Regulatory role of zinc on the biokinetics and biodistribution of (65)Zn during the initiation of experimentally induced colon cancer.

The present study was conducted to evaluate the kinetics of zinc utilization during the formation of colon carcinoma in an animal model of colon carcinogenesis. The rats were segregated into 4 groups: untreated control, 1,2-dimethylhydrazine (DMH) treated, zinc treated, and DMH+zinc treated. Colon carcinogenesis was initiated through weekly subcutaneous injections of DMH (30 mg/kg body weight) for 8 wk. Zinc (in the form of zinc sulphate) was supplemented at a dose level of 227 mg/L in drinking water, ad libitum for the entire duration of study. 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 Tb(1) component showed a significant elevation while the Tb(2) component was significantly diminished in DMH-treated rats, which, however, got normalized following zinc supplementation. The biodistribution and subcellular distribution of (65)Zn was significantly affected in DMH-treated rats when compared to normal control rats. However, zinc significantly reversed the altered (65)Zn uptake in different organs and various fractions of colon. The present study for the first time demonstrated a faster mobilization of zinc during initiation of experimentally induced colon carcinoma and provides a physiological basis for the role of zinc in colon tumorigenesis.

Supplementation of zinc mitigates the altered uptake and turnover of 65Zn in liver and whole body of diabetic rats.

Diabetes is a life threatening disease and its onset is linked with both environmental and genetic factors. Zinc metabolism gets altered during diabetes and results in many complications. The present study was designed to elucidate the effects of zinc supplementation on the biokinetics of (65)Zn in whole body, liver and its biodistribution in diabetic rats. The animals were divided into four groups viz; normal control; diabetic (single intraperitoneal injection of alloxan 150 mg/kg body weight); zinc treated (227 mg/l in drinking water); and diabetic + zinc treated. To carry out biokinetics study, each rat was injected intraperitoneally with 0.74 MBq radioactivity of (65)Zn following 4 weeks of different treatments and the radioactivity was determined by using a suitably shielded scintillation counter. Alloxan induced diabetic rats showed a significant decrease in both the fast (Tb(1)) and slow (Tb(2)) components of biological half-life of (65)Zn which, however, were normalized in whole body (P > 0.05) following zinc supplementation. In case of liver, Tb(2) component was brought back to the normal but Tb(1) component was not increased significantly. The present study indicates that the paucity of zinc in the tissues of the diabetic animals was due to decreased retention of tissue zinc as evidenced by increased serum Zn, hyperzincuria and increased rate of uptake of (65)Zn by the liver. Zinc supplementation caused a significant improvement in the retention of zinc in the tissues and is therefore likely to be of benefit in the treatment of diabetes.

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.

Heavy metal capture from the suspended particulate matter by Morus alba and evidence of foliar uptake and translocation of PM associated zinc using radiotracer (65Zn).

In urban set up, increasing combustion and processing activities have contaminated the air with toxic heavy metals which are generally enriched on atmospheric particulate matter. Vegetation around urban area act as a sink where such metal enriched particles generally deposit on the foliar surfaces, however, role of vegetation in uptake of metals adhered on the atmospheric particulate matter is yet not explored properly and is important to study to evaluate their role as bio-remediator. The undertaken work examines the foliar surface of Morus alba for its potential to deposit and accumulate atmospheric heavy metals. Further, to understand foliar uptake mechanism and translocation of atmospheric metal enriched on particulate matter a simulated experiment was conducted by labeling the known particle size (45 µm and 120 µm) with radio labeled 65Zn, applied on the tagged leaf with two particle loads, 25 mg and 50 mg. The study showed that owing to its rough foliar surface with trichomes and grooves, Morus alba efficiently trap heavy metal enriched particles and was capable of accumulating metals from particulate matter into different plant parts. It was recorded that 65Zn adhered on different size particles was taken up by tagged leaf of mulberry and majorly translocated to the lower stem and roots. It was also inferred from the study that both particle size and particle load significantly affect the foliar uptake and translocation of atmospheric heavy metal. The study focuses on the fact that urban avenue trees are capable of taking up atmospheric heavy metals and can play a crucial role in improving air quality.

Development of a compartmental model of zinc kinetics in mice.

To investigate zinc (Zn) kinetics in mice, tracer ((65)Zn) was administered orally to 9-wk-old female mice in the fed state and tracer and Zn concentration were measured in 21 tissues over the following 8 d. Data were analyzed by compartmental modeling using WinSAAM. A published model for Zn kinetics in rats was modified to fit the data from mice and to calculate transfer rates and pool sizes of Zn. Parallel studies were performed in mice lacking genes for metallothionein (MT), MT-I and MT-II (MT-/-), to quantify differences in Zn kinetics in the absence of these proteins in vivo. We confirmed that tracer time course in most tissues was similar in wild-type mice and those lacking MT, except for the pancreas of MT-/-, which retained less tracer. By fitting tissue and intestinal data simultaneously, we found that intestinal tracer could be explained by unabsorbed isotope and loss of Zn from pancreas went through plasma. Differences in pancreatic data in MT-/- were explained by Zn turning over twice as fast in this tissue (4 h) compared with wild type (9 h). These kinetic studies provide parameter values for normal, fed mice that can be used to assess Zn kinetics in abnormal conditions, as demonstrated by the higher turnover of Zn in the pancreas of MT knockout mice.

Biokinetics and dose estimation of 65Zn in the salivary gland and male reproductive organs.

ICRP is revising its recommendations for radiological protection and has added salivary and secretory glands as new target organs. However, little information is available on the distributions of radionuclides in the salivary gland, secretory glands and male reproductive organs. This study deals with the distribution of 65Zn in the salivary gland and male reproductive organs as a function of time after a single intravenous and oral administration. For the study, 64 Wistar strain male rats, eight weeks of age were used. The rats were periodically sacrificed, the liver, kidney, spleen, pancreas, thymus, salivary gland, testis, epididymitis and prostate gland sampled and the radioactivity of these organs measured with an NaI scintillation counter. The relative concentration of 65Zn was highest in the prostate gland. We estimated the radiation dose in humans using rat data for the salivary and secretory glands as well as reproductive organs after intake of 65Zn.

Laboratory analyses of 60Co2+, 65Zn2+ and (55+59)Fe3+ radiocations uptake by Lemna minor.

The living Lemna minor vascular plant and two different sorbents obtained by chemical treatment of this plant were tested to study the removal process of 60Co2+, 65Zn2+ and (55+59)Fe3+ from low radioactive wastewaters. The most effective sorbent was the protonated biomass, indicating the decisive contribution of the complexation process in the assembly of the uptake mechanisms. The uptake performance of the biosorbent obtained from the L. minor can be increased with approximately 20% by treatment with 0.1 N HNO3. Concerning the metabolically active mechanism, it can be notice the slow elimination of 65Zn2+ and the continuously increase of (55+59)Fe3+ uptake degree. The Na2CO3 generated in situ in systems participates to a double exchange reaction with the metallic cations during the uptake. 60Co2+, 65Zn2+ and (55+59)Fe3+ radiocations prefer for coordination N-donor ligands at the expense O-donors ones.

Zinc-65 imaging of rat brain tumors.

The uptake of zinc, an essential nutrient, is critical for cell proliferation. On the basis of the idea that zinc uptake can be an index of viability in proliferating cells, tumor imaging with (65)Zn was performed using autoradiography. After s.c. implantation of ascites hepatoma (AH7974F) cells into the dorsum, 1 h after i.v. injection of (65)ZnCl(2), (65)Zn uptake in the tumor was higher than in the brain tissue but lower than in the liver, which suggests that brain tumors can be positively imaged with (65)Zn. After implantation of AH7974F cells into the periaqueductal gray, 1 h after i.v. injection of (65)ZnCl(2), (65)Zn uptake in the tumor was approximately 10 times higher than in other brain regions. After implantation of C6 glioma cells into the hippocampus, (65)Zn uptake in the tumor was also much higher than in other brain regions. The present findings demonstrate that brain tumors can be imaged with radioactive zinc. To compare brain tumor imaging with (65)Zn with that of [(18)F]fluorodeoxyglucose (FDG), which is widely used for the diagnosis of brain tumors, (14)C-FDG imaging of the C6 glioma was performed in the same manner. (14)C-FDG uptake in the tumor was approximately 1.5 times higher than in the contralateral region in which (14)C-FDG uptake was relatively high. It is likely that zinc uptake is more specific for brain tumors than is FDG uptake, which suggests that there is great potential for the use of (69m)Zn, a short half-life gamma emitter, in the diagnosis of brain tumors.

Effects of Zn pre-exposure on Cd and Zn bioaccumulation and metallothionein levels in two species of marine fish.

Zinc is an essential trace metal but also a potential toxicant to aquatic organisms. In this study, two juvenile marine fish species, the black sea bream Acanthopagrus schlegeli and the grunt Teraponjarbua, were pre-exposed to Zn either from waterborne (0.74-170 microg L-1) or dietary (39-5926 microg g-1) Zn for 1 or 3 weeks. The concentrations of Zn and metallothionein (MT) in the whole body of the black sea bream and in the gills, viscera and carcass of the grunt were then measured during this pre-exposure. Following the pre-exposure, both fish species were then exposed to 109Cd and 65Zn labeled food or water to quantify the dietary assimilation efficiency (AE) and the uptake rate of dissolved Cd and Zn. Zn concentrations in both fish species were enhanced after pre-exposure, but the increases were much less than the increase of ambient Zn pre-exposure concentration. Following Zn pre-exposure, MT concentrations in the viscera and carcass were significantly elevated, whereas the MT levels were not significantly elevated in the gills. Waterborne and dietary Zn exposure enhanced the uptake rates of dissolved Cd and Zn in both fish. The maximum increases of uptake rate constants of dissolved Cd and Zn were up to 1.9-2.8 and 2.1-2.6 times, respectively, in the seabream and grunt. In contrast, dietary assimilation efficiency of Cd and Zn was not significantly enhanced following Zn pre-exposure. A positive linear relationship was found between the uptake rate constants of dissolved metals and Zn or MT concentrations in the fish. The results suggested that Zn pre-exposure increased the potential of metal uptake from ambient water, but had little effect on dietary metal uptake. Furthermore, the Zn body concentration and metal uptake from the dissolved phase were significantly dependent on the fish body size.

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.

The influence of lithium on biokinetics and biodistribution of 65Zn under different dietary protein regimens in rats.

The present study was designed to evaluate the effect of lithium on the biological half-lives of 65Zn in the rat liver and whole body and also its biodistribution under different dietary protein regimens. To carry out these investigations, each rat after three weeks of different treatments was injected with 1.85 MBq of 65Zn, intraperitoneally (ip) and its radioactivity was recorded in liver as well as in whole body by using gamma ray spectrometer. The radionuclide 65Zn showed two kinetic components indicating its biological half-life: Tb1 the initial fast component and the Tb2 or slow component. Protein deficiency led to a significant elevation of both components in the whole body but not in the liver when compared to the control group. High protein (HP) diet in rats resulted in marked reduction in both components in the liver but not in the whole body. Lithium treatment alone to normal rats caused a significant reduction in the Tb2 but not in the Tb1 component in the whole body as well as in the liver. The administration of lithium normalized the observed increase in both components in the whole body of protein deficient rats. On the contrary, in rats fed with low protein (LP) diet after lithium treatment the Tb(2) component was significantly reduced in the liver. Lithium treatment to HP fed rats, showed a significant decrease in both components in the liver when compared to normal rats: however this decrease was less pronounced in comparison to the HP group that had not been given lithium. Furthermore, one day prior to the end of the treatment period, the rats were injected ip with a tracer dose of 0.37 MBq 65Zn and 24 h later, various organs were excised for 65Zn biodistribution measurements in vitro. Under different dietary protein regimens as well as after administration of lithium the percent uptake values of 65Zn varied in various organs viz: the brain, liver, kidney, small intestine and large intestine as well as in the blood. In conclusion, different protein dietary conditions on co-administration with lithium in the rats seemed to have a dominant role for the retention of 65Zn in the liver the whole body and in other organs.

Release of zinc from the brain of El (epilepsy) mice during seizure induction.

Brain distribution after i.v. injection of 65ZnCl2 into El mice, an animal model of genetically determined epilepsy, was studied by autoradiography to study the utilization of zinc in the brain. The distribution of 65Zn in the brain of El mice 6 days after injection was almost the same as that of ddY (normal) mice, suggesting that the uptake of zinc by the brain of El mice is normal. To study the movement of zinc in the brain in the course of seizure induction, the concentrations of 65Zn in the brain of seizure-afflicted and untreated control El mice were compared 20 days after 65Zn injection. The concentration of 65Zn in the brain of seized El mice was overall lower than that of control El mice; the concentration of 65Zn was decreased notably in the piriform cortex and amygdaloid nuclei complex during convulsion. These results suggest that the release of zinc from the El mouse brain is enhanced during convulsion. The decrease in actively functioning zinc in the brain may be associated with the increase in susceptibility to seizure in the El mouse.

Differential hepatic response of 65Zn distribution between mice bearing experimental tumor and inflammation.

Discriminative detection of tumor and inflammation was tried by radio-imaging of hepatic uptake of 65Zn. This closely related to the level of metallothionein (MT) and reflected the extent of tumor growth in mice and rats transplanted with experimental tumor. The elevation of 65Zn distribution in liver of experimental tumor-bearing mice was inhibited by treatment with Zn-deficient diet, while stimulated by dexamethasone. This stimulation occurred 2 days after tumor transplantation, at which time 67Ga-citrate could not image the tumor. On the other hand, hepatic distribution of 65Zn was also elevated in mice by inducing experimental abscess, although the effect of both treatments on this elevation was different from the case of tumor; the elevation was inhibited by treatment with dexamethasone. These results suggest that radio-imaging of hepatic Zn uptake with a short-life gamma emitting isotope such as 69mZn with use of dexamethasone, if required, may be useful for a preliminary test to detect early-stage malignant disease.

65Zn uptake by liver of rats fed 3'-methyl-dimethylaminoazobenzene.

The uptake of 65Zn, determined by gamma-counting and also by autoradiography, significantly increased with the increasing level of metallothionein in liver 4 weeks after the start of feeding a diet containing 3'-methyl-dimethylaminoazobenzene (3'-Me-DAB), at which time the serum gamma-glutamyl transpeptidase activity was not yet significantly elevated. At this time, furthermore, hepatic uptake of 67Ga-citrate did not increase and that of 99mTc-Sn-colloid decreased in 3'-Me-DAB-fed rats. These results suggest that a short-life gamma-emitting isotope such as 69mZn may be useful for the detection at the early stage of hepatic carcinogenesis.

Preferential uptake of zinc, manganese, and rubidium in rat brain tumor.

The search for radionuclides that can be an index of viability in proliferating cells is important for nuclear medicine diagnosis of brain tumors. On the basis of the finding that 65Zn is preferentially taken up in rat brain tumors, the uptake of various radionuclides was examined in rat brain tumor by using the multitracer technique. Male Fisher rats were intrahippocampally injected with C6 glioma cells. Fourteen days after implantation, the radioactive multitracer solution was injected into the tail vein of tumor-bearing rats. One hour after intravenous injection, the uptake of 65Zn, 83Rb and 54Mn was relatively high in C6 glioma of 15 radionuclides detected, and was much higher than in other brain regions and in the blood. It is likely that rubidium and manganese, in addition to zinc, are preferentially taken up by tumors in the brain.

Biokinetics and dosimetry of chromium, cobalt, hydrogen, iron and zinc radionuclides in male reproductive tissues of the rat.

Following intravenous administration to male rats, the uptake and retention by reproductive tissues of chromium-51, cobalt-57, iron-59, zinc-65 and tritium has been studied for up to 28 days. Chromium-51, 57Co, 59Fe and 3H were not or only transiently accumulated in gonads or accessory sex glands at concentrations greater than whole body concentrations. However, 65Zn was concentrated in the dorsolateral region of the prostate gland and autoradiography showed preferential uptake by epithelial cells and lumen of glands. When combined with other information available from the literature, this data would suggest that current models adequately describe the biokinetics of chromium, cobalt, iron and tritium in the prostate and testes and zinc in the testes. Uptake of zinc by the prostate would appear to be best described by an average value of 0.1% and a conservative value of 0.5%. Allowing for greater uptake of zinc (0.5%) by the prostate, after inhalation of 65Zn in a soluble form increases prostate dose by about 3 fold compared to current models. The pessimistic assumptions of a higher relative biological effectiveness (20) for all Auger emissions from 65Zn in cell nuclei and a heterogeneous distribution of 65Zn to sensitive cells in the prostate increases prostate dose by a further factor of 9. Even on the basis of these cautions estimates, occupational exposures to radioisotopes of these elements do not explain the excess of prostate cancer reported amongst some nuclear workers.

Comparison of cytogenetic damage in cultured cells from cobalt-60 gamma-radiation and the Auger emitter zinc-65.

PURPOSE: To assess the ability of the Auger-emitting nuclide, zinc-65 (65Zn), relative to gamma-irradiation, to cause chromosomal aberrations in cultured rat prostate cells. MATERIALS AND METHODS: Rat prostate adenocarcinoma cells in culture were exposed to doses of 1, 2, 3 or 5 Gy of external gamma-irradiation for 24h or incubated with 0.7, 1.5, 1.8 or 2.8 MBq of 65Zn for 24 h. The uptake by and clearance from cells of 65Zn was measured. Metaphase spreads prepared from washed cells were scored for chromatid- and chromosome-type aberrations. RESULTS: Following exposure to 65Zn or gamma-irradiation, chromatid-type damage was more commonly observed than chromosome-type aberrations. The relationship between induced chromatid damage and gamma dose (to 3 Gy) was best fitted by a second-order polynomial function, while the activity response relationship for chromatid damage caused by 65Zn appeared to be best fitted by a straight line. Measurements of the uptake of 65Zn by cells showed that average concentrations within cells were about 100 times the concentration in the culture medium. Assuming uniform distribution of 65Zn within cells, with 36% in the nucleus, the dose was estimated as 0.70 Gy per MBq added 65Zn, with Auger electrons contributing most (93%) of the dose. Assuming that 20% of cellular zinc was localized in the nucleus, based on previous measurements, the dose to the nucleus was calculated as 0.44 Gy per MBq added 65Zn. RBE values for chromatid damage induced by 65Zn compared to gamma-radiation range from about 1 to 3 based on a uniform dose throughout the cell and from about 2 to 5 based on 20% of 65Zn in the cell nucleus. CONCLUSION: The observed radiotoxicity of 65Zn is consistent with its behaviour as an Auger-emitting radionuclide that is localized to some extent in the nucleus.

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.

Human ZIP1 is a major zinc uptake transporter for the accumulation of zinc in prostate cells.

The prostate gland of humans and other animals accumulates a level of zinc that is 3-10 times greater than that found in other tissues. Associated with this ability to accumulate zinc is a rapid zinc uptake process in human prostate cells, which we previously identified as the hZIP1 zinc transporter. We now provide additional evidence that hZIP1 is an important operational transporter that allows for the transport and accumulation of zinc. The studies reveal that hZIP1 (SLC39A1) but not hZIP2 (SLC39A2) is expressed in the zinc-accumulating human prostate cell lines, LNCaP and PC-3. Transfected PC-3 cells that overexpress hZIP1 exhibit increased uptake and accumulation of zinc. The V(max) for zinc uptake was increased with no change in K(m). Along with the increased intracellular accumulation of zinc, the overexpression of hZIP1 also results in the inhibition of growth of PC-3 cells. Down-regulation of hZIP1 by treatment of PC-3 cells with hZIP1 antisense oligonucleotide resulted in a decreased zinc uptake. Uptake of zinc from zinc chelated with citrate was as rapid as from free zinc ions; however, the cells did not take up zinc chelated with EDTA. The cellular uptake of zinc is not dependent upon an available pool of free Zn(2+) ions. Instead, the mechanism of transport appears to involve the transport of zinc from low molecular weight ligands that exist in circulation as relatively loosely bound complexes with zinc.

Whole-body retention and fetal uptake of 65Zn in pregnant mice fed a Zn-deficient diet.

Whole-body retention and fetal uptake of 65Zn under a Zn-deficient diet were studied in pregnant mice in the late gestational stage after a single oral administration of 65Zn. Whole-body retentions were much greater in mice given a Zn-deficient diet than in those given a Zn-normal diet. Accordingly, the amount of 65Zn transmitted to the offspring in utero was greater in the Zn-deficient diet group. In another experiment, fetal uptake of 65Zn in dams on gestation day 17 was examined over a period of 24 hr after a single intravenous administration of 65Zn to the Zn-deficient and Zn-normal animals. There was no major difference in fetal uptake between the two groups, indicative that approximately a similar proportion of the 65Zn retained in the maternal body was transmitted to the in utero offspring in both groups.