65Zn kinetics as a biomarker of DMH induced colon carcinogenesis.

Dietary factors are considered crucial for the prevention of initiating events in the multistep progression of colon carcinoma. There is substantial evidence that zinc may play a pivotal role in host defense against several malignancies, including colon cancer. The present study was conducted to evaluate the kinetics of (65)Zn utilization following experimental colon carcinogenesis in rat model. Twenty rats were segregated into two groups viz., untreated control and dimethylhydrazine (DMH) treated. Colon carcinogenesis was established through weekly subcutaneous injections of DMH (30 mg/kg body weight) for 16 weeks. Whole body (65)Zn kinetics followed two compartment kinetics, with Tb(1) representing the initial fast component of the biological half-life and Tb(2), the slower component. The present study revealed a significant depression in the Tb(1) and Tb(2) components of (65)Zn in DMH treated rats. Further, DMH treatment caused a significant increase in the percent uptake values of (65)Zn in the colon, small intestine, kidney and blood, whereas a significant decrease was observed in the liver. Subcellular distribution revealed a significant increase in (65)Zn uptake in the mitochondrial and microsomal fractions following 16 weeks of DMH supplementation. In conclusion, the present study demonstrated a slow mobilization of (65)Zn during promotion of experimentally induced colon carcinogenesis and provides a physiological basis for the role of (65)Zn in colon tumorigenesis, which may have clinical implications in the management of colon cancer.

[Soil sorption and transfer from it to plant of the radioactive and stable isotopes of chemical elements: methodology and experimental approach].

The adsorption of pair nuclides 60Co/Co and 65Zn/Zn by solid phase and their availability to plants were studied in soil suspension conditions and in the sod podzolic soil under controlled moisture. The situation when the radionuclide in constant activity is entered to system (surface water, soil) with different heavy metal contamination has been modeled. Was observed that soil contamination with heavy metal Co (Zn) significantly reduce sorption of the radionuclide 60Co (65Zn) by solid phase. As a result, the activity concentration of the 60Co or 65Zn in soil solution and, therefore, their mobility and potential availability to plants increases in 2-4 times with the total metal concentration increasing. The difference between two elements is that high Co concentration increase root uptake of the 60Co, whereas soil contamination with Zn reduce activity of 65Zn in the 14-days barley plants that may reflects diverse plant necessity of two elements and more important role of the isotopic exchange effect in the case of Zn.

Dietary metal bioavailability in razor clam Sinonovacula constricta under fluctuating seston environments.

To understand the potential risks of dietary metals to marine bivalves, it is important to study the interaction between dietary metal bioaccumulation and bivalve feeding behavior. Key processes in affecting the dietary metal influx are the selection of different particles during the ingestion process as well as the differential assimilation of metals during the digestion process. In this study, we quantified the influences of seston quality and quantity on the dietary assimilation of Cd and Zn as well as pre-ingestion particle selectivity in a razor clam Sinonovacula constricta following feeding on radiolabeled diatoms and sediments with different mixtures at four food concentrations. Bioavailability of 109Cd and 65Zn from seston was measured by assimilation efficiency (AE) using a pulse-chase feeding technique. The AEs of Cd and Zn were significantly affected by the seston quantity and quality (higher for Zn than they were for Cd and higher for diatoms than for suspended sediments), but were independent of the presence of other particles during the feeding process. Dual gamma radiotracer technique was further employed to study pre-ingestion particle selectivity. Particle selectivity was weak during pre-ingestion in razor clams, although there was evidence that clams might be able to differentiate particles during the process of pseudofeces production. Our study demonstrated that seston composition and quantity substantially affected the bioavailability of Cd and Zn to the razor clams. The results are important to understand the bioaccumulation of metals in clams living in dynamic food environments of estuary.

Removal of 65Zn from mouse body by isotopic dilution and by DTPA chelation.

Isotopic dilution with stable zinc and chelation with DTPA are recommended for removal of radioactive zinc from the body; however, it is unclear which method is more effective. In the present study, the efficacies of these methods were compared in order to determine which treatment should be selected in case of internal contamination with radioactive zinc. Intraperitoneal administration of stable zinc dose-dependently removed 65Zn from the mouse body. However, the dose could not be increased above 3 mg/kg due to its toxicity. Oral administration of zinc was less effective than intraperitoneal administration at the same dose. Our results suggest that the recommended dose of stable zinc (2-3 mg/kg, p.o.) has little efficacy. The efficacies of Ca-DTPA and Zn-DTPA were strongly dependent on the elapsed time after 65Zn exposure. Zn-DTPA was more effective than Ca-DTPA, and its recommended dose (30 micromol/kg) significantly removed 65Zn. Therefore, chelation therapy with Zn-DTPA should be started as soon as possible after internal contamination with radiozinc.

Comparing single-feeding and multi-feeding approaches for experimentally assessing trophic transfer of metals in fish.

Diet is an important pathway for metal uptake in marine organisms, and assimilation efficiency is one of the most relevant parameters to quantify trophic transfer of metals along aquatic food webs. The most commonly used method to estimate this parameter is pulse-chase feeding using radiolabeled food. This approach is, however, based on several assumptions that are not always tested in an experimental context. The present study aimed to validate the approach by assessing single-feeding and multiple-feeding approaches, using a model species (the turbot Scophthalmus maximus). Using the kinetic data obtained from the single-feeding experiment, the reconstruction of a multi-feeding experiment was tested for consistency with data provided by an actual multi-feeding performed under the same experimental conditions. The results validated the single-feeding approach. Environ Toxicol Chem 2017;36:1227-1234. © 2016 SETAC.

Binding, uptake and efflux of 65Zn by isolated human trophoblast cells.

The isolated trophoblast from human placenta at term has been used as model for placental transport. Binding to the cells showed to be of low specificity and very limited (less than 10%) in presence of serum. Uptake of zinc was 90% reduced at 4 degrees C and depended on the presence of sulfhydryl groups (as suggested with the inhibition produced by N-ethylmaleimide). There is considerable efflux (50%) of zinc from preloaded cells and the process was enhanced by presence of histidine in the medium. The model proved to be useful in studying the mode of zinc transport. The results indicate that the uptake of zinc by the trophoblast is probably the limiting process in the placental transport of this metal.

Influence of zinc on the status of hepatic trace elements and biokinetics of 65Zn in ethanol treated rats.

Whole body counting studies of 65Zn indicated that the Tb1 (the faster component) was significantly decreased while the slower component (Tb2) was increased significantly following ethanol treatment. Interestingly, following zinc treatment to ethanol treated rats, slower component (Tb2) of 65Zn came back to within normal limits while the faster component (Tb1) got significantly elevated in comparison to ethanol treatment. Percent uptake values of 65Zn were found to be increased in liver, intestine, muscle, brain and kidney, and decreased in bone under alcoholic conditions. Interestingly, the uptake values of 65Zn in all the organs except muscle were reverted back to within normal limits upon zinc supplementation to these ethanol intoxicated animals. A significant decrease in zinc contents was noticed in ethanol treated rats, which, however, were raised to normal levels upon zinc supplementation: Copper levels, on the other hand, were significantly enhanced in both ethanol fed and combined ethanol + zinc treated rats. Calcium levels were significantly decreased in both ethanol and zinc treated rats, which however were further reduced upon zinc supplementation to ethanol fed rats. However, no significant change was observed in the concentrations of sodium and potassium in any of the treatment groups. In conclusion, zinc appears to play a protective role by normalizing the turnover of 65Zn in whole body as well as in its uptake in different organs under alcoholic conditions.

65Zinc and endogenous zinc content and distribution in islets in relationship to insulin content.

Uptake of 65Zn and distribution of 65Zn, total zinc, and insulin were measured in rat islets and islet granules under different conditions of islet culture. Specific activity of islet zinc (65Zn/zinc) was less than 15% that of extracellular zinc even after 48 h. In contrast, once in the islet, 65Zn approached 70% of equilibrium with granular zinc in 24 h and apparent equilibrium by 48 h. During a 24-h culture, at either high or low glucose, reduction of both islet zinc and insulin occurred. However, zinc depletion was greater than that predicted if zinc loss was proportional to insulin depletion and occurred only from the granular compartment, which represents only one third of the total islet zinc. Extension of culture to 48 h caused additional insulin depletion, but islet zinc was unchanged. Omission of calcium during the 48-h culture caused a predicted increase in insulin retention, presumably by inhibiting secretion; however, zinc retention was not increased proportionately. Pretreatment of rats with tolbutamide caused a massive depletion of insulin stored in isolated islets, with little change in total islet zinc; subsequent culture of these islets resulted in a greater loss of granular zinc than predicted from the small loss of granular insulin. None of the conditions tested affected the percentage of either 65Zn or total zinc that was distributed in the islet granules. Results show that zinc exists in a metabolically labile islet compartment(s) as well as in secretory granules; and extra-granular zinc, although not directly associated with insulin storage, may act as a reservoir for granular zinc and may regulate insulin synthesis, storage, and secretion in ways as yet unknown.

Absorption of orally administered 65Zn by normal human subjects.

Despite studies by several investigators of human gastrointestinal 65Zn absorption, implications of these data for evaluation of functional zinc status are unclear because limited numbers of normal subjects have been studied. To evaluated zinc absorption in normal humans, 75 subjects (31 women, 44 men, ages 18 to 84 yr) were given 10 micro Ci carrier-free 65Zn orally after an overnight fast. Absorption calculated from total body retention measured 7, 14, and 21 days after administration of tracer was 65 +/- 11% (mean +/- 1 SD), range from 40 to 86%. Comparison of these results with those for patients with a variety of diseases indicate that patients exhibit a wider range of absorption and, in four of six studies patients exhibit decreased mean zinc absorption. These results of gastrointestinal zinc absorption in a large number of normal humans offer a basis for a clearer comparison with data from patients who exhibit abnormalities of zinc absorption.

Location of zinc and 65Zn in spinal ganglia of the rat.

Following the works of Velazquez et al. (1999), Jo-Seung et al. (2000), Wang et al. (2001), Danscher et al. (2001) and the criteria of Zinc-containing neurons established by Frederickson et al.(2000), we have found the presence and localisation of Zinc in the neurons of the dorsal root ganglia of Wistar rat, by using Timm's thecnique and by studying the autoradiographic uptake of 65Zn. The agreement between the results of both techniques allows us to classify these spinal ganglion neurons as Zinc-containing neurons and also, to confirm some of the results of Velazquez et al. (1999).

Inhibitory effects of Estracyt on R-3327 rat prostatic carcinoma.

Estracyt (estramustine phosphate) injected intraperitoneally, 100 mg, per Kg. three days a week for four weeks, retarded growth of the R-3327 tumor in intact rats and in orchiectomized rats given androgen. The growth inhibition was accomplished by reduction of tumor deoxyribonucleic acid concentration and of the activities of acid phosphatase, leucine aminopeptidase, and other hydrolases. Histologic examination revealed cellular necrosis particularly prominent in the orchiectomized, androgen-treated rats. Estracyt did not affect the uptake of 65-Zn in the tumors but markedly reduced the high uptake in the dorsolateral prostate. There was no accumulation of 3H or 14C in the tumors after intravenous administration of 3H, 14C-labeled Estracyt, but the isotope concentrations decreased much in the same way as they decreased in the dorsolateral prostate. The isotopes were retained in the ventral prostate, where their concentrations were approximately twenty times higher than those in the muscle four hours after injection. The results demonstrate the value of the R-3327 tumor in the evaluation of drugs of potential clinical use for the treatment of prostatic cancer. The results also show that Estracyt has an antitumor effect which is not dependent on the antigonadotropic action of the drug.

Transport and subcellular distribution of intranasally administered zinc in the olfactory system of rats and pikes.

Zinc is an essential element, which can act as a neuromodulator and also is bound in zinc proteins in the brain. The olfactory bulb contains high concentrations of zinc. In the present study, 65Zn(2+) was applied on the olfactory epithelium of rats and pikes and the transport of the metal in the olfactory system was then examined. Administration of 65Zn(2+) in the nasal cavity of rats or the olfactory chambers in pikes resulted in an uptake of the metal in the olfactory epithelium and a transport of the metal along the primary olfactory neurons to their terminations in the olfactory bulbs. Low levels of 65Zn(2+) passed these terminals and continued into the interior of the bulbs. In the rats 65Zn(2+) was also detected in the anterior parts of the olfactory cortex. Subcellular fractionations of the olfactory mucosa and olfactory bulb of rats given 65Zn(2+) intranasally showed that the metal is bound both to particulate cellular constituents and to cytosolic components in these tissues. Gel chromatography indicated that some of the zinc in the cytosol is bound to metallothionein in the olfactory mucosa and bulb. Inhalation of zinc-containing dusts or fumes occurs in some work-places and may imply high exposure of the nasal tissues. It is not known whether neurotoxicity may be related to uptake of zinc in the olfactory system. However, this is an issue which deserves attention, since zinc dysregulation has been implied to play a role in Alzheimer's disease. In addition, impairment of the sense of smell and degenerative changes of the olfactory tissues have been seen in early stages of some neurodegenerative disorders.

Age-dependent variation of zinc-65 metabolism in LACA mice.

Mice were gavaged with zinc-65 solution, 8.6-19.3 kBq per mouse, and the whole-body retention and organ content of zinc-65 were measured at different times after administration. The age-dependence of the fractional absorption of zinc-65 from the gastrointestinal tract (f1), the endogenous faecal excretion fraction of zinc-65 (EFEF), tissue distribution and whole-body retention were determined. The f1 values obtained were 0.86 +/- 0.15, 0.64 +/- 0.11, 0.52 +/- 0.07 and 0.39 +/- 0.02 in suckling, adolescent, young adult and older mice, respectively. The EFEF values determined were 0.083 +/- 0.008, 0.099 +/- 0.004, 0.122 +/- 0.018 and 0.144 +/- 0.005 of intraperitoneally injected zinc-65 in suckling, adolescent, young adult and older mice at administration. Zinc-65 mainly distributed in the liver, muscle, lung, kidneys and bone. In some tissues, there was an inverse relationship between the relative content of gavaged zinc-65 and the animal's age at administration. The whole-body biological half-lives of zinc-65 increased with animal age. The influence of the age-dependent variation of zinc-65 metabolism on internal dose and on radiation protection is discussed.

Effects of tetracycline on the absorption of 65zinc in rats.

The effects of tetracycline on the absorption of orally ingested 65Zn was studied on rats by whole body counting assay. 65Zn was given as a single dose to groups of rats, five in each, which were started on tetracycline. Tetracycline was given in daily doses of 25 mg, 50 mg, 100 mg, and 200 mg for 10 days. From the 6th day of the study, 65Zn retention when plotted against time in a semilog plot, approximated linearity. The net absorption of 65Zn in the various groups was determined by extrapolating to zero time the linear curve segment of individual retention curves. Except for the lowest tetracycline dosage, tetracycline significantly impaired 65Zn absorption. The elimination rate of retained 65Zn from the 6th day was significantly higher in the group receiving 200 mg tetracycline as compared with the control group.

Simultaneous determination of (45)calcium and (65)zinc uptake by caco-2 cells.

A simple method for simultaneously determining cell-associated Ca and Zn in Caco-2 cells is described. Calcium and zinc uptake was measured via radioisotopes (45)Ca and (65)Zn. Preliminary studies revealed that (65)Zn, a positron (beta(+)) and gamma emitter, contributed to (45)Ca counts in a liquid scintillation counter (LSC). However, (45)Ca, being a true beta emitter, did not contribute to the counts in a gamma counter (gammaC). To differentiate the counts of (45)Ca from those of (65)Zn, first a (65)Zn-labeled cell suspension was read in a gammaC and an LSC, thus obtaining the relationship between the radioactive counts obtained from the gammaC and LSC. This information defined the linear relationship between gammaC (65)Zn counts per minute (CPM) and LSC (65)Zn CPM. Because the (45)Ca and (65)Zn counts obtained in the LSC are additive, giving total LSC CPM, the value of LSC (45)Ca CPM was obtained by subtracting LSC (65)Zn CPM from total LSC CPM for the dual-labeled cell sample, obtaining then LSC (45)Ca CPM. To determine the absolute activity or disintegrations per minute (DPM) of each isotope in the dual-labeled sample, the linear relationship between DPM and CPM was determined for each isotope. The method is simple and straightforward for the determination of (45)Ca counts from a sample also containing (65)Zn, using gamma and liquid scintillation counters.

Bioavailability of zinc from cooked philippine milled, undermilled, and brown rice, as assessed in rats by using growth, bone zinc, and zinc-65 retention.

The nutritional bioavailability of zinc from cooked milled, undermilled, and brown Philippine rice (variety PSB Rc14) was evaluated in rats, comparing results based on weight gain, tibia zinc incorporation (slope ratio analyses), and zinc radiotracer retention. Milling reduced the phytic acid and mineral content of the rice, resulting in zinc concentrations of 16.5, 19.4, and 27.2 microg/g and phytate/zinc molar ratios of 4, 20, and 28 for milled, undermilled, and brown rice, respectively. Measured zinc bioavailability was similar whether using growth, bone zinc, or radioisotope retention as criteria, at approximately 92, 86, and 77% of zinc sulfate, for milled, undermilled, and brown rice, respectively. However, the higher percent bioavailability of the zinc after milling was insufficient to compensate for the lower zinc content. With respect to zinc, the nutritional value was inversely related to milling, providing approximately 15, 17, and 21 microg bioavailable zinc/g rice, respectively, for milled, undermilled and brown rice of this variety.

Development of zinc deficiency in 65Zn labeled, fully grown rats as a model for adult individuals.

The development of zinc deficiency in adults was studied in a metabolism experiment involving 31 adult, female rats labeled homogenously with 65Zn. The animals were fed restricted amounts (8 g/day) of a semisynthetic diet containing either 58 microgram Zn/g (control, n = 7) or 2 microgram Zn/g (Zn deficiency, n = 24). Control animals were sacrificed at day 0 (n = 3) and day 29 (n = 4). Zinc deficient animals were sacrificed at day 1, 2, 4, 7, 11, 16, 22, and 29 (3 animals per group). The development of zinc deficiency comprised 4 phases: (I) Fecal Zn excretion needed several days to adjust to the low level of Zn intake. The high initial Zn loss via feces was counterbalanced mainly by Zn mobilization from the skeleton. (II) During the 2nd week of deficiency Zn mobilization from tissue storage changed transiently to soft tissues (mainly muscle and fat tissue). (III) After the 2nd week the skeleton resumed to mobilize Zn. (IV) At the end of the study the skeleton Zn storage was exhausted and alkaline phosphatase activity indicated severe Zn deficiency. Urinary Zn excretion was too small to contribute quantitatively to changes in Zn metabolism during any phase of Zn deficiency. In conclusion, adults may compensate a deficient Zn supply by mobilizing tissue Zn for several weeks: The skeleton revealed to be the major short-term as well as long-term source of whole body tissue Zn that can be mobilized.

Influence of ascorbic acid, sodium citrate, and sodium bicarbonate on the uptake of 59Fe-transferrin, 54Mn-transferrin, and 65Zn-transferrin from lactating mouse mammary gland cells.

The effects of ascorbic acid, sodium citrate, and sodium bicarbonate on 59Fe-transferrin, 54Mn-transferrin, and 65Zn-transferrin uptake by the receptors disposed of plasma membrane isolated from lactating mouse mammary gland cells have been investigated. The effect of 10(-2) mol/L ascorbic acid alone and in combination with NaHCO3 on the 59Fe-transferrin uptake is significant and positive. 54Mn-transferrin and 65Zn-transferrin binding to the cell receptors are influenced optimally by 0.5 mol/L sodium bicarbonate. Sodium citrate alone or in combination with other substances always has a negative effect on binding of these three metals. It is suggested that a precise mechanism may exist with large possibilities to rearrange metal uptake and its transport from blood to milk.

Effect of zinc on biological half-lives of 65Zn in whole body and liver and on distribution of 65Zn in different organs of rats following nickel toxicity.

This study was designed to determine the effect of zinc on the biological half-lives of 65Zn in whole body and liver and on distribution of 65Zn in different organs of rats following nickel toxicity. Sprague-Dawley (SD) rats received either nickel in the form NiSO4.6H2O at a dose of 800 mg/L in drinking water, zinc in the form of ZnSO4.7H2O at a dose of 227 mg/L in drinking water, and nickel plus zinc or drinking water alone for a total duration of 8 wk. All of the rats were injected with a tracer dose of 0.37 MBq 65Zn at the end of the treatment period. The effects of different treatments were studied on biological half-lives of 65Zn in whole body and liver and on the distribution of 65Zn in different organs of rats. In the present study, we have noted that nickel treatment to normal rats caused a significant decrease in the slow component (Tb2) in liver, which improved following zinc supplementation. Nickel administration to normal-diet-fed animals caused significant lowering in the percentage uptake of 65Zn values in the brain, liver, and intestine. However, the administration of zinc to nickel-treated rats improved the status of 65Zn in different organs. The Tb2 in the liver and the percentage uptake of 65Zn values elevated following zinc supplementation to nickel-treated rats.

Evaluation of in vivo detection properties of 22Na, 65Zn, 86Rb, 109Cd and 137Cs in plant tissues using real-time radioisotope imaging system.

In plant research, radioisotope imaging provides useful information about physiological activities in various tissues and elemental transport between plant organs. To expand the usage of imaging techniques, a new system was developed to visualize beta particles, x-rays and gamma-rays emitted from plant bodies. This real-time radioisotope imaging system (RRIS) visualizes radioactivity after conversion into light with a CsI(Tl) scintillator plate. Herein, the RRIS detection properties of the gamma-ray emitters (22)Na, (65)Zn, (86)Rb, (109)Cd and (137)Cs were evaluated in comparison with those of radioluminography (RLG) using an imaging plate. The lower quantitative detection limit (Bq mm(-2)) during a 15 min period ranged from 0.1 to 4, depending on the nuclide, similar to that of RLG. When the quantitative ability to detect radiation from various Arabidopsis tissues was analyzed, the quantitative capability in silique and the thick internode tended to be low. In an EGS5 simulation, beta particles were the greatest contributors to RRIS imaging of (22)Na, (86)Rb and (137)Cs, and low-energy x-rays contributed significantly to (65)Zn and (109)Cd detection. Thus, both self-absorption and air space between the sample and scintillator surface could impair quantitative RRIS imaging. Despite these issues, RRIS is suggested for quantitative time-course measurements of radionuclide motion within plants.