Gestational exposure to the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) has been implicated as causative to disparities between ethnic groups with respect to learning disabilities. Dioxin is an extremely toxic environmental pollutant that bioaccumulates in maternal adipose tissue, and is transferred to the developing organism during gestation and lactation. Long-term cognitive deficits have been reported following prenatal exposure to dioxin. N-methyl-D-aspartate (NMDA) receptors in the central nervous system (CNS) have been well known to play an important role in the activity-dependent synaptic plasticity underlying learning and memory and in CNS development including brain cell differentiation. Here, the effects of prenatal exposure to dioxin on the developmental expression profiles of rat hippocampal NMDA receptor subtype 1 mRNA and protein was examined. F-344 rats were exposed to 0 and 700 ng of dioxin/kg on gestational day 15. Real-time PCR and Western blot analysis clearly revealed that dioxin significantly downregulated NMDAR1 mRNA and protein expression during the first postnatal month. The study provides support to the hypothesis that NMDA receptors are important targets for dioxin-induced neurotoxicity in F1 preweaning pups. The results also support the concept that prenatal exposure to dioxin may contribute to the pathogenesis of diseases in the adult.

Animals↗

PubMed2002

Protons activate homomeric Kir6.2 channels by selective suppression of the long and intermediate closures.

The ATP-sensitive K+ channels (KATP) play an important role in regulating membrane excitability. These channels are regulated by H+ in addition to ATP, ADP, and phospholipids. To understand how protons affect the single-channel properties, Kir6.2DeltaC36 currents were studied in excised inside-out patches. We chose to study the homomeric Kir6.2 channel with 36 amino acids deleted at the C-terminal end, as there are ADP/ATP-binding sites in the SUR subunit, which may obscure the understanding of the channel-gating process. In the absence of ATP, moderate intracellular acidosis (pH 6.8) augmented P(open) with small suppression (by approximately 10%) of the single-channel conductance. The long and intermediate closures were selectively inhibited, leading to a shortening of the mean closed time without significant changes in the mean open time. Stronger acidification (<pH6.2) caused channel rundown. Although similar changes in the single-channel properties were observed in the presence of 1 mM ATP, the P(open)-pH relationship curve was shifted by 0.17 pH units toward lower pH levels. ATP had no effect on the inhibition of single-channel conductance during acidosis. Mutation of His175 eliminated the pH effect on the single-channel kinetics, while the single-channel response to acidic pH was retained in the K185E mutant that greatly reduces the ATP sensitivity. These results indicate that Kir6.2DeltaC36 channel gating by protons and ATP relies on two distinct mechanisms opposite to each other, although the pH sensitivity is modulated by ATP.

Adenosine Triphosphate↗

PubMed2002

Effect of humic substances on Cu(II) solubility in kaolin-sand soil.

The type and amount of organic matter present in industrially contaminated soils will influence the risk they pose. Previous studies have shown the importance of humic and fulvic acids (FAs) (important components of soil organic matter) in increasing the solubility of toxic metals but were not carried out using toxic metal levels and the pH range typical of industrially contaminated soils. This study investigated the influence of three humic substances (HSs: humates, fulvates and humins) on the solubility of copper(II) ions in kaolinitic soil spiked with Cu at levels representative of industrially contaminated soil. Humates, fulvates and humin were extracted from Irish moss peat, and controlled pH batch leaching tests were conducted on an artificial kaolin-sand soil that was spiked with each. Further leaching tests were conducted on soil spiked with each HS and copper nitrate. Dissolved organic contents were determined by titration and total and free aqueous copper concentrations in the leachate were measured using AAS and ion selective electrode (ISE) potentiometry respectively (dissolved complexed copper levels were determined by difference). It was found that humates and fulvates are partially sorbed by the soil, probably by chemisorption on positively charged gibbsite (Al-hydroxide) sites in the kaolinite. The addition of 340 mg/kg Cu(II) ions did not significantly affect the amount of humate or fulvate sorbed. Dissolved humates and fulvates form soluble complexes with copper over the pH range 3-11. However, in the presence of kaolinite, soluble copper humates and fulvates are unable to compete with the kaolinite for Cu ions at pH 6-7. Above pH 8, humate and fulvate complexes are the only forms of dissolved Cu. Humin is largely insoluble and has little effect on Cu mobility between pH 2 and 12. The implication of this study is that measurement of total soil organic content and water leaching tests should be a standard part of contaminated site investigation.

Adsorption↗

PubMed2002

Elicitor-like effects of low-energy ultrasound on plant (Panax ginseng) cells: induction of plant defense responses and secondary metabolite production.

In this work we examined the elicitor-like effects of low-energy ultrasound (US) on plant cells with respect to the induction of plant defense responses and secondary metabolite production. Panax ginseng cells in suspension culture were exposed to US (power <or=0.1 W/cm(3) at 38.5 kHz fixed frequency) for short periods of time (30 s-6 min). Two early events in plant defense metabolism and signal transduction pathways, the increased cross-membrane ion fluxes (Ca(2+) influx and K(+) efflux/H(+) influx) and the production of active oxygen species (AOS), were detected in sonicated cultures within 2 min after US exposure. These responses could be induced with small doses of US energy, 0.6-0.8 J/cm(3), and enhanced by increasing US energy within a non-inhibitory range. US treatment stimulated the synthesis of useful secondary metabolites, saponins of ginseng cells, without causing any net loss of the biomass yield of ginseng cell cultures. These results suggest that US can act as a potent abiotic elicitor to induce the defense responses of plant cells and to stimulate secondary metabolite production in plant cell cultures.

Cell Membrane↗

PubMed2002

Genistein, a soybean isoflavone, affects bone marrow lymphopoiesis and prevents bone loss in castrated male mice.

Soybean isoflavones exhibit selective effects on bone metabolism in postmenopausal women as well as in ovariectomized animals. Recently, the role of estrogen in bone metabolism in men has also received attention, because a man with a mutated estrogen receptor-alpha (ER(alpha)) gene will exhibit osteoporotic phenotypes. To examine the possible role of genistein, a soybean isoflavone, in bone marrow hemopoiesis and bone metabolism in men, male mice were orchidectomized (orx) and treated with genistein (0.4-0.8 mg/day) or 17beta-estradiol (E(2); 0.03 microg/day) subcutaneously for 3 weeks. In orx mice, seminal vesicle weight decreased markedly, and it was not affected by the administration of genistein or E(2). The number of bone marrow cells was markedly increased after orx, and the majority was B-220 weakly positive pre-B cells. Increased B-lymphopoiesis was restored completely by E(2) or genistein administration. In orx mice, bone mineral density of the femur decreased markedly, and this bone loss was prevented to a significant extent by treatment with genistein as well as E(2). Histomorphometry showed that the trabecular bone volume in the femoral distal metaphysis decreased markedly after orx, and genistein and E(2) prevented this bone loss. These results suggest that soybean isoflavones prevent bone loss due to androgen deficiency in males.

Animals↗

PubMed2002

Activation of caspase-8 is critical for sensitivity to cytotoxic anti-Fas antibody-induced apoptosis in human ovarian cancer cells.

Two ovarian cancer cell lines named NOS4 and SKOV-3 have been shown to have different sensitivities to a cytotoxic anti-Fas antibody, CH-11. Although both cell lines express Fas molecules on the cell surfaces at the same intensities, apoptosis is induced by CH-11 in NOS4 cells but not in SKOV-3 cells. In this study, the different apoptosis-sensitivities of these cells were assessed. Both cell lines express almost the same levels of FADD, RIP, c-FLIP, FAP-1, Bax, Bcl-2 and Bcl-XL. Evidence of caspase-8, caspase-9 and caspase-3 activation and of cleavage of PARP and Bid was obtained in NOS4 cells but not in SKOV-3 cells. When triggered by FasL protein, DNA fragmentation and caspase-8 activation were observed in SKOV-3 cells, though they were not as clear as in NOS4 cells. All the anti-Fas antibody-mediated signals for apoptosis induction in NOS4 cells were completely blocked by a caspase-8-specific inhibitor, Z-IETD-FMK. These results indicate that the different sensitivities to the anti-Fas antibody are solely dependent on the activation of caspase-8, which could be influenced by yet unknown qualitative or quantitative abnormalities in molecules involved in DISC formation.

Adaptor Proteins, Signal Transducing↗

PubMed2002

Identification and sequencing of HLA-B0714 and B2718 alleles and novel exon 1 sequences of B0709 and B2714 alleles in potential bone marrow donors.

Sequence specific oligonucleotide probe hybridization and sequence specific primer PCR typing of volunteer bone marrow donors suggested the presence of variants of known HLA-B alleles in two individuals. PCR products encompassing HLA-B locus exons 1, 2, and 3 were prepared, subcloned and sequenced. A Hispanic individual had a novel B*07 allele (B*0714) and a Chinese individual had a novel B*27 allele (B*2718). In two other individuals, a previously unknown sequence of exon 1 was determined for HLA-B*0709 (African American) and B*2714 (Native American). These findings further illustrate the substantial genetic variation present at the HLA-B locus within human populations. We discuss the structural variation in the protein sequence for these HLA-B alleles and its potential functional effects.

Asian People↗

PubMed2002

New HLA-B alleles identified and sequences extended in potential bone marrow donors: B5804, B4418, B1558, and B4805.

Nucleic acid-based methods for allele identification have revealed more than 470 polymorphic variants at the HLA-B locus. Screening of potential bone marrow donors with sequence specific primer polymerase chain reactions and sequence specific oligonucleotide probe hybridization assays revealed apparent variants within the B*58, *44, *15, and *48 allele groups. DNA sequencing of cloned DNA identified the new alleles B*5804, B*4418, and B*1558 within these groups and observed new sequence information for the previously reported allele B*4805. These findings further extend our knowledge of the substantial genetic variation present at the HLA-B locus within human populations.

Alleles↗

PubMed2002

A nonsense mutation in exon 3 results in the HLA-B null allele B*5127N.

A new HLA-B null allele has been identified within the B*51 group by combined serological and molecular typing of an Italian Caucasoid family. Serological data indicated that the proband typed homozygous for A2 and B60. Confirmatory typing using sequence specific oligonucleotide hybridization (SSPOH) detected a second B allele within the B*51 group. Allele specific typing (SSP) for B*51 subtypes, including the known B*5111N allele, was performed, and typing results were consistent with B*5101, suggesting the presence of a new null variant. Cloning and sequencing of this allele identified a B*5101 variant with a nonsense mutation in exon 3. This new null allele has been designated B*5127N. The combined use of serologic and DNA-based typing methods facilitates the identification of null and low-expression alleles. An overview of null alleles of class I HLA is presented.

Base Sequence↗

PubMed2002

Mutual passivation of electrically active and isovalent impurities.

The alloy GaN(x) As(1-x) (with x typically less than 0.05) is a novel semiconductor that has many interesting electronic properties because of the nitrogen-induced dramatic modifications of the conduction band structure of the host material (GaAs). Here we demonstrate the existence of an entirely new effect in the GaN(x) As(1-x) alloy system in which the Si donor in the substitututional Ga site (Si(Ga)) and the isovalent atom N in the As sublattice (N(As)) passivate each other's electronic activity. This mutual passivation occurs in Si-doped GaN(x) As(1-x) through the formation of nearest-neighbour Si(Ga) -N(As) pairs and is thermally stable up to 950 degrees C. Consequently, Si doping in GaN(x) As(1-x) under equilibrium conditions results in a highly resistive GaN(x) As(1-x) layer with the fundamental bandgap governed by a net 'active' N, roughly equal to the total N content minus the Si concentration. Such mutual passivation is expected to be a general phenomenon for electrically active dopants and localized state impurities that can form nearest-neighbour pairs.

Alloys↗

PubMed2002

Association between baseline levels of C-reactive protein (CRP) and a dinucleotide repeat polymorphism in the intron of the CRP gene.

Elevation of baseline C-reactive protein (CRP) is associated with increased risk of cardiac disease. This increase might reflect low-grade inflammation, but differences in CRP serum levels might also have a genetic component. To test this possibility, we investigated whether a polymorphic GT-repeat in the intron of the CRP gene contributes to variation in baseline CRP. We found that the polymorphism was associated with differences in baseline CRP in both normal individuals and in patients with the inflammatory disease systemic lupus erythematosus, viz. donors carrying two GT(16) alleles, two GT(21)alleles, or GT(16/21) heterozygotes had two-fold lower serum CRP than those with other genotypes. The frequency of GT(16) and GT(21) was two-fold higher in Caucasians than in African-Americans, but there was no difference in allele distribution between patients and controls. It is not yet known how this genetic polymorphism mediates its effect on CRP expression, and it probably is not a systemic lupus erythematosus susceptibility factor. Rather, the CRP intron polymorphism likely modifies the disease phenotype. On the other hand, the fact that baseline CRP does have a genetic component suggests that in coronary disease, stratification of risk assessment based on CRP levels might be enhanced by consideration of this polymorphism.

Adult↗

PubMed2002

Genomic organization of classical human low-affinity Fcgamma receptor genes.

The classical low-affinity Fcgamma receptor genes (FcgammaRIIA, B, C and FcgammaRIIIA, B) are located on chromosome 1q23, a region that shows strong linkage with human systemic lupus erythematosus (SLE) in several genome-wide scans, and family-based association between FcgammaRIIIA and SLE is now established. High homology among the Fcgamma receptor genes, however, has hampered further study of this region. We have used a human bacterial artificial chromosome (BAC) library to determine the order and orientation of these Fcgamma receptor genes and have sequenced the very highly homologous 5' region (including 3.4 kb of the promoter and the 8 kb from exon 1 to exon 3) of the FcgammaRIIB and FcgammaRIIC genes to enable study of their unique single nucleotide polymorphisms (SNP). We have utilized these data to characterize a linked set of three coding region SNPs in the FcgammaRIIC exon 3 (EC1) that includes the stop codon SNP, which provides an important insight into natural killer cell function. Together, these data provide the basis for the study of additional SNPs in FcgammaR genes in SLE disease susceptibility.

Chromosome Mapping↗