Transfer of Np(V) nitrate from gastrointestinal segments of the adult rat

The transfer of soluble Np(V) nitrate was measured in gastrointestinal segments from adult rats by two procedures: instillation, in segments in which the physico-chemical form of Np might be modified by gastrointestinal factors; and perfusion, in segments in which the luminal state of Np remains constant. These assays allowed accurate measurement of the Np(V) transferred from the intestine to the whole body. The amount measured was proportional to segment length and to the duration of the experiments, which lasted for periods of 0.25 to 2 h. Under these experimental conditions, hourly transfer values were about 2 percent, both per millilitre of Np(V) solution instilled and per 10 cm of jejunum perfused. This flux is very much greater than that which may be deduced from studies in which Np was gavaged into intact rats. Intestinal transfer of Np was constant for Np concentrations ranging from 5 X 10(-12) M to 1 X 10-4) M. Raising the concentration of Np(V) to more than 1 X 10(-4) M reduced its intestinal transfer. Addition of Fe(II) also reduced it. The small intestine was the main site of Np(V) absorption, since the transfer from instilled jejunum was about 20 times that observed from the stomach, and no difference was noted between jejunal and duodenal transfer.     

The absorption of ingested neptunium, plutonium and americium in newborn hamsters

Hamsters aged 1, 4, 7, 22 and 30 days were given oral doses of either plutonium-239 citrate or americium-241 nitrate. The values of gastrointestinal absorption obtained were 3.5, 1.4, 0.04, 0.007 and 0.003 per cent, respectively, for plutonium and 4.5, 1.7, 0.5, 0.006 and 0.02 per cent, respectively, for americium, compared with values in adults of 0.01 per cent for plutonium and 0.05 per cent for americium. The absorption of neptunium was measured in hamsters aged 2 and 4 days and values of 2.3 and 1.7 per cent, respectively, were obtained for 239Np as the nitrate and 5.5 and 2.1 per cent, respectively, for 239Np as the bicarbonate compared with the values in adults of 0.02 per cent for both chemical forms. Thus, the absorption of plutonium, americium and neptunium at 1-2 days of age was about 100 times greater than in adults. The results for plutonium and americium show that absorption decreased rapidly with age over the suckling period. The values of absorption obtained at the time of weaning at 22 days were lower than in adults.     

The gastrointestinal absorption of organically bound forms of plutonium in fed and fasted hamsters

Values of about 0.005-0.01 per cent were obtained for the absorption in fed hamsters of plutonium ingested as Pu4+ citrate, isocitrate, phytate and malate complexes and Pu3+ ascorbate compared with about 0.003-0.004 per cent for Pu4+ nitrate. Replacing drinking water with tea did not affect the result for Pu4+ nitrate. Fasting hamsters for 8 h before the administration of plutonium citrate increased absorption to 0.1-0.2 per cent. An extra period of fasting for 4 h after administration did not lead to a further increase in absorption. Similar values were also obtained when plutonium citrate was administered after a 24 h fast, followed either by immediate access to food or a further 4 h fast. In hamsters fasted for 24 h before administration of either Pu3+ ascorbate or Pu4+ nitrate, about 6-7 per cent of the ingested plutonium was retained in the gastrointestinal tract after one week. At three weeks after ingestion of Pu3+ ascorbate, gastrointestinal retention had fallen 100-fold without an increase in absorption.     

Cellular Proliferation of Intestinal Epithelia in the Rat Two Months after Partial Resection of the Ileum

Sprague-Dawley rats subjected 2 months previously to partial resection (10 per cent) of the small intestine and their controls were injected with tritiated thymidine and sacrificed at 2 and 23 hours. Segments of the duodenum, jejunum, and ileum were autoradiographed, and the migration of the labelled cells during the period between 2 and 23 hours was measured with an eyepiece micrometer. The cells had migrated 35, 42, and 34 per cent of the total distance from the crypts to the tips of the villi in the control segments of duodenum, ileum, and jejunum respectively, and 43, 90, and 82 per cent, respectively, in similar segments from resected animals. The rate of migration in the portion of the intestine remaining after resection was approximately three times the normal rate in the ileum, twice the normal rate in the jejunum, and showed an increase of one-third in the duodenum. These results demonstrate that the rate of cell renewal is considerably greater in the remaining portion of the intestine of resected animals than in normal intestine. The increased rate of migration after resection, together with the increase in the height of the villi, resulted in an increase in the rate of cell renewal amounting to 141 per cent in the ileum, 114 per cent in the jejunum, and 23 per cent in the duodenum when compared with control segments.     

Long term retention of 237Np in rats

This interim report summarizes the results of observations during the first year after a single injection of 237Np nitrate (0.2 or 1.0 mg/kg body weight) into adult female rats and further preliminary data obtained with young animals. The retention of 237Np was followed by whole body counting and serial sacrifice of groups of animals. The retention data could be fitted to three-component exponential equations which show no major differences between the two 237Np dose levels. The half-times and extrapolated initial fractions calculated from the first two exponential terms indicate that one fraction, representing about 40 per cent of the injected 237Np was excreted within the first 5 days and an additional 15 per cent within the first 5 months, while the rest was excreted with a half-time of about 3.5 years. This final long term component is assumed to indicate the rate of loss of 237Np from the skeletal compartment. In young animals both whole-body and skeletal retention of 237Np during the first 5 months of observation was about 50 per cent higher than in the adults. Several soft tissue tumours, mostly mammary tumours, have appeared to approximately the same extent in both control and 237Np treated adult rats but no osteosarcomas were detected up to 15 months after injection of 237Np.     

Effects of fasting and/or oxidizing and reducing agents on absorption of neptunium from the gastrointestinal tract of mice and adult or neonatal rats

Neptunium-237(V) nitrate was administered by gavage to groups of fed or fasted adult and 5-day-old rats. Some groups also received the oxidants quinhydrone or ferric iron, and others received the reducing agent ferrous iron. Adult mice received ferric or ferrous iron and 235Np. When the adult rats were killed at 7 days after gavage, measurements showed that, compared with rats that were fed, a 24-hr fast caused a fivefold increase in 237Np absorption and retention. Both quinhydrone and ferric iron caused an even greater increase in absorption in both fed and fasted rats. Ferrous iron, on the other hand, decreased absorption in fasted rats to values lower than those obtained in fed rats. Similar results were obtained in mice treated with 235Np and either ferric or ferrous iron. The highest absorption obtained after gavage of ferric iron to fasted rats and mice was about two orders of magnitude higher than the value obtained in animals that were fed before gavage. The effects of ferric and ferrous iron on neptunium absorption by neonatal rats were similar to their effects on adult animals but of lesser magnitude. These results are consistent with the hypothesis that Np(V), when given in small mass quantities to fed animals, is reduced in the gastrointestinal tract to Np(IV), which is less well absorbed than Np(V).     

Reduction of Np(V) and precipitation of Np(IV) by an anaerobic microbial consortium

A combination of experimental, analytical, and modeling investigations shows that an anaerobic, sulfate-reducing consortium reduced Np(V) to Np(IV), with subsequent precipitation of a Np(IV) solid. Precipitation of Np(IV) during growth on pyruvate occurred before sulfate reduction began. H₂ stimulated precipitation of Np(IV) when added alone to growing cells, but it slowed precipitation when added along with pyruvate. Increasing concentrations of pyruvate also retarded precipitation. Accumulation of an intermediate pyruvate-fermentation product – probably succinate – played a key role in retarding Np(IV) precipitation by complexing the Np(IV). Hydrogen appears to have two roles in controlling Np precipitation: donating electrons for Np(V) reduction and modulating intermediate levels. That Np(V) is microbially reduced and subsequently precipitated under anaerobic conditions has likely beneficial implications for the containment of Np on lands contaminated by radionuclides, but complexation by fermentation intermediates can prevent immobilization by precipitation.     

Gastrointestinal absorption of neptunium in primates: effect of ingested mass, diet, and fasting

Absorption and retention of neptunium were determined in baboons after intragastric administration of neptunium nitrate solutions at pH 1. The effects of mass, diet, and fasting on absorption were studied. At higher mass levels (400-800 micrograms Np/kg), absorption was about 1%; at lower mass intakes (0.0009-0.005 micrograms Np/kg), absorption was reduced by 10- to 20-fold. The addition of an oxidizing agent (Fe3+) increased gastrointestinal absorption and supported the hypothesis of a reduction of Np (V) when loss masses were ingested. Diets depleted of or enriched with hydroxy acids did not modify retention of neptunium but increased urinary excretion with increasing hydroxy acid content. The diet enriched with milk components reduced absorption by a factor of 5. Potatoes increased absorption and retention by a factor 5, not necessarily due to the effect of phytate. Fasting for 12 or 24 h increased retention and absorption by factors of about 3 and 10, respectively. Data obtained in baboons when low masses of neptunium were administered suggest that the f1 factor used by ICRP should be decreased. However, fasting as encountered in certain nutritional habits is a factor to be taken into consideration.     

VAGINAL SMEARS—A Plea for More Critical Evaluation

To determine whether a conscious effort to reduce the incidence of equivocal (Class III) vaginal cytologic smear readings might have demonstrable clinical value, we have compared a previous study of our predictive efforts with a recent experience in screening a similar number of patients.It was possible to achieve an appreciable reduction in the incidence of Class III smears—from 2.4 per cent to 0.6 per cent. There was a modest increase in the reliability of the Class III smear as an indicator of cancer and a very notable increase in the reliability of the Class IV and V interpretations. While only 66 per cent of the earlier group with Class IV and V smears were shown to have cancer, 86 per cent of the present series with IV and V readings had malignant disease (97 per cent of the Class V group).The ultimate yield of malignancies was similar in the two series, but theoretically the overloading of Classes III, IV and V with women who do not actually have cancer necessarily leads to large numbers of diagnostic surgical procedures that might be avoided. Class III should be used to denote merely a temporary state of inconclusiveness rather than a real suspicion of cancer. With proper collaboration between physician and cytologist, most of the initially confusing situations can be either upgraded or downgraded to the proper rating, and eventually the bulk of patients standing to profit from conization will fall into Classes IV or V.     

A new model of pacing in the mouse intestine

A simple model of pacing in mouse intestine to longitudinal (LM) as well as circular muscle (CM) has been developed. Undissected segments of LM or CM from mouse ileum or jejunum were prepared to record contractions, nerve functions were inhibited, and regular spontaneous contractions were recorded. These had the properties expected of interstitial cells of Cajal (ICC) paced contractions: ileum slower than jejunum, inhibited but not abolished by nicardipine, reduced in frequency by cyclopiazonic acid, abolished by Ca(2+)-free media, and high temperature dependence (Q10 approximately 2.6-3.2). Nicardipine significantly reduced the pacing frequency in LM and CM. Intestinal segments from W/W(V) mice had few irregular contractions in CM but had regular contractions in LM. Other differences were found between LM and CM that suggest that the control of pacing of LM differed from pacing of CM. Moreover, both LM and CM segments in wild-type and W/W(V) and after cyclopiazonic acid responded to electrical pacing (50 V/cm, 50 or 100 ms) at 1 pulse per second. Temperature <26 degrees C inhibited electrically paced contractions in CM. These findings suggest that the current models of ICC pacing need to be modified to apply to intact segments of mouse intestine.     

Changes in metabolites of the energy reserves in individual layers of mouse cerebral cortex and subjacent white matter during ischaemia and anaesthesia

The distribution of glucose, glycogen, ATP, P-creatine and inorganic phosphate was measured in layers I, III, IV, V and VI of cerebral cortex and subjacent white matter of mouse brain. ATP, P-creatine and inorganic phosphate were evenly distributed in all regions examined, whereas levels of glucose and glycogen were higher in white matter than the average for the other layers. Anaesthesia increased levels of glucose and P-creatine in layers I and V and subjacent white matter (other layers were not examined). Anaesthesia doubled the level of glycogen in molecular layer I with lesser increases in layers III, IV, V and VI, but with no change in white matter from the unanaesthetized control value. The metabolic rates in the individual layers were estimated from the rates of expenditure of energy reserves during total ischaemia. In non-anaesthetized mice, white matter had a higher metabolic rate than either layer I or V. Anaesthesia reduced the metabolic rates in all layers; however, the largest reduction occurred in subjacent white matter (86 per cent), with reductions of 54 per cent and 76 per cent respectively in layers I and V.     

A METHOD FOR QUANTITATION OF PROLIFERATIVE INTESTINAL MUCOSAL CELLS ON A WEIGHT BASIS: SOME VALUES FOR C57BL/6

A method for determining the number of intestinal mucosal crypts, S cells, and total proliferative cells, on a weight basis has been presented. The number of crypts was obtained (following injection of tritiated thymidine) by dividing the disintegrations per minute (dpm) per mg intestine by the dpm per crypt. Multiplication of the number of crypts per mg by the number of labeled cells per crypt (determined radioautographically) resulted in the number of S cells per mg intestine. Division of the number of S cells per mg by the fraction of proliferative cells in S (obtained by cell cycle analysis) resulted in the number of proliferative cells per mg intestine. Values for duodenum, jejunum, and ileum of male C57BL/6 mice are given.     

Supplemental sodium phytate and microbial phytase influence iron availability in growing rats.

Five groups of individually housed albino rats (n = 7 each, initial average weight = 42 g) were fed diets based on corn starch and casein over a 4-week period. All diets were supplemented with 35 mg/kg of iron from FeSO4 x 7 H2O. Group I (control) was fed the basal diet free of phytic acid (PA) and phytase. By replacing corn starch by 7.5 g (groups II and IV) and 15 g phytic acid (groups III and V) from sodium phytate per kg diet, molar PA/iron ratios of 18 and 36 were obtained. In groups IV and V, 1000 U phytase from Aspergillus niger per kg diet were added. Food conversion efficiency ratio and growth rate as well as iron in plasma and spleen, hemoglobin, red blood cell count and erythrocyte zinc protoporphyrin were not influenced by the different dietary treatments. Dietary phytate reduced apparent iron absorption in groups II and III. Furthermore hematocrit, transferrin saturation and iron concentration in liver and femur were lowered in rats fed diets with PA, while total and latent iron-binding capacity of plasma increased. Microbial phytase supplementation (groups IV and V) partly counteracted the antinutritive effects of phytic acid on iron availability.     

Processing and transfer of epidermal growth factor in developing rat jejunum and ileum

Using everted sac technique we demonstrated the transfer of ¹²⁵I-mEGF across the jejunal and ileal walls of suckling, weanling and adult rats. The transfer by the suckling rat jejunum and ileum was significantly inhibited by the presence of dinitrophenol and sodium azide or by the replacement of sodium with potassium or choline. RP-HPLC analysis detected carboxy-terminal processing of ¹²⁵I-mEGF in suckling and adult rat jejunum and ileum. Suckling rat jejunum produced ¹²⁵I-des(53)mEGF and ¹²⁵I-des(49–53)mEGF, whereas ¹²⁵I-des(48–53)mEGF was detected in suckling rat ileum or adult rat jejunum and ileum. All three forms of ¹²⁵I-mEGF bound to anti-EGF antibody and EGF receptors. The receptor binding of ¹²⁵I-des(53)mEGF was higher than that of ¹²⁵I-mEGF, but those of ¹²⁵-des(49–53)mEGF and ¹²⁵I-des(48–53)mEGF were greatly diminished. Results indicate a carboxy-terminal processing of mouse EGF during uptake and transfer in the small intestine of developing and adult rats, and the resulting products showed altered receptor binding. An identical amino acid sequence of the C-terminal pentapeptide of EGF from mouse, human and possibly rat may suggest a biological significance of C-terminal processing of EGF in the small intestine.     

Protective effect of lung inflation in reperfusion-induced lung microvascular injury

We used the isolated-perfused rat lung model to study the influence of pulmonary ventilation and surfactant instillation on the development of postreperfusion lung microvascular injury. We hypothesized that the state of lung inflation during ischemia contributes to the development of the injury during reperfusion. Pulmonary microvascular injury was assessed by continuously monitoring the wet lung weight and measuring the vessel wall (125)I-labeled albumin ((125)I-albumin) permeability-surface area product (PS). Sprague-Dawley rats (n = 24) were divided into one control group and five experimental groups (n = 4 rats per group). Control lungs were continuously ventilated with 20% O(2) and perfused for 120 min. All lung preparations were ventilated with 20% O(2) before the ischemia period and during the reperfusion period. The various groups differed only in the ventilatory gas mixtures used during the flow cessation: group I, ventilated with 20% O(2); group II, ventilated with 100% N(2); group III, lungs remained collapsed and unventilated; group IV, same as group III but pretreated with surfactant (4 ml/kg) instilled into the airway; and group V, same as group III but saline (4 ml/kg) was instilled into the airway. Control lungs remained isogravimetric with baseline (125)I-albumin PS value of 4.9 +/- 0.3 x 10(-3) ml x min(-1) x g wet lung wt(-1). Lung wet weight in group III increased by 1.45 +/- 0.35 g and albumin PS increased to 17.7 +/- 2.3 x 10(-3), indicating development of vascular injury during the reperfusion period. Lung wet weight and albumin PS did not increase in groups I and II, indicating that ventilation by either 20% O(2) or 100% N(2) prevented vascular injury. Pretreatment of collapsed lungs with surfactant before cessation of flow also prevented the vascular injury, whereas pretreatment with saline vehicle had no effect. These results indicate that the state of lung inflation during ischemia (irrespective of gas mixture used) and supplementation of surfactant prevent reperfusion-induced lung microvascular injury.     

Effect of protein deficiency in the female rat diet during pregnancy and lactation on the activity of the membrane and soluble forms of digestive enzymes in the offspring small intestine

Protein deficiency in female rats diet during pregnancy and lactation resulted in deceleration of induction of sucrase both forms in the jejunum and ileum; in acceleration of induction of the maltase membrane from in the jejunum; and in suppression of the lactase membrane form in the ileum; in earlier forming of the adult-type distribution of activity of the membrane form of intestinal alkaline phosphatase and in a decrease in activity of the enzyme soluble form. The findings are corroborated by a suppression of activities of the membrane and soluble forms of the small intestine digestive enzymes in 30-day old rat pups fed with a control (adequate) ration starting 21 days after the birth.     

Distribution of 3-hydroxy-3-methylglutaryl-CoA reductase in isolated villus and crypt cells of chick duodenum, jejunum and ileum

3-Hydroxy-3-methylglutaryl-CoA reductase (EC 1.1.1.34), the major rate-limiting enzyme of cholesterogenesis, was studied in epithelial cells isolated in a villus to crypt gradient from chick duodenum, jejunum and ileum, in order to resolve the apparent controversy that exists on the anatomical localization of sterol synthesis in the intestine. Consistent separation was demonstrated by using the marker enzymes alkaline phosphatase, specific to the villus cells, and thymidine kinase, specific to the crypt cells. No relative difference in stability was observed, as shown by the equal distribution of acid phosphatase. Cells were 90-95 per cent viable. The highest specific activity of reductase was located in the microsomal fraction (41 per cent of the total). The mitochondria had lower specific activity (8 per cent of the total). The distribution of reductase activity in epithelial cells of the villus-crypt axis was also studied. The specific activity in each cell fraction from chick duodenum was clearly lower than that in jejunum and ileum. The jejunal and ileal crypt regions showed lower specific activity than the villus cells. About 70 per cent of total reductase activity was found in cells from the upper and the mid villus fraction in each intestinal segment.     

Intestinal absorption and metabolism of a soluble flavonoid, alphaG-rutin, in portal cannulated rats

A highly soluble quercetin glycoside, alphaG-rutin, is a glucose adduct of insoluble rutin, and intestinal absorption and metabolism of alphaG-rutin has not been known. We investigated the intestinal absorption and metabolism of alphaG-rutin by using portal and duodenal cannulated rats and the isolated rat intestinal mucosa. After a duodenal instillation of alphaG-rutin (150 mumol), intact alphaG-rutin, rutin and quercetin were appeared in the portal blood and these concentrations were similarly increased at 15 min. Portal quercetin reached a peak value at 60 min, and the value was higher than those of alphaG-rutin and rutin at that time. Quercetin-conjugates were also increased 30 min after the instillation. The remaining of alphaG-rutin metabolites, mainly rutin, in the intestine were 58% of instilled alphaG-rutin after 150 min. In the experiment by using the isolated mucosa of the jejunum, ileum and cecum, alphaG-rutin and rutin, but not quercetin, appeared in the serosal sides of all segments, and they were increased linearly from 10 to 100 mmol/l of mucosal alphaG-rutin. We also showed portal injected alphaG-rutin was very rapidly cleared from the blood, and appeared a large amount of conjugates. In conclusion, a soluble flavonoid-glycoside, alphaG-rutin, was absorbed as glycosides into the portal blood. A part of alphaG-rutin was hydrolyzed to rutin, but not to aglycone, through the intestine.     

Intestinal absorption and metabolism of a soluble flavonoid, αG-rutin,in portal cannulated rats

A highly soluble quercetin glycoside, αG-rutin, is a glucose adduct of insoluble rutin, and intestinal absorption and metabolism of αG-rutin has not been known. We investigated the intestinal absorption and metabolism of αG-rutin by using portal and duodenal cannulated rats and the isolated rat intestinal mucosa. After a duodenal instillation of αG-rutin (150 μmol), intact αG-rutin, rutin and quercetin were appeared in the portal blood and these concentrations were similarly increased at 15 min. Portal quercetin reached a peak value at 60 min, and the value was higher than those of αG-rutin and rutin at that time. Quercetin-conjugates were also increased 30 min after the instillation. The remaining of αG-rutin metabolites, mainly rutin, in the intestine were 58% of instilled αG-rutin after 150 min. In the experiment by using the isolated mucosa of the jejunum, ileum and cecum, αG-rutin and rutin, but not quercetin, appeared in the serosal sides of all segments, and they were increased linearly from 10 to 100 mmol/l of mucosal αG-rutin. We also showed portal injected αG-rutin was very rapidly cleared from the blood, and appeared a large amount of conjugates. In conclusion, a soluble flavonoid-glycoside, αG-rutin, was absorbed as glycosides into the portal blood. A part of αG-rutin was hydrolyzed to rutin, but not to aglycone, through the intestine.