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Release and absorption of zinc from zinc oxide and zinc sulfate in open wounds.

The pharmacokinetic behaviours of zinc oxide and zinc sulfate when applied as single doses to full-thickness excised rat skin wounds were studied. In the zinc oxide group, the wound fluid zinc concentration increased slightly over the 48-h postoperative period due to increased solubilization of zinc oxide, attributed to increased protein concentration of the wound fluid. When zinc sulfate was applied to the wounds, the wound fluid zinc concentration decreased rapidly during the first 4 postoperative h and then at a slower rate. The changes in the serum zinc level followed essentially the same kinetic pattern as that of the wound fluid zinc levels. The zinc concentration of the wounded tissue remained almost constant in zinc oxide treated wounds whilst it diminished in zinc sulfate treated wounds. In conclusion, zinc oxide delivers zinc ions to wounds over an extended period of time which results in constant wound tissue zinc levels. In contrast, zinc sulfate rapidly delivers zinc ions which results in decreasing tissue zinc levels.

Adult

[Determination of metabolic availability of zinc and estimation of zinc requirements through changes in the activity of metallo-zinc enzymes].

Data on the zinc requirements of animals vary widely with the type of diet or kind of zinc compound considered, as they are dependent on the kind of definition and estimation of mineral requirements used which, in turn, are based on the analytical content of the mineral in the diet. Actually, requirements for a particular trace element are the same for any diet provided differences in the rate of absorption and metabolic availability of the element are taken into consideration. A suggestion is made in the present paper to utilize changes in the activity of different metallo-zinc enzymes as a method for determining the degree of metabolic availability of zinc. Within the framework of a repletion trial young male rats were fed a semisynthetic casein diet containing 1.2 ppm of zinc for a 14-day period of zinc depletion followed by a 15-day repletion period during which the rats received a diet containing 4.5 ppm or 12 ppm of zinc. In this trial the pattern of activity of the alkaline phosphatase in serum and in the femoral bones was studied and the level of pancreatic carboxypeptidase A and B was estimated. The data thus obtained were compared with those of pair-fed control animals and control animals fed a libitum. (96 ppm of dietary zinc). The results of the present trial show the metabolic availability of zinc can be determined by measuring the level of activity of these metallo-zinc enzymes. A suitable model was established. Moreover, attempts have been made to show in which way the zinc demands of growing, full-grown, pregnant and lactating animals may be estimated by measuring changes in the degree of activity of these metallo-zinc enzymes. A definition is, in this case, given by that amount of zinc which allows for an optimum activity of the different metallo-zinc enzymes. Such data are applicable only to a particular diet or are valid only for a particular zinc compound. Principally, it is much better to relate the Zn requirements of the animal to the level of absorbable and metabolically available zinc. The present model has been suggested to provide a possibility for measuring the metabolic availability of zinc in the body.

Alkaline Phosphatase

Effects of dietary zinc upon tissue zinc and percent unsaturated plasma-zinc binding capacity.

Calves were fed 520 ppm zinc for 21 days, then placed on a low zinc diet (20 ppm zinc) for 35 days to determine the biological availability of zinc reserves of tissue. Concentrations of zinc in liver and kidney were elevated greatly by feeding high dietary zinc for 21 days and continued to increase with time. On the low zinc diet, the tissue burden of zinc decreased within 35 days to those comparable to calves initially fed low zinc. Likewise, both zinc in plasma and percent capacity of unsaturated plasma for binding zinc were near negative controls 21 days after removal of calves from high zinc diets. Tissue stores of zinc will not maintain adequate zinc in blood for extended periods, and hence, they probably will not sustain optimal feed intakes and growth rates of calves on low-intakes of zinc.

Animals

Methodology for assessing zinc bioavailability: efficacy estimates for zinc-methionine, zinc sulfate, and zinc oxide.

The bioavailability of zinc-methionine (ZnMET) was compared to that of feed-grade ZnSO4.H2O using three different diets: purified (crystalline amino acid [AA]), semipurified (soy isolate), and complex (corn-soybean [C-SBM]) diet. With the Zn-deficient purified or semipurified diet, weight gain and tibia Zn responded linearly to both ZnSO4.H2O and ZnMET supplementation. Common-intercept, multiple linear regression indicated differences in Zn bioavailability between ZnMET and ZnSO4.H2O for both diets as indicated by bone Zn. With the ZnSO4.H2O standard set at 100%, bioavailability of Zn from ZnMET was 117% (P less than .05) in the AA diet and 177% (P less than .01) in the soy isolate diet. The ZnMET was also compared to ZnSO4.H2O in a C-SBM diet containing 117 mg of Zn/kg. When high levels of Zn were added to this diet (0, 250, 500, and 750 mg/kg of supplemental Zn), consistent tissue Zn responses did not occur beyond the first increment. Addition of lower levels of supplemental Zn (0, 5, 10, 20, 30, 40 and 50 mg/kg) to a Zn-unsupplemented C-SBM basal diet (45 mg/kg of Zn), however, resulted in a broken-line, two-slope response in tibia Zn for both ZnMET and ZnSO4.H2O. Inflection points occurred at 60 and 54 mg of Zn/kg of diet for ZnSO4.H2O and ZnMET, respectively. The ratio of slopes (ZnMET:ZnSO4.H2O) below the inflection points was 206% (P less than .01), indicating that Zn was considerably more bioavailable in ZnMET than in ZnSO4.H2O for chicks consuming C-SBM diets. When feed-grade ZnO was compared to feed-grade ZnSO4.H2O in chicks consuming C-SBM diets, bone Zn slopes below the respective inflection points indicated that Zn was 61% bioavailable in ZnO relative to ZnSO4.H2O.

Animal Feed

Influence of parenteral zinc and actinomycin D on tissue zinc uptake and the synthesis of a zinc - binding protein.

Parenterally administered zinc markedly increased the incorporation of 14-C-cystine and 65-Zn into a low molecular weight zinc binding protein (ZnBP) isolated from liver cytoplasm of rats fed an adequate amount of zinc. This zinc load significantly increased the zinc content in the liver. The increase in hepatic zinc content was inhibited by actinomycin D indicating that DNA-dependent RNA synthesis is required for zinc uptake into liver. Antinomycin D also produced a concomitant decrease in ZnBP synthesis indicating that this protein may be involved in the uptake mechanism in cells. Zinc repletion also stimulated the synthesis of hepatic ZnBP in zinc deficient rats. This stimulation was also prevented by prior administration of actinomycin D. A similar effect was observed in the intestinal mucosal cells. The data collectively indicate that the control of the synthesis of ZnBP which occurs at the transcriptional level of protein synthesis is responsive to zinc status and thus may have a function in zinc metabolism.

Animals

Intracellular distribution of zinc and zinc-65 in calves receiving high but nontoxic amounts of zinc.

The zinc homeostatic control breakdown in cattle fed a high but nontoxic amount of zinc was investigated. Liver copper was decreased by the 600 ppm added dietary zinc indicating altered copper metabolism. However, duodenal copper, liver and duodenal iron and manganese were not affected. Zinc-65 in blood was reduced 90% by the high dietary zinc 48 h following oral zinc-65 dosing. The 600 ppm supplemental zinc increased zinc by 500% in liver, 20 times in pancreas and kidney, and 100% in the duodenum. The increased liver and duodenal zinc was confined largely to the soluble cell fraction. This concentrating of excess tissue zinc in the soluble fraction may be an adaptive mechanism which detoxifies large quantities of zinc and prevents disruption of normal cellular activity.

Animals

Effect of zinc status on salivary zinc concentrations in the rat.

Three experiments were done to determine whether salivary zinc concentration is a more sensitive indicator of zinc status than plasma zinc. Weanling male rats fed a low zinc (less than 1 ppm) diet for 5 weeks with or without zinc (100 ppm) in the drinking water had salivary zinc concentration of 0.19, 0.16, and 0.20 microgram/ml for the zinc-deficient, zinc-supplemented restricted-intake, and zinc-supplemented ad libitum-fed groups, respectively. Combined values for male and female rats after 4 weeks of the same treatments in experiment 2 were 0.60, 1.2 and 0.44 microgram/ml. Saliva collected on day 22 of pregnancy contained 0.30 and 0.24 microgram/ml from zinc-supplemented and zinc-deficient rats, respectively. Salivary zinc concentrations in the deficient rats did not differ from those of the zinc-supplemented ad libitum-fed controls in any of the experiments. Salivary zinc concentration in the zinc-supplemented restricted-intake group in experiment 2 was significantly higher than that in the other two groups. Decreases in serum, bone, and fetal zinc concentrations indicated that the rats were definitely zinc-deficient. Since zinc concentration of mixed saliva in the rat was not decreased by even a severe zinc deficiency, salivary zinc does not appear to be as good an indicator of zinc status as plasma zinc.

Animals

Zinc uptake by blood cells of rats in zinc deficiency and inflammation.

In zinc deficiency, the function of leukocytes is impaired. However, the results of studies on the zinc concentration of blood cells in zinc deficiency are conflicting, probably in part because of technical and analytical problems. The aim of this study was to investigate, under standard conditions, the uptake of 65Zn-labeled zinc by blood cells, taken from zinc-deficient rats and from rats in which an inflammation is induced. In both conditions, the serum zinc concentration is reduced. In clinical practice, this makes it difficult to determine whether the decrease in serum zinc is the result of a real or an apparent zinc deficiency. In stress, like an inflammatory disease, the decrease of zinc reflects an apparent zinc deficiency because of redistribution of serum zinc into the liver and because of decrease in serum albumin concentration. Over 70% of the serum zinc is bound to albumin. Blood cells from zinc-deficient and control rats were isolated using a discontinuous Percoll gradient and incubated under nearly physiological conditions in a 65Zn-containing medium. A significant increase in the in vitro uptake of 65Zn-labeled zinc by the blood cells of zinc-deficient rats was seen: erythrocytes 1.3, mononuclear cells 2.0, and polymorphonuclear cells 2.6 times the control values. During inflammation, no change in 65Zn-labeled zinc uptake by erythrocytes and mononuclear cells was demonstrated after 2 d, although the serum zinc and albumin concentrations were decreased, but a small but significant increase in zinc uptake by polymorphonuclear cells was observed. This study of 65Zn uptake in vitro under standard conditions may prove of value for distinguishing in patients real zinc deficiency from apparent zinc deficiency owing to, e.g., stress, although additional experiments should be performed.

Animals

Dietary protein, as egg albumen: effects on bone composition, zinc bioavailability and zinc requirements of rats, assessed by a modified broken-line model.

The effect of dietary protein concentration on zinc bioavailability, requirements and incorporation into bones was investigated in growing rats. Zinc requirements were determined by the broken-line method. Protein did not affect either absorption or biological half-life of 65Zn added to the diet. Zinc requirements based on weight gain or tibia zinc were generally greater when rats were fed 30% rather than 15% egg white. When fed 30% rather than 15% egg white, zinc-deficient rats gained less weight and tended to incorporate less zinc into bone, whereas zinc-adequate rats gained weight similarly and incorporated more zinc into bone. Dietary protein concentration apparently elevated the maximum amount of zinc incorporated into bones of rats fed adequate zinc. When dietary zinc was adequate (25 mg/kg diet), tibia zinc concentrations increased linearly with dietary concentrations of 15, 25, 35 and 45% egg white. When dietary zinc was adequate, higher protein diets resulted in lower tibia nitrogen, and higher tibia zinc, without substantial changes in tibia size or calcium concentration. These results indicate that high protein diets increase zinc requirements and bone zinc deposition, the latter being a consequence of altered bone zinc metabolism, rather than improved zinc bioavailability.

Administration, Oral

Effects of early wound healing and wound treatment with zinc tape on intestinal absorption and distribution of zinc in rats.

In rats operated with large excisional skin wounds which were not treated with zinc tape, the serum zinc concentration was decreased at every observation time during the first post-operative day and the absorption of zinc from the intestine was increased. Slight decreases in the zinc concentration were seen in some tissues while liver zinc was very high. In rats with wounds treated with zinc tape, serum zinc was markedly increased at 6 hours, lower but above control levels at 7 and 12 hours and markedly increased again at 24 hours. The absorption of zinc from the intestine was low. Zinc concentration were elevated in most tissues sampled; they were high in testis and kidney and very high in liver. The results strongly indicate that the operative trauma and/or acute inflammatory conditions in the wounds in animals not treated with zinc tape initiate a movement of zinc from serum, some tissues and the content of the intestine to the liver. In the zinc tape treated group the increase in liver zinc was supplied mostly by the zinc tape. It is assumed that these large increases in zinc concentration in the liver are the result of the incorporation of zinc into certain zinc containing proteins which are produced in response to the inflammatory process in the wound. An increase was also found in the copper concentration in the liver of zinc tape treated animals and a decrease in the iron concentration in the liver of both groups of operated animals compared to controls.

Absorption

Zinc absorption through skin: correction of zinc deficiency in the rat.

The therapeutic effect of topical administration of zinc was tested in pregnant rats consuming a diet deficient in the element. Four groups of rats were fed a zinc-deficient diet for 24 hr. Half of the animals were treated during this period with a topical application of oil saturated with zinc chloride, for the full 24 hr in one group, and for the last 8 hr in the other. In the two remaining groups, oil without zinc chloride was applied under the same conditions as described above, and in all cases oral ingestion of the supplement was prevented. At the end of the 24-hr period, the animals were killed and plasma zinc was determined. An additional group of animals consuming a diet adequate in zinc was killed without any treatment to provide control values of normal plasma zinc. Rats consuming the deficient diet and without topical zinc supplementation had plasma zinc values significantly lower than all other groups after 24 hr. Animals receiving zinc supplementation for 8 hr had plasma levels similar to those of the control group fed an adequate zinc diet and significantly higher than those of rats that received no zinc application to the skin. In animals in which zinc was applied for 24 hr, plasma zinc values were significantly higher than in any other group, including normal controls. The results indicate that percutaneous transport of zinc may be of sufficient magnitude to be clinically significant and that topical application of this element may be useful in cases of dietary zinc deficiency or diseases producing a zinc deficiency state.

Animals

Smoking alters the relationship between maternal zinc intake and biochemical indices of fetal zinc status.

Maternal smoking impairs fetal zinc status. This study was designed to clarify the effect of smoking on the relationship between maternal zinc intake and zinc status in mother and fetus. Zinc was measured with atomic-absorption spectroscopy. Statistical analyses consisted of descriptive statistics, simple correlations, and stepwise multiple regression. The results suggest that maternal plasma zinc, red blood cell zinc, and alkaline phosphatase at term are not related to maternal zinc intake. In the nonsmoking parturient both cord-vein plasma zinc and cord-vein alkaline phosphatase activity are positively related to maternal zinc intake. In the smoking parturient there is no relationship between maternal zinc intake and fetal zinc status except for a negative relation with cord-vein plasma zinc. Relations between maternal zinc intake and placental zinc can be shown with stepwise-multiple-regression techniques. The data suggests that maternal zinc intake is related not to maternal zinc status but to fetal zinc status in a normal pregnancy. The relation is altered in the pregnancy complicated by smoking.

Alkaline Phosphatase

Zinc metabolism and homeostasis in rats fed a wide range of high dietary zinc levels.

Zinc metabolism and homeostasis were studied in young growing rats fed a 38 ppm zinc diet with added zinc levels ranging from 0 to 8400 ppm for 21 days. High dietary zinc did not cause toxicity symptoms. Stable zinc in feces increased linearly with dietary zinc intake but fecal 65Zn, from a single oral dose, did not increase above the 1200 ppm dietary level. Stable zinc in liver, kidney, and tibia increased two to three times with 2400 ppm added zinc, but was not further elevated until 8400 ppm was fed. Stable zinc in muscle and heart was not affected appreciably by dietary zinc level. In all tissues, 65Zn retention was drastically reduced with 1200 ppm added dietary zinc. Additional dietary zinc reduced 65Zn in muscle and heart but had little effect on liver and kidney 65Zn. The data indicate that rats have fairly effective homeostatic control mechanisms for tissue zinc below about 7200 ppm dietary zinc. Whereas, with dietary zinc up to about 1200 ppm, decreasing absorption is the main route of homeostatic control, above this level, more rapid zinc turnover rates and increasing endogenous zinc excretion appear to have major importance.

Animals

Zinc excretion in young women on low zinc intakes and oral contraceptive agents.

Zinc excretion under conditions of negligible zinc intake (0.17 mg/day) was measured in women taking a combination oral contraceptive agent (+OCA) and in women with normal menstrual cycles (-OCA). A semipurified, constant formula diet, providing negligible amounts of zinc, copper and iron, but adequate levels of other essential nutrients, was fed. During the 35--day study, serum and urinary zinc declined more markedly in the +OCA than -OCA groups. Serum zinc dropped 47% in the +OCA and 21% in the -OCA; urinary zinc declined 83% and 62% in the two groups. After pre-study, zinc was cleared from the gut, fecal zinc decreased about 40% in both groups. Whole body integumental zinc losses were similar for both groups, about 0.7 mg/day. For -OCA, zinc losses via this route were higher during the luteal phase than during menstruation. Menstrual fluid zinc contents were negligible and similar for both groups, about 5 microgram/day. The data suggest that accessible zinc stores are not extensive and that depletion of these stores, as a result of the low-zinc diet, caused the fall in serum zinc. The use of OCA influenced the response to the low-zinc diet, but endogenous zinc losses calculated for day 1 were nearly the same for both groups, about 1.6 mg/day.

Adolescent

Plasma whole blood and urine zinc in the assessment of zinc deficiency in the rat.

Serial measurements of plasma, whole blood, and urine zinc have been made in young adult rats fed zinc deficient and zinc supplemented diets for a total period of 65 days. After commencing a zinc deficient diet plasma zinc fell within 48 hours to 25% of control values and remained at this level throughout the period of study. A substantial diurnal variation was observed in plasma zinc, and the level was significantly lower in the fasting state. Whole blood zinc was unaffected by fasting, showed no diurnal variation, and remained at control values even after 65 days of zinc deficient diet. Urine zinc fell to very low values, 10 days after commencing a zinc deficient diet, but after 30 days rose to control values. Under controlled conditions plasma zinc can be used as an indication of zinc deficiency. Whole blood zinc is of no value in the detection of zinc deficiency. Urine zinc concentration varies with time following a zinc deficient diet.

Animals

Local absorption of zinc from wounds treated with different concentrations of zinc sulphate.

In the present study it was shown in rats that zinc is absorbed from excisional wounds treated with zinc sulphate. Systemic toxic effects were observed in the group treated with 20% zinc sulphate. Local toxic effects were seen in wounds treated with 0.2%, 2% and 20% zinc sulphate. An inhibitory effect of zinc on the migration of granulocytes was suggested on the basis of microscopic observation. In the operated groups which were not treated with zinc and the group treated with 0.02% zinc sulphate a decline was observed in the concentration of zinc in serum. The zinc concentration in serum increased in proportion to the zinc sulphate concentration (0.2%, 2% and 20%) applied to the wounds, while the copper concentration decreased in the groups treated with 2% and 20% zinc sulphate. In all operated groups an increase in zinc and copper concentrations was observed in liver. This was most pronounced in groups treated with higher concentrations of zinc sulphate (0.2%, 2% and 20%). The groups treated with higher concentrations of zinc sulphate also had higher pancreas zinc concentrations than the remaining groups.

Absorption