Effect of Tryptophan Analogs on Derepression of the \u3cem\u3eEscherichia coli\u3c/em\u3e Tryptophan Operon by Indole-3-Propionic Acid

The abilities of 14 tryptophan analogs to repress the tryptophan (trp) operon have been studied in Escherichia coli cells derepressed by incubation with 0.25 mM indole-3-propionic acid (IPA). trp operon expression was monitored by measuring the specific activities of anthranilate synthase (EC 4.1.3.27) and the tryptophan synthase (EC 4.2.1.20) β subunit. Analogs characterized by modification or removal of the α-amino group or the α-carboxyl group did not repress the trp operon. The only analogs among this group that appeared to interact with the trp aporepressor were IPA, which derepressed the trp operon, and d-tryptophan. Analogs with modifications of the indole ring repressed the trp operon to various degrees. 7-Methyl-tryptophan inhibited anthranilate synthase activity and consequently derepressed the trp operon. Additionally, 7-methyltryptophan prevented IPA-mediated derepression but, unlike tryptophan, did so in a non-coordinate manner, with the later enzymes of the operon being relatively more repressed than the early enzymes. The effect of 7-methyltryptophan on IPA-mediated derepression was likely not due to the interaction of IPA with the allosteric site of anthranilate synthase, even though feedback-resistant mutants of anthranilate synthase were partially resistant to derepression by IPA. The effect of 7-methyltryptophan on derepression by IPA was probably due to the effect of the analog-aporepressor complex on trp operon expression

Heat transfer with very high free-stream turbulence and streamwise vortices

Results are presented for two experimental programs related to augmentation of heat transfer by complex flow characteristics. In one program, high free stream turbulence (up to 63 percent) was shown to increase the Stanton number by more than a factor of 5, compared with the normally expected value based on x-Reynolds number. These experiments are being conducted in a free-jet facility, near the margins of the jet. To a limited extent, the mean velocity, turbulence intensity, and integral length scale can be separately varied. The results show that scale is a very important factor in determining the augmentation. Detailed studies of the turbulence structure are being carried out using an orthogonal triple hot-wire anemometer equipped with a fourth wire for measuring temperature. The v' component of turbulence appears to be distributed differently from u' or w'. In the second program, the velocity distributions and boundary layer thicknesses associated with a pair of counter-rotating, streamwise vortices were measured. There is a region of considerably thinned boundary layer between the two vortices when they are of approximately the same strength. If one vortex is much stronger than the other, the weaker vortex may be lifted off the surface and absorbed into the stronger

Games in Agriculture, Food, and Natural Resources Education

Educational games provide learners with team-based, experiential, and problem-centered learning opportunities. Therefore, educational games are recommended to encourage learner success in an increasingly complex and collaborative world. Research exploring interventions to increase teacher affinity toward games is needed to inform expansion of games within classrooms. The current study leveraged the input, environment, and outcomes model to analyze perceptions of games held by school-based agriculture, food, and natural resources (AFNR) educators before and after a professional development experience focused on educational games. Results indicate teachers held a favorable perception of games before engaging in the professional development. Engagement in the professional development was related to only minimal increases in the perceptions held by teachers regarding educational games. Importantly, individual items within the construct illuminate an expanded view of educational games and their utility within AFNR classrooms as a result of the professional development experience. Specifically, respondents saw educational games as being valuable to engage learners in new content, not just as a review tool. Findings suggest a professional development experience related to games may help expand teacher conceptualizations of educational game utility. Specific recommendations are included to expand teacher understanding and use of educational games

Genetic Variation in Concentration of the 33-mer Protein Subcomponent in Wheat

Celiac Disease is a hypersensitive response to gluten caused by HLA-DQ2 or HLA-DQ8 T-cell presentation, initiating destruction of intestinal epithelial cells. Currently, the only remedy for those suffering from celiac disease is elimination of all gluten from the diet. Studies indicate that an indigestible fragment of the gluten molecule, alpha-gliadin subcomponent 33-mer, rich in proline and glutamine, is responsible for the hypersensitivity response. Determination of 33-mer concentration in wheat lines could be beneficial to future development of wheat lines with reduced 33-mer concentration. Protein from wheat flour was extracted and subjected to ELISA techniques in order to quantify the concentration of 33-mer. A technique that quantifies the concentration of 33-mer is a necessary first step for future research efforts focused on identification and development of wheat lines with reduced concentrations of 33-mer. It is possible that wheat with reduced 33-mer may be suitable for consumption by individuals with celiac disease

Large scale analysis of positional effects of single-base mismatches on microarray gene expression data

<p>Abstract</p> <p>Background</p> <p>Affymetrix GeneChips utilize 25-mer oligonucleotides probes linked to a silica surface to detect targets in solution. Mismatches due to single nucleotide polymorphisms (SNPs) can affect the hybridization between probes and targets. Previous research has indicated that binding between probes and targets strongly depends on the positions of these mismatches. However, there has been substantial variability in the effect of mismatch type across studies.</p> <p>Methods</p> <p>By taking advantage of naturally occurring mismatches between rhesus macaque transcripts and human probes from the Affymetrix U133 Plus 2 GeneChip, we collected the largest 25-mer probes dataset with single-base mismatches at each of the 25 positions on the probe ever used in this type of analysis.</p> <p>Results</p> <p>A mismatch at the center of a probe led to a greater loss in signal intensity than a mismatch at the ends of the probe, regardless of the mismatch type. There was a slight asymmetry between the ends of a probe: effects of mismatches at the 3' end of a probe were greater than those at the 5' end. A cross study comparison of the effect of mismatch types revealed that results were not in good agreement among different reports. However, if the mismatch types were consolidated to purine or pyrimidine mismatches, cross study conclusions could be generated.</p> <p>Conclusion</p> <p>The comprehensive assessment of the effects of single-base mismatches on microarrays provided in this report can be useful for improving future versions of microarray platform design and the corresponding data analysis algorithms.</p

Expression of Endogenous Mtv Provirus Transcripts in BALB/c Splenic Lymphocytes

Abstract The presence of antigen(s) related to the exogenous milk-transmitted murine mammary tumor virus on the surface of BALB/c splenic lymphocytes has been documented previously. Since the BALB/c strain lacks murine mammary tumor virus, the presence of murine mammary tumor virus-related antigen(s) on lymphocytes has been ascribed to expression of germinally transmitted Mtv transcripts and proviruses were characterized to evaluate this hypothesis. Transcripts from genomic size Mtv provirus(es) accumulated in the spleen in an age-dependent manner. Two novel Mtv transcripts of 7.8 and 6.4 kb were observed in the spleen. These observations indicate that the transcriptional and translational expression of an endogenous Mtv occurs in normal cells of the lymphoreticular lineage

Murine Mammary Tumor Virus Expression During Mammary Tumorigenesis in BALB/c Mice

Steady-state levels of murine mammary tumor virus (MuMTV) RNA were quantitated during mammary tumorigenesis in BALB/c mice by molecular hybridization with a representative MuMTV complementary DNA (cDNA) probe. Hyperplastic alveolar nodule (HAN) lines are preneoplastic mammary lesions that were induced in BALB/c mice by hormones alone or in combination with 7,12-dimethylbenz(a)anthracene and give rise to mammary tumors. The hormone-induced HAN lines D1 and D2 contained detectable amounts of hybridizable MuMTV sequences. MuMTV RNA sequences were also observed in five of the six transplanted BALB/c mammary tumors that were examined. Similar levels of hybridizable MuMTV RNA were observed between the D1 or D2 HAN line and mammary tumors derived from each HAN line. The D2 HAN line as well as D2, C4, and CD8 mammary tumors accumulated RNA that was apparently homologous to most of the MuMTV genome. Thermal denaturation of hybrids indicated extensive sequence homology between the MuMTV cDNA and hybridizable RNA in the BALB/c HAN lines and mammary tumors. A low level of type C viral RNA was observed in the BALB/c HAN lines and most mammary tumors by molecular hybridization with a cDNA to Moloney murine leukemia virus. These data demonstrate that MuMTV sequences are frequently expressed in hormone-induced BALB/c HAN lines and mammary tumors derived from HAN lines or ductal hyperplasias induced in BALB/c mice by hormones and/or a chemical carcinogen. The transition from the preneoplastic to the neoplastic state in BALB/c mice does not appear to be due to a change in the steady-state levels of MuMTV RNA since the hormone-induced HAN lines and mammary tumors had similar levels of hybridizable MuMTV RNA