Metabolism of administered triethylene tetramine dihydrochloride in humans

Triethylene tetramine dihydrochloride (trien 2HCl) has been used for the treatment of Wilson's disease, which is characterized by the accumulation of copper in various organs. We previously developed an HPLC system for analyzing trien, and found a trien metabolite in the urine when trien was orally given to humans. In this study, the metabolite was identified as 1-N-acetyltriethylene tetramine (acetyltrien) by FAB-MS and 1H-NMR spectroscopy. Trien and acetyltrien were capable of combining with copper, iron and zinc. However, the chelating activity of acetyltrien was significantly lower than that of trien. When trien was given to healthy adults, the amount of trien excreted in the urine was about 1% of the administered trien, whereas that of acetyltrien was about 8%. Most of the trien was excreted within the first 6 hours after the administration, while acetyltrien was excreted for over 26 hours. The urinary copper, iron and zinc levels all increased in parallel with the trien excretion.     

Oral contraceptives, norethindrone and mestranol: effects on tissue levels of minerals

The study involved three levels of dietary zinc (deficient, marginal, and adequate) and four hormonal conditions; namely, no steriods, norethindrone, mestranol, and norethindrone plus mestranol. The steroids were incorporated into diets and fed to 11-wk-old female Sprague-Dawley rats. After 10 wk of treatment, various tissues were excised for mineral assays by atomic-absorption spectrophotometry. Both steroids, reduced weight gain. Mestranol depressed plasma zinc, tibia copper and magnesium, and liver iron, but elevated the zinc levels in liver and erythrocytes, plasma copper, liver magnesium and calcium, and iron content of tibia and heart. In general, the effect was most prominent with adequate zinc but diminished in magnitude with the reduction of zinc intake. In addition, norethindrone increased heart iron and tibia calcium. Mestranol appeared to be the main causative factor and may have induced a possible shift of minerals from one pool to another. As expected, zinc deficiency resulted in the reduction of zinc concentrations of plasma, tibia, kidney, and pancreas, and the elevation of copper, iron, magnesium, and calcium concentrations of various tissues.     

Anxiety and ++phobia in dental patients. Recent findings regarding the "fear of the dentist"

The protein-bound polysaccharide of Coriolus versicolor QUEL (PS-K) expresses the mimetic activity of superoxide dismutase (SOD). Human cancer patients usually suffer from oxidative stress (OS). Examination was made to determine the capacity of this drug with SOD mimetic activity for relieving OS. Rats transplanted with Walker 256 fibrosarcoma showed OS on day 12. After confirming high levels of OS on day 13, PS-K50 mg/kg was intraperitoneally administered, and prompt decrease in O2-release from RBC was noted. The drug ceased to have any effect 24 hours following the first inoculation. Average OS in human cancer patients was found twice that in healthy persons. In human cancer patients perorally administered PS-K3.0 g/day, OS decreased to the normal level one day after the initial administration. Plasma lipid peroxide (LPO) in cancer patients treated with PS-K for 28 days increased and withdrawal of the drug led to decreased LPO.     

Physical map of the Clostridium difficile chromosome

The present study was designed to investigate the effect of mercuric chloride administration on copper, zinc, and iron concentrations in the liver, kidney, lung, heart, spleen, and muscle of rats. The results showed that after dose and time exposure to mercuric chloride, the concentration of mercury in the six tissues was significantly elevated. Data showed that there were no interaction between mercury and tissue iron. There was a considerable elevation of the content of copper in the kidney and liver. The most significant changes in the copper concentration took place in the kidneys. About a twofold increase in the copper content of the kidney was noted after exposure to mercuric chloride (3 mg and 5 mg/kg). Only slight elevations in the copper content occurred in the liver especially in high dose and longer exposure time. In the remaining organs, the copper content was not changed significantly (p > 0.05). The most significant changes in the zinc concentration took place in liver, kidney, lung and heart (5 mg/kg). Marked changes in kidney zinc concentrations were observed at any of the specified doses. Zinc concentrations were significantly increased in kidney of rats sacrificed 9-48 h after s.c. injection of HgCl2 (5 mg/kg); in liver obtained from rats at 18, 24 or 48 h after injection; and in lung after 24 or 48 h of treatment. The heart and spleen zinc concentrations were elevated at 24 and 48 h after injection of HgCl2 (5 mg/kg), respectively. The results of this study implicate that effects on copper and zinc concentrations of the target tissues of mercury may play an important role in the pathogenesis of acute mercuric chloride intoxication.     

Reduced N-acetylaspartate in the brain observed on in vivo proton magnetic resonance spectroscopy in patients with mental retardation

PURPOSE: Mechanisms of postburn immunosuppression are complicated and remain unclear. In the present experiment the effect of linoleic acid hydroperoxide (LOOH) on lymphocytes was evaluated in vitro, and then the changes in postburn lipid peroxide (LPO) levels and lymphocyte functions were measured, in order to investigate the mechanism of immunosuppression following burn. METHODS: Proliferation and IL-2 production and LPO of lymphocytes were assayed after the incubation with LOOH for in vitro study. On day 6 after induction of 11%-12% TBSA full-thickness burn, the animals were sacrificed, and proliferation and IL-2 production of splenic lymphocytes, LPO levels of the plasma, livers and spleens were measured. RESULTS: LOOH inhibited proliferation and IL-2 production and induced lipid peroxidation of lymphocytes in vitro, and Vit E could attenuate the effects of LOOH. LPO levels increased and proliferation and IL-2 production decreased after burn, but those changes in Vit E or SOD group showed no statistical significance compared with normal control group. CONCLUSION: Enhancement of lipid peroxidation and increased LPO after burn may be one of the mechanisms of postburn immunosuppression.     

Effects of different levels of dietary iron on pregnancy superimposed upon growth in the rat

The effects of feeding four levels of dietary iron, 10, 50, 250 and 1.250 mg/kg were studied during pregnancy in growing and adult rats. Hematological measurements, plasma iron and total iron binding capacity, and iron content in liver, spleen and tibia were compared relative to pregnancy, diet and growth. Iron content in fetuses and fetal livers were compared relative to diet and growth of the dams. All parameters were lowest in rats fed the 10 mg Fe/kg diet. The highest level of iron fed, 1,250 mg/kg, resulted in increased iron content in liver, spleen and tibia of all treatment groups but did not alter hematological values or fetal iron content. Pregnant rats fed any of the four levels of iron had significantly lower Hb, Ht, total and storage iron concentration and ferritin and hemosiderin iron in liver than nonpregnant rats fed the same levels. The level of dietary iron needed by growing pregnant rats for maximal iron content in fetuses and fetal livers was between 50 and 250 mg Fe/kg which was fivefold that needed to obtain maximal hemoglobin concentration in dams. However, adult pregnant rats only needed between 10 to 50 mg Fe/kg for both maximal iron stores in fetal tissues and maximal hemoglobin concentration.     

Alaskan malamute chondrodysplasia IV. Concentrations of zinc, copper and iron in various tissues

Trace mineral concentrations in various tissues of the chondrodysplastic (dwarf) Alaskan Malamute are remarkably different as compared to normal. The zinc level in heart tissue was depressed in dwarf animals (26 weeks). Copper concentration in the liver is elevated two to four fold in 26 week old dwarf animals and iron levels are significantly elevated in kidney, liver and pancreas of these animals. These observations suggest that the dwarf Alaskan Malamutes suffer from a genetic defect in trace mineral metabolism. If this is the case, then many of the skeletal lesions reported for these animals may be attributed to disorders in either zinc or copper metabolism.     

Activities of conjugating and antioxidant enzymes following endotoxin exposure

Endotoxin exposure elicits various responses in mammals including the acute phase response that has been shown to cause changes in the activity of several forms of cytochrome P450s and other enzymes. Therefore, the hepatic conjugating enzyme, glutathione S-transferase (GST), and UDP-glucuronosyltransferase (UDPGT), the antioxidant enzymes, glutathione peroxidase (GSHPx), catalase, and superoxide dismutase (SOD), as well as lipid peroxidation were investigated following the administration of endotoxin to male Sprague-Dawley rats (8 mg/kg body weight). Rats were euthanized at various times following endotoxin administration and the livers removed and processed to assess various enzyme activities. Glutathione S-transferase, UDPGT, and GSHPx activity showed statistically significant decreases after 24 hours and remained lower than controls for the duration of the study. Decreases in total SOD and catalase activities were seen at 24, 48, and 72 hours following endotoxin administration; however, only catalase activity showed statistically significant differences between control and treated samples at those time points, and total SOD activity showed a statistically significant decrease at 24 hours. No statistically significant changes were seen in the level of lipid peroxidation in the liver microsomes from endotoxin-treated animals. Changes in the conjugative enzymes and the free-radical scavenging enzymes following endotoxin exposure may alter the host's metabolism and response to free radicals.     

Effects of iron supplementation and discontinuation on serum copper, zinc, calcium, and magnesium levels in women

The purpose of this study was: 1) to establish the prevalence of depleted iron stores, iron deficiency, and low serum levels for copper, zinc, calcium, and magnesium in a healthy female population; and 2) to examine the effects of iron supplementation and discontinuation on the serum levels of the above minerals. One hundred eleven healthy women between the ages of 18 and 40 yr reported for fasted morning blood sampling for iron, copper, zinc, calcium, and magnesium status. Forty-five subjects were either iron-deficient as defined by a hemoglobin level below 120 g.l-1 (four subjects) or iron deplete as defined by a serum ferritin value below 20 micrograms.l-1 (43 subjects). Two subjects fit both criteria. This subgroup continued with the study and were prescribed a normal therapeutic iron dose (320 mg elemental iron per day, taken as two Slow-Fe tablets.d-1 for a period of 12 wk). The subjects then discontinued the iron supplementation for a further 12 wk. The response of the various blood minerals was monitored at 6-wk intervals. Twenty-five subjects completed the full 24-wk treatment. The main conclusions to be made from this study were that: 1) For this sample population of women, iron depletion was quite common (39%), although low hemoglobin values (< 120 g.l-1) were only seen in 3.6%. No subjects fell below the criteria for low serum copper levels (< 13.3 mumol.l-1) nor low serum magnesium levels (< 0.6 mmol.l-1). Seven subjects (6.5%) fell below the criteria for low serum zinc levels (< 11.5 mumol.l-1) while two subjects (1.8%) were below the criteria for low serum calcium levels (< 2.20 mmol.l-1). 2) Therapeutic oral iron supplementation was successful in raising mean serum ferritin values from 15.9 micrograms.l-1 to 36.5 micrograms.l-1 but was not associated with decrements in serum copper or calcium levels. 3) The treatment did not significantly effect serum zinc and magnesium levels during the supplementation period, but a downward trend continued through the discontinuation phase so that at 18 and 24 wk serum zinc and magnesium levels were significantly lower than baseline. 4) Oral contraceptive use was associated with elevated serum copper and ferritin values and lowered serum magnesium levels.     

Surgical procedure for modifying the duodenum in cattle to measure bile flow and the diurnal variation in biliary manganese, iron, copper and zinc excretion

A surgical procedure is described for modifying the duodenum of cattle so that bile can be sampled and its rate of flow measured for long periods. In 12 steers, aged three months to three years and weighing between 50 and 650 kg, bile flow ranged from 2 to 12 ml per minute, the rate of flow increasing with bodyweight. The rate of flow expressed as ml per minute per 100 kg bodyweight decreased as bodyweight increased, the regression equation being in (ml per minute per 10 kg) = 1.65-0.0022 x where x = bodyweight (kg), r = 0.871. The mean concentrations of copper, iron, manganese and zinc in bile were 8.2, 6.2, 17.1 and 3.3 mumol per litre respectively. The concentration of iron was the least variable between the steers. The average total cholate concentration was 100 mmol per litre and total solids ranged from 5.4 to 7.2 per cent. Diurnal patterns in bile flow and trace element excretion were measured in four adult steers during a period of 38.5 hours. Although the rate of excretion of iron, zinc and copper and bile flow varied throughout the period, changes could not be associated with feeding or time of day. The concentration of manganese in bile and its excretion rate varied in a reproducible manner which may be related to feeding activity. The mean output of the four trace elements in bile from the four steers during 24 hours was, copper 37.7 mumol, iron 68.0 mumol, manganese 80.3 mumol and zinc 59.6 mumol.     

Hepatotoxicity induced by iron overload and alcohol. Studies on the role of chelatable iron, cytochrome P450 2E1 and lipid peroxidation

BACKGROUND/AIMS: Clinical experience and studies with experimental animal models indicate a synergistic hepatotoxic effect of dietary iron overload and chronic alcohol ingestion. In order to elucidate the mechanism underlying this synergism, we examined the hepatic levels of ethanol-inducible cytochrome P450 2E1, glutathione and malondialdehyde, and the effect of iron chelation with desferrioxamine, in livers from rats treated with iron and/or ethanol. METHODS: Animals received diets with or without 2.5-3% carbonyl iron for 6-9 weeks, followed by an ethanol-containing diet or a liquid control diet for 5-9 weeks. Desferrioxamine was administered subcutaneously with mini-osmotic pumps. Alanine aminotransferase activity in serum and hepatic contents of glutathione and malondialdehyde were determined. The hepatic level of cytochrome P450 2E1 was determined with Western Blotting using a specific polyclonal antibody. RESULTS: The combination of iron and alcohol led to a marked increase in serum alanine aminotransferase activity as compared with all other treatment groups, and iron chelation with desferrioxamine reversed these increases. Treatment with alcohol alone led to slightly increased aminotransferases compared with controls. The level of cytochrome P450 2E1 was significantly elevated in microsomes isolated from ethanol-treated rats, but neither additional iron supplementation nor desferrioxamine influenced this level significantly. Glutathione contents were increased in the livers of animals treated with iron and/or ethanol. Malondialdehyde values were increased in iron-treated animals, whereas neither ethanol nor desferrioxamine altered malondialdehyde levels significantly. CONCLUSIONS: The toxic effects exerted by the combination of iron overload and chronic ethanol feeding on rat liver are dependent on a pool of chelatable iron. The hepatic level of cytochrome P450 2E1 is markedly induced by ethanol but not further altered by iron overload. Neither increased lipid peroxidation nor depletion of hepatic glutathione levels can explain the synergistic hepatotoxic effects of iron and ethanol in this model.     

The interrelationship of the intensity of free-radical oxidation to the level of serum bilirubin in lesions of the hepatobiliary system]

The level of lipid peroxidation (LPO) products increases in the blood of patients with chronic diseases of the liver and acute cholecystitis with mechanical jaundice. The values normalize after surgery for acute cholecystitis, whereas therapy for chronic diseases of the liver virtually does not correct them. These data demonstrate the relationship between intensity of free-radical oxidation and severity of hepatobiliary process and between plasma content of LPO products and level of serum bilirubin.     

连续光源火焰原子吸收光谱法测定氧化镍中钴铜锌铁钙镁

采用盐酸溶解样品,选择Co 240.72nm、Cu 324.75nm、Zn 213.86nm、Fe 248.33nm、Ca 422.67nm、Mg 202.58nm作为分析谱线,钴、铜、锌、铁选择3个像素点,钙、镁选择9个像素点,建立了连续光源原子吸收光谱法(CS-AAS)同时测定氧化镍中的钴、铜、锌、铁、钙、镁的方法。实验表明:在100mL测定液中加入2mL 200g/L氯化锶溶液,可消除测定介质(体积分数为2%的盐酸)对待测元素的影响;基体镍对测定的干扰可忽略。在优化的实验条件下,钴、铜、锌、铁、钙、镁的校准曲线相关系数均不低于0.999 0,且其方法检出限在0.002~0.092μg/mL之间。按照实验方法对氧化镍样品中钴、铜、锌、铁、钙、镁分别平行测定11次,钙和镁的测定值在0.1%~0.4%之间,其对应的相对标准偏差(RSD)不大于2%;钴、铜、锌、铁的测定值在0.003%~0.04%之间,其对应的相对标准偏差均小于10%。将实验方法应用于电真空镍光谱标准样品(该标样为氧化镍状态)中上述各元素的测定,结果与认定值基本一致。     

Serum concentrations of zinc and copper in bull terriers with lethal acrodermatitis and tail-chasing behavior

OBJECTIVE: To establish similarities or differences in tissue concentrations of zinc, copper, and iron in Bull Terriers with lethal acrodermatitis (LAD) and tail-chasing behavior (TCB) and to confirm the suspicion that copper is involved in the etiopathogenesis of LAD. SAMPLES: Serum samples from 29 Bull Terriers (9 control dogs, 6 dogs with LAD, 14 dogs with TCB), and liver and kidney specimens from 2 dogs and 1 and 4 dogs with LAD or TCB, respectively. PROCEDURE: Serum, liver, and kidney mineral (zinc, copper, and iron) concentrations in Bull Terriers with LAD or TCB and in a group of control dogs were analyzed, using flame atomic absorption after wet ashing technique. RESULTS: Serum zinc and copper concentrations were lower (P < 0.05) in dogs with LAD, compared with values for control dogs and dogs with TCB. Liver zinc and copper concentrations were similar to serum values. Kidney zinc and copper concentrations were similar among the 3 groups. Serum, liver, and kidney iron concentrations had a wide range of variability within all 3 groups. CONCLUSION: Copper deficiency is associated with LAD. The primary cause of LAD may be copper deficiency, with zinc involved secondarily, or combined zinc and copper deficiencies. The role of ion deficiency in TCB was not clarified. CLINICAL RELEVANCE: Serum zinc and copper concentrations should be determined when LAD is suspected.     

N-methyl-D-aspartate receptors in the nucleus accumbens are involved in detection of spatial novelty in mice

Toxic milk mutant (tx) mice accumulate excess copper (Cu) in liver with age and develop symptoms similar to those seen in human Wilson disease. Because metallothionein (MT) is the major Cu-binding protein in tx mouse liver and Cu-MT can enhance lipid peroxidation initiated by an organic hydroperoxide, the potential genotoxicity of Cu-MT in tx mice was assessed in male tx mice (11 to 12 months old) and in age- and sex-matched control wild-type (DL) mice. Toxic milk mutant mice, but not control DL mice, developed regenerative liver nodules (tx-N) with normal histologic appearance. Residual, non-nodular tx mouse liver (tx-R) was microscopically abnormal with large, atypical hepatocytes. The levels of Cu, zinc (Zn), and MT, and the numbers of apoptotic cells (APC) in tx-N, tx-R, and DL livers were measured by atomic absorption spectrophotometry, 109cadmium-heme assay, and the TUNEL method, respectively. Significantly higher levels of MT, Cu, and Zn, as well as increased numbers of APC were found in both tx-N and tx-R compared with DL mouse livers. Intense nuclear and cytoplasmic immunohistochemical staining for MT was observed in both normal and atypical hepatocytes of the tx mouse, whereas only cytoplasmic staining for MT was detected in DL mouse liver tissue. Accumulated Cu could be detected in tx-R and tx-N liver by rhodanine staining but was not detected in other tx mouse organs, or in mouse liver or other organs of DL. The number of APC and level of MT were significantly higher in tx-R liver compared with both tx-N and DL liver. These results suggest that: (a) aged tx mouse accumulate excess Cu in liver accompanied by striking morphologic changes, and (b) although MT binds to Cu in tx mouse liver, the presence of high Cu-MT and Cu in the nucleus can be genotoxic and may lead to enhanced apoptosis.     

滴定法连续测定铅锌矿中铅锌铜

铅锌矿样经盐酸和硝酸溶解后,以硫酸为沉淀剂,使铅与硫酸形成硫酸铅沉淀,锌和铜留在溶液中。过滤后,用乙酸-乙酸铵缓冲溶液溶解硫酸铅沉淀,以二甲酚橙为指示剂,EDTA标准溶液滴定铅。分取两份滤液,一份用于测定锌,另一份用于测定铜。测定锌时以氢氧化物沉淀分离铁、锰等金属离子后,在pH 5.5乙酸-乙酸钠缓冲液中,硫脲存在下,以二甲酚橙为指示剂,用EDTA标准溶液滴定锌。另一份滤液中铜采用碘化钾-硫代硫酸钠滴定法测定。方法用于铅锌矿样品中铅、锌和铜的测定,测定值与国家标准方法的测定值一致,测定结果的相对标准偏差在0.042%～0.088%之间。     

Controlling factors in the development of ceruloplasmin in pigs during the neonatal growth period

The sequence of ceruloplasmin development and limiting factors controlling this process in neonatal piglets was studied in a randomized block design. Guided by the change in serum ceruloplasmin levels, the liver copper concentrations of piglets fed copper-deficient and copper-supplemented diets were compared in three different periods. The plasma of piglets was devoid of both apo- and holo-ceruloplasmin at birth. The system responsible for the synthesis of apoceruloplasmin developed shortly after birth as indicated by a steady increase in serum ceruloplasmin activity during the first week without being affected by the diet. Copper apparently was not the limiting factor at this stage since very high concentrations of copper were found in the livers of newborn piglets. However, the liver copper was depleted quickly within 1 to 2 weeks and became the limiting factor if the diet was deficient in copper. The livers of these piglets were fractionated by differential centrifugation to compare the liver copper distribution in the two dietary groups with and without copper supplementation. The treatments did not affect the copper distribution on a percentage of total liver copper basis. However, fractionation study revealed that, unlike adult pig livers, about 70% to 80% of copper was present in the heavier particle fractions of piglet livers.     

Cerebrospinal fluid levels of transition metals in patients with Parkinson's disease

We compared CSF and serum levels of iron, copper, manganese, and zinc, measured by atomic absorption spectrophotometry, in 37 patients with Parkinson's disease (PD) and 37 matched controls. The CSF levels of zinc were significantly decreased in PD patients as compared with controls (p < 0.05). The serum levels of zinc, and the CSF and serum levels of iron, copper, and manganese, did not differ significantly between PD-patient and control groups. There was no influence of antiparkinsonian therapy on CSF levels of none of these transition metals. These values were not correlated with age, age at onset, duration of the disease, scores of the Unified Parkinson Disease Rating Scale of the Hoehn and Yahr staging in the PD group, with the exception of CSF copper levels with the duration of the disease (r = 0.38, p < 0.05). These results suggest that low CSF zinc concentrations might be related with the risk for PD, although they could be related with oxidative stress processes.     

Cellular distribution of orally and intramuscularly administered iron dextran in newborn piglets

Histochemical studies were performed on tissues from piglets of different ages treated orally with iron dextran soon after birth. The mucosal cells in the distal region of the small intestine were heavily laden with stainable iron granules during the first three days after the iron administration. The absorptive epithelial cells are desquamated within seven to ten days after birth. Consequently, the number of iron granules gradually diminishes during the first seven days after treatment and no iron granules are demonstrated 12 days after the administration of iron. The iron dextran complex is pinocytosed in newborn piglets and then transported via the lymphatic system. Thus the sinusoidal lining cells of the body and mesenteric lymph nodes are already heavily laden with iron granules 24 hours after oral treatment. This iron store is released only slowing during the first weeks of life. Great amounts of iron granules are demonstrated in the liver and spleen macrophages during the first week after the administration of iron. Due to the rapid utilization of iron in growing piglets these iron stores diminish sharply during the weeks following birth. The distribution of stainable iron in the lymph nodes, liver and spleen seven days after intramuscular injection of iron dextran in newborn piglets was comparable to that for oral administration at that stage of the experiment.