Metallothionein-null mice absorb less Zn from an egg-white diet, but a similar amount from solutions, although with altered intertissue Zn distribution.
The influence of metallothionein (MT) on Zn transfer into non-gut tissues was investigated in MT-null (MT-/-) and normal (MT+/+) mice 4 h after oral gavage of aqueous 65ZnSO4solution at doses of 154, 385, 770 and 1540 nmol Zn per mouse. Zn transfer was not significantly different between MT+/+ and MT-/- mice and was directly proportional to the oral dose (slope = 0.127, r = 0.991; 0. 146, r = 0.994, respectively). Blood 65Zn and plasma Zn concentrations increased progressively in MT-/- mice at doses >154 nmol Zn, reaching levels of 2.4% of oral dose and 60 micromol/L, respectively, at the 1540 nmol Zn dose. The corresponding values for MT+/+ mice were approximately half, 1.0% and 29 micromol/L. Intergenotypic differences were found in tissue distribution of 65Zn within the body; MT-/- mice had higher 65Zn levels in muscle, skin, heart and brain, whereas MT+/+ mice retained progressively more Zn in the liver, in conjunction with a linear increase in hepatic MT up to the highest Zn dose. MT induction in the small intestine reached its maximum at an oral dose of 385 nmol Zn and did not differ at higher doses. Absorption of a 770 nmol 65Zn dose from a solid egg-white diet was only one fourth (MT+/+) and one eighth (MT-/-) of the Zn absorption from the same dose of 65Zn in aqueous solution. MT+/+ mice had greater (P < 0.05) Zn absorption from the egg-white diet than did MT-/- mice, indicating that gut MT confers an absorptive advantage, but only when Zn is incorporated into solid food. (+info)
The effect of pregnancy and a chronic, marginal intake of zinc on zinc kinetics was studied in rats. Weanling female rats were fed either a zinc-adequate diet, containing 30 microg Zn/g, (30Zn) (n = 16) or a marginally zinc-deficient diet, containing 6 microg Zn/g, (6Zn) (n = 16). After 6 wk, half of each group was mated (30ZnPG, 6ZnPG). A third group of pregnant rats was pair-fed (PFPG) (n = 6) to the 6ZnPG group. On d 20 of gestation, or at the end of the 9-wk study, 65Zn was injected intravenously. The plasma 65Zn disappearance curve over the next 105 min was used to study the size and fractional turnover rates of two rapidly exchanging zinc metabolic pools (pool a and pool b). Plasma zinc concentrations on d 20 of gestation were significantly lower in the 6ZnPG group compared with the 30ZnPG and PFPG controls, (P < 0.05). The exchangeable pools were also smaller in the 6ZnPG group compared with the 30ZnPg and PFPG groups, (P < 0.02); this reduction was accompanied by a 60% greater fractional turnover rate of pool a, (P < 0.02). Pregnancy outcomes did not differ among the three groups. We conclude that there is an increase in the turnover rate of the exchangeable plasma zinc pool when dietary zinc intake is marginal during pregnancy. This response may help maintain a supply of zinc to the growing fetus when plasma zinc concentrations are reduced. (+info)
Evidence for a zinc uptake transporter in human prostate cancer cells which is regulated by prolactin and testosterone.
The glandular epithelial cells of the human prostate gland have the unique capability and function of accumulating the highest zinc levels of any soft tissue in the body. Zinc accumulation in the prostate is regulated by prolactin and testosterone; however, little information is available concerning the mechanisms associated with zinc accumulation and its regulation in prostate epithelial cells. In the present studies the uptake and accumulation of zinc were determined in the human malignant prostate cell lines LNCaP and PC-3. The results demonstrate that LNCaP cells and PC-3 cells possess the unique capability of accumulating high levels of zinc. Zinc accumulation in both cell types is stimulated by physiological concentrations of prolactin and testosterone. The studies reveal that these cells contain a rapid zinc uptake process indicative of a plasma membrane zinc transporter. Initial kinetic studies demonstrate that the rapid uptake of zinc is effective under physiological conditions that reflect the total and mobile zinc levels in circulation. Correspondingly, genetic studies demonstrate the expression of a ZIP family zinc uptake transporter in both LNCaP and PC-3 cells. The rapid zinc uptake transport process is stimulated by treatment of cells with physiological levels of prolactin and testosterone, which possibly is the result of the regulation of the ZIP-type zinc transporter gene. These zinc-accumulating characteristics are specific for prostate cells. The studies support the concept that these prostate cells express a unique hormone-responsive, plasma membrane-associated, rapid zinc uptake transporter gene associated with their unique ability to accumulate high zinc levels. (+info)
Performance of a 62Zn/62Cu generator in clinical trials of PET perfusion agent 62Cu-PTSM.
The 62Zn/62Cu PET generator can be inexpensively produced and distributed from a single production site operating under typical good manufacturing practice guidelines. It therefore has the potential to greatly facilitate development of clinically practical PET. We report generator performance in a study in which 62Cu-pyruvaldehyde-bis(n4-methylthiosemicarbazone (PTSM) myocardial perfusion imaging is compared with 99mTc-sestamibi in the diagnosis of coronary artery disease. The 62Zn/62Cu generator is an improved version of a previously reported system that employs automated synthesis of 62Cu-PTSM. With this approach, the cumbersome step of 18C purification has been eliminated. METHODS: The 62Zn (9.3 h half-life) parent isotope is prepared by proton bombardment of natural copper at 33 MeV. A typical target irradiated with 37.5 microA/h is delivered by 12:00 PM on the day it is to be processed. Purified 62Zn obtained from the target is loaded onto the generator column in 2 mol/L HCl. The generator is eluted using an internal three-channel peristaltic pump, which delivers 2.25 mL eluant (1.8 mol/L NaCl, 0.2 mol/L HCl) through the generator column to elute the 62Cu in 40 s. The same pump simultaneously pumps an equal volume of buffer (0.4 mol/L NaOAc) and 1 mL ligand solution (2 ppm PTSM, 2% EtOH) passing it through a septum into a 35-cc syringe preloaded with 28 mL sterile water. This solution is thoroughly mixed by agitation of the syringe and injected as a bolus through a 0.2 microm filter. The generator is eluted twice before shipping, providing quality assurance samples, and shipped to the clinical site by overnight delivery. Complete quality assurance testing is performed the evening before the generator reaches the clinical site. RESULTS: A total of 34 generators have been produced and shipped to 2 clinical sites for a phase III Food and Drug Administration study. The load activity on the generators at 8:00 AM the day of clinical use was 1.7+/-0.2 GBq (46.7+/-5.6 mCi), and yield was 72%+/-16%. Breakthrough of 62Zn was undetectable by high-purity germanium spectroscopy for all units. Radiochemical purity was 95.4%+/-2.4%. Volume delivered, pH, sterility, and bacterial endotoxin tests yielded passing results on all generators. The entire process of generator production, from target receipt to generator shipment, took less than 6 h and cost approximately $1000, including shipping charges and cyclotron cost. A total of 68 patients were injected with 2 62Cu-PTSM doses, with a mean injected activity of 0.8+/-0.2 GBq (20.5+/-5.3 mCi) with no adverse side effects. CONCLUSION: Results of this work confirm that the 62Zn/62Cu generator is an easily produced, transportable, and inexpensive source of PET radiopharmaceuticals, which can expand the field of clinical PET imaging by providing radiopharmaceuticals to sites not associated with cyclotrons. (+info)
Zinc transport and metallothionein secretion in the intestinal human cell line Caco-2.
Caco-2, a human cell line, displays several biochemical and morphological characteristics of differentiated enterocytes. Among these is the ability to transport zinc from the apical to the basal compartment. This process was enhanced following exposure by the apical compartment to increasing concentrations of the metal. High pressure liquid chromatography fractionation of the media obtained from cells labeled with radioactive zinc showed that metallothioneins (MTs), small metal-binding, cysteine-rich proteins), were present in the apical and basal media of controls as well as in cells grown in the presence of high concentrations of zinc. Following exposure to the metal, the levels of Zn-MTs in the apical medium increased, while in the basal compartment the greatest part of zinc appeared in a free form with minor changes in the levels of basal MTs. Metabolic labeling experiments with radioactive cysteine confirmed the apical secretion of MTs. A stable transfectant clone of Caco-2 cells (CL11) was selected for its ability to express constitutively high levels of the mouse metallothionein I protein. This cell line showed an enhanced transport of the metal following exposure to high concentrations of zinc and a constitutive secretion of the mouse metallothionein I protein in the apical compartment. Together, these findings strongly support the hypothesis of a functional role between the biosynthesis and secretion of MTs and the transport of zinc in intestinal cells. (+info)
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. (+info)
Remobilization of cadmium in maturing shoots of near isogenic lines of durum wheat that differ in grain cadmium accumulation.
Cadmium accumulation in grain of durum wheat (Triticum turgidum L. var. durum) represents a concern to consumers. In an effort to understand the regulation of Cd accumulation in maturing grain, the remobilization of 109Cd applied to stem and flag leaves was examined in two near-isogenic lines that differ in grain Cd accumulation. Absorbed 109Cd was primarily retained in the labelling flap (50-54% and 65-80% for stem and flag leaves, respectively). Cadmium exported from the stem flap initially (3 d) accumulated in the stem in a declining gradient towards the head. Subsequent remobilization of Cd deposited in the stem was associated with Cd accumulation in the grain. Cadmium exported from the flag leaf flap was primarily directed to the grain. Little (<1%) Cd accumulated in the glumes or rachis, and transport of Cd to shoot tissues below the flag leaf node was low (<1%). On average, 9% and 17% of absorbed 109Cd accumulated in the grain 14 d after labelling the stem and flag leaf, respectively. Irrespective of labelling position, the low Cd-accumulating isoline averaged 1.5-2-fold lower Cd accumulation per grain and Cd concentration in the grain than the high Cd-accumulating isoline. Cadmium accumulation in the grain was inversely correlated with Cd retention in the stem (stem labelled) and labelling flap (flag leaf labelled) for both isolines. Cadmium translocation to the grain was not inhibited by Zn when both were applied simultaneously (50 pM 109Cd; 0.5 microM 65Zn) to the flag leaf. These results show that elevated remobilization of Cd from the leaves and stem to the maturing grain may be partially responsible for the high accumulation of Cd in durum wheat grain. (+info)
Characteristics of cadmium uptake in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens.
Uptake of Cd and Zn by intact seedlings of two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens was characterized using radioactive tracers. Uptake of Cd and Zn at 2 degrees C was assumed to represent mainly apoplastic binding in the roots, whereas the difference in uptake between 22 degrees C and 2 degrees C represented metabolically dependent influx. There was no significant difference between the two ecotypes in the apoplastic binding of Cd or Zn. Metabolically dependent uptake of Cd was 4.5-fold higher in the high Cd-accumulating ecotype, Ganges, than in the low Cd-accumulating ecotype, Prayon. By contrast, there was only a 1.5-fold difference in the Zn uptake between the two ecotypes. For the Ganges ecotype, Cd uptake could be described by Michaelis-Menten kinetics with a V(max) of 143 nmol g(-1) root FW h(-1) and a K(m) of 0.45 microM. Uptake of Cd by the Ganges ecotype was not inhibited by La, Zn, Cu, Co, Mn, Ni or Fe(II), and neither by increasing the Ca concentration. By contrast, addition of La, Zn or Mn, or increasing the Ca concentration in the uptake solution decreased Cd uptake by Prayon. Uptake of Ca was larger in Prayon than in Ganges. The results suggest that Cd uptake by the low Cd-accumulating ecotype (Prayon) may be mediated partly via Ca channels or transporters for Zn and Mn. By contrast, there may exist a highly selective Cd transport system in the root cell membranes of the high Cd-accumulating ecotype (Ganges) of T. caerulescens. (+info)