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At least 91 records · Page 5Linked to original sources
PubMed2006

Cyclooxygenase-1-derived PGE2 promotes cell motility via the G-protein-coupled EP4 receptor during vertebrate gastrulation.

Gastrulation is a fundamental process during embryogenesis that shapes proper body architecture and establishes three germ layers through coordinated cellular actions of proliferation, fate specification, and movement. Although many molecular pathways involved in the specification of cell fate and polarity during vertebrate gastrulation have been identified, little is known of the signaling that imparts cell motility. Here we show that prostaglandin E(2) (PGE(2)) production by microsomal PGE(2) synthase (Ptges) is essential for gastrulation movements in zebrafish. Furthermore, PGE(2) signaling regulates morphogenetic movements of convergence and extension as well as epiboly through the G-protein-coupled PGE(2) receptor (EP4) via phosphatidylinositol 3-kinase (PI3K)/Akt. EP4 signaling is not required for proper cell shape or persistence of migration, but rather it promotes optimal cell migration speed during gastrulation. This work demonstrates a critical requirement of PGE(2) signaling in promoting cell motility through the COX-1-Ptges-EP4 pathway, a previously unrecognized role for this biologically active lipid in early animal development.

Animals↗
PubMed1999

[Deletion analysis and functional studies of the promoter from commelina yellow mottle virus].

Commelina Yellow Mottle Virus(CoYMV) is a double-stranded, circular DNA virus and its promoter could direct GUS gene specifically expressing in phloem tissue of transgenic tobacco plants. To determine the optimal promoter sequence for pholem-specific gene expression, CoYMV promoter was deleted from its 5 end to form promoter fragments with 5 different lengths. Chimeric GUS genes were constructed using the promoter deletion based on the binary vector pBI121. Transgenic tobacco plants evidenced by PCR analysis were obtained with each kind of chimeric GUS gene structure by Agrobacterium mediated transformation. The results of GUS activity assay and histo-chemical staining showed that most of the chimeric GUS genes were expressed in transgenic plants. The GUS activity with the promoter deleted to -870 bp was about 78% higher than that of the full length promoter(1040 bp) and was a little higher than that of the promoter deleted to -585 bp, but the difference is not significant. The GUS activity reduced significantly when the promoter was deleted to -447 bp or -232 bp, whereas the property of phloem-specific expression pattern was still retained. When the promoter was deleted to -44 bp, just upstream adjacent to the TATA box, its tissue-specificity was lost and the activity was reduced to undetectable level. These results suggest that the region between -870 bp-232 bp and downstream of -232 bp of CoYMV promoter could be responsible for promoter activity and tissue specific expression, respectively. A negative regulation sequence might exist upstream of -870 bp of the CoYMV promoter. Therefore, we recommend that the optional CoYMV promoter sequence for phloem specific expression could be downstream from -870 bp or -585 bp. In comparison with CaMV 35S promoter, the GUS activity when driven by -870 bp CoYMV promoter was about 70% of that when driven by the 35S promoter. Considering the fact that 35S promoter-GUS gene is constitutively expressed, while the CoYMV promoter-GUS gene is expressed only in phloem tissues, the activity of the latter in phloem may be the same with or even higher than that of the 35S promoter.

Badnavirus↗
PubMed1983

A comparison of the regulatory effects of human monocytes, pulmonary alveolar macrophages (PAMs) and spleen macrophages upon lymphocyte responses.

Human monocytes, pulmonary alveolar macrophages (PAMs) and spleen macrophages were concentrated by immobilization on cold insoluble globulins. These cell preparations were 90 +/- 3%, 95 +/- 1% and 83 +/- 3% esterase rich, respectively, 87 +/- 4%, 95 +/- 3% and 66 +/- 11% phagocytic and 78 +/- 3%, 79 +/- 9% and 68 +/- 5% reactive with OKM1 monoclonal antibody. Spleen macrophages differed from the other two cell preparations in that significantly fewer reacted with 61D3 or 63D2 monoclonal antibodies. Monocytes and PAMs promoted the mixed leucocyte response by autologous lymphocytes when added at low concentrations, but suppressed this response at high concentrations. Spleen macrophages only promoted the mixed leucocyte reaction but were required in much higher numbers than either monocytes or PAMs for optimal promotion. Likewise, the added presence of monocytes or PAMs in high numbers suppressed Ig synthesis stimulated with pokeweed mitogen, while spleen macrophages were not suppressive in this system. This study shows that the distribution of macrophages that differ in their regulatory effects upon lymphocyte responses varies in different tissues. The human spleen is deficient in macrophage related suppression.

Adult↗
PubMed2006

[Comparison of the relative luciferase activity in secondary CEF by different heterogenous strong promoters, MDV gB promoter and the composed promoters].

To improve the protection efficiency of the recombinant Marek' s disease viruses (MDV) in chickens with or without maternal antibodies,the work of selecting the optimal promoters for the construction of recombinant MDV was carried out. Combined with the efficient genetic manipulation, the composed promoters was constructed by use of the MDV gB core promoter with the regulatory elements from the early immediately promoter and enhancer of hCMV, the promoter and enhancer of SV40 or the partial enhancer of hCMV. And these composed promoters were ligased to the luciferase to construct the eukaryotic expressing vectors and named PhCMV-gB, Psv-gB and Pen-gB, respectively. In vitro, these vectors and internal standard plasmid (pSV-beta-LacZ) were transiently co-transfected into secondary CEF by FuGene 6 Transfection Reagent. Furthermore, cells were harvested 48 hours after transfection. Then the luciferase activity was detected by a luciferase assay kit, at the same time, the beta-galactosidase enzyme activity was detected by a beta-galactosidase enzyme assay kit, and the luciferase activity was corrected by the beta-galactosidase enzyme activity to get the relative luciferase activity. The relative luciferase activity was used as the transcriptional activity. By comparison of the relative luciferase activity of every promoter, it was found that these composed promoters could more effectively drive the reporter gene expression than the full legth of gB promoter did. Among them, PhCMV-gB robustly drove the reporter gene expression. On the other hand, PSV-gB and Pen-gB appeared to have the same strength; But compared with the commercial strong promoters, the transcriptional activity of the composed promoter were less than as or the same as that of the strong promoters. Therefore, at a sense, it can be proposed that these composed promoters have not only the characteristic of MDV gB promoter, but also that of the commercial strong promoters. These provide the choices for further developing the new-type recombinant MDV vaccine.

Animals↗
PubMed1998

Cardiac pacing in Canada in 1998: working towards optimal pacing therapy. Canadian Working Group on Cardiac Pacing.

The Canadian Working Group on Cardiac Pacing (CWGCP) was formed in 1996 with the primary goal of promoting optimal pacing therapy in Canada. In 1997, the CWGCP conducted a survey of pacing practices across Canada. Ninety-two of 125 implanting programs (74%) responded. Implant rates vary by province--from 39 per 100,00 population in Ontario to 63 per 100,000 population in Nova Scotia and Prince Edward Island. Variations in regional implant rates persist even after correcting for the age of the population. Physiological pacing was used for 35% of all implants in Canada in 1996/97. There were marked differences across Canada in the mode of pacing selected. In western Canada, 39.5% of pacing systems implanted were physiological compared with 18.2% in Atlantic Canada and 29% in Quebec. There were also differences in follow-up practices. Approximately 40% of centres follow patients with single chamber pacemakers annually, whereas most other centres still follow these patients every six months. Economic constraints, the size of pacing programs and the involvement of committed pacing physicians are factors that may influence the regional differences in cardiac pacing across Canada.

Canada↗
PubMed1985

Large granular lymphocytes have a promoting activity on human peripheral blood erythroid burst-forming units.

Peripheral blood mononuclear cells were fractionated according to the expression of a variety of surface markers, and the fractions obtained were tested for erythroid burst-forming unit (BFU-E) colony formation. BFU-Es were detected in the HLA-DR+ non-T cell fraction, but gave rise to optimum colony numbers only in the presence of a nonadherent, relatively radioresistant cell. This accessory cell was found among the HLA-DR- non-T, non-B cells, a fraction that was particularly enriched in large granular lymphocytes (LGLs). Experiments carried out to assess directly the surface markers of the accessory cell revealed an FcR+, OKM1+, Leu 7+, Leu 11+, OKT4-, OKT8- surface phenotype, which is consistent with that of the majority of LGLs. Peripheral blood LGLs, purified by Percoll density gradient, proved very efficient in promoting optimal BFU-E colony formation. All of these results indicate that LGLs have a potent erythroid burst-promoting activity. Such activity is probably mediated through the release of soluble factors, as shown by the observation that LGL culture supernatants were as effective as LGLs in sustaining colony formation.

Antigens, Surface↗
PubMed2004

Reducing the risk for preterm birth: evidence and implications for neonatal nurses.

The incidence of preterm birth has been slowly rising despite advances in obstetric health care. Neonatal nurses have ongoing contact with women who have had a preterm birth and, thus, who are at high risk to have another preterm birth. This article discusses current evidence about reducing risk for preterm birth. Although preterm birth cannot always be prevented, preconception care can help identify and modify maternal risk and promote optimal health before conception. Quality prenatal care, as defined by the Institute of Medicine, consists of continuing risk assessment, health promotion, and interventions to modify medical and psychosocial risk. When these 3 components are consistently applied, they may confer some protection against preterm birth. Women at highest risk for preterm birth need to seek prenatal care from an expert in maternal-fetal medicine. Women also need to learn about the common, subtle signs of preterm labor so they can recognize symptoms and quickly seek treatment. Neonatal nurses can provide critical information and anticipatory guidance to women at risk for preterm birth so they can make sound decisions about future pregnancies.

Evidence-Based Medicine↗
PubMed1991

Neural network optimization for E. coli promoter prediction.

Methods for optimizing the prediction of Escherichia coli RNA polymerase promoter sequences by neural networks are presented. A neural network was trained on a set of 80 known promoter sequences combined with different numbers of random sequences. The conserved -10 region and -35 region of the promoter sequences and a combination of these regions were used in three independent training sets. The prediction accuracy of the resulting weight matrix was tested against a separate set of 30 known promoter sequences and 1500 random sequences. The effects of the network's topology, the extent of training, the number of random sequences in the training set and the effects of different data representations were examined and optimized. Accuracies of 100% on the promoter test set and 98.4% on the random test set were achieved with the optimal parameters.

Artificial Intelligence↗
PubMed1993

Gangliosides rescue neuronal cells from death after trophic factor deprivation.

Serum-free cultures of PC 12 cells have been used as a model system for studying neuronal death occurring after neurotrophic factor deprivation. In this system, NGF rescues cells from death and prevents apoptotic DNA fragmentation. We report here that GM1 also promotes long-term survival of naive and NGF-pretreated PC 12 cells in serum-free medium and prevents internucleosomal cleavage of PC 12 cell DNA. In contrast to NGF, GM1 does not promote neurite outgrowth or somatic hypertrophy. The survival effects of GM1 are concentration dependent, with maximal activity at 30-50 microM. Optimal promotion of survival is obtained with multiple additions of GM1. Asialo-GM1 and sialic acid do not mimic these actions, indicating a requirement for the intact GM1 molecule. Prevention of serum-free PC 12 cell death is also obtained with di-, tri-, and tetrasialogangliosides. The ganglioside effects on survival and DNA fragmentation appear to be independent of macromolecular synthesis. GM1 is also effective under conditions in which cellular protein kinase C activity is downregulated by preexposure to high concentration of 12-O-tetradecanoylphorbol-13-acetate. Furthermore, GM1 promotes long-term survival of cultured rat sympathetic neurons after withdrawal of NGF. These findings complement prior observations that gangliosides protect cerebellar granule neurons from neurotoxicity caused by exposure to excitatory amino acids and extend the actions of gangliosides to rescue of neuronal cells deprived of neurotrophic factor support.

Animals↗
PubMed2004

Attitudes and behavior towards contraception among Greek women during reproductive age: a country-wide survey.

OBJECTIVES: The study objectives were to explore current contraceptive behavior of Greek women during reproductive age. STUDY DESIGN: Data were drawn from a country-wide survey, conducted with the use of a self-administered questionnaire. The sample, numbering 797 women of ages 16-45 years, was representative of the Greek female population of reproductive age. RESULTS: The most common contraceptive method reported was the male condom (MC) (33.9%) followed by coitus interruptus (CI) (28.8%), oral pill (4.8%), and coil (3.6%). Other methods counted for a 5% and no use of contraception reached a 23.8%. Attitudes over responsibility of using contraception were also explored. The majority of respondents (52%) stated that contraception use is the responsibility of men. The probability of reporting that women should be responsible in using contraception was higher in women aged 25-34 years, in those with higher level of knowledge over contraception issues and in those with an experience of abortion. CONCLUSIONS: The need for sexual education and easy access to counseling services is apparent in order to promote optimal contraception decision-making. The role of women in taking active responsibility over contraception use should be of great importance in reproductive health promotion projects.

Adolescent↗
PubMed1998

Analysis of a distal cluster of binding elements and other unusual features of the promoter of the highly regulated nit-3 gene of Neurospora crassa.

In Neurospora crassa, the expression of the nit-3 gene (nitrate reductase) is dependent upon nitrogen derepression and nitrate induction and is regulated by two positive-acting transcription factors, NIT2 and NIT4, and a negative regulator, NMR. The presence of a tightly linked cluster of NIT2 and NIT4 binding sites suggested that their close spacing might be required for a synergistic interaction of the NIT2 and NIT4 proteins. We show here that the NIT2 and NIT4 binding sites can be separated without affecting either the expression level or the precise regulation of the nit-3 gene. Studies conducted on the NIT2 site II, which contains only a single GATA element and yet plays a major role in nit-3 gene expression, showed that nucleotides both 5' and 3' of the GATA sequence were important for strong DNA binding in vitro and its activation function in vivo. The nit-3 promoter contains two long AT-rich sequences, one of which is located just upstream of the transcription start sites and is required for optimal promoter function. The nit-3 transcript contains eight TACC repeats in its 5' noncoding region which appear to be involved in mRNA instability. Deletion of these TACC repeats led to a significant increase in the stability of nit-3 mRNA.

Binding Sites↗
PubMed1995

Health promotion in human immunodeficiency virus-infected patients through psychosocial interventions.

The extreme fear, prejudice, and stigmatization associated with human immunodeficiency virus disease make the acquired immune deficiency syndrome epidemic a unique phenomenon of the modern era. A fundamental understanding of the phychosocial issues surrounding the disease is essential, if nurses are to design interventions that promote optimal health among those infected and those at risk for infection.

HIV Infections↗
PubMed2005

Applying evidence to palliative care.

OBJECTIVES: To discuss the evidence-based practice process for palliative care, from defining a clinical problem to implementation and evaluation of an evidence-based practice change. DATA SOURCES: Professional experience, literature, practice guidelines, and web sites related to evidence-based practice and palliative care. CONCLUSION: Nurses caring for patients needing palliative care need evidence from multiple sources. Using a systematic process to find and appraise evidence followed by strategically planning for implementation and evaluation of evidence-based practice changes optimizes the likelihood of enhanced patient care. IMPLICATIONS FOR NURSING PRACTICE: The integration of evidence-based practices into palliative care nursing will empower nurses to promote optimal patient outcomes and decrease practice variability.

Benchmarking↗
PubMed1992

Characterization of the human pleiotrophin gene. Promoter region and chromosomal localization.

The protein (PTN) encoded by the pleiotrophin (PTN) gene belongs to a recently described family of heparin-binding cytokines whose expression is temporally and spatially regulated during development. We have now isolated genomic clones of the human PTN gene, characterized its promoter region, determined its transcription initiation site(s), and established functional activity of the PTN promoter. A fragment -550/+191 that contains a CAAT box, no apparent TATA box, and four consensus sites for the binding of MyoD is sufficient to provide optimal promoter activity. A serum response element is found at -559 to -568. We also have identified the human PTN gene on chromosome 7, band q33 and the mouse Ptn gene on chromosome 6, respectively. The data thus identify and characterize the 5' end of the PTN gene and its promoter region, suggest potential regions that may contribute to the regulation of its transcriptional activity, and localize the PTN gene in human and mouse chromosomes.

3T3 Cells↗
PubMed2001

Effects of mutating different steroidogenic factor-1 protein regions on gene regulation.

The involvement of cyclic adenosine monophosphate cAMP-dependent protein kinase A (PKA) in the regulation of the steroidogenic acute regulatory protein (StAR) and the high-density lipoprotein receptor (HDL-R) genes by steroidogenic factor-1 (SF-1) and cAMP were examined. Cotransfection studies carried out in Kin 8 cells, a Y1 cell line (mouse adrenal) with a mutation in the type I PKA regulatory subunit, demonstrated that an intact PKA is required for maximal activation and that SF-1 participates in cAMP regulation of these genes. Site-directed mutational analysis was performed to examine which SF-1 regions could be involved in SF-1 transcriptional activation of the StAR and HDL-R genes. SF-1 regions protein analyzed were amino acids Thr 60, Ser 203, Ser 431, Thr 462, and the activation function-2 domain (amino acids 449-462). Plasmids encoding each of the mutated SF-1 proteins were cotransfected with the StAR and HDL-R promoter constructs into human bladder carcinoma (HTB-9) cells in the presence or absence of dibutyryl cAMP. The results of these studies suggest that although SF-1 is required for optimal promoter response to cAMP, transcriptional activation of genes by SF-1 and cAMP are promoter dependent, perhaps resulting from gene-specific interactions of this transcription factor with other regulatory proteins.

Amino Acid Sequence↗
PubMed1998

Mechanism of interferon action: identification of essential positions within the novel 15-base-pair KCS element required for transcriptional activation of the RNA-dependent protein kinase pkr gene.

RNA-dependent protein kinase PKR is an important regulator of gene expression in interferon (IFN)-treated and virus-infected cells. The 50-kb gene encoding human PKR kinase (pkr) is inducible by IFN. Transfection analyses, using chloramphenicol acetyltransferase (CAT) as the reporter in constructs possessing various 5'-flanking fragments of the human pkr gene, led to the identification of a functional TATA-less promoter that directed IFN-inducible transcription. Sequence determination and mutational analysis of the pkr promoter region revealed, in addition to a functional copy of the IFN-stimulated response element (ISRE) responsible for inducibility by type I IFN, a novel 15-bp element required for optimal promoter activity mediated by the ISRE. This element (5' GGGAAGGCGGAGTCC 3'), designated KCS for kinase-conserved sequence, is exactly conserved between the human and mouse pkr promoters in sequence and position relative to the ISRE. We have now carried out an extensive mutational analysis of the 15-bp KCS element. Site-directed mutagenesis was performed, whereby every base pair position within the KCS element was replaced by each of the other three alternatives. Forty-five substitution mutants were analyzed for promoter activity by transient transfection analysis of untreated and IFN-treated human cells. The results establish 5' NNRRRGG(C,A,T)GGRGYYN 3', where R stands for purine and Y stands for pyrimidine, as the consensus sequence for the KCS element, both for basal and for IFN-inducible promoter activity. KCS-binding proteins were detected by electrophoretic mobility shift analysis (EMSA). Competition EMSA established that constitutively expressed nuclear proteins bound the KCS element selectively; KCS protein binding activity correlated with promoter activity in the transient transfection reporter assay.

Animals↗
PubMed1994

Clinical significance of coronary vascular adaptations to exercise training.

Coronary vascular adaptations to exercise training have been extensively studied at the microscopic level in animals and correlated with direct and indirect measurements of myocardial blood flow in patients with coronary artery disease. Animals have permitted more extensive study. These findings have generally supported an increased blood flow to the myocardium with exercise training. However, consistent positive structural and functional adaptations to training have not been observed in large animals. Clinical studies have been limited by methodological problems related to techniques for detecting ischemia and measuring myocardial blood flow and the variability in exercise stimulus. Well-established ischemia and high-intensity, long-duration training were the factors that promoted vascular growth in exercising patients with coronary artery disease. Animals studies also have demonstrated the necessity for myocardial ischemia to be present to induce coronary collateral development with exercise training. Optimal promoters of vascular growth in patients with coronary disease may consist of pharmacological interventions combined with exercise training.

Adaptation, Physiological↗
PubMed2001

In vivo and in vitro effects of integration host factor at the DmpR-regulated sigma(54)-dependent Po promoter.

Transcription from the Pseudomonas CF600-derived sigma(54)-dependent promoter Po is controlled by the aromatic-responsive activator DmpR. Here we examine the mechanism(s) by which integration host factor (IHF) stimulates DmpR-activated transcriptional output of the Po promoter both in vivo and in vitro. In vivo, the Po promoter exhibits characteristics that typify many sigma(54)-dependent promoters, namely, a phasing-dependent tolerance with respect to the distance from the regulator binding sites to the distally located RNA polymerase binding site, and a strong dependence on IHF for optimal promoter output. IHF is shown to affect transcription via structural repercussions mediated through binding to a single DNA signature located between the regulator and RNA polymerase binding sites. In vitro, using DNA templates that lack the regulator binding sites and thus bypass a role of IHF in facilitating physical interaction between the regulator and the transcriptional apparatus, IHF still mediates a DNA binding-dependent stimulation of Po transcription. This stimulatory effect is shown to be independent of previously described mechanisms for the effects of IHF at sigma(54) promoters such as aiding binding of the regulator or recruitment of sigma(54)-RNA polymerase via UP element-like DNA. The effect of IHF could be traced to promotion and/or stabilization of open complexes within the nucleoprotein complex that may involve an A+T-rich region of the IHF binding site and promoter-upstream DNA. Mechanistic implications are discussed in the context of a model in which IHF binding results in transduction of DNA instability from an A+T-rich region to the melt region of the promoter.

Bacterial Proteins↗