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Fitness promotion for adolescent girls: the impact and effectiveness of promotional material which emphasizes the slim ideal.

This study looks at techniques for promoting fitness participation among adolescent girls, in particular those which emphasize the "slim ideal." Various promotional posters were designed which systematically used different models (slim, average, and overweight) and different messages (slimness, activity, and health). The relative effectiveness of these posters was tested using a probability sample of 627 female high school students. The slim model was found to be the most effective poster, while slimness was the least effective of the messages. Demographic and self- and body-image factors had relatively little effect on the ratings of the posters. Focus group interviews conducted with the student sample indicated that they were concerned about their body weight, and that they associated slimness with fitness. However, the slimness message was not thought to encourage participation in fitness because overt emphasis on body image may lead to self-consciousness and fear of social rejection. The idea that participation would lead to a slim physique was also thought to be unrealistic. The students were more critical of the slimness message than of the slim model in the posters. Overall, the data indicate that promoting fitness through messages relating fitness to slimness is not an effective approach with adolescent girls. The use of very slim models in promotional material may be effective, but this approach may not be desirable because it reinforces the dominant cultural stereotype of the "ideal" slim female form.

Adolescent↗

Characterization of the promoter region of the human insulin receptor gene. Evidence for promoter activity.

Recombinant clones containing the promoter region of the human insulin receptor gene were isolated from genomic libraries derived from nondiabetic persons. A 1.5-kilobase pair fragment of the 5'-flanking region was sequenced. One transcriptional start site, located at 203 bases upstream from the start of translation was identified by nuclease S1 mapping and the primer extension experiment using the human insulin receptor mRNA. The bacterial chloramphenicol acetyltransferase assay revealed that a 573-base pair fragment immediately preceding the ATG has promoter activity and that the transcript initiates from the normal start site of the insulin receptor gene in the COS cells. The promoter region contains neither a "TATA box" nor a "CAAT box," has an extremely high G + C content, and contains seven central components of potential Sp 1 binding sites (GGGCGG or CCGCCC). These features are common to those found in the regulatory regions of a class of constitutively expressed "housekeeping" genes. A comparison between the promoter sequence of the human insulin receptor and those of other "housekeeping" genes revealed the presence of homologous sequences among these genes, in addition to the potential Sp 1 binding sites.

Base Sequence↗

Characterization of the human histone H2A.X gene. Comparison of its promoter with other H2A gene promoters.

The human gene for the replication-unlinked histone protein H2A.X is a naturally occurring chimera that contains a replication-unlinked promoter yet produces a stemloop mRNA characteristic of replication-linked histone genes. Consistent with the latter attribute, the H2A.X gene was found to lack introns. The promoter of the H2A.X gene was localized to a 120-base pair region upstream of the transcription start site, a region which included a TATA and two CCAAT sequence elements. The proximal of the two CCAAT elements was shown to be an important determinant of H2A.X gene promoter activity. In a comparative study with the CCAAT elements from the replication-linked H2A.1a gene and the replication-unlinked H2A.Z gene, the proximal CCAAT element of the H2A.X gene was found to bind nuclear factors also bound by CCAAT elements in the latter but not in the former. The specificity of the replication-unlinked H2A.X and H2A.Z gene promoters for CCAAT-binding transcription factors appeared to also reside in short homologous sequences about 10 base pairs away on either side of the CCAAT sequence.

Animals↗

Inhibition of protein phosphatases activates P4 promoter of the human insulin-like growth factor II gene through the specific promoter element.

To understand the transcriptional regulation of the human insulin-like growth factor II (IGF-II) gene, we examined the effects of okadaic acid, a potent in vitro inhibitor of protein phosphatases, on the activation of human IGF-II gene expression. Treatment of A-549 human lung adenocarcinoma cells with okadaic acid increased expression of the IGF-II mRNAs. Since the 4.8-kb mRNA is transcribed under the control of human IGF-II P4 promoter, we examined the P4 promoter element responsible for the okadaic acid-mediated transcriptional activation. Transfection of IGF-II P4 promoter-chloramphenicol acetyltransferase constructs demonstrated that the effects of okadaic acid on the induction of IGF-II gene expression are mediated through multiple promoter elements, including an Egr-1 consensus element. We have also shown that okadaic acid induced the expression of the transcription factor Egr-1. Moreover, by using a GAL4-Egr-1 fusion protein, we have directly demonstrated that okadaic acid positively regulates Egr-1 transcriptional activity in vivo. These results indicate that protein phosphatases play an important role in the transcriptional regulation of the IGF-II.

Adenocarcinoma↗

Transactivation of GATA-1 promoter with ETS1, ETS2 and ERGB/Hu-FLI-1 proteins: stabilization of the ETS1 protein binding on GATA-1 promoter sequences by monoclonal antibody.

Ets family proteins activate transcription via binding to the GGAA core sequence located in the promoter/enhancer elements of many cellular and viral genes. GATA-1 is an erythroid-specific transcription factor. The promoter of the chicken GATA-1 gene contains multiple ets binding sites (EBS), two of them are present in palindromic form. The GATA-1 promoter has been shown to be activated by the E26 virus. In this study, we have analysed whether the palidromic EBS of the chicken GATA-1 promoter is a target for binding and activation by members of the cellular ets gene family products. The results herein indicate that both EBS in the palindrome are required for DNA-binding because mutations in either site reduces the activity by at least 95%. Moreover, DNA binding of ETS1 to the EBS palindrome is dramatically stabilized in the presence of a specific monoclonal antibody whose epitope maps between amino acid positions 240-260. Although each of the single sites bind, the efficiency of binding is extremely low. Furthermore, for efficient binding the two sites must be in an inverted configuration because of the fact that the oligonucleotide containing the left and right EBS in the same orientation binds 10-fold less than the oligonucleotide containing the EBS palindrome. Additionally, we show that the transcription of a reporter gene (CAT) either linked to the GATA-1 EBS palindrome or GATA-1 promoter can be activated by cotransfection with ETS1, alternatively-spliced ETS1, ETS2 or ERGB/Hu-FLI-1 expression vectors.

Animals↗

Evaluation of relative promoter strengths of the HIV-1-LTR and a chimeric RSV-LTR in T lymphocytic cells and peripheral blood mononuclear cells: promoters for anti-HIV-1 gene therapies.

Gene therapy approaches for human immunodeficiency virus type 1 (HIV-1) infections focus on the transfer of critical genetic elements into CD4+ T lymphocytes and CD34+ stem cells. Ideally, expression of the anti-HIV-1 gene constructs should be induced during early stages of infection to combat high turnover of the replicating virus. In this study, we investigated the activity of two promoters, HIV-1 long terminal repeat (HIV-1-LTR) and Rous sarcoma virus (RSV) LTR fused with the transactivation response element (TAR) from the HIV-1-LTR (ie RSV-TAR) in presence of Tat, the major HIV-1 transcriptional transactivator and an early gene product in HIV-1 infection. Comparative expression from both of these plasmids was analyzed by measuring expression of a reporter gene, chloramphenicol acetyltransferase (CAT), after transfection of the promoter-CAT constructs and a Tat-expressing plasmid into CEM T lymphocytic cells and peripheral blood mononuclear cells (PBMC). The HIV-1-LTR could be transactivated by Tat in both unstimulated and stimulated cells. Although the RSV-TAR had a relatively high basal level of expression, Tat transactivation of this chimeric promoter occurred only in unstimulated cells. These results suggest that the HIV-1-LTR may be a better promoter for therapeutic gene expression in anti-HIV-1 intracellular immunization approaches.

Avian Sarcoma Viruses↗

Promoter sequences of two homologous pectin esterase genes from Chinese cabbage (Brassica campestris L. ssp. pekinensis) and pollen-specific expression of the GUS gene driven by a promoter in tobacco plants.

The promoter regions of two genomic clones, GBAN215-6 and GBAN215-12 from Chinese cabbage (Brassica campestris L. ssp. pekinensis), were sequenced. The nucleotide sequences of their promoter regions were compared with that of the Bp19 pollen-specific gene of Brassca napus. High nucleotide sequence homologies were observed among these three genes in the region between 210 bp upstream and the putative transcription start site. A sequence motif TGTGGTG, which is similar to that of the PB core motif (TGTGGTT) of two tomato pollen-specific genes, LAT52 and LAT56, was present in these two cloned genes. To determine regulatory sequences responsible for the anther-specific expression of the gene BAN215-6, two recombinant plasmids, pBPE3 (-274- + 109) and pBPE4 (-816- + 109) containing different lengths of the promoter fused with the GUS gene, were constructed and introduced into tobacco plants by Agrobacterium-mediated transformation. The result showed that the 383 bp (-274- + 109) of the BAN215-6 promoter region was sufficient for the anther-specific expression of the GUS gene. The GUS expression in a tobacco plants transformed with these constructs was first detected in uninucleate microspores and persisted at in vitro germinated pollen tubes. The expression level was increased during anther development, reaching the highest level in mature pollens.

Brassica↗

Disrupted IGF2 promoter control by silencing of promoter P1 in human hepatocellular carcinoma.

Previous investigations have supported or indicated a stimulatory role of the insulin-like growth factor II gene (IGF2) in hepatocarcinogenesis. We have studied the transcript levels, promoter usage, and imprinting status of the ICF2 gene and its relationship to H19 in human hepatocellular carcinomas (HCCs) and liver tumor cell lines. The activity of the IGF2 promoter P1 was lost in about 70% of the cases (18 of 25). This is the most prominent abnormality regarding the IGF2 regulation in this study. Total IGF2 as well as promoter P3 transcription were up-regulated in a small group of the tumors. Twenty genetically informative cases were obtained from 26 cases, thus excluding the probability of loss of heterozygosity of the IGF2 gene. Among these, nine showed abnormal monoallelic expression of IGF2. One HCC and one HCC cell line proved loss of functional imprinting of IGF2. H19 and IGF2 were regulated in parallel, and expression levels were variable. Taken together, the disruption of the IGF2 promoter regulation, particularly the loss of P1 activity, is a common feature of human HCCs. The loss of P1 activity explains the frequent loss of biallelic IGF2 expression and may potentially be used as a diagnostic or monitoring marker for human HCC.

Alleles↗

Interventions for promoting adherence to tuberculosis management.

BACKGROUND: Up to half the people with tuberculosis do not complete their treatment. Strategies to improve adherence to treatment can be targeted at the person with the disease or at health workers. OBJECTIVES: The objective of this review was to assess the effects of promoting adherence to anti-tuberculosis treatment and completion of diagnostic protocols for TB. SEARCH STRATEGY: We searched the Cochrane Collaboration Trials, the Cochrane Effective Professional Practice Group trials register, the Cochrane Infectious Diseases Group trials register, Medline, Embase, Lilacs and reference lists of articles. We contacted experts in the field. SELECTION CRITERIA: Randomised and quasi-randomised trials of interventions to promote adherence with curative or preventive chemotherapy and diagnostic protocols for tuberculosis. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. MAIN RESULTS: Eleven trials were included. Reminder cards sent to defaulters, a combination package of a monetary incentive and health education and more supervision of tuberculosis clinic staff increased the number of people completing their tuberculosis treatment. Direct observation by clinic nurses of people swallowing their tuberculosis drugs did not increase the likelihood of treatment success. Return to the clinic for reading of a tuberculin skin test was enhanced by monetary incentives, assistance by lay health workers, contracts and telephone prompts but not by health education. REVIEWER'S CONCLUSIONS: We have found evidence of benefit for a number of specific interventions to improve adherence to anti-tuberculous therapy and completion of diagnostic protocols. These should be implemented by health care providers where appropriate to local circumstances. Future studies in low income countries are a priority and should measure adherence as well as clinical outcomes.

Antitubercular Agents↗

Macrophage oxidative burst and related cytotoxicity. I. Differential activation by tumor-promoting and non-tumor-promoting phorbol esters.

Mouse peritoneal macrophages elicit an oxidative burst (OB) response upon stimulation with the tumor promoter 12-O-tetradecanoyl-phorbol 13-acetate (TPA). In this study we compare the OB-stimulating capacity of phorbol ester derivatives, structurally related to TPA, which differ in their tumor-promoting activity. Non-tumor-promoting derivatives such as phorbol 13-acetate, phorbol 12-myristate, TPA-20-aldehyde and 4-O-methyl TPA were tested. These reagents stimulate macrophages to generate OB products such as O-2 and H2O2, yet the amounts required for stimulation are 1,000 times higher than the amounts of TPA required to elicit a comparable response. It has also been observed that, in the same order of magnitude, the above-mentioned derivatives are less efficient than TPA in rendering macrophages cytolytic toward erythrocytes. Another strong tumor promoter tested, teleocidin, has been found to be as potent as TPA in the activation of macrophage OB and in related activities.

Animals↗

12-O-tetradecanoylphorbol-13-acetate--induced levels of AP-1 proteins: a 46-kDa protein immunoprecipitated by anti-fra-1 and induced in promotion-resistant but not promotion-sensitive JB6 cells.

Neoplastic transformation and transcriptional activation by activator protein-1 (AP-1) complex are stimulated by tumor-promoting agents in promotion-sensitive (P+) but not promotion-resistant (P-) mouse epidermal JB6 cells in culture. This implicates AP-1 as a specific regulator of signal transduction pathways in the promotion phase of neoplastic transformation. We therefore hypothesized that the defective P- responsiveness may be due to limiting levels of AP-1 protein components in those cells. In this investigation, steady-state levels of AP-1 protein components were measured by immunoprecipitating proteins from 12-O-tetradecanoylphorbol-13-acetate (TPA)-treated P+ and P- cells to discern what may limit the AP-1 response. Whereas the AP-1 proteins junB, junD, and fosB did not show differential basal or TPA-inducible levels in P+ and P- cells, a 46-kDa species precipitated by anti-fra-1 antibody was TPA-inducible in P- cells but not in P+ cells, and c-jun protein was present at higher levels in TPA-treated and untreated P+ cells than in P- cells. These data raise the possibility that the 46-kDa fra-1-related protein may be a negative modulator of AP-1 activity and suggest that elevated levels of this 46-kDa species and limiting levels of c-jun may significantly impair AP-1 function or transformation response in P- cells or both.

Amino Acid Sequence↗

Evidence that nitrogen regulatory gene ntrC of Salmonella typhimurium is transcribed from the glnA promoter as well as from a separate ntr promoter.

Previous work has indicated that nitrogen regulatory genes ntrB and ntrC of Salmonella typhimurium are closely linked to glnA, the structural gene encoding glutamine synthetase; proceeding clockwise the order of genes in the 86 U region of the map is polA...ntrC ntrB glnA glnA promoter...rha. To study ntrC transcription we have constructed operon fusions of ntrC to lacZ using the Casadaban Mu d1 (Apr lac) phage so that we can measure beta-galactosidase activity as a reflection of ntrC transcription and we have introduced into fusion strains promoter constitutive mutations at glnA [glnAp(Con)]. The glnAp(Con) mutations, which elevate glnA expression in fusion strains, also elevate beta-galactosidase activity, indicating that ntrC is cotranscribed with glnA. Consistent with this interpretation, polar insertion mutations in glnA decrease beta-galactosidase activity of fusion strains carrying glnAp(Con) mutations. However, glnA insertions do not eliminate beta-galactosidase activity of glnAp(Con) ntrC::Mu d1 strains and they have little effect on beta-galactosidase activity of the original ntrC::Mu d1 fusion strains. The latter results confirm that ntrC can also be transcribed from an ntr promoter downstream of glnA. Polar insertion mutations in ntrB eliminate beta-galactosidase activity of both the original fusion strains and fusion strains carrying glnA(Con) mutations, indicating that the ntr promoter lies between glnA and ntrB.

Base Sequence↗

The non-phorbol ester tumor promoter okadaic acid does not promote morphological transformation or inhibit junctional communication in hamster embryo cells.

Okadaic acid is a potent non-phorbol ester mouse skin tumor promoter. Unlike the phorbol ester tumor promoters, okadaic acid is unable to promote the induction of morphological transformation in Syrian hamster embryo cell colonies. On the contrary, okadaic acid seems to counteract the effect of phorbol esters on transformation. Also unlike phorbol ester tumor promoters, okadaic acid does not inhibit intercellular communication, neither in primary hamster embryo cells, nor in the phorbol ester sensitive cell line BPNi. Furthermore, okadaic acid has no effect on the reoccurrence of communication following removal of 12-O-tetradecanoylphorbol-13-acetate.

Animals↗

Effects of tert-butyl hydroperoxide on promotable and non-promotable JB6 mouse epidermal cells.

Oxidants and agents that induce a cellular prooxidant state can act as carcinogens. We compared the effect of tert-butyl hydroperoxide (Bu-OOH) on DNA strand breakage, poly ADP-ribosylation of chromosomal proteins and the expression of the proto-oncogenes c-fos and c-myc between non-promotable clone 30 and promotable clone 41 of mouse epidermal cells JB6. These pathophysiological effects of oxidants are mechanistically related. Bu--OOH caused more DNA-strand breakage at high concentrations and more extensive poly ADP-ribose accumulation in clone 30 than in clone 41, in reactions which require intracellular free iron. Clone 41 exhibited constitutive c-myc expression while c-fos mRNA was very low in untreated cultures of both clones. Low concentrations of Bu-OOH induced c-myc and more strongly c-fos in clone 41. Both proto-oncogenes were strongly induced in clone 30. Our results allow insights into the mechanisms of action of a typical organic hydroperoxide in JB6 cells. However, they do not uncover the reasons for the differential promotability of the two JB6 clones by oxidants beyond the implication of the constitutive expression of c-myc in promotable clone 41.

Animals↗

Simian virus 40 early promoter mutations that affect promoter function and autoregulation by large T antigen.

A set of nine mutants containing point mutations, and small deletions or insertions, were constructed in the early promoter region of simian virus 40 (SV40) to determine the role of the DNA sequences between the TATA box and the six upstream G + C-rich clusters in early transcription. The mutant templates were tested for transcription activity in vitro in HeLa cell extracts and in vivo in CV-1 and COS cells using the chloramphenicol acetyl transferase gene (CAT) assay. Both in vitro and in vivo results show that the narrow region from nucleotide positions 38 to 41 is an important domain of the early promoter. Deletion and insertion mutations most strongly affect the level of transcription. Specifically a four base-pair deletion in the promoter region enhances the level of transcription four- to sixfold in vitro, but causes a fourfold suppression of CAT gene expression in the in vivo assay. These opposite effects may result from changes in spacing under in vitro and in vivo conditions between the TATA box and the G + C-rich motifs where transcription factors may make simultaneous contact. Of the three T antigen binding sites (I, II and III), sites I and II have already been shown to be involved in the autoregulation of early transcription. Our mutational analyses demonstrate the role of site III, which partially overlaps with nucleotide positions 38 to 41, in the autoregulation of the SV40 early promoter.

Antigens, Viral, Tumor↗

Inhibition of adhesion-promoting activity of a human salivary protein which promotes adhesion of Streptococcus mutans JBP to hydroxyapatite.

The adhesion-promoting proteins (APP) (molecular mass approx. 300 kDa), which promote adhesion of Streptococcus mutans JBP (serotype c) to hydroxyapatite, were isolated from human submandibular-sublingual (SMSL) saliva by gel filtration on a Trisacryl GP2000 M column. The effects of hexoses, pentoses, methyl-pentoses, hexosamines, N-acetylhexosamines, a basic amino acid, polyamines and ammonium chloride on the bacterial adhesion-promoting activity of the APP were examined. Galactosamine, mannosamine, L-lysine, spermine, putrescine, and ammonium chloride inhibited the adhesion-promoting activities of the APP. The other sugars, including the N-acetylhexosamines, were without effect. Thus, compounds containing a primary amino-group appear to have a specific inhibitory effect on adhesion of S. mutans JBP to APP adsorbed onto hydroxyapatite, an activity which is lost if the amino-group is acetylated.

Bacterial Adhesion↗

Effect of tumor-promoting and anti-promoting chemicals on the viability and junctional coupling of human HeLa cells transfected with DNAs coding for various murine connexin proteins.

Gap-junctional intercellular communication is thought to be essential for maintaining cellular homeostasis and growth control. Its perturbation entails toxicological implications and it has been correlated with the in vivo tumor-promoting potential of chemicals. Little is known about the mechanism(s) responsible for the tumor promoters interference with the cellular coupling. Moreover, nongenotoxic carcinogens, as well as connexins (gap-junctional protein subunits), are known to be organ-/tissue-specific; this implies that the effect of different agents should be evaluated on their specific target, that is, connexin. To investigate the role of different connexins in regulating gap-junctional gating and to compare the properties of homotypic junctional channels, we evaluated the effects of tissue-specific tumor promoters and anti-promoters on the viability and intercellular coupling (dye-transfer) of HeLa cells stably transfected with cDNAs coding for connexin(cx)43, cx40, cx26 and cx32. The results demonstrate that the transfectants possess individual junctional permeabilities, differentially affected by the chemicals, they also show different sensitivities to the cytotoxic effect of the compounds. These findings confirm that connexin diversity may be responsible for the different gating properties of gap-junctional channels, being also suggestive for their separate functions and independent regulatory mechanisms.

Animals↗

Histone acetyltransferase p300 promotes the activation of human WT1 promoter and intronic enhancer.

The Wilms' tumor gene-1 (WT1) encodes a zinc finger protein involved in gene regulation during kidney, gonad, and heart development. In addition to its promoter, a 258 bp intronic enhancer is required for tissue-specific expression of WT1 gene. p300 is a histone acetyltransferase (HAT) and exerts essential functions in gene regulation. Here, we show that p300 increased the expression of endogenous WT1 mRNA and promoted the activation of the WT1 promoter and intronic enhancer. The results also revealed that the adenovirus E1A repressed the p300 function, while the p300-binding defective E1A delta 2-36 did not, and p300 HAT activity was important for its function since p300 mutant with the HAT domain deleted partially abrogated its ability to activate the WT1 promoter and intronic enhancer. Furthermore, p300 and c-Myb synergistically activated the expression of WT1 gene. This study revealed that p300 and its HAT activity were involved in regulation of WT1 transcription.

Acetyltransferases↗