Double Isotope Technique Research Articles

Freeze-dried Lactobacillus plantarum 299v increases iron absorption in young females-Double isotope sequential single-blind studies in menstruating women.

BackgroundThe probiotic strain Lactobacillus plantarum 299v has earlier been shown to increase iron absorption when added to foods. However, it is not known if the same probiotic strain in a freeze-dried format included in a capsule increases the iron absorption.ObjectiveThe aim of this study was to test the hypotheses that non-heme iron absorption from a light meal is promoted by a simultaneous intake of freeze-dried Lactobacillus plantarum 299v (Lp299v, DSM 9843).Study designWith a single blinded placebo controlled sequential design, iron absorption from a light breakfast meal administered with or without capsules containing 1010 cfu freeze-dried Lp299v was studied in healthy female volunteers of fertile age. The methodology used was a double isotope technique (59Fe and 55Fe). Two studies were performed using the same protocol.ResultsIn study 1, the absorption of iron from a meal without Lp299v was found to be 17.4 ± 13.4%, and from an identical meal with Lp299v was found to be 22.4 ± 17.3% (mean ± SD). This difference was statistically significant (p = 0.040, n = 14).In study 2, the absorption of iron from a meal without Lp299v was found to be 20.9 ± 13.1%, and from an identical meal with Lp299v found to be 24.5 ± 12.0% (mean ± SD, n = 28), which again was statistically significant (p = 0.003).ConclusionFreeze-dried Lp299v enhances the absorption of iron when administered together with a meal with a high iron bioavailability.Trial registrationClinicalTrials.gov Identifier: NCT02131870

Probiotic strainLactobacillus plantarum299v increases iron absorption from an iron-supplemented fruit drink: a double-isotope cross-over single-blind study in women of reproductive age

Iron deficiency is common, especially among young women. Adding probiotics to foods could be one way to increase iron absorption. The aim of this study was to test the hypothesis that non-haem iron absorption from a fruit drink is improved by adding Lactobacillus plantarum 299v (Lp299v). Iron absorption was studied in healthy women of reproductive age using a single-blind cross-over design in two trials applying the double-isotope (55Fe and 59Fe) technique. In Trial 1, iron absorption from a fruit drink containing 109 colony-forming units (CFU) Lp299v was compared with that from a control drink without Lp299v. Trial 2 had the same design but 1010 CFU were used. The test and control drinks contained approximately 5 mg of iron as ferrous lactate and were labelled with 59Fe (B) and 55Fe (A), respectively, and consumed on 4 consecutive days in the order AABB. Retention of the isotopes was measured with whole-body counting and in blood. Mean iron absorption from the drink containing 109 CFU Lp299v (28·6(sd 12·5) %) was significantly higher than from the control drink (18·5(sd 5·8) %), n 10, P<0·028). The fruit drink with 1010 CFU Lp299v gave a mean iron absorption of 29·1(sd 17·0) %, whereas the control drink gave an absorption of (20·1(sd 6·4) %) (n 11, P<0·080). The difference in iron absorption between the 109 CFU Lp299v and the 1010 CFU Lp299v drinks was not significant (P=0·941). In conclusion, intake of probiotics can increase iron absorption by approximately 50 % from a fruit drink having an already relatively high iron bioavailability.

Evaluating abdominal oedema during experimental sepsis using an isotope technique

Abdominal oedema is common in sepsis. A technique for the study of such oedema may guide in the fluid regime of these patients. We modified a double-isotope technique to evaluate abdominal organ oedema and fluid extravasation in 24 healthy or endotoxin-exposed ('septic') piglets. Two different markers were used: red blood cells (RBC) labelled with Technetium-99m ((99m)Tc) and Transferrin labelled with Indium111 ((111)In). Images were acquired on a dual-head gamma camera. Microscopic evaluation of tissue biopsies was performed to compare data with the isotope technique. No (99m)Tc activity was measured in the plasma fraction in blood sampled after labelling. Similarly, after molecular size gel chromatography, (111)In activity was exclusively found in the high molecular fraction of the plasma. Extravasation of transferrin, indicating the degree of abdominal oedema, was 4·06 times higher in the LPS group compared to the healthy controls (P<0·0001). Abdominal free fluid, studied in 3 animals, had as high (111)In activity as in plasma, but no (99m)Tc activity. Intestinal lymphatic vessel size was higher in LPS (3·7 ± 1·1 μm) compared to control animals (0·6 + 0·2 μm; P<0·001) and oedema correlated to villus diameter (R(2) = 0·918) and lymphatic diameter (R(2) = 0·758). A correlation between a normalized index of oedema formation (NI) and intra-abdominal pressure (IAP) was also found: NI = 0·46*IAP-3·3 (R(2) = 0·56). The technique enables almost continuous recording of abdominal oedema formation and may be a valuable tool in experimental research, with the potential to be applied in the clinic.

Effect of helicobacter pylori infection on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat

IntroductionMicronutrient deficiencies and Helicobacter pylori (HP) infection are prevalent and co‐exist in several countries.ObjectiveThe objective of the study was to determine the effect of HP on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat.Materials/MethodsHP was assessed using the C13‐urea breath test. On day 1 subjects received for breakfast bread fortified, per Kg flour, with either 55 mg ferrous sulfate and 60 mg zinc sulfate, labeled with 59Fe, as sulfate, or same amounts of ferrous fumarate and zinc oxide, labeled with 55Fe, as fumarate. On day 3 they received the other type of bread. On day 17 blood (20 ml) was drawn to assess circulating radioactivity. Iron absorption was determined using Eakins and Brown's double isotopic technique.Results/FindingsPrevalence of HP was 78%. Fourteen HP negative and 16 HP positive individuals completed absorption studies. There were no significant differences in the mean iron absorption of either ferrous sulfate or ferrous fumarate by infection status.ConclusionHP has no effect on iron absorption in asymptomatic adults consuming iron and zinc fortified wheat.Support by FONDECYT grant 1080032

Effect of calcium on iron absorption in women with marginal iron status

We measured non-haem Fe absorption with and without added Ca in a short-term feeding study, in thirteen women with marginal Fe status, by the use of a double stable isotope technique. Supplementing 500 mg Ca as calcium carbonate significantly (P = 0.0009) reduced Fe absorption from a single meal from 10.2 % (range 2.2-40.6) to 4.8 % (range 0.7-18.9). A significant inverse correlation in the absence ( - 0.67, P = 0.010) and presence ( - 0.58, P = 0.037) of Ca, respectively, was found between Fe absorption and Fe stores measured by serum ferritin (SF). Wide variation in Fe absorption was observed between individuals in the absence and in the presence of Ca, despite pre-selection of participants within a relatively narrow range of iron stores (SF concentrations). Correction of Fe absorption data based on group mean SF was not found to be useful in reducing the inter-individual variability in iron absorption. It appears that selecting a study group with a narrow initial range of Fe stores does not necessarily reduce the inter-individual variability in Fe bioavailability measurements. These results support the hypothesis that body Fe stores, although an important determinant of dietary Fe absorption, are not the main factor that determines Fe absorption under conditions of identical dietary intake in subjects with low Fe stores.

Total Iron and Heme Iron Content and their Distribution in Beef Meat and Viscera

To determine the content of total iron (TFe) and heme iron (HeFe) in major cuts of meat and principal viscera of bovine origin. 55Fe (30 mCi) was injected into two 4-month-old calves. Triplicate samples of the 12 basic American cuts of meat and major viscera were obtained from each specimen. Samples were acid digested and their iron content was read by atomic absorption spectrophotometry. Duplicate samples of the basic cuts of meat and major viscera were analyzed to determine the concentration of 55Fe using a double isotopic technique. The mean and standard deviation of TFe for all cuts was 1.4 ± 0.3 mg/100 g of meat. The mean TFe for organs was (per mg/100 g): 0.9 ± 0.1 brain, 3.0 ± 0.05 kidney, 3.2 ± 0.04 heart, 5.7 ± 0.2 lung, 6.0 ± 0.1 liver, and 31.2 ± 0.4 spleen. HeFe was 64% of TFe in meat and 72.8% in spleen, 53.8% in lung, 35.7% in brain, 35.0% in kidney, 27.3% in heart, and only 13.6% in liver. Blood contained 85.5% of the radioisotope and only 1.4% was found in muscle and 1.6% was found in viscera. Results suggest that bovine cuts of meat have a low variation in TFe and that HeFe comprises more than 60% of TFe.

Peripheral Blood Hematocrit in Critically Ill Surgical Patients: An Imprecise Surrogate of True Red Blood Cell Volume

Peripheral blood hematocrit (red blood cell volume/total blood volume) is conventionally used to determine the need for blood transfusions. In critically ill surgical patients, this variable may not accurately approximate true red blood cell volume. We compared peripheral blood hematocrit to (1) plasma volume, (2) estimated circulating blood volume, and (3) a normalized hematocrit to clarify their relationships. Consecutive patients admitted to the surgical intensive care unit were evaluated using the BVA-100 Blood Volume Analyzer (Daxor Corporation, New York City, NY). Plasma volume was directly measured by serial tagged albumin concentration. Red blood cell volume was calculated using plasma volume and the peripheral blood hematocrit result. All volumes were presented as percentage deviation from ideal volumes. These ideal volumes were obtained using a patented formula incorporating ideal body weight as determined by Metropolitan Life tables. The peripheral blood hematocrit was compared with a "normalized" hematocrit, defined as the hematocrit value if plasma volume was adjusted to a normal whole blood volume. Eighty-six data points were recorded for 40 patients with average age 61 +/- 20 yr, APACHE II score 20 +/- 6, and a 13% mortality rate. The primary reasons for admission were severe sepsis/septic shock (n = 11), hemorrhagic shock (n = 7), respiratory failure (n = 20), and cardiac failure (n = 2). Bland-Altman analysis showed a mean difference of 3.4 +/- 7.8 hematocrit percentage points between normalized and peripheral blood hematocrit methods, with a 95% confidence interval of 1.7-5.1 and limits of agreement of +/-15.2 hematocrit percentage points. Peripheral blood hematocrit was lower than the normalized hematocrit in 48% of measurements, higher in 17%, and equivalent in 35%. Peripheral blood hematocrit may not accurately estimate red blood cell volume in a cohort of critically ill surgical patients. This remains to be validated in a larger group of patients, comparing these results with the double isotope technique.

Measurement of gastric emptying after gastric bypass surgery using radionuclides.

The pattern of gastric emptying has been assessed after gastric bypass surgery using a double isotope technique for measuring solid and liquid emptying with a scintillation camera. Eleven control subjects and 12 patients, 12 months post-gastric bypass, were studied. Stomal and gastric pouch size were assessed at endoscopy. Solid emptying was slower and liquid emptying was more rapid after gastric bypass surgery. Initial rapid emptying of solid occurred in the 4 patients with the smallest pouch diameters. There was no correlation between stomal size and rates of solid or liquid emptying nor between the weight loss produced by the operation and the rates of solid or liquid emptying, stoma or pouch size.

Studies on zinc bioavailability from a representative diet in Chinese urban women of childbearing age using a double label stable isotope technique

The objectives of the study were to determine dietary zinc bioavailability by using a double label stable isotope technique, and to identify the zinc content and levels of some possible factors which may affect zinc utilization in a typical representative Chinese urban diet. Twenty urban women of childbearing age were fed a diet extrinsically labeled with 67Zn and received intravenous (i.v.) injections of 70Zn. Fecal monitoring, urine monitoring and chemical balance technique were used simultaneously for comparisons. Isotopic enrichment in feces and urine after oral and i.v. administration of stable zinc isotopes was measured to determine the ways of zinc absorption. The mean (±SD) zinc intake from the representative Chinese diet was 10.22±0.80 mg/day, which is lower than the reference nutrient intake (RNI) level (11.5 mg/day), and mainly came from the subsidiary foods (63%). The mean dietary Fe/Zn weight was 1.1±0.1. The phytate/zinc molar ratio in the diet was about 3:1. The phytic acid×Ca/zinc molar ratio in the diet was about 45:1. The apparent and true absorption rates of 67Zn were 36.2±9.7% and 38.2±10.1%, respectively, which were higher than the apparent absorption determined by balance technique (29.7±10.6%). The mean fractional absorption (FA) determined in urine was 35.7±13.3. In conclusion, the subsidiary food is the main source of dietary zinc in this typical diet. The extrinsic labeling technique by use of stable isotopes was more reliable and accurate to determine zinc absorption. Moreover, the double isotopic tracer ratio (DITR) technique applied to urine samples was a more practical and valuable way to evaluate zinc absorption than other methods. It can be speculated that iron and phytic acid might not be the main inhibitors in the representative Chinese diet.

Local uptake and synthesis of oestrone in normal and malignant postmenopausal breast tissues

Uptake and local formation of oestrone (E1) were studied in vivo by a double isotopic technique in normal and malignant breast tissues from 24 postmenopausal women with breast cancer. Active uptake of radio-labelled E1 beyond plasma was found both in normal and malignant tissue, the effect being significantly greater in non-malignant compared with cancer tissue. The presence of local E1 formation was also demonstrated in most samples. Both uptake and synthesis positively correlated with total amount of radioactive E1 found in the tissues. Uptake appeared to make a greater contribution to E1 levels within the breast than in situ synthesis, although there were marked variations between specimens from different patients and the relative proportion of synthesis to uptake was higher in tumour compared with non-malignant tissue. These results demonstrate quantitative differences in the different compartments by which postmenopausal breasts obtain oestrogen and highlight variations between individual breasts. This may be important in optimising oestrogen deprivation therapy for postmenopausal patients with hormone-dependent cancers.

Can phosphorus limitation inhibit dissolved organic carbon consumption in aquatic microbial food webs? A study of three food web structures in microcosms.

Microcosms with three different food web structures and phosphorus (P) limited growth medium were used to study the interactions between P and organic carbon (C) fractions in pelagic food webs. The cultures were run with low dilution to allow the biological processes to determine the outcome. A double isotope technique was used to follow the C and P compartments. In all systems the primary production was P limited. The measured P:C ratios and the observed accumulation of degradable dissolved organic carbon (DOC) indicated that the growth of heterotrophic bacteria was also P limited. The presence of neither algal grazers nor flagellates feeding on bacteria altered the limitation pattern. A net loss of P from the bacterial fraction was observed after the bloom. Different strategies for nutrient aquisition and growth are proposed as mechanisms enabling simultaneous P limitation of algae and bacteria, and a concomitant accumulation of degradable DOC. The ability of the algae to grow with low P:C ratio keeps the regeneration of P through grazers low enough to cause sustained P limitation of both algae and bacteria. The grazers were important producers of DOC when present. This implies that the usual assumption of carbon limited bacterial growth may lead to wrong conclusions regarding the dynamics of plankton communities and the DOC pool.

Assessment of Intestinal Absorption of Trace Metals In Humans by Means of Stable Isotopes

This study is aimed to demonstrate the feasibility of stable isotopes for the assessment of reliable data on fractional intestinal absorption of trace metals in healthy humans. Among the various methods available, the double isotope technique, i.e. one isotope given orally together with the test substance to be investigated and another isotope injected intravenously to correct for retention and endogenous excretion of the particular trace metal, provides quantitative figures of intestinal absorption at reasonable expenses with regard to costs for materials and number of samples to be evaluated. The trace metals exemplarily included in this study, i.e. iron, cobalt and molybdenum show diverging relations between absorbed fractions and amounts administered which are indicative for different regulatory mechanisms of their body content. Food ligands influence the fractional absorption significantly so that the uptake from a composite meal cannot be derived from results on uptake from particular foodstuffs. Therefore, validated data on the behaviour of intestinal absorption will significantly contribute to a better understanding of human trace metal metabolism.

Role of human liver, kidney, and skeletal muscle in postprandial glucose homeostasis.

Recent studies indicate a role for the kidney in postabsorptive glucose homeostasis. The present studies were undertaken to evaluate the role of the kidney in postprandial glucose homeostasis and to compare its contribution to that of liver and skeletal muscle. Accordingly, we used the double isotope technique along with forearm and renal balance measurements to assess systemic, renal, and hepatic glucose release as well as glucose uptake by kidney, skeletal muscle, and splanchnic tissues in 10 normal volunteers after ingestion of 75 g of glucose. We found that, during the 4.5-h postprandial period, 22 +/- 2 g (30 +/- 3% of the ingested glucose) were initially extracted by splanchnic tissues. Of the remaining 53 +/- 2 g that entered the systemic circulation, 19 +/- 3 g were calculated to have been taken up by skeletal muscle and 7.5 +/- 1.7 g by the kidney (26 +/- 3 and 10 +/- 2%, respectively, of the ingested glucose). Endogenous glucose release during the postprandial period (16 +/- 2 g), calculated as the difference between overall systemic glucose appearance and the appearance of ingested glucose in the systemic circulation, was suppressed 61 +/- 3%. Surprisingly, renal glucose release increased twofold (10.6 +/- 2.5 g) and accounted for ~60% of postprandial endogenous glucose release. Hepatic glucose release (6.7 +/- 2.2 g), the difference between endogenous and renal glucose release, was suppressed 82 +/- 6%. These results demonstrate a hitherto unappreciated contribution of the kidney to postprandial glucose homeostasis and indicate that postprandial suppression of hepatic glucose release is nearly twofold greater than had been calculated in previous studies (42 +/- 4%), which had assumed that there was no renal glucose release. We postulate that increases in postprandial renal glucose release may play a role in facilitating efficient liver glycogen repletion by permitting substantial suppression of hepatic glucose release.

Aromatase regulation and breast cancer.

Aromatase regulation and breast cancer.

Angiotensin converting enzyme inhibition reduces retinal overexpression of vascular endothelial growth factor and hyperpermeability in experimental diabetes.

Angiotensin converting enzyme (ACE) inhibition has been recently suggested to have retinoprotective actions in diabetic patients but the mechanism of this effect is not known. In vitro, angiotensin II stimulates expression of vascular endothelial growth factor (VEGF), a permeability-inducing and endothelial cell specific angiogenic factor which has been implicated in the pathogenesis of diabetic retinopathy in humans and in experimental animals. We sought to determine the effects of ACE inhibition on retinal VEGF expression and permeability in experimental diabetic retinopathy. Streptozotocin-induced diabetic rats and control animals were assigned at random to receive ACE inhibitor treatment or vehicle. At 24 weeks the retinal VEGF protein gene expression was assessed by northern blot analysis and in situ hybridisation. Retinal permeability to albumin was measured using a double isotope technique. Experimental diabetes was associated with cell specific two to fourfold increase in retinal VEGF protein gene expression (p < 0.01) and a 2-fold increase in retinal vascular permeability to albumin (p < 0.01). The localization of VEGF expression in the retina was not altered in animals with experimental diabetes. Angiotensin converting enzyme inhibitor treatment of diabetic rats reduced diabetes-associated changes in VEGF gene expression and vascular permeability. These findings implicate the renin-angiotensin system in the VEGF overexpression and hyperpermeability which accompany diabetic retinopathy and provide a potential mechanism for the beneficial effects of ACE inhibition in diabetic retinal disease.

Simultaneous measurement of 18F-FDG and 99mTc HMPAO activities with a dual head SPECT camera using a double isotope technique

Simultaneous measurement of 18F-FDG and 99mTc HMPAO activities with a dual head SPECT camera using a double isotope technique

Scintigraphic Assessment of Interdigestive Duodenogastric Reflux in Humans: Distinguishing between Duodenal and Biliary Reflux Material

Background: Late duodenal phase III is characterized by retroperistalsis. The physiologic function of this phenomenon is unknown. Our aim was to study the relationship between duodenal motility and the transport of duodenal contents from the biliary tract and the duodenum by using a double-isotope technique. Methods: Manometric analysis of the direction of interdigestive duodenal pressure waves was performed in 12 volunteers. Duodenal marker was infused directly into the proximal duodenum, and bile marker was infused intravenously for 2 h. Radionuclide activity was examined for regions corresponding to the stomach, gallbladder, and duodenum. Results: In phase II, antegrade pressure waves dominated with propulsion of both markers to the jejunum. Retroperistalsis occurred in 90% of the activity fronts and was always (100%) followed by retropulsion of duodenal marker to the stomach. A clear-cut reflux of bile marker was seen in only 17% of the activity fronts. The incidence rate of duodenogastric reflux was highest in phase III (P = 0.008) compared with phase II with an infrequent (P = 0.002) admixture of bile. Bile marker contents increased abruptly in the gallbladder during phase III. Conclusions: Late phase III acts as a retroperistaltic pump, retropelling duodenal contents to the stomach. In this physiologic duodenogastric reflux, bile is avoided by deviation to the gallbladder, probably by a phase III-associated occlusion of the sphincter Oddi.

Correlation between tumour blood flow and fluorouracil distribution in a hypovascular liver metastasis model.

The poor response of colorectal liver metastases to fluorinated pyrimidine chemotherapy may be due to poor drug penetration into the tumour. Chemotherapy delivered by the blood to well perfused areas of tumour must reach less well perfused areas by diffusion. This study examined the relationship between intratumoural blood flow and drug uptake in a hypovascular liver metastasis animal model. We used a double isotope technique to examine the microdistribution of the blood flow tracer [125I]-iodoantipyrine (IAP) and fluorinated pyrimidine 5-[6-3H]-fluorouracil (5-FU) within intrahepatic, hypovascular HSN tumours. There was a significant fall (P < 10(-6)) in both IAP and 5-FU uptake between the liver/tumour edge and tumour centre which resulted in a significant covariation (P < 10(-5)) in tracer uptake with distance. The finding of a close covariation between blood flow and drug uptake in liver metastases suggested that 5-FU diffusion did not compensate for low 5-FU delivery in areas of poor tumour blood flow. The lower 5-FU levels in low compared with high areas of tumour blood flow could reduce the cytotoxic effect and increase the potential for development of drug resistance.

Sentinel node (SN), diagnosed by single and double isotope technique

Sentinel node (SN), diagnosed by single and double isotope technique

Intestinal Absorption of Calcium from Foodstuffs as Compared to a Pharmaceutical Preparation

Only few data are available on intestinal calcium absorption from foodstuffs and composite meals in humans. The aim of the study was to compare intraindividually the calcium absorption from milk and from a breakfast with that from a pharmaceutical calcium preparation of equal calcium content. In 8 healthy volunteers between 44 and 58 years of age, the intestinal calcium absorption was measured in randomized order applying the double isotope technique from: (1) 500 ml of fresh milk (equivalent to 620 mg Ca), (2) a test meal composed of 250 g curd, 150 g yoghurt, 3 slices of pineapple, 2 breakfast rolls, 2 cups of coffee, 10 g of coffee cream, 20 g butter, 50 g jam and 20 g honey (equivalent to 580 mg Ca), and (3) a lactogluconate effervescent tablet (equivalent to 500 mg Ca). All test doses were given on an empty stomach and labelled with 20 mg 44Ca. Simultaneously, 5 mg 42Ca in a sterile isotonic solution were injected intravenously. The mean values of the absorbed fractions are 24.0% ± 5.4% (mean ± SD), 17.9% ± 7.1%, and 28.7% ± 9.1% for the milk, for the meal and for the tablet respectively. The data show that less calcium ist absorbed from foodstuffs as compared to a preparation of optimal bioavailability. But in this study only the difference between absorption from the milk and from the meal was statistically significant. Therefore, it is possible to obtain a sufficient calcium supply of the human body also by properly selected foodstuffs.