Design of reinforced concrete frame structure of exhibition hall

Import 23/08/2017Předmětem bakalářské práce je železobetonová dvoupatrová rámová konstrukce, která má sloužit pro občanskou vybavenost jako výstavní síň. Cílem je navrhnout a posoudit nosné prvky rámové konstrukce, konstrukce schodiště a stropů a založení na základových patkách podle metody mezních stavů, platných norem a konstrukčních zásad. Pro posouzené prvky byly vypracovány výkresy výztuže a stavební výkresy.The subject of Bachelor thesis is two storey reinforced concrete frame structure, which has to serve for civil amenitied as exhibition hall. The aim is to design and assess the support elements of frame structure, the structure of staircase and the ceilings and the foundation on foundation plinths according to the method of limit states, applicable standards and design principles. For assess elements have been created reinforcement drawings and constuction drawings.221 - Katedra konstrukcívýborn

Presumed Little Ice Age glacial extent in the eastern Tian Shan, China

<p>Mountain glaciers across the Tian Shan provide critical freshwater resources for the arid and semi-arid areas in Central Asia. Glacial extent during the Little Ice Age (LIA) has been investigated in individual valleys, but its spatial characteristic across a large region still remains unclear. We delineated the presumed maximum LIA glacial extents in three study regions of the eastern Tian Shan (the Boro-Eren, the Bogeda, and the Barkol-Karlik ranges) using Google Earth and the 30 m Shuttle Radar Topography Mission digital elevation model. The corresponding contemporary glaciers were extracted from the Second Glacier Inventory Dataset of China. The total area of 865 contemporary glaciers was estimated to cover 791.6 km² during a LIA Maximum and decreased to 484.6 km² around 2006–2010, with a relative area loss of 38.8%. The spatial pattern of glacier area loss exhibits a west–east decreasing trend between these three regions. This map provides a data set to investigate the pattern of LIA glacial extents and assess climate impact on water resources in the eastern Tian Shan at a centennial time scale.</p

The <i>yuc8</i> Mutation Reduces the Induction of Hypocotyl Elongation by High Temperature and <i>PIF4</i> Overexpression.

<p>(A) Hypocotyl length showing that the <i>yuc8</i> mutation reduces high temperature-induced hypocotyl elongation. Four-d-old seedlings grown at 22°C were transferred to 29°C in continuous light for additional 2 d before hypocotyl length measurement. Data shown are average±SD. Asterisks represent Student's <i>t</i>-test significance between 29°C and 22°C grown plants for each genotype or between pairs indicated with brackets (**, P<0.01). Shown are representative data from one biological replicate; three biological replicates were conducted, yielding similar results. (B) Hypocotyl length showing that the <i>yuc8</i> mutation partially suppresses the long-hypocotyl phenotype of <i>35S-PIF4</i> plants. The hypocotyl length of 6-d-old seedlings of the indicated genotypes grown at 22°C was measured. Data shown are average±SD. Asterisks represent Student's <i>t</i>-test significance between transgenic/mutant and wild-type plants or between pairs indicated with brackets (*, P<0.05; **, P<0.01). Shown are representative data from one biological replicate; three biological replicates were conducted, yielding similar results.</p

PIF4 Directly Binds to the Promoter Region of <i>YUC8</i>.

<p>(A) Illustration of the <i>YUC5</i>, <i>YUC8</i>, <i>YUC9</i> and <i>YUC10</i> promoter regions showing the presence of G-box DNA motifs. The arrows indicate positions of primers used for ChIP-PCR experiment. Shown are 2-kb upstream sequences of the <i>YUC</i> genes. The translational start site (ATG) is shown at position +1. (B) Gel photographs showing the amplified products from the ChIP assay. The ChIP assays were performed using 6-d-old seedlings expressing the PIF4-HA fusion protein untreated or treated with 29°C for 6 h. Antibody to the HA tag was used to immunoprecipitate PIF4-HA and associated DNA fragments. DNA was amplified by using primers specific to the region containing the G-box element or control regions in <i>ACT2</i> promoter as indicated. Shown are representative data from one biological replicate; this experiment was conducted for three biological replicates, yielding similar results. (C) EMSA assay showing that PIF4 binds the G-box motifs present in the <i>YUC8</i> promoter <i>in vitro</i>. The <i>YUC8</i> promoter fragments containing the G-box motifs were incubated with <i>in vitro</i> TNT-expressed PIF4 protein as indicated. Competition for PIF4 binding was performed with 10×, 20× and 50× cold <i>YUC8</i> probes containing G-box (G-wt, CACGTG) or mutated G-box (G-mut, CACGGG), respectively. FP, free probe. TnT indicates <i>in vitro</i>-expressed luciferase proteins used as a control.</p

Overexpression of <i>PIF4</i> Increases the Expression of <i>YUC8</i> and Elevates Endogenous Free IAA Levels.

<p>(A) <i>YUC8</i> expression in wild type (Col-0) and <i>35S-PIF4</i> plants. Six-d-old Col-0 and <i>35S-PIF4</i> seedlings grown in normal growth conditions (22°C) were harvested at the same time for RNA extraction and qRT-PCR analyses. Transcript levels of <i>YUC8</i> were normalized to the <i>ACTIN7</i> expression and then were relative to those of Col-0 seedlings. Data shown are average and SD of triplicate reactions. Student's <i>t</i>-test between Col-0 and <i>35S-PIF4</i> seedlings was performed (**, P<0.01). Shown are representative data from one biological replicate; three biological replicates were conducted, yielding similar results. (B–C) <i>PIF4</i> and <i>YUC8</i> expression in transgenic plants containing a chemical-inducible construct <i>pMDC7:PIF4</i>. Eight-d-old <i>pMDC7:PIF4</i> seedlings were untreated or treated with 10 µM estradiol for 3 h before harvest for RNA extraction and qRT-PCR analyses. Transcript levels of target genes were normalized to the <i>ACTIN7</i> expression and then were relative to those of untreated seedlings (0 h). Data shown are average and SD of triplicate reactions. Student's <i>t</i>-test between estradiol-treated and untreated plants was performed (**, P<0.01). Shown are representative data from one biological replicate; three biological replicates were conducted, yielding similar results. (D) Overexpression of <i>PIF4</i> leads to increased free IAA levels. Eight-d-old seedlings of wild type and <i>35S-PIF4</i> seedlings grown in normal growth conditions (22°C) were harvested at the same time for free IAA measurement. Data shown are average±SD. Student's <i>t</i>-test between wild-type and <i>35S-PIF4</i> plants was performed (**, P<0.01). Shown are representative data from one biological replicate; this experiment was conducted for three biological replicates, yielding similar results.</p

PIF4 Activates <i>YUC8</i> Expression, as Revealed by Transient Assays of <i>N. benthamiana</i> Leaves.

<p>(A) Transient expression assays showing that PIF4 activates the expression of <i>YUC8</i>. Representative images of <i>N. benthamiana</i> leaves 72 h after infiltration are shown. The right panel indicates the infiltrated constructs. (B) Quantitative analysis of luminescence intensity in (A). Five independent determinations were assessed. Error bars represent SD. Asterisks denote Student's <i>t</i>-test significance compared with control plants: ***, P<0.001. (C) qRT-PCR analysis of <i>PIF4</i> expression in the infiltrated leaf areas shown in (A). Total RNAs were extracted from leaves of <i>N. benthamiana</i> infiltrated with the constructs. Five independent determinations were assessed. Error bars represent SD.</p

Loss of PIF4 Function Disrupts the High Temperature–Induced Elevation of <i>YUC8</i> Transcripts and Free IAA Levels.

<p>(A–B) High temperature-induced expression patterns of <i>PIF4</i> and <i>YUC8</i> in wild type (Col-0) or the <i>pif4</i> mutant. Six-d-old Col-0 and <i>pif4</i> seedlings grown at 22°C in continuous light were transferred to 29°C in continuous light or were continually placed at 22°C for a 24 h time course, respectively. The 22°C-grown and 29°C-grown seedlings for each time point were harvested at the same time for RNA extraction and qRT-PCR analyses. Transcript levels of target genes were normalized to the <i>ACTIN7</i> expression and were relative to those of untreated Col-0 seedlings (0 h). Data shown are average and SD of triplicate reactions. Shown are representative data from one biological replicate; three biological replicates were conducted, yielding similar results. (C) High temperature-induced elevation of free IAA levels in hypocotyls of Col-0 and <i>pif4</i>. The hypocotyls of 6-d-old wild-type and <i>pif4</i> mutant seedlings grown at 22°C and 29°C in continuous light, respectively, were harvested for free IAA measurement. Data shown are average±SD. Student's <i>t</i>-test between 22°C and 29°C grown plants for each genotype was performed (**, P<0.01). Shown are representative data from one biological replicate; This experiment was conducted for three biological replicates, yielding similar results.</p

The <i>shy2-2</i> Mutation Suppresses the Long-Hypocotyl Phenotype of <i>35S-PIF4</i> Seedlings.

<p>(A) Representative images showing that <i>shy2-2</i> suppresses the long-hypocotyl phenotype of <i>35S-PIF4</i>. Shown are 6-d-old seedlings of Col-0, L<i>er</i>, <i>shy2-2</i>, <i>35S-PIF4</i> and <i>35S-PIF4/shy2-2</i> grown at 22°C. (B) Hypocotyl length showing that <i>shy2-2</i> suppresses the long-hypocotyl phenotype of <i>35S-PIF4</i>. Hypocotyl length of six-d-old seedlings of the indicated genotypes grown at 22°C was measured. Data shown are average±SD. Student's <i>t</i>-test between mutant/transgenic and wild-type seedlings was performed (**, P<0.01). Shown are representative data from one biological replicate; three biological replicates were conducted, yielding similar results.</p

DataSheet1_Characterization of active peptides derived from three leeches and comparison of their anti-thrombotic mechanisms using the tail vein thrombosis model in mice and metabonomics.docx

Background and aims: The increasing incidence of cardiovascular diseases has created an urgent need for safe and effective anti-thrombotic agents. Leech, as a traditional Chinese medicine, has the effect of promoting blood circulation and removing blood stasis, but its real material basis and mechanism of action for the treatment of diseases such as blood stasis and thrombosis have not been reported.Methods: In this study, Whitmania Pigra Whitman (WPW), Hirudo nipponica Whitman (HNW) and Whitmania acranutata Whitman (WAW) were hydrolyzed by biomimetic enzymatic hydrolysis to obtain the active peptides of WPW (APP), the active peptides of HNW (APH) and the active peptides of WAW (APA), respectively. Then their structures were characterized by sykam amino acid analyzer, fourier transform infrared spectrometer (FT-IR), circular dichroism (CD) spectrometer and LC-MS. Next, the anti-thrombotic activities of APP, APH and APA were determined by carrageenan-induced tail vein thrombosis model in mice, and the anti-thrombotic mechanisms of high-dose APP group (HAPP), high-dose APH group (HAPH) and high-dose APA group (HAPA) were explored based on UHPLC-Q-Exactive Orbitrap mass spectrometry.Results: The results showed that the amino acid composition of APP, APH and APA was consistent, and the proportion of each amino acid was few different. The results of FT-IR and CD showed that there were no significant differences in the proportion of secondary structures (such as β-sheet and random coil) and infrared absorption peaks between APP, APH and APA. Mass spectrometry data showed that there were 43 common peptides in APP, APH and APA, indicating that the three have common material basis. APP, APH and APA could significantly inhibit platelet aggregation, reduce black-tail length, whole blood viscosity (WBV), plasma viscosity (PV), and Fibrinogen (FIB), and prolong coagulation time, including activated partial thrombin time (APTT), prothrombin time (PT) and thrombin time (TT). In addition, 24 metabolites were identified as potential biomarkers associated with thrombosis development. Among these, 19, 23, and 20 metabolites were significantly normalized after administration of HAPP, HAPH, and HAPA in the mice, respectively. Furthermore, the intervention mechanism of HAPP, HAPH and HAPA on tail vein thrombosis mainly involved in linoleic acid metabolism, primary bile acid biosynthesis and ether lipid metabolism.Conclusion: Our findings suggest that APP, APH and APA can exert their anti-blood stasis and anti-thrombotic activities by interfering with disordered metabolic pathways in vivo, and there is no significant difference in their efficacies.</p

Sensitive Approaches for the Assay of the Global Protein Tyrosine Phosphorylation in Complex Samples Using a Mutated SH2 Domain

Temporal tyrosine phosphorylation (pTyr) plays a crucial role in numerous cellular functions. The characterization of the tyrosine phosphorylation states of cells is of great interest for understanding the underlying mechanisms. In this study, we developed sensitive and cost-effective methods for the assay of the global protein tyrosine phosphorylation in complex samples by using a novel engineered pTyr binding protein, Src SH2 domain triple-point mutant (Trm-SH2). Taking the advantage of the pan-specific interaction of Trm-SH2 to pTyr, a high throughput approach was developed to determine the total protein tyrosine phosphorylation level in a sample. This method allowed the detection of 0.025 ng of tyrosine phosphorylated proteins. The Trm-SH2 was further exploited to develop a method to profile the global tyrosine phosphorylation state. When this approach was applied to analyze the tyrosine phosphoproteome upon stimulation, distinct patterns were observed. This approach is readily used in many research and clinical fields for the analysis of tyrosine phosphorylated proteins in complex samples, including classifying aberrant phosphotyrosine-dependent signaling in cancer