Trypsin activity in the small intestine of rats after application of copper and zinc

In vivo experiments with Sprague-Dawley rats were conducted in order to explore the influence of Cu²⁺, Zn²⁺ as well as of the combinations of both on the activity of trypsin. The solutions of the trace elements were given per os, the animals were killed 30 min after the applications, and the activity of trypsin was determined in the juice of the small intestine by usingN ^α-benzoyl-L-arginine-p-nitroanilide (L-BAPA) as the substrate. The activity of trypsin depends on the concentration of the trace elements. When Cu²⁺ ions are applied, there is a minimum activity at 10⁻⁵ mol Cu²⁺/L and a maximum at 10⁻⁴ mol Cu²⁺/L. When giving Zn²⁺ ions, a minimum of trypsin activity is found at 10⁻⁵ mol Zn²⁺/L and a maximum at 5×10⁻⁶ mol Zn²⁺/L. On the whole, the trypsin activity is lower when the Cu²⁺/Zn²⁺ combinations are applied compared to the addition of the single trace elements. On principle, a good conformity of the in vivo results was found with in vitro results.     

Investigations into combined dietary deficiencies of copper,selenium, and vitamin E in the rat

Interactions between dietary Cu, Se, and vitamin E in ascorbate-induced hemolysis of erythrocytes obtained from rats fed diets deficient or adequate in these elements were investigated. Hemolysis was affected by all three dietary factors, through closely interrelated but distinct mechanisms. In vitamin E-deficient cells, hemolysis was increased and the amount of hemolysis was directly related to the amount of hemoglobin breakdown. Deficiency of Cu or Se decreased hemolysis, but only in vitamin E-deficient cells. Vitamin E did not affect the breakdown of hemoglobin, but Cu and Se did. Hemolysis and hemoglobin breakdown were decreased by the addition of glucose, through mechanisms independent of that involving reduced glutathione metabolism. These results suggest that vitamin E acts within erythrocyte membranes to prevent products of hemoglobin breakdown from initiating peroxidation and subsequent hemolysis. Effects of Cu and Se are linked with that of vitamin E by the involvement of glutathione peroxidase and Cu superoxide dismutase in the cytoplasmic breakdown of hemoglobin, rather than by a direct effect of these enzymes on lipid peroxidation. It is concluded that the erythrocyte, because of its high heme content, probably represents a special system in terms of peroxidative pathways, and these findings may not be directly applicable to other tissues.     

Effects of dietary zinc levels on the activities of enzymes, weights of organs, and the concentrations of zinc and copper in growing rats

Zinc (Zn) is an essential nutrient that is required in humans and animals for many physiological functions, including immune and antioxidant function, growth, and reproduction. The present study was performed to investigate the effects of three Zn levels, including Zn adequate (35.94 mg/kg, as a control), Zn deficiency (3.15 mg/kg), and Zn overload (347.50 mg/kg) in growing male rats for 6 wk. This allowed for evaluation of the effects that these Zn levels might have on body weight, organ weight, enzymes activities, and tissues concentrations of Zn and Cu. The results showed that Zn deficiency has negative effects on growth, organ weight, and biological parameters such as alkaline phosphatase (ALP) and Cu−Zn superoxide dismutase (Cu−Zn SOD) activities, whereas Zn overload played an effective role in promoting growth, improving the developments of organs and enhancing immune system. Hepatic metallothionein (MT) concentration showed an identical increase tendency in rats fed both Zn-deficient and Zn-overload diets. The actual mechanism of reduction of Cu concentration of jejunum in rats fed a Zn-overload diet might involve the modulation or inhibition of a Cu transporter protein by Zn and not by the induction of MT.     

Effect of dietary nickel and iron on the trace element content of rat liver

The level and/or form of dietary iron, dietary nickel, and the interaction between them affected the trace element content of rat liver. Livers were from the offspring of dams fed diets containing 10–16 ng, or 20 μg, of nickel/g. Dietary iron was supplied as ferric chloride (30 μg/g) or ferric sulfate (30 μg, or 60 μg). In nickel-deprived rats fed 60 μg of iron/g of diet as ferric sulfate, at age 35 days, levels of iron and zinc were depressed in liver and the level of copper was elevated. At age 55 days, iron was still depressed, copper was still elevated, but zinc also was elevated. In rats fed 30 μg of iron/g of diet as ferric chloride, liver iron content was higher in nickel-deprived than in nickel-supplemented rats at 30, but not at 50, days of age. Also manganese and zinc were lower in nickel-deprived than in nickel-supplemented rats at age 35 days if their dams had been on experiment for an extended period of time (i.e., since age 21 days). Thus, the levels of copper, iron, manganese, and zinc in liver were affected by nickel deprivation, but the direction and extent of the affects depended upon the iron status of the rat.     

Magnesium,calcium, copper,and zinc in meals

Meals of 12 diets were prepared from conventional foods with precautions against contamination by metallic elements because of epidemiologic associations between ischemic heart disease and the metabolism of magnesium, calcium, copper, and zinc. Magnesium, calcium, copper, and zinc were measured by atomic absorption spectrometry with satisfactory accuracy and precision. The mean daily amount of copper in the diets was less than the apparent adult requirement. Mean amounts of magnesium and zinc were close to apparent requirements; however, adults consuming amounts less than one standard deviation below these means may be depleting body stores and be at risk of pathology. Magnesium and copper were highly correlated (r=0.849,P=0.0001) in meals. This and other significant correlations probably will prevent the relationships of these elements to ischemic heart disease from being elucidated by epidemiology. Metabolic experiments will be necessary to differentiate among several hypotheses.     

Interaction between zinc and calcium in skeletal muscle in young growing rats

The purpose of the present study was to determine whether zinc and calcium could interact at the tissue level. In the first part of the study, adult rats were injected with ZnCl₂ dissolved in a physiological saline solution to determine the effects of Zn on Ca levels in various tissues. In the second part of the study, weaned rats (at day 22 postnatally) were fed a diet supplemented with Zn until day 50 and were then sacrificed. In both instances, blood, brain, heart, liver, and skeletal muscle were taken and analyzed. In the Zn-injected group, the brain, heart, and liver showed no interaction between Zn and Ca. The skeletal muscle, in contrast, showed a decrease in Ca in the homogenate, whereas Zn contents showed a significant increase at the sarcoplasmic reticulum (SR). Likewise, in the Zn-supplemented group, the Zn content of the SR vesicle of the skeletal muscle showed an increase, whereas Ca content of the pellet (14,000 g), which contains cell debris, nucleus, mitochondria, and SR vesicles of this group, showed a decrease. Current findings suggest antagonistic effects between Zn and Ca on this tissue. Zn may play a critical role in cellular function through the alteration of itnracellular distribution of Ca in skeletal muscle.     

Rat liver metabollothionein interactions of silver,zinc, and cadmium

Weanling rats were fed diets either alone or with combinations of silver, elevated zinc, or elevated cadmium for 7 weeks. The rats were then killed, and the silver, zinc, and cadmium proteins isolated from the livers by gel filtration and ion exchange chromatography. When silver was fed alone in diet, low levels of this metal were eluted as two peaks from the ion exchange column. In contrast, when silver was fed with cadmium or elevated zinc, three metal-containing peaks were eluted from this ion exchange column. Amino acid analysis revealed that the major proteins binding these metals are metallothioneins, as judged by high cysteine content.     

The effect of ethanol on zinc and copper metabolism in rats

Studies performed on adult female rats over a period of 10 weeks indicated that the consumption of alcohol (20% v/v) did not appear to disturb the zinc or copper balance, nor did it adversely affect tissue zinc or copper levels, even in zinc-restricted animals. On the contrary, higher plasma zinc levels were consistently observed in animals receiving alcohol together with the experimental diets.     

Increased nitration and diminished activity of copper/zinc superoxide dismutase in placentas from diabetic rats

Abstract

Nitration-induced protein damage in the placenta leads to impaired blood flow and deficient feto–placental exchange in diabetic pregnancies. This work studied the effect of nitric oxide and peroxynitrite on Cu/Zn SOD activity in rat placentas and evaluated whether Cu/Zn SOD is nitrated in the placenta from diabetic rats at mid-gestation. Protein nitration was evaluated by EIA, Cu/Zn SOD activity by inhibition of the epinephrine auto-oxidation, Cu/Zn SOD expression by western blot and specific nitration by immunoprecipitation. This study found higher levels of protein nitration (p < 0.001), diminished Cu/Zn SOD activity and enhanced protein expression (p < 0.01) in placentas from diabetic rats. Placental Cu/Zn SOD activity was inhibited by peroxynitrite (p < 0.01). Besides, nitration of Cu/Zn SOD was elevated in placentas from diabetic rats (p < 0.01). These results show that rat Cu/Zn SOD can be nitrated, a modification that could lead to the depressed activity of this enzyme found in placentas from diabetic rats.     

The role of dietary copper,manganese, selenium,and vitamin E in lipid peroxidation in tissues of the rat

The role of dietary Cu and Mn in maintaining tissue integrity, through the effects of these metals on activity of the superoxide dismutase (SOD) enzyme, and their interactions in peroxidative pathways involving Se and vitamin E was investigated. Weanling rats were fed diets deficient in Mn, Cu, Se, and/or vitamin E for 35 days, in a factorial experimental design. Dietary effects on peroxidation, measured in mitochondrial fractions prepared from liver and heart tissue, were compared with changes in the activities of glutathione peroxidase and the Cu and MnSOD enzymes. Decreased heart MnSOD and CuSOD activities, resulting from dietary Mn and Cu deficiencies, were both associated with increased peroxidation. Adequate Se (and glutathione peroxidase activity) prevented the peroxidation associated with either of these deficiencies, but was ineffective with a combined Cu−Mn deficiency. These effects of Se were only observed in tissue lacking glutathione transferase activity. Effects of Cu, Mn, and Se on peroxidation appeared to be present at both levels of vitamin E, although in both tissues, vitamin E deficiency greatly increased the overall peroxidation. Comparison of these in vitro peroxidation results with the deficiency associated lesions observed in vivo indicates that changes in SOD activities and peroxidation pathways may be the dominant cause of these lesions in only some cases. In others, the roles of Cu and Mn in different metabolic pathways appear to be of greater importance.     

Testicular atrophy, zinc concentration, and angiotensin-converting enzyme activity in the testes of vitamin a-deficient rats

Angiotensin-converting enzyme (ACE) as a part of the renin angiotensin system (RES) regulates blood pressure and fluid and electrolyte homeostasis, and the enzyme is considered to have a function in reproduction. Reduced enzyme activities have been observed in atrophied testes as a results of zinc and pituitary deficiencies. Vitamin A deficiency causes atrophy of testes. The present study was conducted on three groups of male, 3-wk-old, Wistar rats. After 54 d of the experimental period, testicular weights of the vitamin A-deficient rats (Agroup, allowed free access to vitamin Adeficient diet) was significantly lower than its pair-fed, PF (given restricted amount control diet) and A+ (allowed free access to control diet) groups. Zinc concentrations and both soluble and particulate ACE activities in the testes of vitamin A-deficient rats (Agroup) were significantly lower than the other two groups. No significant differences were observed regarding zinc concentration, particulate ACE, and total ACE activities in the testes of PF and A+ groups. Vitamin A deficiency did not significantly affect the enzyme activity in the lung. From the observations of the present study, we speculate that testicular atrophy in vitamin A deficiency may have resulted from lower zinc concentration and decreased ACE activity in that organ.     

Influence of sodium fluoride and caffeine on the concentration of fluoride ions, glucose, and urea in blood serum and activity of protein metabolism enzymes in rat liver

The aim of the study was examining the effect of fluoride ions and caffeine administration on glucose and urea concentration in blood serum and the activity of protein metabolism enzymes and selected enzymes of the urea cycle in rat liver. The study was carried out using 18 male Sprague-Daowley rats (4.5 mo old). Rats were divided into three groups. Group I received distilled water ad libitum. Group II received 4.9 mg F⁻/kg body mass/d of sodium fluoride in the water, and group III received sodium fluoride (in the above-mentioned dose) and 3 mg/kg body mass/d of caffeine in the water. After 50 d, the rats were anesthetized with thiopental and fluoride ions, glucose, and urea concentration in blood serum were determined. Also determined were the activities of aspartate aminotransferase, alanine aminotransferase glutamate dehydrogenase, ornithine carbamoylotransferase and arginase in liver homogenates. Liver was taken for pathomorphological examinations. The applied doses of F⁻ (4.9 mg/kg body mass/d) and F⁻+ caffeine (4.9 mg F⁻/kg body mass/d+3 mg caffeine/kg body mass/d) resulted in a statistically significant increase of fluoride ion concentration in blood serum, a slight increase of the glucose concentration, and no changes in the concentration of urea in blood serum. This might testify to the absence of kidney lesions for the applied concentrations of F⁻. No change in the functioning of hepatocytes was observed; however, slight disturbances have been noted in the functioning of the liver, connected with the activation of urea cycle, increase of arginase activity, and accumulation of F⁻ in this organ. There was no observed significant influence of caffeine supplementation on the obtained results.     

Interactions among nickel,copper, and iron in rats

In two fully crossed, three-way, two by three by three, factorially arranged experiments, female weanling rats were fed a basal diet supplemented with iron at 15 and 45 μg/g, nickel at 0, 5, and 50 μg/g and copper at 0, 0.5, and 5 μg/g (Expt. 1) or 0, 0.25, and 12 μg/g (Expt. 2). Expt. 1 was terminated at 11 weeks, and Expt. 2 at 8 weeks because, at those times, some rats fed no supplemental copper and the high level of nickel began to lose weight, or die from heart rupture. The experiments showed that nickel interacted with copper and this interaction was influenced by dietary iron. If copper deficiency was neither very severe or mild, copper deficiency signs of elevated levels of total lipids and lipid phosphorus in liver and plasma, and cholesterol in plasma, were made more severe by supplemental dietary nickel. Rats in which nickel supplementation exacerbated copper deficiency did not exhibit a depressed level of copper in liver and plasma. Also, although iron deprivation enhanced the interaction between nickel and copper, iron deprivation did not significantly depress the level of copper in liver and plasma. The findings confirmed that, in rats, a complex relationship exists between nickel, copper, and iron, thus indicating that both the iron and copper status of experimental animals must be controlled before data about nickel nutriture and metabolism can be compared among studies.     

Changes in the total content of iron,copper, and zinc in serum,heart, liver,spleen, and skeletal muscle tissues of rats infected withTrypanosoma cruzi

The effects of theTrypanosoma cruzi infection on the total content of the essential trace elements iron (Fe), copper (Cu), and zinc (Zn) in serum, heart, liver, spleen, and skeletal muscle were determined in “Wistar” rats inoculated with reticulotropic “Y” strain trypanosoms (Tryps) in their slender blood form. The 250 rats were divided in two groups of 80 rats (L-1 and L-2) and one of 90 (C) used as controls. L-1 and L-2 were inoculated with 2×10⁵ and 5×10² Tryps, respectively. Ten rats of the C group were killed the inoculation day (i), and ten rats of each group chosen at random were killed and blood parasitemia determined at 5, 10, 15, 20, 25, 30, 60, and 90 post-i days covering the infection acute-phase myocarditis. Previously cryohomogenized and lyophilized tissues were digested in an HNO₃ ⁻ H₂O₂ mixture with the aid of a microwave oven, and the elements Fe, Cu, and Zn were determined by Flame Atomic Absorption Spectrometry. Generally, more intense changes were observed in the L-1 group. Serum Fe and Zn levels are lower and Cu levels higher in groups L-1 and L-2 than in C. However, Fe is not significantly sequestered in the liver during the acute phase of the infection as expected, but of the tissues studied, the spleen was the main site of Fe binding. Zn tended to increase in all tissues, except in the spleen, where during the acute phase of the infection, the total content of Zn in groups L-1 and L-2 was lower than in group C. Cu increased mainly in the spleen and muscle. In general, each tissue presented its own pattern of redistribution related to its nature, functions, and number of parasites inoculated, and these patterns may have been altered by the tropism of the parasite.     

The effect of exercise training and water extract from propolis intake on the antioxidant enzymes activity of skeletal muscle and liver in rat

[Purpose]

In this study, the authors have intended to investigate the effects that the exercise training and the intake of the water extract from propolis have on the activity of antioxidant enzymes.

[Methods]

For this purpose, the exercise training (70% VO₂max treadmill running exercise for 60min)of 5 times per week for six weeks and the intake (50mg/kg/day) of the water extract from propolis were performed by separating the experimental animals (SD rats, n=32) into CON(n=8) group, CON+Ex(n=8), PA(n=8), and PA+Ex(n=8).

[Results]

As a result, the following conclusions were obtained: The concentration of the blood glucose and insulin of the CON+Ex group and PA+Ex group which are the exercise parallel group were significantly decreased in comparison with the control group, whereas if comparing the glycogen concentration in skeletal muscle and liver tissue between the exercise parallel group and the CON group, the former showed significantly high value in comparison with the latter (p < .05). In the case of the activity of the antioxidant enzyme in the skeletal muscle and the liver tissue, the activities of SOD, GPX and CAT in the gastrocnemius muscle tissue of the experimental animals showed significantly high value in PA+Ex group in comparison with other experimental groups (p < .05). In addition, the SOD activity in the liver tissue showed that only PA+Ex group was significantly increased, whereas GDX activity showed significantly higher value in CON+Ex group and PA group than CON group (p < .05). However, the activity of CAT in the liver tissue showed that there is no difference between the experimental groups. As a result that measured the concentration of MDA in order to evaluate the damage level of the tissue by oxygen free radicals, the difference between the groups in the liver tissue was not shown, while it was shown that only PA+Ex group in the skeletal muscle tissue was significantly decreased in comparison with other experimental groups (p < .05).

[Conclusion]

Taken together the above findings, it is considered that the parallel treatment of the exercise training and the water extract from propolis can not only increase the use of glycogen of the skeletal muscle and liver tissue, but also it can give the effect to suppress the creation of active oxygen by inducing the activity of the antioxidant enzyme in the body, and in the future, the possibility as the exercise supplements and the antioxidant of the water-soluble propolis are expected.     

锰对葡萄根中离子吸收及抗氧化酶系统的影响

以2个葡萄品种(金手指、康拜尔)为材料,采用温室沙培实验,研究不同浓度Mn处理对葡萄根中离子吸收及抗氧化酶活性的影响.结果表明,随着Mn2+浓度的增大,葡萄根中元素含量呈现不同的变化,总体上看Ca和Mg的含量降低,Mn、Cu和Zn的含量增加,Fe含量则随锰处理浓度增加呈先下降后略有升高的趋势.在抗氧化系统中POD活性随 Mn浓度的升高而逐渐降低,而CAT和APX酶活性呈先升高后降低的趋势,SOD活性变化不大,说明保护酶系统形成了一定的适应高锰胁迫的机制,这些抗氧化酶活性的增强能够提高葡萄适应和抵抗重金属胁迫的能力.     

The effect of various dietary zinc concentrations on the biological interactions of zinc,copper, and iron in rats

Three groups (14 rats each) were fed one of the following diets for 8 wks: a control purified basal diet containing 12 ppm zinc, 5 ppm copper, and 35 ppm iron; the basal diet with less than 2 ppm zinc; or the basal diet supplemented with 1000 ppm zinc. Rats fed the zinc-deficient diet had decreased weight gain, moderate polydipsia, and intermittent mild diarrhea. The zinc-supplemented rats had a cyclical pattern of food intake and weight loss from weeks 5 to 8. Tissue concentrations suggest that zinc and copper were not mutually antagonistic with chronic dietary imbalances. If tissue element concentrations reflected intestinal uptake, then competition and/or inhibition of intestinal uptake occurred between zinc and iron. The fluctuations in tissue element concentrations that occurred with increased duration of the study were at variance with previous studies of shorter time periods. The dietary proportions of zinc, copper, and iron appear to influence zinc, copper, and iron metabolism at the intestinal and cellular transport levels over a given period of time.     

重金属Cd、Zn、Cu、Pb对土壤微生物和酶活性的影响

采用室内培养实验(25℃),研究了不同培养时间下重金属Cd、Zn、Cu、Pb(浓度分别为50,800,400,800mg.kg-1)污染对土壤微生物和酶活性的影响。结果表明,土壤蔗糖酶、过氧化氢酶和脱氢酶活性随着培养时间的增加而显著下降,在培养20d的时候达到最小值,然后酶活性缓慢升高。Cu对脲酶活性以及Cd对酸性磷酸酶和脲酶活性的抑制作用随时间增加而增加。土壤微生物生物量碳、细菌、真菌和放线菌数量随培养时间的增加均表现出先降低后升高的变化趋势。Cd和Cu对微生物生物量氮的抑制作用则随着培养时间的增加而增强,在培养30d时微生物生物量氮到达最低值,分别较培养10天减少了12.6%和16.5%。     

铅、锌及其交互作用对鱼腥草(Houttuynia cordata)叶绿素含量及抗氧化酶系统的影响

采用营养液培养方法研究铅、锌及其交互作用对鱼腥草叶绿素含量及抗氧化酶系统的影响。实验结果显示，随着Pb浓度的增加，鱼腥草叶绿素含量逐渐降低但无显著变化。Zn在一定浓度下能提高鱼腥草叶绿素含量，而在高浓度Zn胁迫下，叶绿素含量急剧下降。鱼腥草叶片中SOD、POD和CAT3种酶活性都是随着Pb浓度的增加先上升后下降。随着Zn浓度的增加，SOD和CAT也是先上升后下降，POD则是逐渐上升。Pb—Zn交互作用增加了鱼腥草叶绿素含量，对SOD和POD活性具有抑制作用，对CAT活性影响不明显。同时研究结果还表明，单一Pb、Zn对鱼腥草叶绿素含量和抗氧化酶系统的影响大于Pb、Zn二者的共同作用，其中高浓度Zn对鱼腥草的伤害作用最大，而当溶液Pb处理浓度达到400mg／L时，鱼腥草仍能正常生长，说明鱼腥草具有较强的耐Pb能力。     

Effects of rutin supplementation on antioxidant status and iron,copper, and zinc contents in mouse liver and brain

The effect of rutin on total antioxidant status as well as on trace elements such as iron, copper, and zinc in mouse liver and brain were studied. Mice were administrated with 0.75 g/kg or 2.25 g/kg P. O. of rutin for 30 d consecutively. Following the treatment, the activity of total antioxidant status, catalase, Cu,Zn-superoxide dismutase, Mn-superoxide dismutase, zinc, copper, and iron were measured in mouse liver and brain. The results showed that rutin significantly increased the antioxidant status and Mn-superoxide dismutase activities in mouse liver, but it had no effect on these variables in the brain. Treatment with a higher concentration of rutin significantly decreased catalase activity and iron, zinc, and copper contents in mouse liver; it also resulted in a slower weight gain for the first 20 d. These results indicate that rutin taken in proper amount can effectively improve antioxidant status, whereas at an increased dosage, it may cause trace element (such as iron, zinc, and copper) deficiencies and a decrease in the activities of related metal-containing enzymes.