Abstract: This study aimed to investigate a possible connection between removal of dental amalgam restorations supported by antioxidant therapy and indicative changes of clinical chemistry parameters. A group of 24 patients, referred for complaints related to amalgam restorations, underwent a removal of their amalgams. All patients were treated with antioxidants (vitamin B-complex, vitamin C, vitamin E, and sodium selenite). An age- and sex-matched control group of 22 individuals was also included. The mercury (Hg) and selenium (Se) concentration in plasma, Hg concentration in erythrocytes, and 17 clinical chemistry variables were examined in three groups: patients before amalgam removal (Before), patients after amalgam removal (After), and control individuals (Control). The Hg and Se values decreased (p < 0.05) in plasma, and the Hg concentration decreased (p < 0.05) in erythrocytes after amalgam removal. The variables serum lactate dehydrogenase (serum LDH) and serum sodium differed significantly both when comparing Control with Before (p < 0.01) and Before with After (p < 0.01). The variables white blood cell count (WBC), blood neutrophil count, blood eosinophil count, blood basophil count, blood lymphocyte count, blood monocyte count, serum potassium, and serum creatinine differed in the Before/After test (p < 0.05). Multivariate statistics (discriminant function analysis) could separate the groups Before and After with only one misclassification.

Abstract:

Abstract: Amalgam separators are used to physically remove dental amalgam from waste water in dental clinics. They are thereby supposed to reduce mercury (Hg) emissions to the municipal waste water system to acceptable levels. We here present results from a comparative study in situ of three amalgam separators available on the market, all with a claimed efficiency of 99% according to Danish and ISO protocols, and using sedimentation as the principle of separation. We also present corresponding data for an investigational prototype of an improved separator. The obtained efficiency of the three commercial separators is far below what is stated by the manufacturer and by authorities assumed to be the efficiency in clinical conditions. They reduced Hg emissions by 79 - 91%, leaving an average Hg content in outgoing waste water of 1.5 mg L(-1). However, the prototype separator participating in this study retained 99.9% of the waste water Hg emissions, leaving an average Hg content in outgoing waste water of 0.004 mg L(-1). Physical restrictions prohibit sedimentary type separators to recover the Hg fractions causing the largest damages in wastewater treatment plants. This fraction is not considered in the ISO protocol for testing amalgam separators, which therefore needs to be revised. Abolishing the use of dental amalgam and cleaning the tubing systems is the most efficient long-term solution to reduce Hg emissions from dental clinics. Until then, Hg emissions originating from placing, polishing or removing existing amalgam fillings, should be counteracted by the use of low-emission amalgam separators, already on the market or presently being developed for use alone or together with sedimentary type amalgam separators.

Abstract: Cadmium (Cd) is a potentially toxic metal widely distributed in the environment and known to cause adverse health effects in humans. During coxsackievirus infection, the concentrations of essential and nonessential trace elements (e.g., iron (Fe), copper (Cu), and Cd) change in different target organs of the infection. Fe and Cu are recognized cofactors in host defence reactions, and Fe is known to be associated with certain pathological conditions of the brain. However, whether nonessential trace elements could influence the balance of essential trace elements in the brain is unknown. In this study the brain Fe, Cu, and Cd contents were measured through inductively coupled plasma mass spectrometry and their distributions determined by nuclear microscopy in the early phase (day 3) of coxsackievirus B3 (CB3) infection in nonexposed and in Cd-exposed female Balb/c mice. In CB3 infection the brain is a well-known target that has not been studied with regard to trace element balance. The brain concentration of Cu compared with that of noninfected control mice was increased by 9% (P < 0.05) in infected mice not exposed to Cd and by 10% (not significant) in infected Cd-exposed mice. A similar response was seen for Fe, which in infected Cd-exposed mice, compared to noninfected control mice, tended to increase by 16%. Cu showed an even tissue distribution, whereas Fe was distributed in focal deposits. Changes in Cd concentration in the brain of infected mice were less consistent but evenly distributed. Further studies are needed to define whether the accumulation and distribution of trace elements in the brain have an impact on brain function.

Abstract: OBJECTIVES: This study was done to evaluate the results and clinical relevance of an optimized lymphocyte proliferation test, MELISA, for metal-induced inflammation in patients with CFS-like symptoms. The treatment of patients consisted of the replacement of incompatible dental materials (RID) together with supportive anti-oxidant therapy. DESIGN OF THE STUDY: 513 patients were tested by MELISA at the beginning of the study. Out of this group, 248 patients were available for follow-up MELISA after RID. METHODS: In MELISA, lymphocytes are isolated from the blood and cultivated with different metal salts in tissue culture medium containing 10% inactivated human AB+ serum or autologous serum. After 5 days, the presence of metal-reactive lymphocytes are measured by isotope labelling of newly formed DNA in growing lymphoblasts and evaluated by calculating the Stimulation Index. RESULTS: Nickel was the most common sensitizer, followed by inorganic mercury, thimerosal, lead, cadmium, palladium and gold. After RID treatment, a decrease of metal-specific lymphocyte responses in patients who reacted to metals at the beginning of the study could be observed. The cultivation of lymphocytes in autologous and homologous serum did not significantly affect the results. Simultaneous, the health status of patients improved as well. CONCLUSIONS: Replacement of incompatible dental materials resulted in down-regulation of metal-induced lymphocyte sensitivity in vitro, as well as in the improvement of health status of majority of patients with unspecific CFS-like symptoms.

Abstract: The objective of this study was to determine the concentration changes of 13 elements in erythrocytes and plasma after the removal of dental amalgam and other metal alloys. Blood samples from 250 patients were collected, separated into erythrocytes and plasma, and analyzed by inductively coupled plasma-mass spectrometry. The 250 patients were divided into 3 groups (Negative, Zero, and Positive) depending on their estimation of quality of life in an earlier study. Magnesium in plasma, selenium and mercury in plasma, and erythrocytes showed decreased concentrations after amalgam removal in all groups (p < 0.05). Titanium in plasma, copper in plasma, and erythrocytes and zinc in plasma exhibited decreased concentrations after amalgam removal in the Negative and Positive groups (p < 0.05). Silver in plasma and gold in erythrocytes decreased in the Zero and Positive groups after amalgam removal (p < 0.05). Copper in erythrocytes and silver and gold in plasma showed higher concentrations after amalgam removal in the Negative compared to the Positive group (p < 0.05), suggesting that patients in the Negative group excrete metals slowly. Moreover, the cobalt levels in plasma were lowest in the Negative group and only this group showed a significant increase in vitamin B12 levels in blood after amalgam removal.

Abstract: Arsenic (As), a potentially toxic trace element, has been shown to influence viral replication and resistance to microbial infection. However, the impact of infection on the normal As status in target organs involved in the disease process has not been studied to date. In the present study, As was measured through inductively coupled plasma mass spectrometry in the plasma, liver, spleen, kidney, heart, pancreas and brain at days 1 and 3 of coxsackievirus B3 infection in female Balb/c mice. The severity of the infection was assessed from clinical signs of disease. The infection changed plasma As in a biphasic pattern with a small increase (n.s.) at day 1 that turned into a decreasing trend (13%, p<0.05) by day 3. In the liver, spleen, heart, pancreas and kidney As was unchanged at day 1 but, at day 3, it had decreased by 71% (p<0.01), 64% (p<0.01), 55% (p<0.01), 63% (p<0.01) and 73% (p<0.01), respectively. In the brain, As went unchanged. The pathophysiological interpretation of these findings requires further research.

Abstract: As part of the general host response to coxsackievirus B3 (CB3) infection, the concentration of essential and nonessential trace elements changes in different target organs of the infection. Essential (e.g., Se) and nonessential (e.g., Hg) trace elements are known to interact and affect inflammatory tissue lesions induced by CB3 infection. However, it is unknown whether these changes involve the brain. In the present study, the brain Hg and Se contents were measured through inductively coupled plasma-mass spectrometry and their distribution investigated by means of nuclear microscopy in the early phase (d 3) of CB3 infection in normally fed female Balb/c mice. Because of the infection, the concentration of Hg (4.07 +/- 0.46 ng/g wet wt) and Se (340 +/- 16 ng/g wet wt) in the brain increased twofold for Hg (8.77 +/- 1.65 ng/g wet wt, p < 0.05) and by 36% for Se (461 +/- 150 ng/g wet wt, ns). Nuclear microscopy of brain sections from mice having elevated Se and Hg concentrations failed to find localized levels of the elements high enough to make detection possible, indicating approximately homogeneous tissue distribution. Although the pathophysiological interpretation of these findings requires further research, the increase of Hg in the brain during infection might have an influence on the pathogenesis of the disease.

Abstract: The pathogenesis of some heart diseases has been associated with changes in the balance of certain trace elements. However, whether blood trace element changes exist that are related to changes in the cardiovascular system are, in most cases, unknown. In this study, blood trace element levels were analysed in 46 patients with non-rheumatic aortic valve sclerosis that were previously shown to have a disturbed trace element balance in their valve tissue, including 11/15 elements. Results showed significant changes of blood levels of 8/15 trace elements in these patients when compared with blood levels in 46 healthy controls. Of these elements, Cd and Mg were the only elements that increased in both blood and valves. Cu and Se were increased in blood but decreased in valves, whereas Co and Zn were decreased in blood but increased in valves. Several elements (As, Ca, Fe, Pb, and V) were unchanged in blood although changed in valves. Although Mn and Hg showed changes in blood, this was not evident in the valves. Al and Ag were the only elements that did not change in both blood and valves. Significant covariation in blood and valve levels was only observed for Al and Pb. The recorded pattern of trace element changes indicates a complex competition/exchange between body compartments in this disease, where the increased blood Cu/Zn ratio suggests an ongoing infectious/inflammatory process.

Abstract: In experimental studies on the common human coxsackievirus B type 3 (CB3) infection, administered cadmium (Cd) is known to accumulate in the liver and kidneys. CB3 adapted to Balb/c mice was used to study whether infection affects the Cd-binding protein, metallothionein (MT) and if this alters the normal physiological trace element balance in the liver, kidney, spleen and brain. On day 3 of infection, degradation of liver proteins (44%, P<0.01) occurred, whereas in the spleen, protein increased (63%, P<0.05). The infection increased MT five-fold (P<0.01) in liver and kidneys, and in spleen by 34% (P<0.05). A redistribution of Cd and copper (Cu) from the liver to the kidney was associated with this increase in MT, resulting in an increased (P<0.01) kidney/liver ratio for both elements. The infection increased the zinc (Zn) concentration more in the kidney than in the liver, but the kidney/liver ratio was not significantly affected. Results show that MT is increased in several organs during the early phase of infection and is associated with redistribution of both essential and non-essential trace elements. This may be a normal response in common infections that could adversely influence the pathogenesis when the host is concomitantly exposed to potentially toxic trace elements, even at levels in the physiological range.

Abstract: The objective was to study toxin-induced effects on physiological parameters in the rabbit and whether these parameters show dose-response and co-variation after administration of a recombinant fusion protein between staphylococcal enterotoxin (SE) and the Fab fragment of an antibody. Rabbits are very sensitive to SE toxins and the cardiovascular and immune effects are similar to those observed in septic shock in man. The test compound, r-C242 Fab-SEA, was administered intravenously to anaesthetised New Zealand white rabbits at doses in the range of 0.00005-50 microg/kg. All rabbits were checked for titres of anti-SEA antibodies before entering the experiment, since they could neutralise the effect of the test compound. Heart rate, blood pressure and body temperature were continuously monitored before and during 6 h after dosing. Immediately before the start of administration and 3 and 6 h during the experiment, blood gases (pO(2) and pCO(2)), pH, haematology, clinical chemistry, cytokine response (TNF-alpha) and trace elements (Mn, Cu, Zn, Se, Ag, Cd, Hg and Pb) were measured. No mortality occurred, but at 50 microg/kg severe adverse clinical signs developed. The decrease in blood pressure was weakly dose-related. Heart rate, ECG, body temperature, pCO(2) and pH were not affected by the treatment. pO(2) tended to increase as a function of time, but not in relation to dose. WBC and PLT decreased dose dependently. TNF-alpha was not affected by the treatment. The major effects on clinical chemistry were a dose-dependent increase in AST and creatinine. Potassium and urea showed dose dependent increases, mainly at higher doses, though these changes were of less value for drug selection purposes. Trace element changes were observed, including an increase in Mn and a decrease of Zn at all doses. The Cu/Zn ratio decreased below normal at low doses, whereas at high doses in which adverse effects developed, it increased above normal. Post mortem examination revealed minimal to moderate dose-related granulocytic infiltrate in the lungs. The present study showed dose-response and co-variation between several changes in cardiovascular, haematology, clinical chemistry and trace element parameters during the initial phase of toxin-induced effects preceding a possible lethal endpoint and associated patho-physiological changes.

Abstract: Selenium and mercuric chloride (MC) interactions regarding effects on cell growth and cell death have been studied. Human K-562 cells were pretreated or simultaneously treated with either selenite (5 or 50 microM) or selenomethionine (10 or 50 microM) and with MC (35 or 50 microM). The 35-microM MC treatments resulted in a clear inhibition of cell growth with no obvious difference between mercury-treated and mercury-selenium-treated cells. Furthermore, the apoptotic frequency was similar at all observations for all selenium treatments with 35 microM MC. In the simultaneously treated selenite and 50- microM MC combinations, a selenite-dependent protection was shown both by increased cell growth and by lower apoptotic frequency at 48 and 96 h of exposure. Both treatments with selenomethionine showed protection observed as an increased cell growth at 48 and 96 h and as decreased apoptotic frequency at 96 h of exposure.

Abstract: INTRODUCTION: The trigger for some cases of juvenile diabetes has been suggested to be an interaction between a virus and various trace elements. Infection with human coxsackievirus B3 (CB3) in the murine model results in viral replication and inflammation in the pancreas. AIM: To determine how infection affects the trace element balance in the pancreas. METHODOLOGY: Concentrations of the following trace elements were measured in the serum and pancreas during the early phase (days 1 and 3) of CB3 infection in female Balb/c mice: aluminium, arsenic, cadmium (Cd), calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), lead (Pb), magnesium (Mg), manganese (Mn), mercury (Hg), selenium, silver, vanadium (V), and zinc (Zn). The trace element concentrations were measured through inductively coupled plasma mass spectrometry. The histopathology was established by hematoxylin-eosin techniques and immunohistochemical staining of both CD4 and CD8 cells of the pancreas. RESULTS: Infected mice developed expected clinical signs of disease. The only changes at day 1 occurred in the serum, with a pronounced decrease in the Zn concentration and a small increase in the V concentration. At day 3, concentrations of several trace elements, including Cu, Zn, Fe, Ca, V, and Mn, showed pronounced changes in both the serum and the pancreas. Ca, Cu, Mg, Mn, and V, but none of the potentially toxic elements, accumulated in the pancreas. Cu and V concentrations increased in the serum as well. CONCLUSION: Several trace element changes, preceding the development of pancreatitis, occurred in the pancreas in this viral infection, the exact pathogenic interpretation of which warrants further studies.

Abstract: An association between Chlamydia pneumoniae and atherosclerotic cardiovascular diseases has been suggested. However, other factors may interact in the pathogenesis of valve sclerosis. Therefore, trace elements important for C. pneumoniae growth and host defense and markers of C. pneumoniae infection were studied in sclerotic valves and serum. Forty-six patients undergoing surgical valve replacement due to advanced aortic sclerosis were prospectively studied. Valves from 15 forensic cases with no heart valve disease and plasma from 46 healthy volunteers served as controls. C. pneumoniae was detected in 16/46 (34.8 %) sclerotic valves and in 0/15 forensic controls. IgG and IgA antibodies to C. pneumoniae were present in 54.3% and 26.1 % patients, respectively. In the patients' valves, iron, magnesium, and zinc each correlated to calcium, a marker of the histopathological severity of disease. Patients showed 10- to 70-fold increases of these trace elements in valves and an increased copper/zinc ratio in serum. In a majority of aortic sclerosis patients, one of several markers of C. pneumoniae infection were detected and all patients had a disturbed trace element balance in valves and serum suggestive of active immune process and infection. The pattern of trace element changes was essentially similar regardless of positive makers of C. pneumoniae, suggesting a similar etiopathogenesis in both subgroups. The 20-fold increase in iron, essential for C. pneumoniae growth, in sclerotic valves suggests a new possible link to this infection in aortic sclerosis.

Abstract: In Coxsackievirus B3 (CB3) infection, the heart and pancreas are major target organs and, as a general host response, an associated immune activation and acute phase reaction develops. Although iron (Fe), copper (Cu), and zinc (Zn) are involved in these responses, sequential trace element changes in different target organs of infection have not been studied to date. In the present study, Fe, Cu, and Zn were measured through inductively coupled plasma mass spectrometry (ICP-MS) in the plasma, liver, spleen, heart, and pancreas during the early phase (d 1 and 3) of CB3 infection in female Balb/c mice. The severity of the infection was assessed through clinical signs of disease and histopathology of the heart and pancreas, including staining of CD4 and CD8 cells in the pancreas. During infection, the concentrations of Fe, Cu, and Zn changed in the plasma, liver, and pancreas, but not in the spleen and heart. The changes in plasma Cu, Zn, and Fe seemed to be biphasic with a decrease at d 1 that turned into increased levels by d 3. Cu showed similar biphasic changes in the liver, spleen, and pancreas, whereas, for Zn and Fe, this pattern was only evident in the liver. In the pancreas, the reverse response occurred with pronounced decreases in Fe (23%, p < 0.05) and Zn (64%, p < 0.01) at d 3. Although the pathophysiological interpretation of these findings requires further research, the sequential determination of these elements may be of clinical value in enterovirus infections in deciding the stage of disease development.

Abstract: A myocarditic coxsackievirus B3 (CB3) infection in adult male A/J mice was used to investigate the effects of 2,3,7,8-tetrachlorodibenzo-para-dioxin (TCDD) exposure on mortality and on inflammatory lesion, virus and trace element contents of the heart. The mice were injected with four weekly intraperitoneal (i.p.) injections of TCDD (a loading dose of 5 microg/kg followed by three maintenance doses of 1.4 microg/kg). To reach a steady-state body burden of TCDD the mice were allowed a 90-day recovery period before infection with CB3 virus. TCDD increased the infection-induced mortality rate, whereas in TCDD-exposed mice, heart lesions at day 7 after the virus inoculation (median value 0.67% of the tissue section area; interquartile range 0.28; not statistically significant) were one-third of that in non-exposed infected mice (2.07% of the tissue section area; interquartile range 3.06). The size of the inflammatory heart lesion correlated to the amount of virus (r(s) = 0.829, P < 0.01) as well as to the calcium (Ca: r(s) = 0.725, P < 0.01) and the magnesium (Mg: r(s) = -0.615, P < 0.05) contents. In TCDD-exposed mice in situ hybridisation of viral RNA in the myocardium at day 7 showed a tendency to decreased amounts of virus, as well as a less pronounced increase in myocardial Ca content, both supporting a milder myocardial disease after TCDD exposure. No effect of TCDD exposure was seen on the zinc (Zn) or selenium (Se) levels in the myocardium. In conclusion, although TCDD seemed to have a limiting effect on viral replication and the development of the inflammatory lesion in the myocardium, mortality was increased by TCDD in this infection model. However, TCDD had no significant effects on the selected trace elements that could be of importance for the severity of the inflammatory lesion (Ca, Se), for the local host response activation (Zn) or for the development of myocardial disease complications (Mg). Accordingly, the increased mortality may be a result of an infection-induced increase in TCDD toxicity to vital organs other than the heart, and/or a TCDD-induced change in the tissue affinity and virulence of the virus, possibly causing involvement of other target organs in the infectious process and changed pathogenesis.

Abstract: Several trace elements are essential nutrients for an optimal functioning of organs and tissues, including the immune system and the heart. The pathogenesis of some heart diseases has been associated with changes in the balance of certain trace elements. The etiology of nonrheumatic aortic valve sclerosis is unknown, however. A prospective study was performed on trace element changes in the sclerotic valves of 46 patients undergoing surgical aortic valve replacement because of aortic stenosis. Valves from 15 individual forensic cases without known cardiac disease served as controls.The contents of 15 trace elements (Al, As, Cd, Ca, Co, Cu, Fe, Pb, Mg, Mn, Hg, Se, Ag, V, and Zn) were measured by inductively coupled plasma - mass spectrometry (ICP-MS) of aortic valve tissue from both patients and forensic autopsy controls. Some trace elements showed similar concentrations in sclerotic and control valves (Al, Ag, Hg, Mn), whereas a few were moderately changed in the sclerotic as compared with the control valves, including an increase in Cd by 52% (p < 0.05) and decreases in Se by 14% (p < 0.05), in V by 42% (p < 0,001), and in Cu by 45% (p < 0.001). However, there were pronounced increases (p < 0.001) in the concentrations of As (5-fold), Ca (70-fold), Co(10-fold), Fe (20-fold), Pb (8-fold), Mg (20-fold), and Zn (10-fold) in the sclerotic valves. Thus, sclerotic aortic valve disease is associated with a pronounced imbalance in several trace elements of well-known importance for cardiovascular and immune function as well as in trace elements with hitherto unknown significance.

Abstract: The interaction between selenium and cadmium was studied in relation to cellular uptake and expressions of selenium-cadmium interaction. Human K-562 cells were pre-treated or simultaneously treated with (5 or 50 microM) selenite or (10 or 50 microM) selenomethionine and with (60 or 75 microM) cadmium nitrate. Cells pre or simultaneously treated with selenite revealed increased cadmium concentration with increased doses of selenite, particularly pronounced in the simultaneous treatments. In both treatments, selenium protection was observed during the exposure period, but not during the growth period. In cells simultaneously treated with selenomethionine and 60-microM cadmium, an increase in cadmium concentration was observed after increased selenium dose. In addition, it was found that simultaneous selenomethionine treatment with 60-microM cadmium resulted in selenium protection during the exposure period, although protection was not observed during the growth period.

Abstract: OBJECTIVES: The purpose of this study was to evaluate treatment of patients suffering from chronic ill health with a multitude of symptoms associated with metal exposure from dental amalgam and other metal alloys. SETTING AND DESIGN: We included 796 patients in a retrospective study using a questionnaire about symptom changes, changes in quality of life as a consequence of treatment and assessment of care taking. METHODS: Treatment of the patients by removal of offending dental metals and concomitant antioxidant therapy was implemented according to the Uppsala model based on a close co-operation between physicians and dentists. RESULTS: More than 70% of the responders, remaining after exclusion of those who had not begun or completed removal, reported substantial recovery and increased quality of life. Comparison with similar studies showed accordance of the main results. Plasma concentrations of mercury before and after treatment supported the metal exposure to be causative for the ill health. MAIN FINDINGS: Treatment according to the Uppsala model proved to be adequate for more than 70% of the patients. Patients with a high probability to respond successfully to current therapy might be detected by symptom profiles before treatment. CONCLUSIONS: The hypothesis that metal exposure from dental amalgam can cause ill health in a susceptible part of the exposed population was supported. Further research is warranted to develop laboratory tests to support identification of the group of patients responding to current therapy as well as to find out causes of problems in the group with no or negative results.

Abstract: Twenty-seven consecutive patients with health problems associated with dental amalgam were recruited. In spite of thorough medical examinations, there were no diagnoses available. The patient group was dominated by women. A healthy age- and sex-matched control group with dental amalgams without symptoms was also recruited. Metal level monitoring in plasma and nuclear microscopy of isolated individual blood cells were carried out. Significant increases of copper, iron, zinc and strontium were found in patient plasma. There was no significant difference in plasma selenium between the groups. Mercury was significantly increased in patient plasma, although there was overlap between the groups. In erythrocytes a significant increase in calcium and a significant decrease in magnesium, copper, manganese and zinc were found. Calcium, magnesium, manganese and copper increased in patient neutrophil granulocytes. A significant decrease was found for zinc. A conspicuous finding was the presence of measurable mercury in a few of the cells from the patient but not in the control group. Thus, nuclear microscopy of isolated individual blood cells might provide a better diagnostic tool for metal exposure than blood plasma measurements.

Abstract: The diverse genetic backgrounds of lupus-prone murine models, which produce both quantitative and qualitative differences in disease expression, may be a valuable resource for studying the influence of environmental exposure on autoimmune disease in sensitive populations. We tested this premise by exposing autoimmune-prone BXSB and the nonautoimmune C57BL/6 mice to the heavy metal mercury. Although both strains express a nonsusceptible H-2 haplotype, exposure to mercury accelerated systemic autoimmunity in both male and female BXSB mice, whereas the C57BL/6 mice were resistant. The subclasses of antichromatin antibodies elicited in BXSB mice by mercury exposure were more consistent with the predominant Th1-type response of idiopathic disease than with the Th2-type response found in mercury-induced autoimmunity (HgIA). The appearance and magnitude of both humoral and cellular features of systemic autoimmunity correlated with the mercury dose. Furthermore, environmentally relevant tissue levels of mercury were associated with exacerbated systemic autoimmunity. These studies demonstrate that xenobiotic exposure can accelerate spontaneous systemic autoimmunity, and they support the possibility that low-level xenobiotic exposure enhances susceptibility to systemic autoimmunity in genetically susceptible individuals.

Abstract: Selenium and methylmercuric chloride (MMC) interactions regarding cellular uptake and selenium protection on MMC toxicity have been studied. Human K-562 cells were pretreated or simultaneously treated with either selenite (5 or 50 microM) or selenomethionine (10 or 50 microM) together with (3.5 or 5 microM) MMC. Cells simultaneously treated with selenite or selenomethionine and 3.5 microM MMC showed a decreased mercury concentration with increased selenium dose especially seen in the selenite combinations. The simultaneous selenite and MMC 3.5 microM combinations showed growth curves with an increasing number of viable cells with increased selenite dose. All combinations with 5 microM MMC were toxic to the cells. Interactions between selenite or selenomethionine and MMC regarding cellular uptake of mercury and selenium were observed and indications of selenite protection against MMC toxicity in human K-562 cells were noticed.

Abstract: Selenium and mercuric chloride (MC) interactions regarding cellular uptake and selenium protection on MC toxicity have been studied. Human K-562 cells were pretreated or simultaneously treated with either selenite (5 or 50 microM) or selenomethionine (10 or 50 microM) together with MC (35 or 50 microM). Both treatments with selenite showed an increase of mercury uptake with increased selenium dose. In the pretreated or simultaneously treated selenite and 35 microM MC combinations, no inhibition of growth was seen, whereas all 50-microM MC combinations were toxic to the cells. A selenite-dependent protection was obtained for both exposure protocols when considering the cellular uptake of mercury. The cells died when the accumulation on d 4 reached more than about 0.8 x 10(-15) mol/cell of mercury, whereas they survived up to twofold more mercury uptake when exposed to selenite. Selenomethionine gave, with a few exceptions, similar effects as selenite on MC uptake and toxicity.

Abstract: The present study was undertaken in order to study the effects of the broad-acting chelating agent CaNa2-EDTA on plasma trace elements and cardiovascular function in anesthetised New Zealand White rabbits. Trace elements are important for cardiovascular and immune functions and the rabbit is a well-accepted species in cardiovascular studies. The test compound CaNa2-EDTA was administered intravenously to rabbits at single doses of 4, 20, and 100 mg/kg. In addition, at 20 mg/kg, the effects of a second dose after 3 h were also investigated. Heart rate, blood pressure and body temperature were continuously monitored during a 6-h interval after injection of CaNa2-EDTA. Immediately before administration (-1 min) and at 3 and 6 h over the period of the experiment, the plasma cytokine response (tumor necrosis factor-alpha) and trace elements (Mn, Cu, Zn, Se, Ag, Cd, Hg, Pb) were measured. Regardless of dose, blood pressure was found to decrease, but no corresponding changes in heart rate were observed. Both a repeated administration of 20 mg/kg and a single dose of 100 mg/kg were detrimental and caused severe cardiovascular effects and lethality. alpha-TNF tended to increase, though only at 100 mg/kg. The electrocardiogram and body temperature were not affected by the treatment. The most pronounced trace element change was a dose-dependent increase in Mn that was equally pronounced at all time-points after the dose. There was an initial decrease in Cd at low dose levels (4 and 20 mg/kg) that turned into an increase after 6 h at 20 mg/kg and from 2 h at 100 mg/kg. A similar pattern with pronounced decreases at low dose levels was observed for Zn. Cu decreased similarly at all dose levels. For the other trace elements, no or inconsistent effects were observed. This model allows the study of concomitant cardiovascular and trace element changes during treatment with drugs and chelating agents preceding a possible lethal end point and associated pathophysiologic changes.

Abstract: Effects of some metals on the growth of cultured human erythroleukemia K-562 cells were investigated when grown in two different types of media based upon RPMI-1640 or Ham's F-10. The study on proliferation, using RPMI-1640 supplemented with sodium selenite, selenomethionine, mercuric chloride, methylmercuric chloride and cadmium nitrate showed no inhibition of growth at concentrations of 2.5, 25, 25, 2.5 and 25 microM, while at 75, 250, 50, 5 and 50 microM toxicity was apparent. Selenite at 5-50 microM and selenomethionine at 50-100 microM inhibited the growth. In Ham's F-10 supplemented with the same compounds no inhibition was found at concentrations of 5, 10, 25, 1 and 50 microM, while at 50, 100, 50, 5 and 75 microM toxic effects were noted. Selenite 10 microM and selenomethionine 25-50 microM inhibited the proliferation. Measurements of trace element levels in pellets of K-562 cells grown in RPMI-1640 or Ham's F-10 unveiled higher cell contents of cadmium and selenium in cells grown in RPMI-1640, being consistent with higher concentrations of these elements in that medium. Manganese and mercury concentrations were higher in cells grown in Ham's F-10 correlating with a higher medium concentration of these elements. The growth responses and cellular uptake differed between the metals and the selenocompounds and although extrapolating the results to humans is difficult the selenium exposures were in approximately the same order of magnitude as in human exposures. The compounds could be ranked according to decreasing toxicity as: methylmercuric chloride > mercuric chloride, cadmium nitrate, sodium selenite > selenomethionine.

Abstract: The aluminium content in bone has been related in several ways: to the weight of wet bone, to the weight of dry bone, to the weight of bone-ash and to the calcium content of bone. We determined the accuracy and precision of measurement (using an inductively coupled mass-spectrometer) in 30 bone samples taken from one patient. The coefficient of variation of the aluminium/weight-quotient was 12.4 per cent for wet bone, 4.7 for dry bone and 5.0 for bone ash; and the coefficient of variation of the aluminium/calcium-weight-quotient was 7.5 per cent. Thus, the aluminium content in bone seems to be best related to the weight of dry bone.

Abstract: During most infections plasma, concentrations of trace elements change, but it is unclear if this reflects changes in infected target tissues. In coxsackievirus B3 (CB3) infection, the myocardium is a target in both humans and mice. The concentrations of 12 trace elements were analyzed by inductively coupled plasma-mass spectrometry (ICP-MS) in the myocardium of sham-inoculated controls and infected A/J mice 4 and 7 d postinoculation. The size of the inflammatory lesion was positively correlated to the virus content of the heart, as estimated by histopathology and in situ hybridization, respectively. Iron, cobalt, vanadium, and selenium showed transient changes, whereas for the other elements, tendencies on d 4 were manifest on d 7. A three-fold increase in calcium on d 7 suggests prestages of calcification, whereas increases in zinc, selenium, and copper may be the result of the accumulation of immune cells. The magnesium decrease may contribute to the increased sensitivity to cardiac arrhythmias in myocarditis.

Abstract: The selenium uptake and retention have been studied in K-562 cells exposed to selenite or selenomethionine. In the uptake experiments the cells were exposed to two doses of selenite (5 or 50 microM) or selenomethionine (10 or 50 microM). In the retention study the cells were treated for 2 h with the above mentioned doses of the selenocompounds before being observed at different times. The selenium uptake in cells exposed to selenite 5 microM began to saturate at 8 h, but increased again between 48 and 96 h. In cells exposed to selenite 50 microM the selenium uptake never reached a maximum, however, at 48 and 96 h the cell viability decreased strongly. The two doses of selenite showed different retention patterns, with a relatively small cellular decrease of selenium after treatment with selenite 5 microM compared to treatment with 50 microM of selenite. The selenium uptake in cells exposed to selenomethionine 10 microM or selenomethionine 50 microM began to saturate at 24 h and 48 h, respectively. The retention patterns were similar for both selenomethionine doses with a continuous decrease of the selenium concentration during the whole observation period. The results indicated a more controlled uptake and retention pattern of selenomethionine compared to selenite.

Abstract: Methyl mercury (MeHg) has been shown to change Coxsackie virus type B3 (CB3) myocarditis in a direction compatible with the development of chronic disease. Murine models of CB3 myocarditis closely mimic the pathogenesis in humans. There are also indications that metals, such as mercury, and trace elements may interact and adversely affect viral replication and development of inflammatory lesions. The effects of low-dose MeHg exposure on myocardial trace element distribution, as determined by means of nuclear microscopy, was studied in CB3 myocarditis. Balb/c mice were fed a MeHg-containing diet (3.9 mg/kg diet) for 12 wk prior to infection. Areas of inflammatory lesions in the myocardium were identified by traditional histologic examination, and serial tissue sections in these selected areas were used for immune histology (macrophages), in situ hybridization of virus genomes, and nuclear microscopy of tissue trace element distribution. Areas with no inflammation or virus were compared with areas of ongoing inflammation and viral replication. In the inflammatory lesions of MeHg-exposed mice as compared to nonexposed mice, the myocardial contents of calcium (Ca), manganese (Mn), and iron (Fe) were significantly increased, whereas the zinc (Zn) content was decreased. The increased Ca and decreased Zn contents in the inflamed heart may partly explain a more severe disease in MeHg-exposed individuals. Although not significant in the present study, with a limited number of mice, the inflammatory and necrotic lesions in the ventricular myocardium on d 7 of the infection was increased by 50% (from 2.2% to 3.3% of the tissue section area) in MeHg-exposed mice and, also, there was a tendency of increased persistence of virus with MeHg exposure. No increased MeHg uptake, either in the inflammatory lesions or in the areas of noninflamed heart tissue in infected mice, could be detected. The present results indicate that a "competition" exists between potentially toxic heavy metals from the environment/diet and important trace elements in the body and that a disturbed trace element balance adversely influences the development of pathophysiologic changes in inflammatory heart disease.

Abstract: During most infections the plasma levels of trace elements change, but it is not clear if this reflects changes in the infected tissues. Coxsackievirus B3 (CB3) infection may result in viral replication, subsequent inflammation and changed trace element levels in the myocardium. In the present study, the trace element levels in the plasma and heart of adult male A/J mice were determined during the pre-inflammatory stage (day 4) of CB3 myocarditis for the following trace elements: aluminium (Al), arsenic (As), calcium (Ca), cobalt (Co), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), selenium (Se), silver (Ag), vanadium (V) and zinc (Zn). The severity of the infection was assessed through clinical signs of disease and trace element levels were measured through inductively-coupled plasma mass-spectrometry (ICP-MS). In the heart, the levels decreased for V (59%; p < 0.01), Co (38%; p < 0.01), Al (81%; p < 0.01), As (66%; p < 0.01) and Se (16%; p < 0.01). Increased levels were detected for Mn (13%; p < 0.05), Fe (48%; p < 0.01), Cu (34%; p < 0.01) and Ag (46%; p < 0.01). In the plasma, decreases were detected in the level of Zn (32%; p < 0.05), whereas increases were seen in Mn (362%; p < 0.05), Fe (272%; p < 0.01), Co (71%; p < 0.05), Cu (25%; n.s.) and Mg (43%; p < 0.01) levels. A correlation was found between the levels in plasma and myocardium for Co (r(s) = -0.636; p < 0.05), Fe (r(s) = 0.764; p < 0.05), Mn (r(s) = 0.682; p < 0.05) and Mg (r(s) = -0.791; p < 0.05). Thus, determination of some of these trace elements in the plasma may be useful to indicate target tissue involvement in the early pre- inflammatory stage of an infectious disease. Some of these elements are important nutrients for the immune system, while others may be associated with the development of disease complications, such as cardiac arrhythmias.

Abstract: Many patients attribute their health problems to amalgam and other dental metals. In genetically susceptible indviduals, mercury and gold may function as haptens and elicit allergic and autoimmune reactions. The frequency of metal-induced lymphocyte responses was examined in 3,162 patients in three European laboratories using MELISA(R), an optimized lymphocyte proliferation test. The patients suffered from local and systemic symptoms attributed to dental restorations. The effect of dental metal removal was studied in 111 patients with metal hypersensitivity and symptoms resembling Chronic Fatigue Syndrome (CFS). After consultation with a dentist the patients decided to replace their metal restorations with non-metallic materials. The changes in health and in vitro lymphocyte reactivity were studied by inquiries and follow-up MELISA(R). Lymphocyte reactivity was also analyzed in 116 healthy subjects with no complaints of metal allergy. A significant number of patients had metal-specific lymphocytes in the blood. Nickel was the most common sensitizer, followed by inorganic mercury, gold, phenylmercury, cadmium and palladium. As compared to lymphocyte responses in healthy subjects, the CFS group had significantly increased responses to several metals, especially to inorganic mercury, phenylmercury and gold. Following dental metal removal, 83 patients (76%) reported long-term health improvement. Twenty-four patients (22%) reported unchanged health and two (2%) reported worsening of symptoms. Following dental metal replacement, the lymphocyte reactivity to metals decreased as well. We propose that an inflammatory process induced by metals may modulate the hypothalamic-pituitary-adrenal axis (HPA axis) and trigger multiple non-specific symptoms characterizing CFS and other chronic conditions like myalgic encephalitis (ME) and multiple chemical sensitivity (MCS).

Abstract: Dental amalgam restorations are a significant source of mercury exposure in the human population, but their potential to cause systemic health effects is highly disputed. We examined effects on the immune system by giving genetically mercury-susceptible Brown Norway (BN) rats and mercury-resistant Lewis (LE) rats silver amalgam restorations in 4 molars of the upper jaw, causing a body burden similar to that described in human amalgam-bearers (from 250 to 375 mg amalgam/kg body weight). BN rats with amalgam restorations, compared with control rats given composite resinous restorations, developed a rapid activation of the immune system, with a maximum 12-fold increase of the plasma IgE concentration after 3 wks (p < 0.001; Mann-Whitney's test). LE rats receiving amalgam restorations showed no significant increase of plasma IgE (p > 0.05). After 12 wks, BN rats with amalgam restorations showed significantly increased (p < 0.05) titers of immune-complex (IC) deposits in the renal glomeruli and in the vessel walls of internal organs. These rats also showed a significant (p < 0.05), from six- to 130-fold, increase in tissue mercury concentration in the concentration order kidney > spleen > cerebrum occipital lobe > cerebellum > liver > thymus, and the tissue silver concentration was significantly (p < 0.05) increased from three- to 11-fold. Amalgam-implanted BN rats showed a significant (p < 0.05) increase in copper concentration in the kidney and spleen, and in kidney selenium concentration. We conclude that dental amalgam restorations release substantial amounts of their elements, which accumulate in the organs and which, in genetically susceptible rats, give rise to activation of the immune system and systemic IC deposits.

Abstract: In order to elucidate the question of free radical involvement in acute porphyric crisis, antioxidants were administered to two acute intermittent porphyria patients with long-standing recurrent attacks. Clinical condition and urinary excretion of porphyrins and porphyrin precursors were monitored before, during and after an eight week therapy with daily doses of vitamin E, beta-carotene, ascorbic acid, selenium, vitamin Q, acetylcysteine, mannitol and carnitine. Blood cell trace element profiles were followed. The administration of the compound antioxidant formula was found not to further impair the clinical or biochemical conditions of the patients but the incidence of the recurrent crises or the severity of the symptoms were not positively affected. Aberrant blood cell trace element profiles with increased granulocyte manganese were normalized during treatment, on cessation of the therapy again resuming the abnormal pretreatment patterns, which may suggest an origin in oxidative stress. No correlation was observed between the concentration of granulocyte manganese and the excretion of 5-aminolaevulinic acid. Indications for participation of this porphyrin precursor in a radical generating process leading to generalized mitochondrial superoxide dismutase induction, as conceivably signalled by increased intracellular manganese, were thus not obtained. The failure to note a clinical response to antioxidant therapy may be due to factors dependent upon dosage of, or interaction between, the antioxidant compounds given, or on restricted bioavailability of the antioxidants at critical anatomical sites, and does not per se invalidate the model of acute porphyria as a hyperoxidative condition.

Abstract: The significantly increased concentrations of granulocyte manganese in subjects with AIP may be an indication of overexpression of manganese-associated enzymes. In this study we present further observations related to this phenomenon and speculate that this may provide a rational basis for hypotheses attempting to explain the pathogenesis of the acute attack of porphyria. Such hypotheses are advanced with regard to pyruvate carboxylase, mitochondrial superoxide dismutase and glutamine synthetase, three manganese-dependent enzymes associated with either ALA-generating or ALA-dependent processes. The metabolic impacts in acute porphyria of these enzymes would be functions of the current energy charge of the organism, and would thus explain the protecting and ameliorating effects of glucose in these conditions. Although granulocytes from AIP subjects have elevated manganese concentrations, this did not appear to be associated with increased activities of two enzymes assayed, pyruvate carboxylase or mitochondrial superoxide dismutase. However, enzyme activities in white blood cells do not necessarily represent the levels of catalytic activity in cell types involved in the phenotypic expression of porphyria. Thus it proposed that hypotheses along these new lines of thinking are not flawed by the apparently missing correlations, and should not be therefore discarded. The possible roles of manganese-associated enzymes in the mechanisms behind the acute porphyric attack are discussed in some detail in the paper.

Abstract: The incidence of fragility fractures has increased epidemically. Especially patients with senile dementia (including Alzheimer's disease) have a greatly increased risk of fragility fractures. Aluminum inhibits bone mineralization; the greater the aluminium exposure, the higher the risk of an early fracture. Aluminium is neurotoxic and may, in addition to genetic factors, play a role in the development of Alzheimer's disease by contributing to the formation of the characteristic beta-amyloid and neurofibrillary tangles. Thus, a common denominator between Alzheimer's disease and bone fragility may be a chronic low-grade aluminum intoxication. The epidemic of fragility fractures may be caused by increased aluminium exposure--due to the use of aluminum cooking pots or the pollution acidification of our environment. In our pilot study of 26 hip-fracture patients (13 patients with Alzheimer's disease and 13 individually age- and gender-matched non-demented patients), the aluminum content, determined mass-spectro-metrically, was higher in trabecular bone biopsies from the patients with Alzheimer's disease than from the non-demented patients (p = 0.005). The aluminum content was also higher in the younger of the 26 patients (p = 0.02). Our findings agree with the hypothesis that aluminum plays a role in the development of Alzheimer's disease and bone fragility.

Abstract: Effects of Pb2+, Ni2+, Hg2+ and Se4+ on cultured human glioma U-343MG cells were investigated considering uptake, toxicity and, in combination with radiation, clonogenic cells survival. The cells were exposed to 0-100 microM of the metals for a week before the evaluation. The tests showed a tendency to toxicity with 10 microM nickel although not significant (P > 0.05). Selenium, lead and mercury exerted a significant toxicity (P < 0.05) at 2.5 microM, 10 microM and 1 microM, respectively. To challenge the clonogenic cell survival capacity, the cells were irradiated with 60Co photons after being exposed to the highest nontoxic concentration of the different metals. The clonogenic cell survival tests, after irradiation, showed no significant change if the cells were exposed to 5 microM nickel, 0.5 microM selenium or 5 microM lead compared with those not exposed. Mercury, 0.1 microM, gave a relative reduction in survival compared with only irradiated cells of 58 +/- 17%. Thus, only mercury affected the radiation-induced damage and/or repair. When exposed to the highest nontoxic concentrations of the different metals, the cultures did not display a significant uptake ratio (metal concentration ratio of exposed cells to control cells) of nickel (3.1 +/- 3.3), only a small uptake ratio of selenium (4.0 +/- 0.4), while there was a large uptake ratio of both lead (2.6 +/- 1.7) x 10(2) and mercury (1.5 +/- 0.2) x 10(1). The results indicated that nickel was neither especially toxic nor influenced the clonogenic cell survival after irradiation. Mercury was more toxic and also influenced the radiation sensitivity. Lead was taken up strongly but did not influence the radiation sensitivity. Selenium accumulated but gave no detectable effect on the radiation sensitivity.

Abstract: Interaction between selenium and the heavy metals cadmium and mercury was studied in an experimental rat model (Sprague-Dawley). The rats were administered either one single trace element or combinations of selenium and cadmium as well as selenium and mercury. Salts of these trace elements were administered intraperitoneally daily during thirty days. Thereafter the animals were sacrificed and kidneys and livers excised rapidly. Thin sections were produced by a cryotome and subsequently freeze-dried. Nuclear microscopy of the sections showed that in the combination groups there was a co-localization of selenium and the heavy metals. None of the expected pathological signs of cadmium and mercury toxicity were observed. The conclusion was that selenium exerted a protective effect against the toxicity of cadmium and mercury through mechanisms still to be unveiled.

Abstract: Genetic factors are major contributors in determining the susceptibility to systemic autoimmune diseases. We studied the influence of genotype on systemic autoimmunity by treating female mice of the H-2s strains SJL/N, SJL/J, A.SW, and B10.S with mercuric chloride (HgCl2) for 10 weeks and then following autoantibody and tissue immune deposits during the subsequent 12 months. All strains developed antinucleolar antibodies (ANoA) of the IgG class which reacted in immunoblotting with a 34-kDa nucleolar protein identified as fibrillarin. The titre of ANoA attained after 10 weeks' treatment varied from 1:1,280 to 1:20,480 in the order: A.SW > SJL > > B10.S. Following cessation of HgCl2 treatment ANoA and antifibrillarin antibodies (AFA) persisted for up to 12 months, although some B10.S mice showed pronounced reduction not only of their autoantibody titres, but also systemic immune deposits when compared to other H-2s strains. A second set of autoantibodies targeted chromatin and in some mice specifically histones, and were distinguished from the ANoA by a rapid decline after treatment and a susceptibility linked to the non-H-2 genes of the SJL. Tissue levels of mercury remained elevated above untreated controls throughout the study period, suggesting that the mercury detected in lymphoid tissues may provide stimulation of lymphoid cells specific for fibrillarin for a considerable period after exposure has ceased. We conclude that H-2 as well as non-H-2 genetic factors distinctly influence not only the susceptibility to induction of autoimmunity, but also the specificity and magnitude of the response.

Abstract: X-ray microanalysis was used to detect elemental changes in the insulin-producing tumor cell-line RINm5F. To improve discrimination between mobile ions and ions bound to macromolecules a new approach was employed, consisting of multivariate statistical analysis of correlations between the concentrations of Na, Mg, P, S, Cl, K, and Ca. RINm5F cells, cultured on Formvar-coated titanium grids, were stimulated with high K+ or ATP, that are both known to stimulate insulin release. The buffers used contained Ca2+ or one of the Ca(2+)-analogues Sr2+ and Ba2+, to represent Ca2+ uptake in response to stimulation. After stimulation the cells were shock-frozen and freeze-dried overnight. Incubation for 10-20 seconds in a Ca(2+)-containing buffer did not significantly affect elemental composition, whereas cellular Mg, P and K decreased in a Sr(2+)-containing buffer. Depolarization with high K+ concentration caused an increase in the cellular Na content, both in Ca(2+)- and Sr(2+)-containing buffers, but not in the buffer where Ca2+ had been replaced by Ba2+. X-ray microanalysis is useful for detection of elemental changes subsequent to stimulation of cultured cells. Moreover, multivariate statistical analysis strengthens the idea that stimulation of RINm5F cells causes redistribution of ions possibly due to changes in the state of binding of some elements to cellular proteins.

Abstract: Previously symptomatic and permanently asymptomatic carriers of a gene mutation for acute intermittent porphyria as well as matched controls were screened with regard to a series of variables of possible relevance to the development of porphyric symptoms. The basis for the study was a concept of acute porphyria as a condition of a permanent system overload of oxidative stress, with long term effects on hepatic and renal tissue, and with instances of periodic overload of free radicals giving rise to acute neurologic involvement. Leukocyte concentrations of manganese, calcium, iron and zinc, as well as erythrocyte calcium differed between the groups, acute intermittent porphyria gene carriers, irrespective of previous porphyric illness, showing significantly higher levels than the controls. Manganese was found to be the most discriminative component of all the 78 variables investigated, accounting for about 98 per cent of the variance between the groups. An increment, by a factor of four, in cellular manganese is suggestive of an increase, in acute intermittent porphyria, of a manganese associated enzyme, e.g. glutamine synthetase, pyruvate carboxylase or mitochondrial superoxide dismutase. The best fit into the model considered is provided by a theory focused on superoxide dismutase, induced in response to superoxide anion radical produced from aminolaevulinic acid. In porphyria gene carriers seemingly resistant to porphyric manifestations, an increase in potentially prooxidant cellular iron is matched by a proportional increment in manganese, i.e. presumably by a corresponding mitochondrial superoxide dismutase induction. This mechanism is not operative in porphyric individuals prone to development of neuropsychiatric symptoms. In acute intermittent porphyria with a history of porphyric illness there is a positive correlation between erythrocyte manganese and serum folate and a negative correlation between leukocyte ferrochelatase activity and serum cobalamin concentration. This may mirror a role of the cobalamin-folate system in the acute porphyric process.

Abstract: The magnitude of inflammatory lesions in the hearts of coxsackie B3 (CB3)-virus infected mice can be affected by the potentially toxic heavy metals cadmium (Cd), nickel (Ni) and methyl mercury (MeHg). The infection is associated with a changed distribution, such as Cd accumulation in the spleen and kidneys. New target organs for Ni during the infection were the heart, pancreas and lungs in which inflammatory lesions were present. This increased uptake was correlated with the disturbed function of immune cells and an increased inflammatory reaction. Ni and MeHg appeared to have a direct effect on immune cells that resulted in changed natural killer cell activity and decreased mobilization of macrophages, CD4+ and CD8+ cells into the inflammatory lesions. Although MeHg increased spleen T cell activity and gamma-interferon (IFN-gamma) levels, the inflammatory lesions in the heart increased. Another detrimental effect of MeHg treatment was evident by an increased calcium and decreased zinc content in the inflamed heart, which may partly explain the more severe inflammatory lesion. The host's response, CB3 infection, changed the distribution of each metal in a specific way, a fact which may subsequently result in altered target organ toxicity and resistance to the infection.

Abstract: A comparative study of some aspects of freeze-substitution as a preparative method for X-ray microanalysis of diffusible ions in biological specimens was carried out. Four substitution fluids were compared with each other and with cryosections. As criteria, elemental ratios as determined by X-ray microanalysis of thin sections of mouse pancreas were used. Freeze-substitution for 3 weeks in diethyl ether or for 2 days in tetrahydrofurane gave results comparable to those obtained in cryosections. Slow warming to room temperature after freeze-substitution in tetrahydrofurane gave slightly better results than fast warming. Freeze-substitution in acetone resulted in noticeable loss and redistribution of ions and freeze-substitution in methanol gave very poor results.

Abstract: Elemental redistribution induced by insulin secretion, was investigated by electron and proton probe X-ray microanalysis. In particular, ion fluxes following immediately upon stimulation were studied. As the sensitivity of the electron probe was insufficient, the proton microprobe was employed. In order to see whether the cell is asymmetric with respect to Ca2+ influx, the cells were stimulated in the presence of Sr2+ (as a Ca2+ analog). Insulin-secreting cells (RINm5F cells and isolated mouse beta-cells) were cultured on grids and shock-frozen at 2-30 seconds after stimulation. In a large number of cells, the major elements and and large fluxes were analyzed by the electron microprobe. In the proton microprobe, selected cells were analyzed and elemental maps were compared with electron micrographs of the same cells. The proton microprobe, but not the electron microprobe, could detect an influx of Sr in response to K+-stimulation for 2 seconds, in RINm5F cells. No polarization of Sr2+ uptake in RINm5F-cells could be detected, and the beta-cells did not respond to high K+ by uptake of Sr. Momentary stimulation of beta-cells also resulted in a significant increase in Na, detected by the electron probe. Spreading of the beta-cells on the substrate appears to influence the subcellular elemental distribution. Thus, the proton probe has potential to detect small changes in elements such as those occurring after short-time stimulation.

Abstract: Dental amalgam fillings are the most important source of mercury exposure in the general population, but their potential to cause systemic health consequences is disputed. In this study, inbred mice genetically susceptible to mercury-induced immune aberrations were used to examine whether dental amalgam may interfere with the immune system and cause autoimmunity. Female SJL/N mice were implanted in the peritoneal cavity with 8-100 mg silver amalgam or silver alloy for 10 weeks or 6 months. Chronic hyperimmunoglobulinemia, serum IgG autoantibodies targeting the nucleolar protein fibrillarin, and systemic immune-complex deposits developed in a time- and dose-dependent manner after implantation of amalgam or alloy. Splenocytes from mice implanted with amalgam or alloy showed an increased expression of class II molecules. The functional capacity of splenic T and B cells was affected in a dose-dependent way: 10 weeks of low-dose and 6 months of high-dose amalgam implantation strongly increased mitogen-induced T and B cell proliferation, whereas 10 weeks of high-dose implantation decreased the proliferation. Not only mercury but also silver accumulated in the spleen and kidneys after amalgam implantation. In conclusion, dental amalgam implantation in a physiological body milieu causes chronic stimulation of the immune system with induction of systemic autoimmunity in genetically sensitive mice. Implantation of silver alloy not containing mercury also induced autoimmunity, suggesting that other elements, especially silver, have the potential to induce autoimmunity in genetically susceptible vertebrates. Accumulation of heavy metals, from dental amalgam and other sources, may lower the threshold of an individual metal to elicit immunological aberrations. We hypothesize that under appropriate conditions of genetic susceptibility and adequate body burden, heavy metal exposure from dental amalgam may contribute to immunological aberrations, which could lead to overt autoimmunity.

Abstract: To clarify whether elemental changes are present before the onset of diabetes, freeze-dried pancreas sections from young (18-19 days old), genetically prediabetic Chinese hamsters were subjected to proton bombardment and the concentrations of 15 elements (Na, Mg, Al, P, S, Cl, K, Ca, Mn, Fe, Cu, Zn, Rb, Cd, and Pb) in B cells and exocrine pancreas were calculated from the x-rays emitted. We have previously shown that islet B cells and exocrine pancreas from adult, overtly diabetic Chinese hamsters contain subnormal levels of Al (-61%, -88%) and excess levels of Cu (+92%, +59%), Rb (+13%, +13%), and Mg (+6%, +6%) in B cells and exocrine pancreas, respectively (Juntti-Berggren et al., Biosci Rep 1976;7:33-41). In the present study the prediabetic B cells contained normal levels of all 15 elements, whereas the prediabetic exocrine pancreas contained a subnormal level of Fe (-10%; p less than 0.005). Hence, the development of diabetes in the Chinese hamster does not seem to be associated with an early change in the elemental composition of the pancreatic B cells. In fact, the overt diabetic condition may cause changes in the body's handling of some elements.

Abstract: The concentrations of electrolytes and trace elements in erythrocytes were determined in eight children with cystic fibrosis (CF) and the parents of four of the children and compared with age- and sex-matched healthy controls. The children with CF had significantly lower median erythrocyte concentrations of sodium, magnesium and zinc and a higher median concentration of calcium than both the healthy control children and the parents of the CF children. Although the sodium and magnesium median values were higher in CF parents than in CF children they were nevertheless significantly lower than in adult controls. These data indicate a system for heterozygote detection.

Abstract: Mice were injected intravenously with lead chloride, 75 micrograms/g body weight. The mice were in an experimental delay of implantation, which offered the functionally steady conditions required for developing the testing procedures. One day after the administration of lead, blastocysts were flushed from the uterine cavity, placed on thin foils of Kapton, and air-dried. The dried blastocysts were analyzed in a nuclear microprobe with a particle intensity of 10 nanoampere (nA) for 300 s using a spatial resolution of 3 microns. The average lead concentration of blastocysts at implantation was 3.90 micrograms/g dry weight. We judge this technique to be useful for evaluating the transport of heavy metals from mother to preembryos, not only under the present experimental conditions but also in normal pregnancies and in other species.

Abstract: By using the proton microprobe technique we have investigated the elemental composition of both pancreatic beta-cells and exocrine pancreas from fed and 24 h or 48 h starved obese hyperglycemic mice. Among the 15 elements measured in the beta-cells both Ca and Fe increased while Mg and S decreased significantly after 24 h of starvation, the effects being more pronounced after 48 h. When animals were starved for 48 h there was a decrease in the contents of Cl, Rb and Cu, whereas that of Al and Mn increased with 152 and 55%, respectively. There was an initial decrease in Na after 24 h of starvation, which was followed by an increase after 48 h. This is in contrast to Cd, which first increased and then decreased to a value lower than that obtained in the fed animal. The content of K showed a small decrease and that of Pb showed an increase only in the 24 h starved group. In the beta-cells the contents of Zn and P did not change subsequent to starvation. In the exocrine pancreas Na, Cl and P decreased after 24 h of starvation and except for Na, the decrease was maintained when the starvation period was increased to 48 h. After 24 h there was a significant, though transient, increase in K, Mg and Rb. With regard to the contents of Zn, Cu and S there was a progressive decrease as the starvation continued. In contrast to the endocrine pancreas the content of Al in the exocrine pancreas did not change after 48 h of starvation. There was no change in islet insulin content subsequent to starvation. The extent to which the observed changes in beta-cell elemental composition is involved in the impaired insulin release associated with starvation, merits further investigations.

Abstract: Female rats were given single intraperitoneal doses of fluoride (F) (0, 4, 7, or 14 mg F/kg body weight). Plasma F levels returned to predose values within 24 h. Incisors from animals killed 35 or 70 days after the F doses had been given were analyzed for mineralization defects by microradiography, and for F and P concentrations by nuclear microprobe. At 35 days, all F-injected rats had enamel fluorosis. At 70 days, by which time the incisors would have been renewed nearly twice, fluorosis was still evident in the 14 mg/kg body weight group. The enamel and dentin F concentrations at each time point were proportional to the administered F doses that had been given weeks earlier. The F concentrations at 35 days were similar to those found at 70 days. The results support the hypothesis that, following pulse loading, F can be mobilized from the bone adjacent to the enamel organ and result in local F concentrations sufficiently large to adversely affect amelogenesis.

Abstract: Freeze-dried pancreas sections from 7-, 17- and 27-week-old genetically diabetic (db/db) and normal (+/-/+/-) mice were subjected to proton bombardment and the concentrations of 15 elements in B cells and exocrine pancreas were calculated from the characteristic X-rays emitted. In the 7-week-old diabetic animals, B cells contained significantly above-normal levels of Na and S, while exocrine pancreas contained subnormal levels of Ca, and excess Mn. The B cells from the 17-week-old diabetic animals contained subnormal levels of Cu and the exocrine pancreas of the 27-week-old diabetic animals was deficient in Cd. The 7-, 17- and 27-week-old, genetically diabetic (db/db) mice were hyperglycemic, hyperinsulinemic and heavier than age-matched normal (+/-/+/-) mice. Although significant changes were found in elemental composition when comparing both B cells and exocrine pancreas at different ages, the changes were not consistent. Therefore, it appears as if the measured elemental changes were random and not related to the onset of diabetes.

Abstract: Twenty Swedish Landrace lambs were divided randomly into two equal groups, group I and group O. Group I received approximately 0.1 mg selenium as sodium selenite per day and group O 0.1 mg selenium as selenium in yeast per day. The animals were fed per day 1.2-1.5 kg of hay and 0.3 kg of a commercial concentrate supplemented with vitamins and minerals. The trial lasted 3.5 months. As monitored by micro particle induced X-ray emission (mu-PIXE), the number of selenium bearing erythrocytes increased faster in group O than in group I indicating some difference in the metabolism of inorganic and organic selenium. During the trial the concentration of sulphur decreased in the erythrocytes in group O. The reason for this is not known. The zinc and copper concentrations of individual erythrocytes were unaltered. The blood GSH-Px activity increased at the same rate in the two groups indicating that selenium as sodium selenite and selenium in yeast was about equally available. The rate of growth in the two groups was almost the same and a significant difference in the final weight was not observed.

Abstract: The elemental profiles of thrombocytes and mononuclear cells were investigated in five patients with Infantile and eight with Juvenile Neuronal Ceroid Lipofuscinosis. The patients with the infantile form had suffered from the disease for a year and those with the juvenile form for some six years. The thrombocytes exhibited increased concentrations of calcium and magnesium, but the same concentrations of iron and zinc as found in healthy subjects. The mononuclear cells exhibited an increased concentration of iron and a reduced concentration of zinc. The elevated concentrations of magnesium, calcium and iron in the thrombocytes and mononuclear cells may represent the end products of ceroid pigmentation. Five patients with Juvenile and one with Infantile Neuronal Ceroid Lipofuscinosis were treated with antioxidants along with vitamins E, B2 and B6, but this treatment did not affect significantly the concentration of iron in the mononuclear cells. However, selenium was detected in some mononuclear cells in all the patients so treated. This was unexpected since iron (III), being antagonistic to selenium in the form of selenite--which was the antioxidant given--forms a stable complex which cannot be broken down biologically.

Abstract: The distribution of lead in the cerebellum of suckling Sprague-Dawley rats was examined using a nuclear microprobe for elemental mapping of tissue sections (particle-induced X-ray emission, 3-microns beam of 2.5 MeV protons; micro-PIXE). The rats were injected intraperitoneally with a lead-containing vehicle or vehicle only from ages 1 to 14 days. The calculated doses were 7.8 (low-dose) and 15.6 (high-dose) micrograms lead/g body weight. The rats were killed at 20 days of age. The vascular system was rinsed quickly with 0.15 M ammonium acetate to obtain determinations of intra-parenchymal lead with minimal influence of lead bound to erythrocytes and plasma proteins. Brains were frozen in propane/propylene in liquid nitrogen. Cryostat sections, 15 microns thick, were air dried on formvar coats that covered a hole, 15 mm in diameter, in a plastic disc, and were used for lead analysis by micro-PIXE. Very low concentrations of lead were found in the brain of controls. Lead levels in homogenates from cerebrum and cerebellum measured by atomic absorption spectrometry (AAS) were: low-dose 1.2-2.2 micrograms/g wet weight and high-dose 1.4-2.4 micrograms/g wet weight. The lead levels measured with the micro-PIXE method were in good agreement with the levels found with AAS. Lead was present in the cerebellar white matter in two to three times higher amounts than in the cortical grey (low-dose white matter 11-18 micrograms/g dry weight, grey matter 2.0-5.5 micrograms/g dry weight). This was true for both low and high dose exposed rats.(ABSTRACT TRUNCATED AT 250 WORDS)

Abstract: The mass fraction of certain elements was measured in isolated granulocytes and isolated granulocyte granule fractions from patients with active inflammatory arthritides (N = 6) and healthy controls (N = 6). The patients had significantly increased amounts of Ca in the granulocytes, in the specific and light azurophil granules, but normal Ca amounts in the dense azurophil granules. Sr was below the detection limit in the granulocytes and granule fraction from controls, but it appeared in high concentrations in the granulocytes and all granule fractions from the patients. The patients had considerably increased granulocyte amounts of Mn but only slightly increased Mn concentrations in the specific granules. Mn was not detectable in azurophil granules from patients and controls. A prominent accumulation of Fe was seen in the granulocytes from the patients, together with an Fe accumulation in the specific granules. Fe was below the detection limit in azurophil granules from patients and controls. The patients had reduced granulocyte Zn and reduced amounts of Zn in the dense and light azurophil granules but normal Zn amounts in the specific granules. The results obtained indicate that the granulocyte accumulation of Ca, Sr, and Fe observed during chronic inflammation is associated with corresponding granule accumulation of these metals; the considerable Mn accumulation in granulocytes during inflammation is not localized in their granules; and the granule subpopulations differ in their capacity to store certain metals.

Abstract: Subjectively healthy HLA-B27 positive 1st degree relatives (n = 14) of patients with ankylosing spondylitis (AS) were investigated concerning the mass fraction of calcium (Ca), magnesium (Mg), manganese (Mn), iron (Fe), zinc (Zn), strontium (Sr) and copper (Cu) in isolated blood cells using the nuclear microprobe technique. No relative had laboratory signs of inflammatory disease defined by acute phase plasma proteins. An accumulation of Mg, Ca, Mn and Fe was found in granulocytes compared with healthy controls. In platelets there was an accumulation of Fe and a reduction of the Cu content. In erythrocytes Ca was accumulated and the levels of Mg, Mn and Cu were reduced compared with the controls. Five of the relatives had radiological signs of sacroiliitis and 1 of these had sacroiliac tenderness. Relatives with and without radiological sacroiliitis showed no differences in the cellular metal amounts. When the alterations were compared with those previously found in patients with AS, a striking similarity was noted, although the changes were quantitatively less pronounced. In contrast B27 negative 1st degree relatives (n = 11) had normal mineral amounts in their cells. However, it seems less likely that altered metal handling could play a primary role for the disease susceptibility linked to HLA-B27 since B27 positive healthy controls (n = 12) without AS in the family had normal cellular stores of the measured elements. Rather our findings indicate that redistribution of cellular metals is an extremely sensitive marker of an inflammatory process not evident by clinical symptoms or increase of acute phase plasma proteins.

Abstract: Diabetes mellitus spontaneously develops in certain sublines of non-obese Chinese hamsters, and the diabetic L-subline is known for subnormal pancreatic insulin release in vitro. The cause of the secretory defect is unknown. Freeze-dried pancreas sections from genetically diabetic Chinese hamsters and normal controls were subjected to proton bombardment and the concentration of 15 elements in B cells and acini was calculated from the X-rays emitted. Diabetic B cells contained significantly less Al (-61%) and significantly more Cu (+92%), Mg (+6%) and Rb (+13%) than their normal counterparts. The diabetic acini showed similar, significant changes. The molar ratio between K and Na was about 10 in endocrine as well as exocrine pancreas from both groups of animals, implying that neither sample preparation nor irradiation had induced significant diffusive changes. In conclusion, the high K/Na ratio suggests that the diabetic B cell has a well-functioning Na+/K+ pump. However, significant and parallel changes in Al-, Cu-, Mg- and Rb-levels were found in both the B cells and acinar portion of the diabetic pancreas. It is not clear whether these elemental changes cause the islet secretory defect or result from it.

Abstract: The mass fraction of calcium (Ca), magnesium (Mg), manganese (Mn), iron (Fe). zinc (Zn), strontium (Sr) and copper (Cu) in isolated granulocytes, erythrocytes and platelets was measured using the nuclear microprobe technique. The cellular profile of metal variation was conspiciously altered in patients (n = 29) with ankylosing spondylitis compared with the profile found in age and sex matched healthy controls. Ca was accumulated in erythrocytes and granulocytes but decreased in platelets (p less than 0.001). Mg was increased in granulocytes but appeared in reduced concentrations in platelets and erythrocytes (p less than 0.001). The cellular amounts of Mn was increased in granulocytes (p less than 0.001), normal in platelets (p greater than 0.05) and subnormal in erythrocytes (p less than 0.001). Zn was reduced in all 3 cell types (p less than 0.001). Fe accumulation was evident in granulocytes and platelets (p less than 0.001). Sr was only measurable in granulocytes from patients. Cu was below the detection limit in the different cell types isolated from patients, but appeared in measurable amounts in erythrocytes and platelets from controls. The granulocyte amounts of Ca, Mg, Mn and Sr were strongly related to the acute phase reaction. Negative correlations were found between erythrocyte Mg and Zn and the inflammatory activity. The patients had increased serum levels of Cu, normal levels of Ca, Mg and Sr and decreased levels of Zn, Fe and Mn. No relationship was found between the serum concentrations of these elements and their respective cellular stores, except for a weak negative correlation between granulocyte and serum Fe.(ABSTRACT TRUNCATED AT 250 WORDS)

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Abstract: The cellular stores of iron in granulocytes and platelets isolated from 29 patients with ankylosing spondylitis were measured by the nuclear microprobe technique. The mean iron content in polymorphonuclear cells (PMNs) was 32 (SD 3) micrograms/g dry weight and in platelets 11 (2.6) micrograms/g dry weight. Corresponding values for age and sex matched healthy controls were 5.2 (1.9) and 4.6 (0.8) micrograms/g (p less than 0.001). Significant correlations were found in the patient group between (PMN) iron and the circulating levels of transferrin, total iron, and lactoferrin (p less than 0.05). PMN iron was not related to serum ferritin. Platelet iron correlated with transferrin (p less than 0.01) but not with the other iron binding proteins. Significant relationships were also found between the PMN iron stores and the inflammatory activity defined by erythrocyte sedimentation rate (ESR) and the immunoglobulins A and G. These data further illustrate the altered iron kinetics in chronic inflammatory disease and record the fact that the redistribution of iron associated with the inflammatory process also includes granulocytes and platelets.

Abstract: The concentrations of trace elements in three different types of blood cells--erythrocytes, thrombocytes and neutrophils--were determined by nuclear microprobe analysis in 11 children with Down's syndrome (DS) and compared with those in 13 controls. The median concentration of copper in all three types of blood cells and that of calcium in erythrocytes were significantly higher in DS children than in controls. In contrast, the median levels of zinc in erythrocytes, of manganese and iron in erythrocytes and neutrophils, and of magnesium in erythrocytes and thrombocytes were significantly lower in DS than in control children. In 10 of the 11 DS children, detectable erythrocyte levels of titanium were found, which is remarkable, since this was not observed in any of the cells from the control children.

Abstract: Freeze-dried pancreas sections from obese-hyperglycemic mice were subjected to proton bombardment and the elemental contents in the beta-cells and the exocrine part were obtained from the characteristic X-rays emitted. Quantitative data were provided for 18 different elements. The mole ratio between K and Na exceeded 10, implying that neither the sample preparation nor the irradiation had induced significant diffusive changes. With the demonstration of this high K/Na ratio it seems likely that also the beta-cells are equipped with an efficient Na+/K+ pump. The beta-cells contained about 70 mmoles Cl per litre cell water. Observed amounts of Ca and Mg were equivalent to those previously recorded by electrothermal atomic absorption spectroscopy. The significant role of Zn for the storage of insulin was emphasized by the demonstration of 3 times as much of this element in the beta-cells as compared with the exocrine pancreas. In addition, the sensitivity of the proton microprobe enabled measurements of various trace elements such as Rb, Cr, Cu, Al and Pb not previously demonstrated in the pancreatic beta-cells.

Abstract: The mass fraction of Ca and Mg in isolated erythrocytes and granulocytes was measured using the nuclear microprobe technique. Conspicuous abnormalities were observed in cells from patients with rheumatoid arthritis and other inflammatory arthritides. Compared with the normal cellular content, total Ca was increased an average of 3 times in erythrocytes and 5 times in granulocytes. Total granulocyte Mg was increased about 3 times, whereas erythrocyte Mg was reduced to as much as 60% of normal. These abnormalities were less prominent or were absent in scleroderma patients, except for levels of granulocyte Ca, which were increased more than 3 times beyond normal in this patient group. A significant positive correlation was found between serum haptoglobin and erythrocyte or granulocyte Ca content among these patients, but not between haptoglobin and erythrocyte or granulocyte Mg values. During corticosteroid treatment, a significant increase in erythrocyte Mg and a significant reduction in erythrocyte Ca were noted, but normalization of these levels was not achieved. Granulocyte Ca was also significantly reduced, while granulocyte Mg remained unaltered. Serum levels of Ca and Mg were within normal ranges and were not influenced by corticosteroid therapy. The results indicate that at least Ca abnormalities in erythrocytes and granulocytes are associated with the intensity of the inflammatory process and that the amounts of Ca and Mg in these cells are influenced by potent antiinflammatory therapy.

Abstract: By the use of the nuclear microprobe technique, the concentrations of zinc in isolated erythrocytes, platelets, and granulocytes were measured in patients with rheumatoid arthritis, other inflammatory arthritides, and scleroderma. Markedly reduced cellular zinc values were found compared to those measured in healthy subjects. No relation was found to inflammatory activity or disease duration. Plasma zinc was reduced in the majority of the patients and was negatively correlated to the inflammatory activity estimated by ESR and serum orosomucoid. No relation was found between total zinc values in plasma or cells or disease duration. Corticosteroid therapy was instituted in a number of the patients with inflammatory arthritides and induced a significant elevation of total zinc in all cell types, although normalization was not achieved. Plasma zinc values remained unchanged during the treatment.

Abstract: Higher than normal amounts (p less than 0.001) of manganese were found in granulocytes isolated from patients with rheumatoid arthritis, other inflammatory arthritides, and scleroderma, while erythrocytes and platelets contained normal amounts of manganese. The cause of manganese accumulation is not known but evidently is linked to the intensity of the inflammatory process since significant positive correlations (p less than 0.001) were seen between erythrocyte sedimentation rate or serum haptoglobin and granulocyte manganese. During corticosteroid therapy granulocyte manganese started to return to normal levels but the relative changes were modest. Strong positive correlations (p less than 0.001) were found between granulocyte manganese and granulocyte stores of calcium and iron, suggesting a close relationship in the cellular regulation of these elements in chronic inflammatory conditions.

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Abstract: Total cellular strontium and calcium were measured by the nuclear microprobe technique. Increased mass fraction of both elements was found in granulocytes isolated from patients with active rheumatoid arthritis and other kinds of inflammatory arthritides. Increased granulocyte calcium but only marginally elevated granulocyte strontium was demonstrated in patients with scleroderma. The granulocyte accumulation of strontium and calcium seems to be linked to the degree of inflammatory activity, because the granulocyte content of both elements was positively correlated to the plasma concentration of acute-phase proteins. Corticosteroid therapy induced a marked reduction of granulocyte strontium but a more modest decrease of granulocyte calcium. The serum levels of strontium and calcium were within the normal ranges in all patients and were not significantly altered by corticosteroids.

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Abstract: Proton microprobe analysis was used to determine the fluoride concentrations in enamel and dentin surfaces exposed to a fluoride-containing and a conventional amalgam in vitro. After seven weeks of exposure, a considerable fluoride uptake was demonstrated from the fluoride-containing amalgam. The fluoride deposition was greater in the dentin than in the enamel. The greatest fluoride concentrations in enamel surfaces, about 4000 ppm, were found in the outer 0.05 micrometers of the tissue. In the dentin the greatest concentrations, about 9000 ppm, were found at a depth of 11.5 micrometers. This study demonstrates the ability of the fluoride-containing amalgam to deposit fluoride into dental hard tissues.

Abstract: The contents of the elements antimony, arsenic, cadmium, copper, lead, mercury, selenium, silver, tin and zinc in bone tissue from autopsy specimens of the femur of workers who had been exposed to a large number of metals in a smeltery and refinery in Northern Sweden as well as of a control group have been quantitatively assayed. The analytical techniques used were atomic absorption spectrophotometry, neutron activation analysis and particle induced X-ray emission analysis (PIXE) in a proton microprobe. Increased levels of lead in the bone tissue of exposed workers compared to a non-exposed control group were observed. The median level of lead in the group of exposed workers exceeded the corresponding value of the control group about 5 times. Using the proton microprobe in the PIXE-mode, concentration profiles of copper, lead and zinc were examined within the Haversian system of the bone samples.

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Abstract: A method is described for the measurement of the distribution of lead in bone tissue. Two cases of human femur have been analyzed; one worker was exposed to lead in heavy metal industry, the other case presents a reference worker from the same environment not excessively exposed to lead. Lead was determined through proton induced X-ray emission using microprobe technique. The mean lead concentration in the poisoned and the reference case was 70 and 30 ppm, respectively. The reference case showed an even lead distribution in the femur, while the poisoned case exhibited peaks in the distribution indicating that exposure to lead was not evenly distributed. The described technique has potential for microanalysis within the Haversian system for recording the history of exposure.