Absorption and elimination of trivalent and hexavalent chromium in humans following ingestion of a bolus dose in drinking water

These studies investigate the magnitude and valence state of chromium absorbed following plausible drinking water exposures to chromium(VI). Four adult male volunteers ingested a single dose of 5 mg Cr (in 0.5 liters deionized water) in three choromium mixtures: (1) Cr(III) chloride (CrCl3), (2) potassium dichromate reduced with orange juice (cr(III)-OJ); and (3) potassium dichromate [Cr(VI)]. Blood and urine chromium levels were followed for 1-3 days prior to and up to 12 days after ingestion. The three mixtures showed quite different pharmacokinetic patterns. CrCl3 was poorly absorbed (estimated 0.13% bioavailability) and rapidly eliminated in urine (excretion half-life, approximately 10 hr), whereas Cr(III)-OJ was absorbed more efficiently (0.60% bioavailability) but more slowly (half-life, approximately 17 hr), and Cr(VI) had the highest bioavailability (6.9%) and the longest half-life (approximately 39 hr). All three chromium mixtures caused temporary elevations in red blood cell (RBC) and plasma chromium concentrations, but the magnitude and duration of elevation showed a clear trend (Cr(VI) > Cr(III)-OJ > CrCl3). The data suggest that nearly all the ingested Cr(VI) was reduced to Cr(III) before entering the bloodstream based on comparison to RBC and plasma chromium patterns in animals exposed to high doses of Cr(VI). These findings support our prior work which suggests that water-soluble organic complexes of Cr(III) formed during the reduction of Cr(VI) in vivo explain the patterns of blood uptake and urinary excretion in humans at drinking water concentrations of 10 mg/liter or less.     

Modification of pentasaccharide core of surface N-glycans during differentiation of HL-60 cells

Graves' disease (GD) is generally thought of as a multifactorial disorder in which genetic susceptibility interacts with environmental and endogenous factors to cause disease. The importance of genetic factors is suggested by the clustering of GD within families and by a higher concordance rate for disease in monozygotic than dizygotic twins. This has, however, recently been shown to be less pronounced than previously thought. During the last decade, much effort has been put into characterization of the genetic background of GD. Until recently most studies have examined associations between GD and the human leukocyte antigen (HLA) region, but recent advances in molecular techniques have opened the way for whole-genome screening. A number of HLA and non-HLA candidate genes have been proposed, but despite several large investigations within multiplex families no major susceptibility genes have been identified. This brief review discusses relevant articles published from 1940 through 1997 regarding the influence of genetic factors in the etiology of GD. Ongoing studies will focus on whole genome screening in multiplex families as well as population based twin studies. However, the possibility of GD being a heterogeneous disease without a single well-defined genotype and phenotype should be left open.     

Transient poly(ADP-ribosyl)ation of nuclear proteins and role of poly(ADP-ribose) polymerase in the early stages of apoptosis

A transient burst of poly(ADP-ribosyl)ation of nuclear proteins occurs early, prior to commitment to death, in human osteosarcoma cells undergoing apoptosis, followed by caspase-3-mediated cleavage of poly(ADP-ribose) polymerase (PARP). The generality of this early burst of poly(ADP-ribosyl)ation has now been investigated with human HL-60 cells, mouse 3T3-L1, and immortalized fibroblasts derived from wild-type mice. The effects of eliminating this early transient modification of nuclear proteins by depletion of PARP protein either by antisense RNA expression or by gene disruption on various morphological and biochemical markers of apoptosis were then examined. Marked caspase-3-like PARP cleavage activity, proteolytic processing of CPP32 to its active form, internucleosomal DNA fragmentation, and nuclear morphological changes associated with apoptosis were induced in control 3T3-L1 cells treated for 24 h with anti-Fas and cycloheximide but not in PARP-depleted 3T3-L1 antisense cells exposed to these inducers. Similar results were obtained with control and PARP-depleted human Jurkat T cells. Whereas immortalized PARP +/+ fibroblasts showed the early burst of poly(ADP-ribosyl)ation and a rapid apoptotic response when exposed to anti-Fas and cycloheximide, PARP -/- fibroblasts exhibited neither the early poly (ADP-ribosyl)ation nor any of the biochemical or morphological changes characteristic of apoptosis when similarly treated. Stable transfection of PARP -/- fibroblasts with wild-type PARP rendered the cells sensitive to Fas-mediated apoptosis. These results suggest that PARP and poly(ADP-ribosyl)ation may trigger key steps in the apoptotic program. Subsequent degradation of PARP by caspase-3-like proteases may prevent depletion of NAD and ATP or release certain nuclear proteins from poly(ADP-ribosyl)ation-induced inhibition, both of which might be required for late stages of apoptosis.     

Intestinal nutrient-gene interaction: the effect of feed deprivation and refeeding on cholecystokinin and proglucagon gene expression

We tested the hypothesis that dietary components reaching the bovine small intestine influence the expression of genes that encode the gastrointestinal neuropeptides cholecystokinin (CCK) and glucagon-like peptide-1 (GLP-1). The amount of digesta reaching the intestine was manipulated during the experiment by withholding feed from five heifers fitted with ruminal, duodenal, and ileal cannulas for 48 h and then subsequent refeeding. Duodenal and ileal biopsies were collected using a fiber-optic endoscope. A Northern hybridization procedure was used to evaluate changes in gene expression. Blood concentrations of CCK and GLP-1 were determined with RIA. The data indicate that CCK blood concentration and mRNA abundance decreased during the period of feed deprivation, but they returned to predeprivation values within 16 to 24 h of refeeding. The GLP-1 blood concentration also decreased during feed deprivation and returned to predeprivation values within 4 to 8 h of refeeding, despite the fact that proglucagon mRNA abundance did not change significantly during feed deprivation and refeeding. These findings provide evidence that CCK and GLP-1 are released in response to nutrients that reach the small intestine and may be involved in the physiological process of digestion and possibly play a role in regulating feed intake in ruminants.     

Inhibition of recombinant human mitochondrial aldehyde dehydrogenase by two intermediate metabolites of disulfiram

Disulfiram is used in aversion therapy for alcoholism. S-Methyl-N,N-diethylthiocarbamate (MeDTC) sulfoxide, a potent inhibitor of the target enzyme mitochondrial aldehyde dehydrogenase (ALDH2), is thought to be the principal active metabolite of disulfiram in vivo. We examined the effects on recombinant human ALDH2 of two intermediate metabolites of disulfiram, S-methyl-N,N-diethyldithiocarbamate (MeDDC) sulfoxide and MeDDC sulfine. MeDDC sulfoxide was a potent inhibitor of ALDH2 with an IC50 of 2.2 +/- 0.5 microM (mean +/- SD, N = 4) after preincubation with enzyme for 30 min. MeDDC sulfine was a relatively weak inhibitor of ALDH2 under the same conditions with an IC50 value of 62 +/- 14 microM. The inhibition of ALDH2 by both compounds was irreversible and did not require the cofactor NAD. The latter finding demonstrates that inactivation of ALDH2 is independent of the dehydrogenase activity of the enzyme. GSH blocked almost completely the inhibition by 20 microM of MeDDC sulfoxide and greatly diminished the inhibition by 200 microM of MeDDC sulfine. Inactivation by MeDDC sulfoxide was time dependent. MeDTC sulfoxide was a more potent inhibitor of recombinant human ALDH2 (IC50 = 1.4 +/- 0.3 microM after preincubation for 15 min) than either of the intermediate metabolites, and its inhibition was unaffected by GSH. Our results suggest that these newer intermediate metabolites of disulfiram, especially the more potent MeDTC sulfoxide, have the potential to inhibit the target enzyme ALDH2 in patients receiving disulfiram. However, until the significance of the interactions of the inhibitors with GSH is more fully understood, the contribution of MeDDC sulfine and MeDDC sulfoxide to the pharmacological effects of disulfiram in vivo is uncertain.     

Cost-effectiveness of reducing sulfur emissions from ships

We model cost-effectiveness of control strategies for reducing SO2 emissions from U.S. foreign commerce ships traveling in existing European or hypothetical U.S. West Coast SO(x) Emission Control Areas (SECAs) under international maritime regulations. Variation among marginal costs of control for individual ships choosing between fuel-switching and aftertreatment reveals cost-saving potential of economic incentive instruments. Compared to regulations prescribing low sulfur fuels, a performance-based policy can save up to $260 million for these ships with 80% more emission reductions than required because least-cost options on some individual ships outperform standards. Optimal simulation of a market-based SO2 control policy for approximately 4,700 U.S. foreign commerce ships traveling in the SECAs in 2002 shows that SECA emissions control targets can be achieved by scrubbing exhaust gas of one out of ten ships with annual savings up to $480 million over performance-based policy. A market-based policy could save the fleet approximately $63 million annually under our best-estimate scenario. Spatial evaluation of ship emissions reductions shows that market-based instruments can reduce more SO2 closer to land while being more cost-effective for the fleet. Results suggest that combining performance requirements with market-based instruments can most effectively control SO2 emissions from ships.     

Lower pregnancy losses in lactating dairy cows fed a diet enriched in alpha-linolenic acid

The objectives were to determine if a diet enriched in α-linolenic acid (ALA) would influence ovarian function, early embryo survival, conception rates, and pregnancy losses in lactating dairy cows. Beginning 28 d before breeding, Holstein cows (55 ± 22 d postpartum; mean ± SD) were assigned to diets supplemented with either rolled flaxseed (FLAX; 56.7% ALA, n = 62) or rolled sunflower seed (SUNF; 0.1% ALA, n = 59) to provide approximately 750 g of oil/d. Diets continued for 32 d after timed artificial insemination (TAI, d 0) following a Presynch/Ovsynch protocol. Barley silage- and barley grain-based TMR were formulated to meet or exceed National Research Council requirements. Metabolizable protein and net energy for lactation concentrations were similar in the 2 diets. Based upon a mean dry matter intake of 22 kg/d, cows fed FLAX or SUNF consumed > 410 g or < 1 g of ALA, respectively. Pregnancy was confirmed by ultrasound 32 d after TAI. Nonpregnant cows were placed on a second Ovsynch regimen and reinseminated 42 d after first TAI, and received oilseeds for 32 d after second TAI. Relative to prediet levels, FLAX increased the ALA content of milk by 187%. Ovarian ultrasonography was performed in 8 cows per diet; the mean diameter of ovulatory follicles was larger in cows fed FLAX compared with SUNF (16.9 ± 0.9 vs. 14.1 ± 0.9 mm), but follicle number, corpus luteum size, and plasma progesterone concentrations remained unaffected. Presumptive conception (progesterone < 1 ng/mL on d 0 and > 1 ng/mL on d 21) rates to first TAI were greater in FLAX than in SUNF (72.6 vs. 47.5%). Pregnancy losses were lower in cows fed FLAX (9.8%) compared with those fed SUNF (27.3%). Including flaxseed in the ration of dairy cows increased the size of the ovulatory follicle and reduced pregnancy losses.     

Prevalence of Mycobacterium avium ssp. paratuberculosis infections in Canadian dairy herds

Johne's disease is a progressive, chronic disease with inflammation of the small intestine of ruminants caused by Mycobacterium avium ssp. paratuberculosis (MAP). Accurately estimating prevalence of MAP infections is important when controlling spread of infection or monitoring effectiveness of control programs. In the absence of a consistent test method used in prevalence studies across Canada, prevalence estimates among regions and programs cannot be compared. The aim of the current study was to estimate and compare prevalence of MAP infection in Western Canada, Ontario, Québec, and the Atlantic provinces, as well as among varying herd sizes and housing types. On 362 dairy farms located in all 10 provinces of Canada, environmental samples were collected and cultured for detection of MAP. For each herd, 1 sample was collected from the lactating cow area and manure storage. An additional environmental sample was collected from the area where breeding-age heifers were housed. Using prior distributions from previous research, diagnostic sensitivity and specificity were calculated to assess the ability of only 2 environmental samples (manure storage and lactating cow area) to identify MAP-positive farms, resulting in a sensitivity and specificity of 38 and 100%, respectively. We found no difference in sensitivity and specificity when including breeding-age heifers environmental samples. Test characteristics were applied to environmental culture results from the 362 participating farms in all 4 regions, resulting in true prevalence estimates of 66% for farms in Western Canada, 54% in Ontario, 24% in Québec, and 47% in Atlantic Canada. Herds housed in tiestalls had lower prevalence than freestall-housed herds, and herds with 101–150 and >151 cows had higher prevalence than herds with ≤100 cows. This was the first time MAP prevalence was determined using 1 detection method, performed in 1 laboratory, and within a single year across Canada, enabling direct comparisons of prevalence among regions, housing types, and herd sizes.