Copper-silicon interaction studies in young, rapidly growing turkeys fed semipurified starter diets.

A 3x3 factorial experiment was conducted to study the interactions between Cu and Si in growing turkeys fed a basal dextrose-casein semipurified diet supplemented at 0, 270, and 540 ppm Si, and 2, 8, and 75 ppm Cu levels of inclusion. There were no significant (p>0.05) Cu, Si, or Cu-Si interaction effects on turkey growth. However, there were significant (p<0.05) Si effects on hemoglobin concentrations, and Cu-Si interaction effects on hematocrit and plasma Mg levels (p<0.01). Plasma Ca, P, Zn, Cu, total cholesterol, and serum alkaline phosphatase activity were not affected (p>0.05) by Cu, Si, or Cu-Si interaction. Heart and liver weights were affected (p<0.05) by the Cu-Si interaction. In general, a 8-ppm Cu supplemental level prevented cardiac hypertrophy only in the presence of 270 or 540 ppm Si inclusion in the diets. There were significant (p<0.05) Cu effects on liver Cu and Mn and heart Zn concentrations, Si effects on liver Mn levels, and Cu-Si interaction on liver Zn concentrations. Within the 0- and 270-ppm Si groups, Cu-deficient turkeys (2 ppm Cu) had higher liver Zn levels but not within the 540-ppm Si group. The Cu-Si interaction did not affect (p>0.05) concentrations of Cu in the heart and liver tissues. It is postulated that the Cu-Si interactions demonstrated in this study could have profound implications on the cardiovascular and skeletal health of birds.

The effect of subclinical selenium toxicosis on pregnant beef cattle.

A field investigation conducted by the South Dakota Animal Disease Research and Diagnostic Laboratory suggested that subclinical selenium toxicosis in pregnant cows may have contributed to an outbreak of aborted/stillborn calves in a high-selenium region of South Dakota. This study was undertaken to evaluate the relationship between abortion and subclinical selenium toxicosis in the dam and to assess the effects of subclinical selenium toxicosis on the bovine immune system. Fifteen pregnant cows were fed diets containing 0.25 (control), 6.0, and 12.0 ppm selenium beginning at 80-110 days gestation. Although selenium toxicosis has been reported to cause abortion, this study failed to reproduce abortions. A single cow in the 12-ppm selenium treatment group gave birth to a weak calf, which subsequently died. This calf had myocardial lesions consistent with those described for selenium toxicosis and had hepatic selenium levels of 9.68 ppm (wet weight). Elevated dietary selenium resulted in the depression of several leukocyte function parameters in pregnant cows. A statistically significant depression in forced antibody response was identified in both selenium-supplemented groups. A significantly diminished mitogenic response to concanavalin A and pokeweed mitogen was also observed in the 12-ppm selenium group. Although a similar pattern of depression was also observed with phytohemagglutinin, differences were not significant. These findings indicate that even in the absence of clinical alkali disease, elevated selenium levels may adversely affect both pregnancy outcome and the bovine immune system.

Plasma selenium in patients with cirrhosis.

Plasma selenium concentration is decreased in patients with cirrhosis and, based on this finding, it has been suggested that patients with cirrhosis are selenium deficient. We measured plasma selenium concentration and the two plasma selenoproteins, glutathione peroxidase (GSHPx-3) and selenoprotein P, in the plasma of patients with cirrhosis of Child classes A, B, and C and in control subjects. Plasma selenium declined in proportion to the severity of the cirrhotic condition, as indicated by the Child class. Selenoprotein P, which originates largely in the liver, declined in a similar manner. Plasma glutathione peroxidase activity increased, and GSHPx-3 originates in the kidney. Selenium in the non-selenoprotein pool, shown by others to be largely selenomethionine in albumin, declined. Thus, although plasma selenium is decreased in patients with cirrhosis, the increase in plasma glutathione peroxidase activity, which occurs in them, suggests that patients with cirrhosis do not have selenium deficiency.

Improving emergency obstetric care at a district hospital, Makeni, Sierra Leone. The Freetown/Makeni PMM Team.

PRELIMINARY STUDIES: A facility review and focus group discussions revealed poor capacity to manage obstetric complications. INTERVENTIONS: In response, a physician with obstetric skills was posted, and a second physician was trained. Courses in life-saving obstetric skills were held for nurses and midwives. An unused operating theater was made functional with simple modifications. A generator and blood bank were installed. Drugs and supplies were made available through a revolving fund. Subsequently, community interventions focused on improving utilization. RESULTS: The number of women seeking treatment for major obstetric complications at the district hospital increased from 31 in 1990 to 98 in 1995, while the case fatality rate (CFR) among these women dropped from 32% to 5%. Cesarean sections increased from two in 1990 to 38 in 1995. In 1995, 444 abortion-related procedures were performed--almost all of them for unwanted pregnancy--compared with only 22 in 1990. COSTS: The cost of material improvements and training was approximately US$39,000, of which 46% was from project funds, 41% from non-governmental organizations and 13% from government. CONCLUSIONS: Women with obstetric complications will seek hospital care if services are available. Government hospital services can be improved by building on existing resources. Obstetric CFR can be dramatically reduced. The need for safe abortion services, which are currently illegal in Sierra Leone, is demonstrated.

Determination of selenium in feeds and premixes: collaborative study.

A total of 17 laboratories participated in a collaborative study for the determination of selenium in feeds and premixes using either a fluorometric or a continuous hydride generation atomic absorption (HGAA) method. Each collaborator analyzed 16 blind duplicate samples of feed and premixes from various feed manufacturers. The amount of Se in these materials ranged from 0.2 to 5500 micrograms/g. Six laboratories used only the fluorometric procedure, 8 laboratories used only the hydride generation atomic absorption procedure, and 3 laboratories used both procedures. One laboratory in the fluorometric study and 3 laboratories in the HGAA study were initially excluded because of invalid data. Poor agreement between the blind duplicates indicated probable sample interchange and/or dilution error. The data from 8 laboratories were submitted to statistical analysis, including data from 2 laboratories participating in both studies. The repeatability standard deviation (RSDr) for samples analyzed by the fluorometric procedure ranged from 5.9 to 33%, and the reproducibility standard deviation (RSDR) ranged from 12 to 33%. RSDr for samples analyzed by HGAA ranged from 2.8 to 18%, and RSDR ranged from 4.0 to 36%. Both fluorometric and continuous hydride generation atomic absorption methods for the determination of Se in feeds and premixes have been adopted first action by AOAC INTERNATIONAL.

Selenium detoxification by methylation.

The major detoxification process for selenium is methylation. The major pathway for the formation of methylated compounds in biological systems is transmethylation catalyzed by methyltransferases. In this study methyltransferase activity was assayed in the liver cytosol prepared from different strains of rats using HPLC method by measuring the formation of epinephrine from norepinephrine. The methyltransferase activity in Fischer, Wistar, and Sprague-Dawley rats were 1.65 x 10(-12), 0.74 x 10(-12), and 1.2 x 10(-12) moles epinephrine formed/mg protein/hr respectively. Volatilization of injected Se (2.5 mg/kg, i.p.) was found to be higher in Wistar rats compared to Fischer rats. LD50 for Wistar and Fischer rats were 2.58 and 3.15 mg Se/kg respectively. Our results suggest that the methylation of dimethyl selenide to trimethhyl selenide appears to be an important step in the detoxification of selenium.

Human kidney proximal tubules are the main source of plasma glutathione peroxidase.

The sites of synthesis of extracellular (E) glutathione peroxidase (GPX), a unique selenoglycoprotein present in plasma, are not known. To investigate the possibility that the kidney is the main source for the plasma GPX, we examined GPX activities and selenium concentrations in the plasma of patients with renal failure on dialysis and nephrectomized patients before and after kidney transplantation. Plasma GPX activities in these patients were 42, 22, and 180% of normal EGPX activity, respectively, whereas plasma Se levels were within the normal range. Twenty-four hours after nephrectomy of anesthetized rats, plasma GPX activity was 30.0 +/- 6.4% of the activity at zero time. Northern hybridization analysis of eight human tissues probed with EGPX and cellular glutathione peroxidase (CGPX) cDNA revealed that the ratio of EGPX to CGPX was highest in the kidney. cRNA in situ hybridization studies on kidney slices showed that only proximal tubular epithelial cells and parietal epithelial cells of Bowman's capsule contained EGPX transcripts. Caki-2, a proximal tubular renal carcinoma cell line, makes and actively secretes EGPX. Taken together, these results strongly suggest that kidney proximal tubular cells are the main source for GPX activity in the plasma.

Partial sequence of human plasma glutathione peroxidase and immunologic identification of milk glutathione peroxidase as the plasma enzyme.

Plasma glutathione peroxidase (p-GSHPx) is a unique selenoglycoprotein. A hepatic cell line synthesizes both this extracellular form for secretion and the cellular form that remains within the cells. Because the two forms could be a result of post-translational modifications of a product of a single gene, we partially sequenced p-GSHPx. Purified p-GSHPx was trypsin digested, and three of the peptides were sequenced. Only one of the peptide sequences was partially homologous to a sequence found in human cellular glutathione peroxidase. Because p-GSHPx is a secreted enzyme, we determined whether GSHPx in milk (another extracellular fluid) is due to this form of the enzyme. Ninety percent of human milk GSHPx activity could be precipitated by anti-p-GSHPx-immunoglobulin G. Thus, most, if not all, GSHPx activity in milk is due to the plasma selenoprotein form of the enzyme. In milk of two North American women, 3.6% and 14.3% of selenium was associated with GSHPx.

Selenium and amino acid composition of selenoprotein P, the major selenoprotein in rat serum.

Selenoprotein P is the second plasma selenoprotein to be purified. It is a glycoprotein and has been shown to be distinct from plasma glutathione peroxidase. This study characterizes selenoprotein P further. Deglycosylation of the protein shifts its migration on sodium dodecyl sulfate-polyacrylamide gel electrophoresis from Mr 57,000 to Mr 43,000, indicating it has a substantial carbohydrate component. Measurement of selenium indicates a selenium content of 7.5 +/- 1.0 atoms/molecule based on a polypeptide weight of 43,000. Amino acid analysis accounts for all the selenium as selenocysteine. The protein is also rich in cysteine (17 residues) and histidine (23 residues). Fragmentation of selenoprotein P by trypsin and by cyanogen bromide produces peptides with varying selenium content. This indicates that selenium-rich regions of the protein exist. The concentration of selenoprotein P determined by radioimmunoassay in serum from control rats is 26.3 +/- 4.5 micrograms/ml and in serum from selenium-deficient rats it is 2.7 +/- 0.8 micrograms/ml. Depletion of selenoprotein P from control serum using an immunoaffinity column indicates that over 60% of serum selenium in the rat is contained in this protein. These results demonstrate that selenoprotein P is the major form of selenium in rat serum. It is the first selenoprotein described which has more than one selenium atom/polypeptide chain.

Porcine focal symmetrical poliomyelomalacia: test for an interaction between dietary selenium and niacin.

Experiments were conducted to test the hypothesis that dietary supplementation with nicotinamide would retard or eliminate the signs of selenium induced porcine focal symmetrical poliomyelomalacia (PFSP). Mixed-sex feeder pigs, approximately five weeks old, were divided into four groups and daily received, by oral capsule, the following treatments: no supplementation (control); 2.86 mg sodium selenite per kg body wt (selenium only); 44 mg nicotinamide per kg body wt (niacin only); or both the niacin and selenium (niacin + selenium). Over the ten day treatment body weights and behavior scores were recorded, as well as collection of fluid (blood, serum, urine) samples. Upon death, tissue samples (kidney, liver, brain, spinal cord and muscle) were obtained. All of these samples were analyzed for total selenium and bioactive niacin compounds. After gross pathological analysis, 11 samples from specific brain and spinal cord regions were taken for fixation and processing for histological analysis by light microscopy. The selenium only group showed behavior signs related to PFSP after two days of treatment with the average time of death at 6.5 days. Tissue levels of selenium were elevated and histological analyses established the expected lesions of PFSP. No disorders were noted in the control and niacin only groups. The niacin + selenium groups had slightly retarded changes in behavior scores (first differences from controls on day 4) but their mean day of death (7.5 days of treatment) did not differ from that of the selenium only groups. Histological analyses of these tissues revealed similar lesions to the selenium only group, but they may have been of lesser magnitude. The data were consistent with, but only partially supportive of, the above hypothesis.

Antihuman plasma glutathione peroxidase antibodies: immunologic investigations to determine plasma glutathione peroxidase protein and selenium content in plasma.

Plasma glutathione peroxidase (GSHPx) (glutathione: H2O2 oxidoreductase) is a unique selenoglycoprotein. Treatment of this enzyme with glycopeptidase F partially deglycosylates it and establishes the presence of N-linked sugar moieties. Antibodies raised in a rabbit against the purified enzyme from plasma were found to be specific, noninhibitory, and capable of precipitating the enzymatic activity. The antibodies precipitated greater than 90% of the GSHPx activity of normal plasma, thus indicating that the selenoenzyme is the main if not the sole GSHPx activity of plasma. The antibodies did not precipitate RBC GSHPx. A slight cross-reactivity of the antibodies was found with rat plasma GSHPx. A GSHPx activity precipitation assay of normal plasma in the presence of selenium (Se)-deficient plasma indicates that no cross-reactive protein in the Se-deficient plasma interferes with the precipitation of the GSHPx activity from normal plasma. Thus, GSHPx protein as well as activity is deficient in plasma in the absence of Se. Antibodies against GSHPx either from RBCs or from plasma were used to specifically immunoprecipitate most of the GSHPx activity from RBCs or plasma, respectively, in healthy individuals to determine the amount of Se associated with the protein. GSHPx accounts for approximately 15% of the Se in RBCs and 12% of the Se in plasma. Thus, in normal individuals, these proteins account for only a fraction of plasma and RBC Se.

Porcine focal symmetrical poliomyelomalacia: experimental reproduction with oral doses of encapsulated sodium selenite.

Sodium selenite (encapsulated as doses of 1.4 mg, 2.6 mg and 4.2 mg per kilogram of body weight) was given orally on a daily basis to male weaner pigs, and features of these animals were compared to a control group. Porcine focal symmetrical poliomyelomalacia was produced in all experimental groups between 3 and 20 days after initiation of the treatment. Analysis of blood and several tissues revealed an elevated selenium content for all pigs. Histological lesions in the brain and the cervical lumbar/sacral spinal cord enlargements included endothelial proliferation, neuronal degeneration, microcavitation and glial cell reaction.

Distribution of serum selenium, copper, and zinc in normal human pregnancy.

The distribution of the serum levels of selenium, zinc, and copper in human pregnancy at various gestational ages were determined from two ethnically and geographically different populations (Rosebud Indian Reservation and southeastern South Dakota) of 410 normal subjects. As gestation age increased, there was a significant increase and a slight decrease in the mean levels of copper and zinc, respectively. No change in the levels of selenium was observed. Significantly higher levels of both pregnancy and non-pregnancy serum copper were observed in the Rosebud population compared to that in southeastern South Dakota, possibly due to the significantly higher level of copper in the Rosebud water. No differences were observed in the zinc or selenium levels between the two populations. Serial measurements of these trace metals during the third trimester of pregnancy were performed on 18 subjects, and supported the trends described for copper and selenium. No decrease in zinc was observed in the individual subjects.

Toxic effects of selenium on growing swine fed corn-soybean meal diets.

A total of 96 crossbred pigs received various levels of sodium selenite to determine the effect of dietary selenium (Se) on growing swine fed corn-soybean meal diets. Levels of supplemental Se were 0, 4, 8, 12, 16 and 20 micrograms/g. There were linear decreases (P less than .01) in both gain and feed intake with increasing levels of dietary Se. Feed/gain increased numerically as dietary Se increased. Hair Se increased quadratically (P less than .01) and blood Se increased linearly (P less than .01) with increasing level of dietary Se. Cell volume and hemoglobin were not affected by dietary treatment. Increasing dietary Se significantly increased glutathione peroxidase (GSH-Px), glutamic-oxalacetic transaminase (GOT). and glutamic-pyruvic transaminase (GPT). External signs of selenosis were noted in some pigs fed 12 or 20 micrograms/g of Se. The toxic level of Se in a corn-soybean meal diet for crossbred pigs appears to be between 4 and 8 micrograms/g. Of variables studied, growth rate was the most sensitive indicator of chronic selenosis in swine.

Effects of seleniferous grains and inorganic selenium on tissue and blood composition and growth performance of rats and swine.

Two experiments were conducted to determine the effect of varying dietary selenium (Se) levels and Se source on growing swine. In Exp. 1, seleniferous wheat and oats were used to formulate diets containing .47, 2.58, 5.60 or 8.40 micrograms/g organic Se. Dietary Se level had no effect on pig performance during the 6-wk experiment as measured by daily gain, daily feed intake or feed/gain. Blood composition and enzyme activity were not affected by dietary treatment. Selenium concentrations of blood, hair, liver, kidney, heart, spleen and diaphragm muscle were increased linearly (P less than .01) as dietary Se increased. In addition, liver weight as a percentage of body weight was increased linearly (P less than .01) as dietary Se level increased. No signs of chronic Se poisoning were observed. Dietary treatments in Exp. 2 were similar to Exp. 1 with the exception that sodium selenite was utilized as the Se source and the diets were fed for 17 wk. Inorganic Se levels of .54, 2.63, 5.69 or 8.33 micrograms/g had no effect on pig performance as measured by daily gain, daily feed intake or feed/gain. Selenium concentrations of blood, hair, liver, kidney, spleen and diaphragm muscle were significantly increased as dietary Se level increased. Liver weight as a percentage of body weight was increased at the two highest dietary Se levels. Blood glutathione peroxidase activity was significantly increased by dietary treatment, while other blood variables were not affected. No signs of chronic Se poisoning were observed.(ABSTRACT TRUNCATED AT 250 WORDS)