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PubMed2001

Copper in disorders with neurological symptoms: Alzheimer's, Menkes, and Wilson diseases.

Copper is an essential element for the activity of a number of physiologically important enzymes. Enzyme-related malfunctions may contribute to severe neurological symptoms and neurological diseases: copper is a component of cytochrome c oxidase, which catalyzes the reduction of oxygen to water, the essential step in cellular respiration. Copper is a cofactor of Cu/Zn-superoxide-dismutase which plays a key role in the cellular response to oxidative stress by scavenging reactive oxygen species. Furthermore, copper is a constituent of dopamine-beta-hydroxylase, a critical enzyme in the catecholamine biosynthetic pathway. A detailed exploration of the biological importance and functional properties of proteins associated with neurological symptoms will have an important impact on understanding disease mechanisms and may accelerate development and testing of new therapeutic approaches. Copper binding proteins play important roles in the establishment and maintenance of metal-ion homeostasis, in deficiency disorders with neurological symptoms (Menkes disease, Wilson disease) and in neurodegenerative diseases (Alzheimer's disease). The Menkes and Wilson proteins have been characterized as copper transporters and the amyloid precursor protein (APP) of Alzheimer's disease has been proposed to work as a Cu(II) and/or Zn(II) transporter. Experimental, clinical and epidemiological observations in neurodegenerative disorders like Alzheimer's disease and in the genetically inherited copper-dependent disorders Menkes and Wilson disease are summarized. This could provide a rationale for a link between severely dysregulated metal-ion homeostasis and the selective neuronal pathology.

Alzheimer Disease↗

PubMed1994

Target cell death triggered by cytotoxic T lymphocytes: a target cell mutant distinguishes passive pore formation and active cell suicide mechanisms.

The role of the target cell in its own death mediated by cytotoxic T lymphocytes (CTL) has been controversial. The ability of the pore-forming granule components of CTL to induce target cell death directly has been taken to suggest an essentially passive role for the target. This view of CTL-mediated killing ascribes to the target the single role of providing an antigenic stimulus to the CTL; this signal results in the vectoral degranulation and secretion of pore-forming elements onto the target. On the other hand, by a number of criteria, target cell death triggered by CTL appears fundamentally different from death resulting from membrane damage and osmotic lysis. CTL-triggered target cell death involves primary internal lesions of the target cell that reflect a physiological cell death process. Orderly nuclear disintegration, including lamin phosphorylation and solubilization, chromatin condensation, and genome digestion, are among the earliest events, preceding the loss of plasma membrane integrity. We have tested directly the involvement of the target cell in its own death by examining whether we could isolate mutants of target cells that have retained the ability to be recognized by and provide an antigenic stimulus to CTL while having lost the capacity to respond by dying. Here, we describe one such mutant, BW87. We have used this CTL-resistant mutant to analyze the mechanisms of CTL-triggered target cell death under a variety of conditions. The identification of a mutable target cell element essential for the cell death response to CTL provides genetic evidence that target cell death reflects an active cell suicide process similar to other physiological cell deaths.

Animals↗

PubMed2002

[Patient with COPD. Comprehensive assistance in home care].

In order to respond to the objectives of a Home Care Patient Program bearing in mind the patient as the fundamental element from his/her individual and human dimensions, considering his/her family and family environs, it is necessary to establish and integrated, interdisciplinary therapeutic method based on a coordinated program involving all the professional involved in the process. The work of nurses, an essential element in this program, should be based on a conceptual model. It is important to elaborate an integral care plan based on the detection of the needs, objectives to achieve, team interventions both on the patient as the caretaker and to evaluate the results in order to achieve the highest degree of effectiveness and efficiency in home care of patients.

Home Care Services↗

PubMed1996

Trace metals in populations of freshwater isopods: influence of biotic and abiotic variables.

Trace metal levels in water, sediments and freshwater isopods from 28 different water systems in the Netherlands were measured during the period of 1986 to 1989. Distinct element-specific internal distribution patterns were present, with Cd and Cu stored mainly in the hepatopancreas (30-60% of total body burden) and Pb and Zn in the hindgut and exoskeleton with hemolymph. Mean whole-body concentrations of the non-essential elements Cd and Pb in individually analyzed isopods varied over three orders of magnitude between populations. The variability of Zn and Cn were within one order of magnitude difference. The variability of trace metal levels between populations exceeded within-population variability. Within-population variability was related to seasonal and biological factors such as body weight, sex, reproductive state, and species abundance. The highest concentrations were found in small juvenile animals compared to adults, females compared to males, and Proasellus meridianus compared to Asellus aquaticus. However, after correction for size effects using a power-curve regression model no significant differences remained between sex and species. Seasonal fluctuations accounted for 33-79% of the within population variability. Trace metal levels in isopods were predicted from concentrations in water and sediments in combination with aqueous Ca, Cl-, DOC, and sediment characteristics (Org-C, clay, CaCO3) using a multiple regression model. With this predictive model 42-63% of the variance could be explained. In situ determined partitioning coefficients (apparent BCF, biota-sediment BSAF, and sediment-water distribution coefficient Kd) varied between locations and covaried with factors related to trace metal bioavailability (aqueous Ca, Cl- and DOC, sediment Org-C, clay, and CaCO3). Especially for Cd and Cu field-derived BCF values were in agreement with previously reported experimental studies. It is concluded, that A. aquaticus may be a suitable candidate-organism for biomonitoring available trace metal levels in littoral freshwater systems. Finally, some practical recommendations are given for future field surveys with freshwater isopods with respect to sample size, allometric standardization, period of sampling and statistical design.

Analysis of Variance↗

PubMed2005

The impact of the hyperacid Ijen Crater Lake. Part I: Concentrations of elements in crops and soil.

In Asembagus (East Java, Indonesia) irrigation water is contaminated with effluent from the hyperacid Ijen Crater Lake resulting in a low pH and high levels of various elements. As a first step towards a risk assessment, locally produced food items (rice, maize, cassava leaf, cassava root, peanuts) were collected and concentrations of As, B, Ca, Cd, Co, Cu, Fe, Mg, Mn, Mo, Ni, Pb, V, Zn were compared to samples from a reference area and with literature values. Further, concentrations in rice were compared to total soil concentrations in paddy fields. Compared to the reference area, food items produced in the contaminated area had increased levels of Cd, Co, Ni and Mn in particular, while levels of Mo were lower. In contrast, total soil concentrations of Cd and Mn in particular have decreased whereas especially Mo was increased. In combination with the observed soil acidification, it is likely that the bioavailable concentration of most elements in the contaminated soil is higher (except for Mo) due to an increased weathering rate and/or input via the contaminated irrigation water. In terms of human health, concentrations in foods were generally within normal literature values. However, it was observed that essential elements (in particular Fe) known for their inhibitory effects on e.g. Cd and Mn toxicity did not accumulate in crops whereas Cd and Mn did.

Crops, Agricultural↗

PubMed1984

Reference values for elemental concentrations in some human samples of clinical interest: a preliminary evaluation.

Over 60 investigators, some of them with an outstanding international reputation as analysts, were requested to identify reliable data from their countries for elemental concentrations in normal adult human samples of clinical interest by contributing their own data as well as screening the literature information. A set of samples consisting of whole blood and its components, urine, milk, liver and hair were chosen and considered for 15 elements of biological significance: Zn, Cu, Fe, Se, Mn, Pb, Cd, Hg, As, Mo, Cr, Co, I, Ni and F. The results partly cover over 40 countries from the global regions of Africa, Asia, Europe, North, South and Central America, Australia and New Zealand. This survey has been useful in demonstrating certain trends of trace element picture around the world, at least qualitatively. Both diet and environment have a strong influence on the distribution pattern of several elements such as As, Cd, Mn, Pb, Se and Zn. A limited comparison of the available information on soil status of different countries revealed interesting associations for elements such as Mn and Zn. Importantly, this study revealed that only a few countries were in a position to supply a reasonable amount of data on samples requested here. In particular, for a number of countries, reliable data for even very essential elements such as Cu, Zn and Fe were not available. In view of the nutritional importance of several elements the time is ripe for international organizations to intervene and help produce some reference data for selected global regions which lack data of any kind.

Diet↗

PubMed1994

Conserved regulatory elements in the type I DNA topoisomerase gene promoters of mouse and man.

The gene for mammalian type I DNA topoisomerase is constitutively expressed, but also regulated by a number of external stimuli. We compared the nucleotide sequences of the human and the mouse topoisomerase I gene promoters because promoter elements, essential for basic as well as regulated gene expression, should be conserved during evolution. We found that proximal upstream sequences are highly conserved and include potential binding sites for ubiquitous transcription factors, a regulatory CRE site as well as two novel promoter elements that have been shown to be important for the expression of the human gene. The more distal parts of the upstream sequences are less well conserved but include two regions that are almost identical in the human and the mouse gene. One of these regions contains a binding site for a basic-helix-loop-helix/leucine-zipper protein, and the other contains an AT-rich element with the potential for DNA bending.

Animals↗

PubMed2005

Hepatic and renal concentrations of 10 trace elements in crocodiles (Crocodylus niloticus) in the Kafue and Luangwa rivers in Zambia.

Hepatic and renal concentrations of the elements arsenic, cadmium, cobalt, copper, lead, manganese, mercury, molybdenum, selenium and zinc were determined in samples collected from four crocodiles from the Kafue River, Kafue National Park and five crocodiles from the Luangwa River, Luangwa National Park, Zambia. The concentrations of the essential elements were similar to those reported in other vertebrates. Arsenic and cadmium concentrations were low (medians below 0.05 microg As/g and below 0.16 microg Cd/g, wet wt.). Mercury and lead concentrations were several orders of magnitude higher (medians up to 3.7 microg Hg/g, and up to 8.7 microg Pb/g, all wet wt.) than in hippopotami from the same rivers, probably as a result of food-chain biomagnification. Judging by the results obtained in this study, pollution from the mining activity around the Kafue River drainage area in the Copperbelt region has not significantly influenced the trace element concentrations in tissues of the crocodiles in the Kafue National Park. The trace element concentrations measured may serve as reference values in future studies on crocodilians.

Alligators and Crocodiles↗

PubMed2000

Clusterin gene in rat sertoli cells is regulated by a core-enhancer element.

Clusterin is a ubiquitous glycoprotein that is promiscuously expressed at a low basal level but can be highly induced by a variety of stress conditions. In contrast, in some secretory cells associated with tissue-fluid interfaces such as the Sertoli cells in the testis, clusterin demonstrates high constitutive expression. In this study, we address the mechanisms that regulate the constitutive expression of the clusterin gene by using primary cultures of immature rat Sertoli cells. We have identified a region of the rat clusterin gene promoter that activated transcription only in Sertoli cells and that mapped between positions -426 and -311. Sequence analysis of this region revealed a high concentration of potential regulatory elements. Using gel-shift assays combined with hydroxyl radical footprinting, we identified the elements recognized by the Sertoli cell nuclear factors. Comparison of the interactions with this region of the nuclear factors from different cell types demonstrated that recognition of the core-enhancer element is specific for the Sertoli cells, and in vitro, the core region was recognized by the transcription factor CBF. Transient transfections showed that a core enhancer is responsible for more than a half of the total promoter activity and is an essential element for the cell-specific activity of the Sertoli-specific region. In addition to the core enhancer, tandem Sp1 sites are also required for maximal activity of this region.

Animals↗

PubMed2006

The inner cavity of Escherichia coli DegP protein is not essential for molecular chaperone and proteolytic activity.

The Escherichia coli DegP protein is an essential periplasmic protein for bacterial survival at high temperatures. DegP has the unusual property of working as a chaperone below 28 degrees C, but efficiently degrading unfolded proteins above 28 degrees C. Monomeric DegP contains a protease domain and two PDZ domains. It oligomerizes into a hexameric cage through the staggered association of trimers. The active sites are located in a central cavity that is only accessible laterally, and the 12 PDZ domains act as mobile sidewalls that mediate opening and closing of the gates. As access to the active sites is restricted, DegP is an example of a self-compartmentalized protease. To determine the essential elements of DegP that maintain the integrity of the hexameric cage, we constructed several deletion mutants of DegP that formed trimers rather than hexamers. We found that residues 39 to 78 within the LA loops, as well as the PDZ2 domains are essential for the integrity of the DegP hexamer. In addition, we asked whether an enclosed cavity or cage of specific dimensions is required for the protease and chaperone activities in DegP. Both activities were maintained in the trimeric DegP mutants without an enclosed cavity and in deletion DegP mutants with significantly reduced dimensions of the cage. We conclude that the functional unit for the protease and chaperone activities of DegP is a trimer and that neither a cavity of specific dimensions nor the presence of an enclosed cavity appears to be essential for the protease and chaperone activities of DegP.

Chromatography, Gel↗

PubMed1990

Transcription of osmB, a gene encoding an Escherichia coli lipoprotein, is regulated by dual signals. Osmotic stress and stationary phase.

The osmB gene, which encodes an outer membrane lipoprotein, can be induced by both osmotic and growth phase signals. Construction of two transcriptional fusions, an osmB-lacZ fusion in single copy on the bacterial chromosome and an osmB-cat fusion carried on a multicopy plasmid, demonstrated that induction of osmB by hyperosmolarity and during the stationary phase of growth occurred at the level of transcription. Two transcription initiation sites were identified by RNase protection of in vivo message. The downstream P2 promoter is the primary site for regulation; the basal level of expression is initiated at P2 and transcription from P2 is induced by elevated osmolarity or upon reaching stationary phase. Transcription from the P1 promoter, 150 base pairs (bp) upstream of the P2 promoter, occurred only when both osmotic and growth phase signals were present simultaneously; that is, when cells growing in high osmolarity medium have reached stationary phase. Deletion analysis narrowed the sequences necessary for P2 regulation to the 42-bp region upstream from the transcription start site. A 7-bp sequence just upstream from the -35 region was identified as a cis-acting regulatory element essential for osmotic stimulation of osmB expression. A hexanucleotide sequence within this segment could form the left arm of a region of dyad symmetry, flanking the -35 region of the promoter. Stationary phase induction at P2 does not require the 7-bp element.

Bacterial Outer Membrane Proteins↗

PubMed1989

Interactions of thorium and cerium with adenine nucleotides.

The present study demonstrates an effect of the actinide, Thorium (Th), and the lanthanide, Cerium (Ce), on the non-enzymatic hydrolysis of adenine and guanine nucleotides. Thorium was found to be a more effective catalyst of the reaction than Cerium. Further, both the elements promoted the binding of Creatine kinase (CK) to Cibacron blue F3 GA, the substrate analogue of the enzyme, even in the absence of Mg2+, the physiological cofactor. These observations imply that toxic heavy metals can mimic the functional effects of the essential elements despite differences in their physicochemical properties.

Adenine Nucleotides↗

PubMed2003

An ascidian homolog of vertebrate iodothyronine deiodinases.

In all classes of vertebrates, the deiodination of the prohormone T(4) to T(3) represents an essential activation step in thyroid hormone action. The possible presence of iodothyronine deiodinase activity in protochordates has been demonstrated in vivo. Recent molecular cloning of the genomes and transcripts of several ascidian species allows further investigation into thyroid-related processes in ascidians. A cDNA clone from Halocynthia roretzi (hrDx) was found to have significant homology (30% amino acid identity) with the iodothyronine deiodinase gene sequences from vertebrates, including the presence of an in-frame UGA codon that might encode a selenocysteine (SeC) in the active site. Because it was not certain that the 3' untranslated region (UTR) contained a SeC insertion sequence (SECIS) element essential for SeC incorporation, a chimeric expression vector of the hrDx coding sequence and the rat deiodinase SECIS element was produced, as well as an expression vector containing the intact hrDx cDNA. COS, CHO, and HEK cells were transfected with these vectors, and deiodinase activity was measured in cell homogenates. Outer-ring deiodinase activity was detected using both T(4) and reverse T(3) as substrates, and activity was enhanced by the presence of the reductive cofactor dithiothreitol. The enzyme activity was optimal during incubation between 20 and 30 C (pH 6-7) and was strongly inhibited by gold-thioglucose. The Halocynthia deiodinase appears to be a high Michaelis-Menten constant (K(m)) enzyme (K(m) reverse T(3), 2 microM; and K(m) T(4), 4 microM). Deiodinase activity was completely lost upon the substitution of the SeC residue in the putative catalytic center by either cysteine or alanine. Transfection of the full-length hrDx cDNA produced deiodinase activity confirming the presence of a SECIS element in the 3'UTR, as revealed by the SECISearch program. In conclusion, our results show, for the first time, the existence of an ascidian iodothyronine outer-ring deiodinase. This raises the hypothesis that, in protochordates, the prohormone T(4) is activated by enzymatic outer-ring deiodination to T(3).

3' Untranslated Regions↗

PubMed2001

Mineral status of female rats affects the absorption and organ distribution of dietary cadmium derived from edible sunflower kernels (Helianthus annuus L.).

The intake of food cadmium (Cd) in microg/day over time can increase the body burden of this element. Some human populations that consume subsistence rice-based diets low in calcium (Ca), iron (Fe), and zinc (Zn) are more susceptible to Cd poisoning than populations that consume more nutritious diets. This study determined the effects of marginal deficiencies of these essential elements on the absorption and organ retention of Cd from a natural food that contains Cd, edible sunflower kernels (Helianthus annuus L.; SFK). Weanling female rats were fed diets containing 20% SFK in a 2x2x2 factorial design with marginal and adequate amounts of Ca, Zn, and Fe. Marginal Zn (11 mg/kg) and Fe (13 mg/kg), and Cd (0.18 mg/kg) were derived solely from 20% SFK. These amounts of Fe and Zn represented 39 and 90% of the NRC requirement for the rat, respectively. The marginal dietary Ca concentration (2.5 g/kg) was one-half the NRC requirement. After 5 weeks on the experiment, rats were fed 1 g of their respective diets containing SFK extrinsically labeled with 37 kBq 109Cd, and absorption was determined by whole-body counting techniques. Rats were then killed and organs collected for 109Cd assays. No effect of treatment on weight gain was observed; however, when dietary Zn was low, feeding marginal Ca elevated Cd absorption by 50% (P<0.05) over those fed adequate Ca and Zn. Feeding marginal Fe elevated Cd absorption >2.5-fold (P<0.001) over those fed adequate Fe. In contrast, the naturally occurring Zn in SFK that provided 90% of the rat's requirement was enough to deter excessive absorption of Cd and enough to alone prevent significant elevation of organ Cd. Organ content of 109Cd and Cd followed the same general pattern as whole-body absorption. These data show that marginal nutritional deficiencies of Ca and Fe can readily enhance the body burden of Cd that comes from the diet. Also, some natural competitors of Cd, such as Zn, contained in foods can independently minimize Cd absorption.

Absorption↗

PubMed2004

Competition and stoichiometry: coexistence of two predators on one prey.

The competitive exclusion principle (CEP) states that no equilibrium is possible if n species exploit fewer than n resources. This principle does not appear to hold in nature, where high biodiversity is commonly observed, even in seemingly homogenous habitats. Although various mechanisms, such as spatial heterogeneity or chaotic fluctuations, have been proposed to explain this coexistence, none of them invalidates this principle. Here we evaluate whether principles of ecological stoichiometry can contribute to the stable maintenance of biodiverse communities. Stoichiometric analysis recognizes that each organism is a mixture of multiple chemical elements such as carbon (C), nitrogen (N), and phosphorus (P) that are present in various proportions in organisms. We incorporate these principles into a standard predator-prey model to analyze competition between two predators on one autotrophic prey. The model tracks two essential elements, C and P, in each species. We show that a stable equilibrium is possible with two predators on this single prey. At this equilibrium both predators can be limited by the P content of the prey. The analysis suggests that chemical heterogeneity within and among species provides new mechanisms that can support species coexistence and that may be important in maintaining biodiversity.

Animals↗

PubMed1995

Transcriptional regulation of the human T cell receptor delta gene.

T cell receptor delta gene expression is regulated by a T cell-specific transcriptional enhancer located within the J delta 3-C delta intron. An essential element of the enhancer was localized to a small 30 bp segment denoted delta E3. Two specific factors, CBF/PEBP2 and c-Myb, bind to adjacent sites within delta E3 and cooperate functionally to mediate transcriptional activation. These factors are likely to play essential roles in the developmental activation of the TCR delta gene in vivo.

Gene Expression Regulation↗

PubMed2001

CCAAT/enhancer-binding proteins (C/EBP) beta and delta activate osteocalcin gene transcription and synergize with Runx2 at the C/EBP element to regulate bone-specific expression.

CCAAT/enhancer-binding proteins (C/EBP) are critical determinants for cellular differentiation and cell type-specific gene expression. Their functional roles in osteoblast development have not been determined. We addressed a key component of the mechanisms by which C/EBP factors regulate transcription of a tissue-specific gene during osteoblast differentiation. Expression of both C/EBPbeta and C/EBPdelta increases from the growth to maturation developmental stages and, like the bone-specific osteocalcin (OC) gene, is also stimulated 3-6-fold by vitamin D(3), a regulator of osteoblast differentiation. We characterized a C/EBP enhancer element in the proximal promoter of the rat osteocalcin gene, which resides in close proximity to a Runx2 (Cbfa1) element, essential for tissue-specific activation. We find that C/EBP and Runx2 factors interact together in a synergistic manner to enhance OC transcription (35-40-fold) in cell culture systems. We show by mutational analysis that this synergism is mediated through the C/EBP-responsive element in the OC promoter and by a direct interaction between Runx2 and C/EBPbeta. Furthermore, we have mapped a domain in Runx2 necessary for this interaction by immunoprecipitation. A Runx2 mutant lacking this interaction domain does not exhibit functional synergism. We conclude that, in addition to Runx2 DNA binding functions, Runx2 can also form a protein complex at C/EBP sites to regulate transcription. Taken together, our findings indicate that C/EBP is a principal transactivator of the OC gene and the synergism with Runx2 suggests that a combinatorial interaction of these factors is a principal mechanism for regulating tissue-specific expression during osteoblast differentiation.

Animals↗

PubMed2006

Rho-dependent terminators and transcription termination.

Rho-dependent transcription terminators participate in sophisticated genetic regulatory mechanisms, in both bacteria and phages; they occur in regulatory regions preceding the coding sequences of genes and within coding sequences, as well as at the end of transcriptional units, to prevent readthrough transcription. Most Rho-dependent terminators have been found in enteric bacteria, but they also occur in Gram-positive bacteria and may be widespread among bacteria. Rho-dependent termination requires both cis-acting elements, on the mRNA, and trans-acting factors. The only cis-acting element common to Rho-dependent terminators is richness in rC residues. Additional sequence elements have been observed at different Rho termination sites. These 'auxiliary elements' may assist in the termination process; they differ among terminators, their occurrence possibly depending on the function and sequence context of the terminator. Specific nucleotides required for termination have also been identified at Rho sites. Rho is the main factor required for termination; it is a ring-shaped hexameric protein with ATPase and helicase activities. NusG, NusA and NusB are additional factors participating in the termination process. Rho-dependent termination occurs by binding of Rho to ribosome-free mRNA, C-rich sites being good candidates for binding. Rho's ATPase is activated by Rho-mRNA binding, and provides the energy for Rho translocation along the mRNA; translocation requires sliding of the message into the central hole of the hexamer. When a polymerase pause site is encountered, the actual termination occurs, and the transcript is released by Rho's helicase activity. Many aspects of this process are still being studied. The isolation of mutants suppressing termination, site-directed mutagenesis of cis-acting elements in Rho-dependent termination, and biochemistry, are and will be contributing to unravelling the still undefined aspects of the Rho termination machinery. Analysis of the more sophisticated regulatory mechanisms relying on Rho-dependent termination may be crucial in identifying new essential elements for termination.

Gene Expression Regulation, Bacterial↗