[Dr. H. Hill on his 65th birthday].
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Biomedical subjects
Publications and source records attributed to J Harmeyer.
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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.
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.
In goats and other ruminants, urea functions as a source of nitrogen for protein biosynthesis in the digestive tract. Ammonia can be absorbed in the digestive system when formed in excessive quantitites and enhance formation of urea, or it can be derived from urea of blood plasma when its formation from feed sources is small. Entry rates of urea into plasma may vary from 4 to 80 mumol/min per kg.75 body weight depending on dietary conditions. Urea formation is related to nitrogen intake of which approximately 70% passes into the urea pool of plasma. Irreversible losses of urea of plasma into the digestive tract vary between 10 and 90% depending on the protein to energy ratios of the diet. Entry of urea from plasma into the rumen appears to be a passive process which is sensitive to short-term changes of urea concentrations in plasma. Permeability of ruminal epithelium to urea may be altered by fermentation products of rumen (ammonia, carbon dioxide, volatile fatty acids). The influx of nitrogen into the rumen is related to needs for nitrogen of microbial populations and is associated with changes of renal excretion and tubular reabsorption of urea. Combined gastrointestinal and renal responses exert a synergistic effect on improved utilization of urea of plasma when uptake of dietary nitrogen is limited in goats and other ruminants.
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The effects of continuous intravenous infusions (6 h) of ammonium chloride (5.6; 11.2; and 16.8 mumol.kg-1.min) on plasma glucose and immunoreactive insulin (I.R.I.) levels were studied in three adult sheep. Infusions of 5.6 and 11.2 mumol.kg-1.min elevated ammonia levels in circulating blood from 100 to 150 and 300 microgram.100 ml-1, respectively, but showed no appreciable effect on plasma glucose and I.R.I. concentrations. Infusion of 16.8 mumol.kg-1.min-1 resulted in a blood ammonia concentration of about 400 microgram.100 ml-1 after six hours of infusion. Blood ammonia returned to normal 1 to 2 hours after the end of infusion. Plasma glucose concentration tended to increase slightly from 65 to 75 mg . 100 ml-1 when 16.8 mumol of NH4Cl were infused kg-1.min-1 and remained at the elevated level at least for two additional hours when ammonia infusions were stopped. Plasma I.R.I. tended to decrease from 48 to 38 microunits . ml-1 during the time of the NH4Cl infusion and increased continually to 82 microunits . ml-1 when NH4Cl infusions were stopped. It is concluded from the time courses of plasma glucose and plasma I.R.I. that the effect of ammonia infusion of these parameters cannot entirely be explained by a regulatory release of adrenaline.
A radioimmunoassay for porcine parathyroid hormone has been developed and applied to measure immunoreactive parathyroid hormone (PTH) in plasma of pigs with hereditary vitamin D dependency rickets (VDDR) (pseudovitamin D deficiency rickets). Levels of 25-hydroxycholecalciferol (25-(OH)-D3) in plasma were measured by a protein binding assay. Both plasma concentrations of PTH and 25-(OH)-D3 showed an approximately 4-fold increase compared to normal pigs. PTH levels increased with duration of the disease. Daily dosing of the animals with 1--4 micrograms of 1,25-dihydroxycholecalciferol (1,25-(OH)2-D3) reduced PTH concentrations and resulted in clinical healing. Iv administration of 10 micrograms of 25-(OH)-D3/day did not alter PTH concentrations nor the clinical symptoms. The results suggest that these animals suffer from regulatory hyperparathyroidism. The metabolic defect could be due to a failure of the kidney to convert 25-(OH)-D3 to 1,25-(OH)2-D3.
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The mechanism of transfer of magnesium across isolated rumen epithelium was studied in vitro using 28Mg. Isolated pieces of rumen epithelium developed a transmural potential difference of 8.2 +/- 2.3 mV, lumen side negative, during the first hour of incubation when mounted between the two halves of an incubation chamber. A significant net transport of magnesium from lumen to blood was measured when the initial concentration of magnesium in the solutions bathing both surfaces of the epithelium was 2.5 mM. Net flux of magnesium took place against an electrical gradient and was dependent on the presence of the transmural potential difference. The process was saturable at magnesium concentrations of about 5 mM and was markedly reduced by lowering the temperature of the bathing solution, by addition of 10(-4) M ouabain and by extending the time of incubation. The findings strongly support the view that magnesium absorption from the rumen is mediated by an active transport process.
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Growth of a mixed ruminal population taken from a sheep on a protein-free (all urea) purified diet was estimated by an in vitro fermentation technique including precipitation of microbial protein by trichloracetic acid. Volatile fatty acid production in vitro was determined, and the associated a denosine triphosphate was estimated as moles volatile fatty acids X 2.4. On this basis, the quantity of microbial protein synthesized per mole of adenosine triphosphate increased at higher microbial growth rates.
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Since structure and specifity of amino acid transport systems are basically similar in kidney and intestines it was suspected that rickets associated with hyperaminoaciduria was linked with defective intestinal amino acid uptake. Therefore amino acid absorption from everted sacs of small intestines was studied in 2 piglets suffering from vitamin D dependent rickets, and 7 healthy piglets using 14-C labelled amino acids. The experiments revealed an unaffected intestinal absorption in the rachitic animals, whereas the uptake decreased significantly with increasing age of the experimental animals. The results from these studies suggest that regulatory factors in the blood system, e.g. increased level of PTH or lack of specific vitamin D metabolites, do not directly affect amino acid transport systems in kidney and intestines. It is concluded that hyperaminoaciduria has to be mediated through specific and thus far unidentified effects at the tubular cell level.