Incorporation of tritium into the DNA of hematopoietic tissues during prolonged administration of tritium oxide

With long-term (90 days) administration of tritium oxide (0.37 MBq/g body weight) to ras the carbon-bound tritium accumulated in DNA of haemopoietic tissues during two-month administration of the isotope (the accumulation half-time of 15-25 days); during the next month, the isotope level remained nearly constant (about 20 X 10(6) decay/min/g DNA). Elimination of tritium from DNA started 3 days after termination of its administration and proceeded with two half-times (4-8 days and 12-18 days). The ratio of the tritium content per 1 M hydrogen of DNA to tritium content per 1 M hydrogen of tissue water increased up to 0.5-0.7 during the uptake of tritium oxide, and up to 4-7 after the administration of the isotope had ceased.     

Relative biological effectiveness of tritium oxide based on nucleic acid metabolic indices

Relative biological effectiveness (RBE) of tritium oxide, as compared to gamma-rays (137Cs), with regard to LD50/30 is 2,32 +/- 0,69 for rats. The RBE coefficients for tritium oxide are obtained with regard to some indices of nucleic acid metabolism in the thymus and spleen during the dose formation (0-14 days). The RBE of tritium oxide increases with a decrease in radiation dose as determined according to the concentration and content of DNA per organ and activity of thymus DNAases.     

Comparison of the structure and catabolism of rat thymus DNA exposed to tritium oxide and gamma-radiation in equal doses

A decrease in DNA concentration and in the molecular mass of single-stranded DNA, an increase in the PDN content, and activation of acid DNAses in rat thymus were observed after a single administration of tritium oxide in a dose of 22 mBq/g (a cumulative dose of 7.8 Gy) and gamma-irradiation at a corresponding dose-rate and value of the cumulative dose. These changes were most pronounced during the period of dose accretion, i.e. during 14-30 days after the beginning of irradiation. The degree to which the indices under study varied from the controls was 2-3 times in rats given tritium oxide than in those exposed to gamma-irradiation.     

Nucleic acid metabolism in rat hematopoietic tissues during and after prolonged administration of different doses of tritium oxide

During the long-term administration to rats of tritium oxide in doses of 0.37, 0.925 and 1.85 MBq/g body mass the content of karyocytes and nucleic acids in the bone marrow and spleen was decreased, the rate of their biosynthesis changed, the DNA structure impaired, the content of salt-soluble polydeoxynucleotides increased, and DNAases activated. The observed changes were function of dose. After the end of the administration of the isotope the animals which had received a lesser tritium dose exhibited a more rapid and complete recovery.     

The influence of early nutrition on growth and the circulating levels of immunoreactive somatomedin A.

The effect of nutrition during early development on growth and serum immunoreactive somatomedin A was examined in rats by rearing in small or large liters. At weaning, body weight, brain weight, liver weight and DNA content was significantly reduced in rats whose nutrition was restricted by rearing in large litters. Brain DNA content was significantly altered. Serum somatomedin A was significantly reduced in the growth-retarded rats as compared to those whose growth was enhanced by rearing in small litters. Similarly, maternal serum somatomedin A was significantly reduced in rats nursing large litters.     

Effect of hypophysectomy on estrogen conjugation and on plasma and tissue concentrations of tritium after administration of 3H-estradiol-17beta.

Estradiol-17beta-6,7-3H was injected into ovariectomized (control) and ovariectomized, hypophysectomized (hypox) rats in order to study the binding of estradiol in the brain. Hypophysectomy resulted in a significant increase in the concentration of tritium in the hypothalamus, and preoptic area, as well as cortex, muscle, plasma and liver. However, since the liver was lighter in hypox rats, the total tritium content in the liver was unchanged from controls. Part of the weight reduction in the liver was due to a loss of stainable glycogen, which took place within 24 hours of hypox. The increase in circulating tritiated estradiol after hypox led us to investigate hepatic metabolism of estradiol. In vitro studies on liver slices from control and hypox rats demonstrated a significant reduction in the formation of conjugates of estrogen. Specifically, estradiol glucuronide and estrone sulfate formation were reduced in hypox rats and conversely the unmetabolized estradiol concentration was higher. Hypophysectomy for 24 hours results in a significant decrease in hepatic metabolism of estradiol-17beta.     

Evaluation of the Effects of Pentagastrin, Gastrin and Pancreatic Glucagon On Cell Proliferation In the Rat Gastrointestinal Tract

ABSTRACT The effects of six injections of a range of doses (100–1000 μg/kg bodyweight) of pentagrastrin and single injection of a range of doses of porcine gastrin (10–40 μg/kg bodyweight) and pancreatic glucagon (25–100 μg/kg bodyweight) on cell proliferation in the intestine of fasted rats has been investigated. the end-point employed included the measurement of ¹⁴C leucine incorporation and thymidine-derived tritium content of the body of the stomach, duodenum, jejunum, ileum and colon. the carbon 14 and tritium content per μg of tissue in triplicate samples of fifty individually dissected crypts of glands were determined. From these data and the wet weight of the washed, blotted, intestinal segments, values for crypts/μg tissue and crypts/segment were calculated. the results demonstrated that pentagastrin at physiological doses decreased cell proliferation slightly in stomach, while gastrin and glucagon were without effect. In the small intestine, pentagastrin and gastrin were without significant effect with the exception that they increased the weight of the duodenum. In contrast, a high physiological dose of glucagon increased DNA and protein synthesis throughout the small bowel, but particularly in the ileum. Pharmacological doses of pentagastrin and all doses of gastrin appeared to increase cell proliferation in the colon although the possibility could not be excluded that this was due to stimulation of precursor uptake. Gastrin also increased colonic weight. Glucagon had no effects in the colon. These observations are compatible with the hypothesis that (i) the primary effects of gastrin and pentagastrin on the proximal intestine are as secretogogues and effects on cell proliferation may be secondary, (ii) gastrin and pentagastrin at physiological levels do not stimulate small intestinal cell proliferation, however glucagon does, and (iii) gastrin at physiological levels and pentagastrin at pharmacological levels may stimulate cell proliferation in the colon.     

Evaluation of the effects of pentagastrin, gastrin and pancreatic glucagon on cell proliferation in the rat gastrointestinal tract

The effects of six injections of a range of doses (100-1000 micrograms/kg bodyweight) of pentagrastrin and single injection of a range of doses of porcine gastrin (10-40 micrograms/kg bodyweight) and pancreatic glucagon (25-100 micrograms/kg bodyweight) on cell proliferation in the intestine of fasted rats has been investigated. The end-point employed included the measurement of 14C leucine incorporation and thymidine-derived tritium content of the body of the stomach, duodenum, jejunum, ileum and colon. The carbon 14 and tritium content per microgram of tissue in triplicate samples of fifty individually dissected crypts of glands were determined. From these data and the wet weight of the washed, blotted, intestinal segments, values for crypts/micrograms tissue and crypts/segment were calculated. The results demonstrated that pentagastrin at physiological doses decreased cell proliferation slightly in stomach, while gastrin and glucagon were without effect. In the small intestine, pentagastrin and gastrin were without significant effect with the exception that they increased the weight of the duodenum. In contrast, a high physiological dose of glucagon increased DNA and protein synthesis throughout the small bowel, but particularly in the ileum. Pharmacological doses of pentagastrin and all doses of gastrin appeared to increase cell proliferation in the colon although the possibility could not be excluded that this was due to stimulation of precursor uptake. Gastrin also increased colonic weight. Glucagon had no effects in the colon. These observations are compatible with the hypothesis that (i) the primary effects of gastrin and pentagastrin on the proximal intestine are as secretogogues and effects on cell proliferation may be secondary, (ii) gastrin and pentagastrin at physiological levels do not stimulate small intestinal cell proliferation, however glucagon does, and (iii) gastrin at physiological levels and pentagastrin at pharmacological levels may stimulate cell proliferation in the colon.     

Postnatal development of uterine response to estradiol-17β in the rat

The gradual acquisition of uterine responsiveness to a single injection of estradiol-17β was studied in rats aged 5–30 days with respect to wet weight and the content of DNA, RNA, and protein. No significant response in any of these parameters was found in 5- or 10-day-old rats. In 15- and 20-day-old rats all the parameters, except DNA content, were elevated by the hormone treatment; only in 30-day-old rats was there a significant increase in DNA content.     

Effect of tritium oxide in a wide range of doses on the nucleic acid metabolism of the rat spleen

A single injection of tritium oxide in a dose of 1.1 MBq/g (0.5 Gy for 30 days) was shown to impair the nucleic acid metabolism in the rat spleen. The changes in the indices under study (e.g. mass, nucleic acid content and biosynthesis) increased with the dose, and the recovery started later and was incomplete. Qualitative differences were found in the effects of tritium oxide and gamma radiation with regard to the rate of DNA biosynthesis: 24 h following the injection of the radionuclide specific activity of DNA increased with dose, whereas this function was inverse in the case of gamma irradiation as it was reported in the literature.     

Cell proliferation in organs of rats bearing hepatoma 3924A: effects of X-rays of surgery

For inbred rats with Morris hepatoma 3924A, increases in tumor size were accompanied by increases in weight and DNA content of spleen, DNA content of tibial marrow, and peripheral white cell concentrations of blood. White blood cell concentrations of rats with tumors weighing more than 5 g were approximately two-fold greater than for rats without tumors. Neutrophils were primarily responsible for the increase in white cells. Local x-radiation of 3750R to the tumor when the tumor was small prevented tumor growth and the increases in spleen weight, incorporation of 3H-thymidine into spleen DNA, white blood cell count, and tibial marrow DNA content related to tumor growth. Surgical removal of large tumors resulted in a return of spleen weight and DNA content to near normal values within 1 week. Despite the evidence for increased cell proliferation in hematopoietic tissues of rats with hepatoma 3924A, no systematic relationship has been observed between tumor size and animal survival following treatment with the cell cycle specific agent 5-fluorouracil when tumors have varied in size from 0.5 g to 5 g at the time of drug treatment.     

An evaluation of tritium and fluorescence labelling combined with multi-detector SEC for the detection of carbonyl groups in polysaccharides

The carbonyl content of a pectic polysaccharide from Sphagnum papillosum (sphagnan) and periodate oxidised alginates was investigated using three different carbonyl labelling strategies combined with size-exclusion chromatography (SEC) with multi-angle laser light scattering (MALLS) and on-line fluorescence or off-line tritium detection. The labelling strategies were tritium incorporation via NaB³H₄ reduction, and fluorescent labelling with carbazole carbonyl oxyamine (CCOA), or 2-aminobenzamide (2-AB), respectively. Carbonyl quantification was based on labelled pullulan, dextran and alginate standards possessing only the reducing end carbonyl group. As a result the carbonyl distribution in the polysaccharides could be determined. In sphagnan it was found that the carbonyl content increased with increasing molecular weight, whereas in periodate oxidised alginate the carbonyl content was as expected independent of the molecular weight. The methods proved useful for carbonyl detection in water soluble polysaccharides in general. The tritium incorporation method was preferred for alkali stable polysaccharides, while the CCOA method was most suitable for acid stable polysaccharides with low carbonyl content. The 2-AB method is applicable for all polysaccharides tested with varying carbonyl content; however, it lacks the ability to detect ketone functionalities.     

Variable effect of ghrelin administration on pancreatic development in young rats. Role of insulin-like growth factor-1.

Ghrelin, an endogenous ligand of the growth hormone secretagogue receptor, has been primarily isolated from the human and rat stomach. Ghrelin has been shown to stimulate appetite and fat deposition in adult rats and humans. The aim of this study was to investigate the effect of ghrelin administration on pancreatic growth in suckling, weaned and peripubertal seven week old rats. Rats were treated with saline or ghrelin (4, 8 or 16 nmol/kg/dose) intraperitoneally twice a day: suckling rats were treated for 7 or 14 days starting from the first postnatal day, three week old weaned rats and seven weeks old rats were treated for 5 days. Treatment with ghrelin did not affect animal weight in suckling or weaned rats, whereas in young seven week old rats, ghrelin caused a significant increase in body weight. Ghrelin decreased food intake in weaned rats; whereas in seven week old rats, food intake was enhanced. In suckling rats, ghrelin decreased the pancreatic weight, pancreatic amylase content, DNA synthesis and DNA content. In contrast, ghrelin increased pancreatic weight, DNA synthesis, DNA content and amylase content in weaned or young seven week old rats. Pancreatic blood flow was not affected by ghrelin in any group of rats tested. Ghrelin increased serum level of growth hormone in all rats. This effect was weak in suckling rats, higher in weaned and the highest in seven week old animals. Ghrelin did not affect serum level of insulin-like growth factor-1 (IGF-1) in suckling rats. In weaned and in seven week old rats, treatment with ghrelin caused increase in serum level of IGF-1. We conclude that ghrelin reduces pancreatic growth in suckling rats; whereas in weaned and young seven week old animals, treatment with ghrelin increases pancreatic growth. This biphasic effect of ghrelin in young animals on pancreatic growth seems to be related to age-dependent changes of the release of anabolic IGF-1.     

The lack of cardiac hypertrophy in newborn Prague hypertensive rats.

Newborn rats of four different strains with spontaneous hypertension show heart enlargement mainly due to cardiac hyperplasia. To determine whether this anomaly is common in all genetically hypertensive rats, we have compared newborns of Prague hypertensive rats (PHR) with their respective normotensive controls (PNR). The heart ventricles, kidneys and livers of newborn animals were analyzed for their weight, protein and DNA content. The total heart weight and the heart/body weight ratio were significantly lower in PHR than in PNR. On the other hand, there were no differences in total or relative kidney weight and in total liver weight. The relative protein content was significantly lower in kidney and liver of PHR but there were no differences between hypertensive and normotensive animals in relative DNA content of all organs studied. Our results suggest a possible dissociation of genes which determine organ weights from those responsible for blood pressure determination.     

Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat

This study was conducted to investigate the effects of a moderate dose of purified soybean agglutinin on performance and nitrogen digestibility in rats as well as to determine its effects on the protein, DNA and RNA content of the small intestine and pancreas. Twenty-four Sprague - Dawley rats were randomly allotted into one of four groups for a 10-day nitrogen balance experiment. The four groups of rats were fed 7 g of a casein-cornstarch based diet or a similar diet supplemented with 0.1, 0.2 or 0.4 mg/g purified soybean agglutinin. All experimental diets were adjusted to an identical nutrient level. Dose of soybean agglutinin had no significant effect on rat performance. Incorporation of soybean agglutinin in the diet reduced apparent protein digestibility and the utilization of dietary protein by increasing nitrogen loss from the faeces and urine. Fresh pancreatic weight increased in rats fed soybean agglutinin at a level of 0.4 mg/g in the diet compared to the control, but the dry pancreatic weight and the protein content of the pancreas did not differ among the four groups. However the DNA and RNA content of the pancreas had a tendency to increase with a higher level of soybean agglutinin. The weight of the jejunum and its protein, DNA and RNA content were not significantly affected by soybean agglutinin, but the dry weight and the RNA of the jejunum tended to increase with higher levels of soybean agglutinin in the diet. In conclusion, purified soybean agglutinin, at moderate levels in the rats diet, had negative effects on digestive function, such as nitrogen digestibility, nitrogen retention and nitrogen balance. As the level of soybean agglutinin increased, the effects became more pronounced. Meanwhile, hypertrophy of the pancreas was observed with higher doses of soybean agglutinin incorporation in the diets.     

Lung mechanics and connective tissue proteins in diabetic Bio-Breeding/Worcester Wistar rats

We studied lungs of spontaneously diabetic Bio-Breeding/Worcester (BB/W) Wistar rats which resemble human insulin-dependent diabetes mellitus. Compared with the age-matched control group, the body weight of the diabetic rats tended to be smaller and lung wet and dry weight were similar, but lung dry weight, relative to body weight and to lung wet weight, was significantly larger. Both air and saline lung volumes were reduced in the diabetic rats, and volume-pressure (V-P) curves expressed as a percent of maximal lung volume were significantly shifted downward and to the right of those in the control group over the midportion. Total DNA and RNA contents were similar in both groups, whereas protein content and concentration and protein/DNA and RNA/DNA ratios were significantly reduced in the diabetic rats. In contrast, content and concentration of 4-hydroxy-L-proline, elastin, and crude connective tissue were significantly higher in the diabetic group. We conclude that the increase in connective tissue proteins in the BB/W rats is most likely responsible for the shift in the V-P curves.     

Age dependence of cardiac growth in the normal and carbon monoxide-exposed rat.

“Young” rats, 5 days of age, and “old” rats, 25 days of age, inhaled 500 ppm carbon monoxide in air until 50 days of age. Heart weight (HW) relative to same-age controls was maximal at 25 days of age in the young rats, a time at which hemoglobin and hematocrit reached minimal values. Weight of the right ventricle (RV), left ventricle (LV), and combined atria showed a similar pattern. HW and weights of LV and RV of the old rats were intermediate to those of the young rats and controls at 40, 45, and 50 days of age. DNA content of the LV and RV increased sharply in the young rats after initial CO exposure, departing significantly from equal-age controls. DNA content in both ventricles in both young and control rats reached plateaus after 20–25 days of exposure, with the former more than 50% above the latter. DNA content of both ventricles of the old rats was similar to the controls at 50 days of age. In addition, both DNA concentration and protein to DNA ratios were examined in the three groups. The study supports the notion that the potential for cardiac DNA synthesis and probably muscle cell hyperplasia in the rat comes to an end during the 5th through 25th days of postnatal life.     

Relative biological effectiveness of tritium oxide by the criterion of death of the germ cells in mice

It was shown that low-level beta-radiation of tritium is much more effective than gamma-radiation of 137Cs with respect to reduction of the mouse testis weight. The RBE coefficient increases from 1.8 at a dose of 1 Gy to 2.2-2.3 at 0.1 Gy. On the basis of the data obtained by the authors and those reported in the literature a quality factor is proposed for tritium: QF = 2. Using THO and Na36Cl labels a mean water content of the testis cells, necessary for the estimation of a tritium-radiation dose absorbed, has been determined: gamma ct = 0.70 +/- 0.02 ml/g.     

Applications of tritium NMR to macromolecules: A study of two nucleic acid molecules

Summary We have tritium labeled two nucleic acid molecules, an 8 kDa DNA oligomer and a 20 kDa hammerhead RNA for tritium NMR investigations. The DNA sequence studied has been previously used in homonuclear studies of DNA-bound water molecules and tritium NMR was expected to facilitate these investigations by eliminating the need to suppress the water resonance in tritium-detected ³H-¹H NOESY experiments. We observed the anticipated through-space interactions found in B-form DNA in the NOESY experiments and an unexpected antiphase cross-peak at the water frequency. T₁ measurements on the tritiated DNA molecule indicated that relaxation rates were also accelerated for tritium and protons. Tritium NMR spectra of the hammerhead RNA molecule indicated conformational dynamics in the conserved region of the molecule in the absence of Mg²⁺ and spermine, two components necessary for cleavage. The dynamics were also investigated by ¹⁵N-correlated ¹H spectroscopy and persisted after the addition of Mg²⁺ and spermine.     

Value of the tritium test for determining the fat content in the body of rats

An indirect method for estimation of the fat percentage in the animal organism, a tritium test, was studied on laboratory male rats aged 4 and 12 months. Results obtained from the tritium test and direct chemical analysis were compared. With age a mean absolute error of the tritium test increased (from 1 to 8%) as against actual values of the water and fat percentage in the organism obtained by a direct chemical analysis. The data obtained testify to the relative insolvency of the tritium test, as well as the necessity to carry additional investigations in order to obtain adequate data.