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PubMed1993

A DNA unwinding element and an ARS consensus comprise a replication origin within a yeast chromosome.

We have defined a replication origin, ORI305, within chromosome III of Saccharomyces cerevisiae by means of mutational analysis. cis-acting elements required for origin activity in the chromosome, as assayed by two-dimensional gel electrophoresis of replication intermediates, are the same as those required for the function of an autonomously replicating sequence, ARS305, in a plasmid. Essential elements include (i) an 11 bp sequence that is a near match to the ARS consensus and (ii) a broad sequence directly 3' to the consensus near match. Origin function is inactivated by point mutations in the essential near match sequence, suggesting that the sequence contributes to specifying the origin in the chromosome. Other consensus near matches with different sequences are present but are not required. The essential 3'-flanking sequence exhibits DNA helical instability and is sensitive to deletion mutations that stabilize the DNA helix. The wild-type 3'-flanking sequence can be functionally substituted by dissimilar sequences that also exhibit helical instability. The requirement for DNA helical instability indicates that the essential 3'-flanking sequence serves as a DNA unwinding element in the chromosome.

Amino Acid Sequence↗

PubMed1993

Multiple pathways of reversion in viroids for conservation of structural elements.

From site-directed mutagenesis of potato spindle tuber viroid (PSTVd) it had been concluded earlier that the formation of a thermodynamically metastable structure containing hairpin II (HP II) is critical for infectivity. In order to differentiate between structural and sequence effects, in the present work base pairs in HP II were exchanged by site-directed double mutations without significant alterations in the native rod-like structure of PSTVd. The mutants were viable and genetically stable in the first generation, but one of the two mutations reverted to the wild-type nucleotide in the second generation. Single-site mutations in the stem of HP II, which had been described as revertants to the wild-type sequence earlier, were analysed with respect to the time course of reversion and the sequence variation during reversion. All replicating sequence variants were separated by gel electrophoretic techniques and the sequences and their relative frequencies were determined. From both types of studies it can be concluded (i) that HP II is a functional element in the (-)strand replication intermediate, generated due to sequential folding during synthesis, and that it is essential for template activity of (+)strand synthesis; (ii) that G:U pairs are tolerated transiently in (-)strand HP II; the lower stability of such a HP II is compensated by additional mutations outside HP II which suppress the competition of a rod-like structure; and (iii) that the reversions are generated spontaneously during (-)strand synthesis. Furthermore, the double-stranded structure of HP II is the essential element for short term replication of PSTVd but the exact sequence of the wild-type proves to be superior with regard to fitness and replicability of PSTVd.

Base Sequence↗

PubMed1993

Expression and localization of elements of the plasminogen activation system in benign breast disease and breast cancers.

The malignant potential of solid tumors is related to the ability to invade adjacent tissue and to metastasize. These properties of cancer cells depend on the synthesis of proteolytic enzymes which are able to digest adjacent connective tissue and basement membranes. We hypothesized that all elements of the plasminogen activation system might be overexpressed in malignant human breast tumors, functioning as an essential element in tumor invasion and metastasis. As determined by histopathological methods, the malignant tumors showed statistically significantly higher expression of urokinase plasminogen activator (uPA), type-1 plasminogen activator inhibitor (PAI-1), and especially urokinase plasminogen activator receptor (uPAR) than benign tissues. All those elements were present in higher amounts in the cancer cells than in the cells of benign or normal breast tissues. High exhibition of tissue plasminogen activator (tPA) found in cancer seems to be random and not related to the malignant or benign state, since benign and malignant tumors show overexpression of tissue plasminogen activator with similar frequency. When the tumors express high amounts of uPA, they express a high amount of uPAR in 50% of cases and PAI-1 in 57.3% of cases. When urokinase is expressed in low amount, the receptor is low in 28.6% and inhibitor in 21.4% of malignant breast tumors. This statistically significant consensus, 78.6% in the case of urokinase and its receptor and 78.6% in case of urokinase and its inhibitor, suggests that these activities may be the result of a unique mechanism of control, activated in the last steps of malignant transformation.

Breast Diseases↗

PubMed2000

The role of glycosylation in ionotropic glutamate receptor ligand binding, function, and trafficking.

Members of the ionotropic glutamate receptor (iGluR) family have between 4 and 12 consensus asparagine (N)-linked glycosylation sites. They are localized on the extracellular N-termini, and the loop between the penultimate and last transmembrane domains. These regions also contain the essential elements for formation of the ligand binding site. N-linked glycosylation does not appear to be essential for formation of the ligand binding site per se, but there are demonstrated interactions between glycosylation state and ligand binding affinity, receptor physiology, susceptibility to allosteric modulation and, in some cases, trafficking. There is no indication of a general role for N-linked glycosylation in iGluRs; instead the effects of glycosylation vary among glutamate receptor subtypes and splice variants, with specific effects on structure or function with different subunits.

Animals↗

PubMed2004

Contamination with nonessential metals from a solid-waste incinerator correlates with nutritional and immunological stress in prefledgling black kites (Milvus migrans).

We assessed whether levels of heavy metals in blood have detrimental effects on the health of prefledgling black kites (Milvus migrans) exposed to emissions from a solid waste incinerator near Madrid, central Spain. As health indexes, we considered body mass corrected by structural size as a general indicator of nutritional condition and white blood cell counts (especially the heterophil:lymphocyte ratio) as an indirect measure of immune function. Nutritional condition was positively related to levels of Zn, an essential element with a major role in many physiological pathways, while the opposite trend was found for the essential Cu, the concentration of which tends to increase with decreasing nutritional condition. We found an interaction between Zn and Cd, indicating that high levels of Zn produce smaller effects of Cd on body mass. Potential immunological stress, expressed as an increase in the heterophil:lymphocyte ratio, caused by the toxic Cd increased as levels of Cu decreased. Thus, nestlings with low levels of Cu and poor nutritional condition suffered more immunological stress due to Cd contamination than nestlings with higher levels of Cu. We also found a negative interaction between Zn and Cd, indicating a response of essential Zn to increasing levels of Cd, which may be indicative of the negative effects of Cd on the immune system. These results suggest that the participation of metallothioneins in detoxification and metal regulation may also indirectly enhance the adrenal stress response to contaminants, probably through a competition of Cd and Zn for Zn binding sites on metallothioneins, reducing the ability of Cd to stress the immune system.

Air Pollutants↗

PubMed2002

Ingredients for change: revisiting a conceptual framework.

Finding ways to deliver care based on the best possible evidence remains an ongoing challenge. Further theoretical developments of a conceptual framework are presented which influence the uptake of evidence into practice. A concept analysis has been conducted on the key elements of the framework--evidence, context, and facilitation--leading to refinement of the framework. While these three essential elements remain key to the process of implementation, changes have been made to their constituent sub-elements, enabling the detail of the framework to be revised. The concept analysis has shown that the relationship between the elements and sub-elements and their relative importance need to be better understood when implementing evidence based practice. Increased understanding of these relationships would help staff to plan more effective change strategies. Anecdotal reports suggest that the framework has a good level of validity. It is planned to develop it into a practical tool to aid those involved in planning, implementing, and evaluating the impact of changes in health care.

Delivery of Health Care↗

PubMed2001

Awareness of dying: an experience of Chinese patients with terminal cancer.

Despite the effort to provide an alternate way to dying, there seems to be little effort to examine the real situation of hospice care. For example, the patients' desired outcomes are seldom addressed. It is recently that studies began to investigate the features of "good death." However, work in eastern cultures, such as Chinese, remains scarce. The purpose of this study is to gain an understanding of what it means to die a "good death" from the perspective of Chinese patients. Thirty-three Chinese hospice patients with terminal cancer were interviewed. Grounded in the analysis of qualitative data, seven elements that contribute to dying a good death emerged. Awareness of dying was identified as the foremost essential element of a good death. From the findings of this study, most respondents considered receiving a diagnosis of cancer as one of the significant points of psychological distress in the course of their illness. Half of the patients openly talked about their cancers and more than half of them discussed various issues related to death. That means, about one-third of the respondents talked about death and cancer with openness. These preliminary findings may reflect the progress of hospice work in Hong Kong. Meanwhile, 13 respondents did not mention the word "death" and 16 of them did not mention that they had cancer. They talked about their illness and future using the expressions that they preferred. It is important that health care professionals also respect people who show awareness of dying as much as those who do not present clear evidence of this awareness. Both groups of people can have a positive experience when they die.

Adult↗

PubMed2005

Transcriptional regulation of nanog by OCT4 and SOX2.

Nanog, Sox2, and Oct4 are transcription factors all essential to maintaining the pluripotent embryonic stem cell phenotype. Through a cooperative interaction, Sox2 and Oct4 have previously been described to drive pluripotent-specific expression of a number of genes. We now extend the list of Sox2-Oct4 target genes to include Nanog. Within the Nanog proximal promoter, we identify a composite sox-oct cis-regulatory element essential for Nanog pluripotent transcription. This element is conserved over 250 million years of cumulative evolution within the eutherian mammals. A Nanog proximal promoter-EGFP (enhanced green fluorescent protein) reporter transgene recapitulates endogenous Nanog mRNA expression in embryonic stem cells and their differentiated derivatives. Sox2 and Oct4 interaction with the Nanog promoter was confirmed through mutagenesis and in vitro binding assays. Electrophoretic mobility shift assays indicate that the Sox2-Oct4 heterodimer forms more efficiently on the composite element within Nanog than the similar element within Fgf4. Using chromatin immunoprecipitation, we show that Oct4 and Sox2 bind to the Nanog promoter in living mouse and human embryonic stem cells. Furthermore, by specific knockdown of Oct4 and Sox2 mRNA by RNA interference in embryonic stem cells, we provide genetic evidence for a link between Oct4, Sox2, and the Nanog promoter. These studies extend the understanding of the pluripotent genetic regulatory network within which the Sox2-Oct4 complex are at the top of the regulatory hierarchy.

Animals↗

PubMed1988

Element localisation and distribution in the fungi Aureobasidium pullulans and Aspergillus niger, measured using the Oxford scanning proton microprobe.

The distribution and relative concentrations of essential elements were determined in germ tubes of Aureobasidium pullulans and hyphae of Aspergillus niger using the Oxford scanning proton microprobe. In both fungi, K, P and S were the major constituents, with Ca, Na and trace metals like Cu, Mn, Fe and Zn present at lower levels. In A. pullulans, elements were not distributed uniformly throughout the cells and, in general, the highest elemental concentrations occurred at the tip and in the older parts of the germ tube, particularly where there was yeast-like cell or branch development. In A. niger, elemental distribution was more uniform and there was a general gradient of increasing concentration away from the hyphal tip followed by a drop in levels in older regions. The scanning proton microprobe appears to have considerable potential for the investigation of fungal differentiation and morphogenesis.

Aspergillus niger↗

PubMed1994

Promoter analysis of the PDA1 gene encoding the E1 alpha subunit of the pyruvate dehydrogenase complex from Saccharomyces cerevisiae.

The location and sequence of the PDA1 gene, encoding the E1 alpha subunit of the pyruvate dehydrogenase (PDH) complex from Saccharomyces cerevisiae, were determined. The PDA1 gene was located on a 6.2 kb fragment of chromosome V, approximately 18 kb centromere distal to RAD3. Consistent with this, the PDA1 gene was genetically mapped at 4 cM from RAD3. A part of the 6.2 kb fragment of chromosome V was sequenced. The nucleotide sequence contained the PDA1 open reading frame and the entire putative promoter. Computer analysis revealed a putative GCN4 binding motif in the PDA1 promoter. The presence of transcriptional elements was experimentally determined by deletion analysis. To this end, ExoIII deletions were constructed in the 5' to 3' direction of the PDA1 promoter and effects on transcription were determined by Northern analysis. Transcription was unaffected upon deletion to position -190 relative to the ATG start codon. Deletions from position -148 and beyond, however, reduced promoter activity at least 40-fold. Apparently the 42 bp between nucleotides -190 and -148 contain an element essential for transcription. Inactivation of the PDA1 promoter could not be attributed to deletions of a recognizable TATA element or any known yeast regulatory motifs. The possible role of the CCCTT sequence present in the 42 bp region and also in the promoters of the other genes encoding subunits of the PDH complex is discussed.

Amino Acid Sequence↗

PubMed1985

Transient accumulation of c-fos RNA following serum stimulation requires a conserved 5' element and c-fos 3' sequences.

Transcription of the c-fos gene is transiently activated to generate large amounts of unstable c-fos RNA when quiescent fibroblasts are stimulated by polypeptide mitogens or whole serum. A cloned human c-fos gene (c-fosH) transfected into mouse fibroblasts is regulated in a similar manner. An element essential for transcription activation is located between nucleotides -332 and -276 relative to the mRNA cap site. This element has properties similar to those of previously characterized transcription enhancer elements. However, replacement of the 5' activating element by enhancers from SV40 or Moloney murine leukemia virus does not allow regulated c-fosH expression. The study of fusion genes showed that in addition to the 5' activating element, transient accumulation of c-fosH RNA following serum stimulation requires sequences at the 3' end of the c-fosH gene.

Animals↗

PubMed2006

Epithermal neutron activation analysis of Cr(VI)-reducer basalt-inhabiting bacteria.

Epithermal neutron activation analysis (ENAA) has been applied to study elemental composition of Cr(VI)-reducer bacteria isolated from polluted basalts from the Republic of Georgia. Cr(VI)-reducing ability of the bacteria was examined by electron spin resonance, demonstrating that the bacteria differ in their rates of Cr(VI) reduction. A well-pronounced correlation between the ability of the bacteria to accumulate Cr(V) and their ability to reduce Cr(V) to Cr(III) observed in our experiments is discussed. Elemental analysis of these bacteria also revealed that basalt-inhabiting bacteria are distinguished by relative contents of essential elements such as K, Na, Mg, Fe, Mn, Zn, and Co. A high rate of Cr(III) formation correlates with a high concentration of Co in the bacterium. ENAA detected some similarity in the elemental composition of the bacteria. The relatively high contents of Fe detected in the bacteria (140-340 microg/g of dry weight) indicate bacterial adaptation to the environmental conditions typical of the basalts. The concentrations of at least 12-19 different elements were determined in each type of bacteria simultaneously starting with the major to ultratrace elements. The range of concentrations spans over 8 orders of magnitude.

Arthrobacter↗

PubMed1993

Activation of the tyrosine aminotransferase gene is dependent on synergy between liver-specific and hormone-responsive elements.

Tyrosine aminotransferase (TAT; L-tyrosine:2-oxoglutarate aminotransferase, EC 2.6.1.5) gene activity is stimulated by glucocorticoids and glucagon and is repressed by insulin. Expression and responsiveness to the different signal transduction pathways are restricted to the liver, in which the gene is activated shortly after birth. Here we provide a model for the basis of this tissue specificity of the hormonal control. In the two enhancers mediating hormone induction of TAT gene activity we find the hormone response elements in combination with binding sites for constitutive liver-enriched transcription factors: proteins of the hepatocyte nuclear factor 3 family bind in the vicinity of the glucocorticoid response element located 2.5 kb upstream of the transcription start site, while hepatocyte nuclear factor 4 interacts with an essential element in the cAMP-responsive enhancer at -3.6 kb. By juxtaposing the liver-specific element and the target sequence of the signal transduction pathway the regulatory properties of either enhancer can be reconstituted. Thus, the interdependence of the respective enhancer motifs restricts the hormonal activation of the TAT gene to the liver. The coincidence of the onset of TAT gene expression around birth with the perinatal changes in the concentrations of glucocorticoids, glucagon, and insulin suggests cooperation of signal transduction pathways and cell type-specific transcription factors in the developmental activation of the TAT gene.

Animals↗

PubMed2003

Faculty's perceptions of humanistic teaching in nursing baccalaureate programs.

In an attempt to investigate faculty's perceptions of humanistic teaching in nursing baccalaureate programs, a qualitative study was conducted by interviewing 10 faculty members (including one dean, and five associate deans) who all had experience in teaching nursing baccalaureate students. The participants were from five universities located in the southwestern, southeastern, and southern parts of the USA. The data were collected from 6/28-7/12, 2002. All the interviews were audio-taped and transcribed verbatim. Data analysis was inductive, seeking understanding of individuals' perceptions. When a number of coded contexts were considered to have common elements, they were merged to form categories and presented as results. The findings showed that there are five essential elements in faculty's perceptions of humanistic teaching in nursing baccalaureate programs: availability, caring, authenticity, empowerment, and a transformative curriculum.

Attitude of Health Personnel↗

PubMed1993

Characterization of functional Sp1 transcription factor binding sites in the hepatitis B virus nucleocapsid promoter.

The hepatitis B virus nucleocapsid minimal promoter contains sequence elements which are similar to the Sp1 transcription factor binding site consensus sequence. The interaction of these regulatory elements with Sp1 was examined by DNase I footprinting with purified Sp1 protein and DNase I footprinting and gel retardation analysis with nuclear extracts from human cell lines and was examined functionally with transient transfection assays in human hepatoma and Drosophila melanogaster Schneider line-2 cells. DNase I footprinting identified two regions of the nucleocapsid promoter, representing three recognition elements, that bound purified Sp1. Gel retardation analysis with Huh7 nuclear extracts demonstrated that each of the three recognition elements bound the same or similar transcription factor(s) as that recognized by the Sp1 consensus sequence recognition element. The function of the nucleocapsid promoter elements was examined by transient transfection assays in D. melanogaster Schneider line-2 cells by using these binding sites cloned into a minimal promoter element. Each of these regulatory regions transactivated transcription from the minimal promoter element in response to exogenously expressed Sp1. In addition, the second Sp1 site was shown to be an essential element of the nucleocapsid promoter in human hepatoma cells. This demonstrates that the hepatitis B virus nucleocapsid promoter contains three functional Sp1 binding sites which may contribute to the level of transcription from this promoter during viral infection.

Animals↗

PubMed1991

Effect of a platinum chemotherapy drug on intracellular elements during the cell cycle, using X-ray microanalysis.

Intracellular elemental concentrations were measured in the cytoplasm, nucleus and nucleolus of cultured Chinese hamster ovary (CHO) cells, using energy dispersive electron probe X-ray microanalysis and transmission electron microscopy. Synchronous CHO cell populations were analyzed at different times during the growth cycle. The elements K, P, Mg and Zn were all more concentrated in the nucleus and nucleolus than in the cytoplasm, while no specific subcellular compartmentalization was evident for the elements Na, Cl, Ca, Fe and S. Significant changes in intracellular elemental concentrations were associated with the progression of cells from G1 phase to S phase of the growth cycle. Most significant were the effects on the monovalent ions, Na, K and Cl. The effect of a second generation platinum chemotherapy agent, cis-dichloro-bis (isopropylamine) trans-dihydroxy platinum IV (iproplatin) on intracellular elements was investigated by analyzing subcellular compartments of drug-treated synchronous CHO cell populations. Changes in intracellular elemental levels occurred, most notably in the nucleus of G1 phase cells, when a general depletion of most essential elements was evident. Attempts to analyze cells for their Pt content proved disappointing, since quantitation for Pt was not possible. However, measurement of Pt peak/background ratio yielded significant Pt peaks on analyzing lysosomes of cells treated with high concentrations of iproplatin. These Pt peaks were associated with high levels of S and Fe.

Animals↗

PubMed1989

Non-conventional indigenous mineral supplements used for cattle feeding in the pastoral rangelands of Ethiopia.

Salt licks, soil licks and well waters from various parts of Ethiopia used by pastoral nomads for cattle feeding were sampled and assessed for composition and quality. Most of the licks were alkaline and had low solubility both in water and acid. All the salt licks and some of the soil licks contained a considerable amount of sodium and these could constitute a reasonable and cheap source of the element. Except for iron which had a concentration in most of the supplements much in excess of requirements the rest of the essential elements assayed occurred in the licks in quantities too small to satisfy estimated requirements of cattle. This particularly applied to phosphorus and copper which are below critical ranges in the pastures of the region. The excessive amount of Fe could have an antagonistic effect on P and Cu. The well waters contained elements in safe limits but water from Medecho contained total soluble salts exceeding safe limits. Except as sources of Na the licks especially soil licks can be considered unsatisfactory sources of other essential minerals. The widespread use of soil licks in the region should be discouraged until thorough assessment has been done since these licks may have deleterious direct and indirect effects on performance of cattle.

Animal Feed↗

PubMed1997

A typology of simulators for medical education.

The growth of simulation technology has brought about rapid innovation in medical education systems. The task of developing and programming these systems is complex. To simplify the process, we developed a typology of medical simulators which allows critical elements to be identified and characterized. The analysis identifies the patients, the procedure, the physician, and the professor as essential elements in any medical education simulator. The level of interactivity of these elements determines the "realism" of the simulator. The development of a fully interactive simulator for ultrasound education will be presented.

Computer Simulation↗