.95). These data suggest that pyridoxal 5'-phosphate may be related to 1,25-dihydroxyvitamin D retention in the nuclei, possibly through interaction of the pyridoxal 5'-phosphate with the vitamin D receptor protein in the nuclei.

Alkaline Phosphatase↗
PubMed1992

Reconstitution and characterization of a Na+/Pi co-transporter protein from rabbit kidney brush-border membranes.

A protein with Na+/Pi co-transporter activity has been extracted from rabbit brush-border membranes with chloroform/methanol and purified by hydroxyapatite chromatography. The protein has been incorporated by the dilution method into liposomes formed from different types and ratios of lipids. The greatest reconstitution has been achieved into liposomes prepared from cholesterol (20%), phosphatidylcholine (20%), phosphatidylethanolamine (30%) and phosphatidylserine (30%) (CH/PC/PE/PS). Pi uptake by these proteoliposomes had the following characteristics: (i) the initial rate was markedly greater in the presence of an inwardly directed Na+ gradient (600 pmol/10 s per mg) than with a K+ gradient (65 pmol/10 s per mg); (ii) maximal uptake was increased 8-fold above the equilibrium value ('overshoot') when a Na+ gradient was applied; (iii) Pi was not merely bound to proteoliposomes but was transported intravesicularly; and (iv) Na(+)-dependent Pi uptake was sensitive to the known phosphate transport inhibitors. This first successful attempt of reconstitution of Na+/Pi transport activity into proteoliposomes led us to isolate and characterize physico-chemically the protein responsible. Its isoelectric point was about 5.8, and urea/SDS gel electrophoresis revealed a broad band of molecular mass ranging from 63 to 66 kDa under both reducing and non-reducing conditions. In the native form, the molecular mass analysed by gel filtration was estimated to be 170 +/- 10 kDa, suggesting that the protein is a polymer, probably stabilized by hydrophobic bonds. Endoglycosidase F treatment decreased the molecular mass to approx. 50 kDa. It is postulated that this acidic glycoprotein might represent a subunit of the intact Na+/Pi co-transporter from rabbit kidney brush-border membranes.

Animals↗
PubMed1992

[Biochemical markers of bone remodelling and their usefulness in the diagnosis of osteoporosis].

In 65 subjects (42 women and 23 men) aged from 40 to 80 years, at risk for primary or secondary osteoporosis development without concomitant diseases affecting renal and hepatic function and vitamin D metabolism, alkaline phosphatase activity (F.A.s) and total calcium level (Cas) were determined in the blood serum, and also the excretion in the morning urine portion of hydroxyproline and calcium was measured in relation to creatinine excretion (FUHpr/kr; FUCa/kr). In each patient an X-ray was done of the thoracolumbar spine, and on its basis the X-ray vertebral index was calculated. The study has been undertaken in order to elucidate whether the above stated compounds regarded as biochemical markers of bone remodelling, and the X-ray vertebral index are useful in the diagnosis of osteoporosis. An increased F.A.s activity in the blood serum, and higher values of the X-ray vertebral index were found both in the group of patients with primary and secondary osteoporosis, in relation to normal values. It was found that FUHpr/kr and FUCa/kr were increased in patients with secondary osteoporosis in comparison to patients with primary osteoporosis. A statistically significant correlation was also noted between FUHpr/kr and FUCa/kr, and F.A.s in patients with primary and secondary osteoporosis. In both studied groups no relationship was found between the value of the X-ray vertebral index, and the levels of biochemical markers of bone remodelling.

Adult↗
PubMed1992

Growth rate in children with vitamin-D-dependent rickets in relation to 1-alpha-hydroxyvitamin D3 dosage.

Growth rate of five children with vitamin D-dependent rickets was analyzed during the long-term treatment with an active analog of vitamin D3. Considerable increase in growth rate together with the improvement of biochemical values and radiological pattern took place during the initial phase of administration of 1-hydroxyvitamin D3. During the maintenance treatment of long duration with 1-hydroxyvitamin D3 both the acceleration of growth and catch-up growth persisted. However, in 4 among 5 children studied an inhibition of growth was observed during different periods of time. Only in one boy was this connected with the conclusion of the process of physiological growth. In three remaining children a slow-down in growth rate appeared during the pre-pubertal period or was the effect of lowering the dose of 1-hydroxyvitamin D3 as an countermeasure to hypercalciuria. In such cases inhibition of growth was caused by the administration of too small a dose of 1-hydroxyvitamin D3 in relation to the requirement. In all cases the appearance of biochemical features of rickets aggravation, such as low blood serum phosphate concentration and elevated alkaline phosphatase activity, preceded the observable inhibition of growth. The results obtained allow us to conclude that the inhibition of growth observed during the long-term treatment of rickets with 1-hydroxyvitamin D3 may be regarded as the first signal of inadequate dosage of 1-hydroxy vitamin D3.

Adolescent↗
PubMed1988

Identification of Na+,Pi-binding protein in kidney and intestinal brush-border membranes.

An Na+, Pi-binding protein has been extracted from kidney and intestinal brush-border membranes with an organic solvent and has been purified by Kieselghur and Sephadex LH-60 chromatography. The molecular mass of this protein has been estimated to be about 155 kDa as determined by gel-filtration chromatography on Sepharose 2B. Under denaturing conditions, polyacrylamide-gel electrophoresis revealed a monomer of molecular mass about 70 kDa. The protein has high specificity and high affinity for Pi [K0.5 (concentration at which half-maximal binding is observed) near 10 microM]. Na2+ binding also exhibits saturation behaviour, with a K0.5 near 7.5 mM. Pi binding is inhibited by known inhibitors of Pi transport in brush-border membrane vesicles. It appears that this protein could be involved in Na+/Pi co-transport across the renal and intestinal brush-border membranes.

Animals↗
PubMed1988

Application of the rat thymus receptor in a specific, sensitive and simplified assay of 1,25-dihydroxyvitamin D in blood serum.

A simple method has been employed to prepare crude nuclear extract from rat thymus, using hypertonic buffer after previous treatment with hypotonic buffer. The preparation is free from serum vitamin D-binding protein and contains a 3.7 S receptor molecule, which specifically binds 1,25-dihydroxyvitamin D-3 (1,25-(OH)2D3). The receptor is of high affinity (KD = 0.85 X 10(-11) M at O degrees C) and low capacity (260-460 fmol/g tissue). The Scatchard analysis of ligand binding results in a concave downward curve. The Hill analysis of the same data gives good linear fitting (r = +0.971) with the Hill coefficient nH = 1.63. These facts indicate positive cooperativity between two ligand binding sites of the rat thymus 1,25-(OH)2D3 receptor. The preparation was used in a competitive protein binding assay of 1,25-(OH)2D in serum extracts, purified on Sep-Pak C18 followed by silica Sep-Pak cartridges. The method was sensitive to 0.5 pg/tube (2.0 ng/l) when 1 ml of serum was extracted. Intra- and interassay coefficients of variation were 9% and 14%, respectively. The serum 1,25-(OH)2D concentration estimated in 33 children (mean age 6.5 +/- 3 years) was 46.6 +/- 18.4 ng/l (mean +/- S.D.).

Animals↗
PubMed1988

Influence of lactose on phosphate metabolism in rats.

1. Lactose, sucrose or maltose were administered to rats by gavage together with 32PO4, and blood 32P was determined. 2. PO4 uptake into blood was increased only when lactose and PO4 were administered into the gut. 3. Weaned male rats were fed on a control diet or a diet containing 30, 60 or 120 g lactose/kg. After a 5 d period of adaptation, a 6 d P balance was carried out. After a further 24 d blood levels of PO4, and calcium and PO4 uptake by brush-border membrane vesicles prepared from the renal cortex, were determined. 4. The absorption and retention of P were higher in lactose-fed rats. Faecal P excretion decreased with increasing dietary lactose content. Urinary P excretion was lower in the rats fed on 30 and 60 g lactose/kg whereas the 120 g lactose/kg diet increased urinary P. 5. Membrane vesicles isolated from rats fed on the diets containing 30 and 60 g lactose/kg showed a higher uptake of PO4, but with the 120 g lactose/kg diet uptake was lower than with membrane vesicles isolated from rats fed on the control diet. 6. A statistically significant increase in the serum PO4 level was observed in the rats fed on the diet with 120 g lactose/kg. 7. The presence of lactose in the diet caused a stimulatory effect on PO4 absorption and retention.

Animals↗