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Identification of a promoter region for 3.6-kilobase mRNA of hepatitis B virus and specific cellular binding protein.

The promoter region for transcription of the 3.6-kilobase mRNA of hepatitis B virus was identified by the chloramphenicol acetyltransferase assay by using HuH-7 hepatoma cells and was found to function directly in virus production by way of the transient expression system of HBV. The 5'-upstream sequence from nucleotides 1573 to 1657 (the transcription start site) was indispensable for promoter function, while the AT-rich sequence (from nucleotides 1581 to 1604) containing a directly repeated sequence TGTT connecting the same flanking sequence PyAAAGAC (where Py is a pyrimidine) at both sides was an essential element within this promoter region. A specific cellular factor which interacted with the essential element was detected in the HuH-7 cell extract. A similar binding factor was also observed in HepG2 and huH2-2 hepatoma cells. This factor may thus be responsible for regulating 3.6-kilobase mRNA, pregenome RNA transcription, or both.

Base Sequence↗

Streamlining outpatient geriatric assessment: essential social, environmental and economic variables.

There are no known standardized instruments that encompass all of the essential elements of a streamlined outpatient geriatric assessment. A process of arriving at a consensus on essential elements of streamlined geriatric assessment in 3 domains was initiated and described. Two rounds of the Delphi technique were conducted to identify social, environmental and economic variables essential for such an assessment. Consensus among an expert panel was achieved and a list of 27 variables was reduced to 16. This result is discussed in terms of assessment context and standardizing assessment, using case examples.

Aged↗

Essential and toxic element concentrations in fresh and formalin-fixed human autopsy tissues.

The concentrations of five essential elements and six potentially toxic elements were determined in seven organs collected at autopsy from 30 human subjects. Elemental analyses were carried out by graphite furnace atomic absorption spectroscopy, and inductively coupled plasma emission spectroscopy, and inductively coupled plasma mass spectroscopy, and concentrations in fresh and formalin-fixed tissues were compared. Formalin-fixation long-term storage has little effect on most element concentrations in tissue, except for Al and Mn, which changed with prolonged storage in formalin. The kidney and liver contained the greatest concentrations of toxic elements compared with other organs, whereas the essential elements were uniformly distributed among all organs. There was no more than a 10-fold difference in the tissue concentration of the elements studied among the organs, except for the concentration of Fe in liver, and Ca and Mg in bone. We also demonstrate that these elements are homogeneously distributed in tissues.

Aluminum↗

Collaborative practice: myth or reality?

In response to demands of internal constituencies and pressures from external forces, many healthcare institutions have been experimenting with collaborative practice, wherein professionals function interdependently in patient-centered healthcare. In this article, the authors move beyond philosophical constructs and focus on the essential elements of this important innovation. Making collaborative practice a reality in institutions requires an understanding of the essential elements, persistent and continuing efforts, and rigorous evaluation of outcomes. Satisfaction, quality, and cost effectiveness are essential factors on two dimensions: outcomes for patient care providers; and outcomes for patients. Ultimately, collaborative practice can be recognized by demonstrated effective communication patterns, achievement of enhanced patient care outcomes, and efficient and effective support services in place. If these criteria are not met, collaborative practice is a myth and not a reality in your institution.

Hospitals↗

Molybdenum: an essential trace element.

Molybdenum is found in most foods, with legumes, dairy products, and meats being the richest sources. This metal is considered essential because it is part of a complex called molybdenum cofactor that is required for the three mammalian enzymes xanthine oxidase (XO), aldehyde oxidase (AO), and sulfite oxidase (SO). XO participates in the metabolism of purines, AO catalyzes the conversion of aldehydes to acids, and SO is involved in the metabolism of sulfur-containing amino acids. Molybdenum deficiency is not found in free-living humans, but deficiency is reported in a patient receiving prolonged total parenteral nutrition with clinical signs characterized by tachycardia, headache, mental disturbances, and coma. The biochemical abnormalities in this acquired molybdenum deficiency include very low levels of uric acid in serum and urine (low XO activity) and low inorganic sulfate levels in urine (low SO activity). Inborn errors of isolated deficiencies of XO, SO, and molybdenum cofactor are described. Although XO deficiency is relatively benign, patients with isolated deficiencies of SO or molybdenum cofactor exhibit mental retardation, neurologic problems, and ocular lens dislocation. These abnormalities seem to be caused by the toxicity of sulfite and/or inadequate amounts of inorganic sulfate available for the formation of sulfated compounds present in the brain. XO and AO may also participate in the inactivation of some toxic substances, inasmuch as studies suggest that molybdenum deficiency is a factor in the higher incidence of esophageal cancer in populations consuming food grown in molybdenum-poor soil.

Deficiency Diseases↗

Acid precipitation--effects on trace elements and human health.

Environmental pollution by acid precipitation increases the solubilization and mobilization of toxic metals. Through the food chain, this may alter the intake of toxic and essential elements in man. Potential adverse health effects could follow after increased human exposure. For the general population, the exposure pattern and health effects caused by aluminium, cadmium, lead and mercury are of particular concern. Although there are several indications that the exposure to toxic elements (e.g. aluminium, cadmium, lead and methylmercury), as well as the intake of essential elements (e.g. selenium), may be affected by acid precipitation, there is presently no firm evidence of adverse health effects in man. However, the present data clearly indicate that the safety margins are small. Thus, the ongoing acidification in many areas must be stopped before such effects become evident. The effects on trace element status and human health by acid precipitation were discussed at the ISTERH (International Society for Trace Element Research in Humans) Conference in Stockholm, May, 1992. The main findings are briefly summarized here.

Acid Rain↗

A global perspective on cadmium pollution and toxicity in non-occupationally exposed population.

Cadmium is a non-essential element that has high rates of soil to plant transference compared with other non-essential elements, and certain plant species accumulate large amounts of cadmium from low cadmium content soils. In this paper, levels of cadmium found in major food groups are highlighted together with cadmium levels found in liver and kidney samples from non-occupationally exposed populations. Data on human kidney cadmium levels identified recently, including the study in our own laboratory, are compared with older studies. Human-tissue cadmium contents showed large variations among individuals, but sources of the variation remain unknown. Exposure levels of 30-50 microg per day have been estimated for adults and these levels have been linked to increased risk of bone fracture, cancer, kidney dysfunction and hypertension. Increased mortality was found among individuals showing signs of cadmium renal toxicity compared with those without such signs, suggesting that renal toxicity may be an early warning of complications, sub-clinical or clinical morbidity.

Cadmium↗

Analysis of transcriptional regulation of human breast aromatase by in vitro and in vivo studies.

Aromatase mRNA in normal breast tissues is mainly transcribed from exon 1b (I.4) of the gene. However, in breast cancer tissues, it is often transcribed from exon 1c/1d (I.3/PII). Such a switching from exon 1b to exon 1c/1d has often found concomitantly with elevated levels of aromatase mRNA. To elucidate the molecular mechanism of a switching of multiple exons 1 and enhanced expression of aromatase in the breast tissues, we identified essential elements responsible for transcription from exon 1b. Gel shift assays indicated that there are two essential elements for transcription from exon 1b between -300 and -500 bp in the promoter region. The two unique elements have homologous consensus DNA sequences, and competed for binding of the specific nuclear protein with each other. We next generated transgenic mouse expressing a reporter gene controlled by multiple promoters in the human aromatase gene. The mouse gave tissue-specific expression of the reporter gene and tissue-specific utilization of the alternative promoter regions. These results show that this transgenic mouse is a good model animal for the study of tissue-specific regulation of human aromatase gene.

Alternative Splicing↗

Secondary structure of the human T-cell leukemia virus type 1 rex-responsive element is essential for rex regulation of RNA processing and transport of unspliced RNAs.

Rex protein of human T-cell leukemia virus type 1 (HTLV-1) induces cytoplasmic expression of unspliced gag/pol mRNA and singly spliced env mRNA and thus is essential for replication of the virus. This regulation requires a cis-acting rex-responsive element (RXE), located in the 3' region of the viral RNA. By external deletion, we have identified RXE composed of 205 nucleotides. The secondary structure of RXE was confirmed by studies on its susceptibility to nuclease digestions to consist of four stem-loops and a long stretch of stem structure. Substitution and deletion mutations revealed that two regions of the stem-loops and their secondary structures are essential for rex regulation. Similar secondary structures were found in the corresponding regions of HTLV-2, bovine leukemia virus and human immunodeficiency virus. Furthermore, a sequence of 11 nucleotides in the RXE was found to be conserved in the secondary structures of HTLV-1, HTLV-2, and bovine leukemia virus. These observations suggest that the secondary structure as well as the conserved sequence may be important in expression of unspliced RNA even with diverged sequences as observed in these viruses.

Base Sequence↗

[Inherited neurological disease--relationship between an essential trace elements and Wilson/Menkes disease].

Because copper is an integral component of various enzymes in every cells, it has been known to be essential for cell activities. Copper is also a very toxic ion, so a specific series of copper transport process must exist to carry copper to the sites where it is required, and to ensure copper homeostasis without allowing toxic accumulation. Wilson disease and Menkes disease are the inherited diseases caused by genetic defects in copper metabolism. Wilson disease is related to the toxic effects of copper accumulation in liver, which leads to progressive liver damage and subsequent overflow to brain causing a loss of coordination and involuntary movement. Menkes disease is caused by the deficiency of serum copper and of copper-dependent enzymes, in various tissues except liver which characterized by neurologic degeneration and mental retardation, connective tissue and vascular defects, characteristic brittle and depigmented hair, and death in early childhood.

Copper↗

Transgenic regulation of moth chorion gene promoters in Drosophila: tissue, temporal, and quantitative control of four bidirectional promoters.

Bidirectional chorion gene promoter regions from three silkmoth species, Bombyx mori, Antheraea pernyi, or Antheraea polyphemus (members of two different moth families), were tested for their ability to transcriptionally activate a bacterial marker gene (chloramphenicol acetyltransferase) in transformant Drosophila. Relatively short 5' flanking DNA fragments (272-367 bp) of chorion gene pairs are sufficient to confer a high degree of tissue and choriogenic stage specificity of expression to the marker gene. Thus, significant conservation of molecular interactions controlling transcription during choriogenesis is observed between the distantly related orders, Lepidoptera and Diptera. However, quantitative and fine temporal regulation in the Drosophila host does not fully parallel the in situ regulation in moths, indicating that some regulatory protein-DNA interactions have diversified in the approximately 250 million years since the last common ancestor of these insect groups. Limited in vitro mutagenesis of a B. mori promoter DNA has shown that a central 189-bp region includes elements sufficient for the qualitative specificity of chorion-specific expression. The same experiments have shown that a previously identified essential element, centered on the TCACGT hexamer, is not sufficient for chorion-specific expression: an additional essential element or elements are found farther upstream, within a 112-bp DNA region. Comparisons of silkmoth and Drosophila chorion gene promoter sequences have identified some candidates for cis-acting elements involved in the developmental regulation of chorion gene expression.

Animals↗

Identification of an essential core element and stimulatory sequences in a Kluyveromyces lactis ARS element, KARS101.

A Kluyveromyces lactis chromosomal sequence of 913 bp is sufficient for replication in Saccharomyces cerevisiae and K. lactis. This fragment contains a 12 bp sequence 5'-ATTTATTGTTTT-3' that is related to the S. cerevisiae ACS (ARS consensus sequence). This dodecamer was removed by site-directed mutagenesis and the effect on K. lactis and S. cerevisiae ARS (autonomous replicating sequence) activity was determined. The dodecamer is essential for S. cerevisiae ARS function but only contributes to K. lactis ARS activity; therefore, its role in K. lactis is unlikely to be the same as that of the essential S. cerevisiae ACS. A 103 bp subclone was found to retain ARS activity in both yeasts, but the plasmid was very unstable in S. cerevisiae. Deletion and linker substitution mutagenesis of this fragment was undertaken to define the DNA sequence required for K. lactis ARS function and to test whether the sequence required for ARS activity in K. lactis and S. cerevisiae coincide. We found a 39 bp core region essential for K. lactis ARS function flanked by sequences that contribute to ARS efficiency. The instability of the plasmid in S. cerevisiae made a fine-structure analysis of the S. cerevisiae ARS element impossible. However, the sequences that promote high-frequency transformation in S. cerevisiae overlap the essential core of the K. lactis ARS element but have different end-points.

Base Sequence↗

A novel 14-base-pair regulatory element is essential for in vivo expression of murine beta4-galactosyltransferase-I in late pachytene spermatocytes and round spermatids.

During murine spermatogenesis, beginning in late pachytene spermatocytes, the beta4-galactosyltransferase-I (beta4GalT-I) gene is transcribed from a male germ cell-specific start site. We had shown previously that a 796-bp genomic fragment that flanks the germ cell start site and contains two putative CRE (cyclic AMP-responsive element)-like motifs directs correct male germ cell expression of the beta-galactosidase reporter gene in late pachytene spermatocytes and round spermatids of transgenic mice (N. L. Shaper, A. Harduin-Lepers, and J. H. Shaper, J. Biol. Chem. 269:25165-25171, 1994). We now report that in vivo expression of beta4GalT-I in developing male germ cells requires an essential and previously undescribed 14-bp regulatory element (5'-GCCGGTTTCCTAGA-3') that is distinct from the two CRE-like sequences. This cis element is located 16 bp upstream of the germ cell-specific start site and binds a male germ cell protein that we have termed TASS-1 (transcriptional activator in late pachytene spermatocytes and round spermatids 1). The presence of the Ets signature binding motif 5'-GGAA-3' on the bottom strand of the TASS-1 sequence (underlined sequence) suggests that TASS-1 is a novel member of the Ets family of transcription factors. Additional transgenic analyses established that an 87-bp genomic fragment containing the TASS-1 regulatory element was sufficient for correct germ cell-specific expression of the beta-galactosidase reporter gene. Furthermore, when the TASS-1 motif was mutated by transversion, within the context of the original 796-bp fragment, transgene expression was reduced 12- to 35-fold in vivo.

Animals↗

Transcription of the Schizosaccharomyces pombe U2 gene in vivo and in vitro is directed by two essential promoter elements.

As compared to the metazoan small nuclear RNAs (snRNAs), relatively little is known about snRNA synthesis in unicellular organisms. We have analyzed the transcription of the Schizosaccharomyces pombe U2 snRNA gene in vivo and in the homologous in vitro system. Deletion and linker-scanning analyses show that the S.pombe U2 promoter contains at least two elements: the spUSE centered at -55, which functions as an activator, and a TATA box at -26, which is essential for basal transcription. These data point to a similar architecture among S.pombe, plant and invertebrate snRNA promoters. Factors recognizing the spUSE can be detected in whole cell extracts by DNase I footprinting and competition studies show that the binding of these factors correlates with transcriptional activity. Electrophoretic mobility shift assays and gel-filtration chromatography revealed a native molecular mass of approximately 200 kDa for the spUSE binding activity. Two polypeptides of molecular masses 25 and 65 kDa were purified by virtue of their ability to specifically bind the spUSE.

Binding, Competitive↗

Brain trace element concentrations in aging.

Trace element concentrations were determined in various human brain regions over the complete life span using instrumental neutron activation analysis. Several different patterns of trace element alteration were observed with age. Brain Al, Cl and Na concentrations increase with advancing age, while K, P and Rb decline. Ag, Co, Fe, Sb and Sc concentrations increase up to the 40 to 79 age range then decline. Br, Se and Zn remain relatively constant throughout adult life. Hg, Mn and Cs show no consistent trend with age. In infant brains Br and Cl increase and Al, Cr, Cs, Fe, Mn, P, Rb, Sc, Se and Zn decrease compared to adults. The essential elements that remain within narrow concentration limits throughout adult life suggest the presence of an efficient homeostatic mechanism for their regulation in the brain, while those that are altered with age suggest modifications in control mechanisms or altered relationships with other elements. Increased concentrations of non-essential elements may reflect accumulation from our environment, impaired removal or altered balance with other elements.

Adult↗

Effects of weight loss and exercise on the distribution of lead and essential trace elements in rats with prior lead exposure.

We studied the effects of weight loss and non-weight-bearing exercise (swimming) on blood and organ lead and essential metal concentrations in rats with prior lead exposure. Nine-week-old female Sprague-Dawley rats (n = 37) received lead acetate in their drinking water for 2 weeks, followed by a 4-day latency period without lead exposure. Rats were then randomly assigned to one of six treatment groups: weight maintenance with ad libitum feeding, moderate weight loss with 20% food restriction, and substantial weight loss with 40% food restriction, either with or without swimming. Blood lead concentrations were measured weekly. The rats were euthanized after a 4-week period of food restriction, and the brain, liver, kidneys, quadriceps muscle, lumbar spinal column bones, and femur were harvested for analysis for lead, calcium, copper, iron, magnesium, and zinc using atomic absorption spectrophotometry. Both swimming and nonswimming rats fed restricted diets had consistently higher blood lead concentrations than the ad libitum controls. Rats in the substantial weight loss group had higher organ lead concentrations than rats in the weight maintenance group. Rats in the moderate weight loss group had intermediate values. There were no significant differences in blood and organ lead concentrations between the swimming and nonswimming groups. Organ iron concentrations increased with weight loss, but those of the other metals studied did not. Weight loss also increased hematocrits and decreased bone density of the nonswimming rats. The response of lead stores to weight loss was similar to that of iron stores because both were conserved during food restriction in contrast to decreased stores of the other metals studied. It is possible that weight loss, especially rapid weight loss, could result in lead toxicity in people with a history of prior excessive lead exposure.

Animals↗

Importance of international cooperation in food safety.

All countries need to ensure that national food supplies are safe, of good quality and available in adequate amounts at affordable prices to ensure good nutrition and health for all population groups. The enforcement of food standards by efficient national food control authorities in domestic markets and at the points of import and export has been increasingly recognized as a means of raising the value of exported goods by reducing the number of rejected or reconditioned consignments, and of ensuring the safety of the food and its acceptability by the final consumer. However, those national efforts have sometimes induced some non-tariff barriers to food trade and distribution. In addition, new developments in the technologies of food production, processing and marketing pose a new challenge to ensure safety of food. The strengthening of national food control infrastructures in particular in developing countries including the strengthening of staff capabilities, the need for harmonization of food at international levels, the need for collection and exchange of data on food control and food contamination issues are essential elements to ensure food safety in the world. International cooperation has an important role to play in achieving these essential elements.

Food Additives↗

Developmental patterns of aluminum and five essential mineral elements in the central nervous system of the fetal and infant guinea pig.

Al is found in the developing conceptus, but little information is available concerning its tissue distribution and its changes in concentration with age. Because Al has affinity for many of the same biological ligands as the essential mineral cations Ca, Mg, Zn, Fe, and Mn, we hypothesized that Al might show a pattern of developmental concentrations that was similar to one or more of these elements in the brain, a major target of Al toxicity. Concentrations of Al, Ca, Mg, Zn, Fe, and Mn were measured in spinal cord, brainstem, cerebellum, and forebrain of guinea pig fetuses on gestation day (GD) 30 and 45, at birth, and on postnatal day (PND) 3, 6, and 12. Dams were fed commercial guinea pig chow, which contained 47 micrograms Al/g. Tissue Al and Mn were measured with electrothermal atomic absorption spectrophotometry (ETAAS), and the other elements with inductively coupled axial plasma spectroscopy (ICAP-AES). Al concentrations in the brain regions were highest in spinal cord, brainstem, and cerebellum, and decreased during late gestation and lactation. Al did not show marked increases in regional brain concentrations during the final third of gestation as did Fe, Mg, and Zn. In contrast to Fe and Ca, Al did not accumulate in placenta. Al was the only element to show higher concentrations in spinal cord than in any other tissue at birth. In summary, the tissue distribution of Al did not follow that of essential cations as examined in this study.

Aluminum↗