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Biomedical subjects

J Harmeyer

Publications and source records attributed to J Harmeyer.

At least 37 records · Page 2Linked to original sources
PubMed1987

Phosphate transport in brush-border membranes from control and rachitic pig kidney and small intestine.

1. Na-Pi co-transport was analysed using renal cortical and small intestinal brush-border membrane vesicles which were isolated from control (normal, heterozygotes) and rachitic piglets (homozygotes). 2. A kinetic analysis of Na-dependent initial linear uptake of Pi was performed using vesicles obtained from control animals. The results suggest similar kinetic properties for the renal and small intestinal co-transport system. (i) A sigmoidal dependence on Na concentration of Pi uptake suggests the involvement of more than one Na ion in the co-transport. (ii) Increasing Na concentration leads to an increase in the apparent affinity of the transport system for Pi and has minimal effect on the apparent Vmax (maximum velocity of uptake). (iii) Increasing pH leads to an increase in Pi transport rate. 3. The kinetic characteristics of the Na-Pi co-transport system in vesicles obtained from rachitic animals were similar to those in controls. The apparent Vmax, but not the apparent Km (Michaelis constant) for Na and Pi, is reduced in intestinal and renal brush-border membranes isolated from rachitic animals as compared to control animals. Injection of vitamin D3, three days prior to killing of rachitic litter-mates, increased the Na-Pi uptake rate in the brush-border membrane vesicles isolated from these piglets. 4. It is concluded that intestinal and renal brush-border membranes from piglets contain a similar Na-Pi co-transport system and that in vitamin-D-dependent rickets the number of operating transport units is reduced in both membranes.

Animals↗
PubMed1987

Vitamin D3 metabolism in a pig strain with pseudo vitamin D-deficiency rickets, type I.

Vitamin D metabolism was studied in the 'Hannover Pig', a strain which suffers from pseudo vitamin D-deficiency rickets, type I. Animals of this strain are known to be devoid of renal 25-hydroxyvitamin D3-1 alpha-hydroxylase and -24-hydroxylase activities. Pigs with florid rickets and hypocalcaemia were treated with single im injections of 0.25 to 1.25 mg of vitamin D3, doses that have been shown in previous studies to be effective in producing transient healing of rachitic symptoms. The levels of vitamin D3 and its most relevant physiological metabolites in plasma were estimated at intervals before and after this vitamin D3 treatment. Vitamin D3 rose from 14.8 +/- 8.1 to 364 +/- 190 nmol/l (mean +/- SD) 2 to 3 days post injectionem, 25-hydroxyvitamin D3 from 131.0 +/- 46.2 to 1068 +/- 160 nmol/l within 7 days post injectionem. The 1 alpha, 25-dihydroxyvitamin D3 concentration in plasma was elevated from 73.9 +/- 25.0 to 281 +/- 168 pmol/l 2 to 3 days post injectionem and declined continually from that time. 24R,25-dihydroxyvitamin D3 and 25S,26-dihydroxyvitamin D3 levels after treatment showed different responses in different animals being either elevated or unchanged. Clinical healing of the pigs with these doses of vitamin D3 was attributed to the transient rise of 1 alpha,25-dihydroxyvitamin D3 in plasma. It was assumed that 1 alpha,25-dihydroxyvitamin D3 synthesis takes place under these circumstances in extrarenal tissues.

24,25-Dihydroxyvitamin D 3↗
PubMed1986

Absence of renal 25-hydroxycholecalciferol-1-hydroxylase activity in a pig strain with vitamin D-dependent rickets.

Cholecalciferol-1-hydroxylase activities were estimated in renal cortex homogenates and mitochondrial preparations from three groups of 6- to 12-week-old pigs. Five animals suffering from an inherited form of vitamin D-deficiency rickets showed symptoms of florid rickets when used for this study. Six pigs were normocalcemic heterozygous litter mates of the rachitic strain and three pigs were normal controls (German land-race and wild pigs). The renal cortex homogenates and mitochondrial preparations were incubated for 5-30 min at 37 degrees C with 25-(26-27-methyl-3H) OHD3 as substrate. 1,25(OH)2D3 was subsequently identified in normal phase (Zorbax-Sil) and reversed phase (Zorbax-ODS) HPLC eluates. 1-hydroxylase activities were demonstrated in both normal controls and heterozygote offspring and were ten times above minimum detectability of the assay. The km was 255 +/- 74 (SD) and 278 +/- 85 nmol X 1(-1) in controls and heterozygote offspring, respectively. The Vmax in the two groups was between 0.11 and 0.794 and decreased with age of the animals. Km and Vmax did not differ between the two groups. In homozygous, hypocalcemic rachitic animals no 1-hydroxylase activity was detectable in either homogenates or mitochondrial preparations. Addition of kidney homogenate from a rachitic animal to a homogenate from a normal pig did not specifically depress 1-hydroxylase activity in the mixture. Treatment of rachitic pigs with 1.0 microgram/day of 1,25(OH)2D3 for 4 weeks also had no effect on 1-hydroxylase activity. It is concluded that the rachitic pigs suffer from an inborn error of renal 1,25(OH)2D3 production.(ABSTRACT TRUNCATED AT 250 WORDS)

24,25-Dihydroxyvitamin D 3↗
PubMed1986

The passage of protozoa from the reticulo-rumen through the omasum of sheep.

1. Protozoa in rumen contents and omasal effluent of growing wethers were counted. The wethers were equipped with rumen and abomasal cannulas, and omasal sleeves attached to the omasal-abomasal orifice. Rumen fluid dilution rates were elevated by continuous infusions of hypertonic mineral solutions (3-4 litres/d) for 24 d. Rumen contents and omasal effluent were sampled between 9 and 21 h during the last 10 d of each experiment. 2. Protozoal concentrations in omasal effluent were only 0.2-0.3 those found in the rumen under normal conditions. The ratio of protozoal concentrations in rumen: those in omasal effluent was for small Diplodinium spp. 4.6 (SD 0.9), for Ophryoscolex spp. 4.3 (SD 1.0), for Dasytricha ruminantium 4.0 (SD 0.5), for Isotricha spp. 3.8 (SD 0.8), for Entodinium spp. 3.6 (SD 0.9) and for Polyplastron multivesiculatum 2.6 (SD 0.5). 3. Elevation of rumen fluid dilution rate by 20 and 55% respectively, increased protozoal concentrations in omasal effluents from 22 to 33% and from 31 to 47% those in rumen contents. The apparent residence times of protozoa in the rumen were decreased 50% by the infusion of a mineral-salt solution. The increase in rumen fluid dilution rate had no significant effect on concentrations of protozoa in the rumen or on the differences of the apparent residence times between different species. The apparent residence time of holotrichs remained the same before and after infusion of the mineral-salt solution. 4. Apparent residence times of individual species of protozoa in the rumen were, under normal feeding conditions, 2.55 d, and were four to six times longer than the mean residence time of CrEDTA in the rumen.

Animals↗
PubMed1986

[The quantitative determination of vitamin D3 and its metabolites in plasma].

A method is described which enables determination of vitamin D3 and its physiologically most important metabolites, i.e. 25-OHD3, 24,25-(OH)2D3, 25,26-(OH)2D3 and 1,25-(OH)2D3 in a plasma sample of about 2 to 4 ml. The whole procedure involves two preparative and one analytical steps: Extraction with methanol/methylene chloride (2:1), chromatographic separation on Lipidex 5000 using a stepwise gradient of n-hexane and chloroform and finally HPLC separation on Zorbax-Sil columns with n-hexane isopropanol mixtures and subsequently reversed phase separation on RP 18-columns and mixtures of methanol and water. Except for 1,25-(OH)2D3 all D compounds were quantified by UV-detection with 1.4 ng of substance being the lowest detectable amount. 1,25-(OH)2D3 was measured by radioimmunoassay. Prior to HPLC analysis the extract was separated into three fractions on Lipidex 5000 which contained 1) vitamin D3, 2) 25-OHD3 and 3) the dihydroxy metabolites. The three fractions were separated by HPLC using different mixtures of isopropanol/n-hexane and methanol/water, respectively. Retention times of the individual D-components longer than 10 min appeared to be essential to separate these compounds from accompanying material. Overall recoveries of the individual metabolites were for vitamin D3 48.9%, for 25-OHD3 54.2%, for 24,25-(OH)2D3 50.9% and for 1,25-(OH)2D3 52.5%. Application of the methods to plasma samples from pigs with pseudovitamin D deficiency rickets, typ I, revealed a reduced concentration of 1,25-(OH)2D3 and 24,25-(OH)2D3 and an elevated level of 25-OHD3 in these animals. The results obtained by this method contributed substantially to a better understanding of the aetiological factors associated with this disease.

Biotransformation↗
PubMed1980

Glucocorticoid levels in blood plasma of normal and preketotic cows.

Twenty pregnant cows were kept in two feeding groups. Feeding regimens were designed to induce high milk production (16 kg milk/day) in group I and low production (2 kg/day) in group II. Milk weights were corrected for fat content. After onset of lactation, each cow was fed according to actual production level. Four cows from group I and two from group II developed clinical ketosis during the first few weeks of lactation. Blood taken weekly from all animals from about 2 weeks before until an average of 7 weeks after parturition was assayed for glucocorticoids, glucose and ketone bodies. Average plasma cortisol concentration for both groups was 4.5 +/- 2.6 ng/ml (range from 0 to 13 ng/ml). Plasma cortisol levels in cows which later developed clinical ketosis were not different from those in cows that remained healthy. There was, however, a positive correlation between blood glucose and plasma cortisol, and a negative correlation between blood ketone bodies and plasma cortisol. The findings suggest that adrenal cortical activity is interrelated with onset of ketosis although plasma cortisol levels appear unsuitable for identifying ketotic cows prior to clinical manifestation of the disorder.

Acidosis↗
PubMed1980

Thiamin balance in the gastrointestinal tract of sheep.

Six 5- to 6-month-old sheep fitted with rumen fistulas and reentrant cannulas in the duodenum and in the ileum were adapted to two dies low in thiamin and containing different percentages of urea nitrogen. The sheep were subjected to seven thiamin balance experiments, with continuous feeding and total collection of duodenal and ideal contents and feces for periods of 6 day each. Thiamin and dry matter were determined in aliquots of collected digesta and feces. Daily thiamin intake was always less than .3 mg, but average daily flow of thiamin into the duodenum was between 1.53 and 3.46 milligrams. Microbial net synthesis of thiamin in the forestomach system was between 1.44 and 3.23 mg/day, so 90 to 96% of thiamin entering the duodenum was of microbial origin. Disappearance of thiamin from the small intestines approximately equaled thiamin net synthesis in the forestomachs, indicating high absorption of microbially produced thiamin. In five experiments, thiamin balance in the large intestines was positive, but no measurements were made of thiamin breakdown within, and absorption from, the large intestines.

Animals↗