Experimental investigation of wall thickness and hole shape variation effects on full-coverage film cooling performance for a gas turbine vane

The effects of wall thickness and hole shape variation on a full-coverage film cooled turbine vane are investigated in a stationary and linear cascade utilizing the pressure sensitive paint technique. The varied wall thickness produces hole length-to-diameter ratio (L/D) in a range from L/D = 2 to 5, and holes tested include simple angle hole, compound angle hole, and fan-shaped hole. Five rows of holes are provided on the pressure side while three rows of holes are provided on the suction side, with six rows of cylindrical holes drilled on the leading edge to construct showerhead film cooling. The tested blowing ratios for the showerhead, pressure side, and suction side range from 0.25 to 1.5, with a density ratio of 1.5. The freestream Reynolds number is 1.35 × 105, based on the axial chord length and the inlet velocity, with a freestream turbulence intensity level of 3.5% at the cascade inlet. The results indicate that the wall thickness variation produces significant influence on the pressure side film cooling effectiveness, while only marginal effect on the showerhead and suction side film cooling. Also observed is that the fan-shaped hole generates the highest film cooling effectiveness on pressure or suction side. Also discussed is the surface curvature effect, combining with effects of wall thickness and hole shape variations, on the film cooling effectiveness in comparison to the flat-plate data

An Anomaly-Based Method for Identifying Signals of Spring and Autumn Low-Temperature Events in the Yangtze River Valley, China

June 2015 QIAN ET AL. Vol. 54 1216-123

Length to diameter ratio effect on heat transfer performance of simple and compound angle holes in thin-wall airfoil cooling

Heat transfer coefficients on a flat plate surface downstream a row of simple and compound angle cylindrical holes are investigated using high-resolution thermographic liquid crystal technique. A variation of flow parameters including blowing ratio, and geometry parameters including compound angle and length-to-diameter ratio are examined. Blowing ratios (M) ranging from 0.3 to 2, length to diameter ratios (L/D) from 0.5 to 5, and two compound angle (β: 0°, 45°) are employed composing a test matrix of 70 test cases. Detailed local, spanwise averaged, and area averaged heat transfer coefficients hf/h0 are presented to illustrate the effect of length-to-diameter ratio and compound angle. The film cooling performance is also evaluated using NHFR method and Δφ method by combining adiabatic film effectiveness and heat transfer coefficient data. Results indicate that Δφ method has superiority in evaluating film cooling performance due to its direct reflection of temperature reduction by film protection

Knowledge-Based Reconstruction of mRNA Transcripts with Short Sequencing Reads for Transcriptome Research

While most transcriptome analyses in high-throughput clinical studies focus on gene level expression, the existence of alternative isoforms of gene transcripts is a major source of the diversity in the biological functionalities of the human genome. It is, therefore, essential to annotate isoforms of gene transcripts for genome-wide transcriptome studies. Recently developed mRNA sequencing technology presents an unprecedented opportunity to discover new forms of transcripts, and at the same time brings bioinformatic challenges due to its short read length and incomplete coverage for the transcripts. In this work, we proposed a computational approach to reconstruct new mRNA transcripts from short sequencing reads with reference information of known transcripts in existing databases. The prior knowledge helped to define exon boundaries and fill in the transcript regions not covered by sequencing data. This approach was demonstrated using a deep sequencing data set of human muscle tissue with transcript annotations in RefSeq as prior knowledge. We identified 2,973 junctions, 7,471 exons, and 7,571 transcripts not previously annotated in RefSeq. 73% of these new transcripts found supports from UCSC Known Genes, Ensembl or EST transcript annotations. In addition, the reconstructed transcripts were much longer than those from de novo approaches that assume no prior knowledge. These previously un-annotated transcripts can be integrated with known transcript annotations to improve both the design of microarrays and the follow-up analyses of isoform expression. The overall results demonstrated that incorporating transcript annotations from genomic databases significantly helps the reconstruction of novel transcripts from short sequencing reads for transcriptome research

CRISPR-Cas9D10A nickase-assisted base editing in the solvent producer Clostridium beijerinckii

Clostridium beijerinckii is a potentially important industrial microorganism as it can synthesize valuable chemicals and fuels from various carbon sources. The establishment of convenient to use, effective gene tools with which the organism can be rapidly modified is essential if its full potential is to be realized. Here, we developed a genomic editing tool (pCBEclos) for use in C. beijerinckii based on the fusion of cytidine deaminase (Apobec1), Cas9 D10A nickase and uracil DNA glycosylase inhibitor (UGI). Apobec1 and UGI are guided to the target site where they introduce specific base‐pair substitutions through the conversion of C·G to T·A. By appropriate choice of target sequence, these nucleotide changes are capable of creating missense mutation or null mutations in a gene. Through optimization of pCBEclos, the system derived, pCBEclos‐opt, has been used to rapidly generate four different mutants in C. beijerinckii, in pyrE, xylR, spo0A, and araR. The efficiency of the system was such that they could sometimes be directly obtained following transformation, otherwise only requiring one single restreaking step. Whilst CRISPR–Cas9 nickase systems, such as pNICKclos2.0, have previously been reported in C. beijerinckii, pCBEclos‐opt does not rely on homologous recombination, a process that is intrinsically inefficient in clostridia such as C. beijerinckii. As a consequence, bulky editing templates do not need to be included in the knockout plasmids. This both reduces plasmid size and makes their construction simpler, for example, whereas the assembly of pNICKclos2.0 requires six primers for the assembly of a typical knockout plasmid, pCBEclos‐opt requires just two primers. The pCBEclos‐opt plasmid established here represents a powerful new tool for genome editing in C. beijerinckii, which should be readily applicable to other clostridial species

Continuous and complete conversion of high concentration p-nitrophenol in a flow-through membrane reactor

Here, we report on a green and effective method for the continuous and complete conversion of high concentrations of p-nitrophenol (PNP) using a flow-through membrane reactor and less NaBH4. The catalytic membrane was successfully fabricated by loading Pd nanoparticles onto the surface of a branched TiO2 nanorod-functionalized ceramic membrane. The modification with branched TiO2 nanorods can significantly improve the loading amount of Pd nanoparticles onto ceramic membranes, resulting in enhanced catalytic performance. With 6 mg of Pd, 93 L m−2 hr−1 of flux density and 8.04 cm2 of membrane surface area in the flow-through membrane reactor, PNP at a concentration of 4,000 ppm can be converted to high-value p-aminophenol using less NaBH4 (using a molar ratio of NaBH4:PNP of 9.6) within 24 s at 30°C. More importantly, the conversion can be continuously and stably performed for 240 min

Metabolic engineering of the L-phenylalanine pathway in Escherichia coli for the production of S- or R-mandelic acid

<p>Abstract</p> <p>Background</p> <p>Mandelic acid (MA), an important component in pharmaceutical syntheses, is currently produced exclusively via petrochemical processes. Growing concerns over the environment and fossil energy costs have inspired a quest to develop alternative routes to MA using renewable resources. Herein we report the first direct route to optically pure MA from glucose via genetic modification of the L-phenylalanine pathway in <it>E. coli</it>.</p> <p>Results</p> <p>The introduction of hydroxymandelate synthase (HmaS) from <it>Amycolatopsis orientalis </it>into <it>E. coli </it>led to a yield of 0.092 g/L S-MA. By combined deletion of competing pathways, further optimization of S-MA production was achieved, and the yield reached 0.74 g/L within 24 h. To produce R-MA, hydroxymandelate oxidase (Hmo) from <it>Streptomyces coelicolor </it>and D-mandelate dehydrogenase (DMD) from <it>Rhodotorula graminis </it>were co-expressed in an S-MA-producing strain, and the resulting strain was capable of producing 0.68 g/L R-MA. Finally, phenylpyruvate feeding experiments suggest that HmaS is a potential bottleneck to further improvement in yields.</p> <p>Conclusions</p> <p>We have constructed <it>E. coli </it>strains that successfully accomplished the production of S- and R-MA directly from glucose. Our work provides the first example of the completely fermentative production of S- and R-MA from renewable feedstock.</p

Reconstruction of xylose utilization pathway and regulons in Firmicutes

<p>Abstract</p> <p>Background</p> <p>Many Firmicutes bacteria, including solvent-producing clostridia such as <it>Clostridium acetobutylicum</it>, are able to utilize xylose, an abundant carbon source in nature. Nevertheless, homology searches failed to recognize all the genes for the complete xylose and xyloside utilization pathway in most of them. Moreover, the regulatory mechanisms of xylose catabolism in many Firmicutes except <it>Bacillus </it>spp. still remained unclear.</p> <p>Results</p> <p>A comparative genomic approach was used to reconstruct the xylose and xyloside utilization pathway and analyze its regulatory mechanisms in 24 genomes of the Firmicutes. A novel xylose isomerase that is not homologous to previously characterized xylose isomerase, was identified in <it>C. acetobutylicum </it>and several other Clostridia species. The candidate genes for the xylulokinase, xylose transporters, and the transcriptional regulator of xylose metabolism (XylR), were unambiguously assigned in all of the analyzed species based on the analysis of conserved chromosomal gene clustering and regulons. The predicted functions of these genes in <it>C. acetobutylicum </it>were experimentally confirmed through a combination of genetic and biochemical techniques. XylR regulons were reconstructed by identification and comparative analysis of XylR-binding sites upstream of xylose and xyloside utilization genes. A novel XylR-binding DNA motif, which is exceptionally distinct from the DNA motif known for <it>Bacillus </it>XylR, was identified in three Clostridiales species and experimentally validated in <it>C. acetobutylicum </it>by an electrophoretic mobility shift assay.</p> <p>Conclusions</p> <p>This study provided comprehensive insights to the xylose catabolism and its regulation in diverse Firmicutes bacteria especially Clostridia species, and paved ways for improving xylose utilization capability in <it>C. acetobutylicum </it>by genetic engineering in the future.</p