Project description:RATIONALE: The use of cholecalciferol and calcium carbonate may keep colon cancer from coming back in patients with colon cancer that has been removed by surgery.
PURPOSE: This randomized clinical trial is studying two different doses of cholecalciferol to compare how well they work when given together with calcium carbonate in treating patients with colon cancer that has been removed by surgery.
Project description:Microbially induced carbonate precipitation (MICP) refers to the biogeochemical process in which calcium carbonate is precipitated by altering the local geochemical environment (Mortensen et al. 2011). These alterations occur as a by-product of common microbial metabolic activities by increasing the local carbonate content as well as pH thereby saturating the solution in respect to carbonate. To better understand the microbial ecology of MICP on a community level in natural environments, we chose to evaluate microbial communities derived from travertine adjacent to Crystal Geyser (CG), Utah. CG is a cold-driven, CO2 rich geyser which is surrounded by colorful travertine that has been suggested to be generated through microbial processes. We used a cultivation-independent, multi-omics approach combined with geochemical measurements to investigate metabolic pathways and physiologies potentially involved in MICP at CG. We collected samples from the top 20 cm of travertine adjacent to Crystal Geyser, Utah in November 2019 and June 2021 (38.9384° N, 110.1354° W) wearing gloves at all times. We sampled 1 m away from the borehole (CG-1) and 10 m away from the borehole (CG-10). We preserved all collected samples in RNAlater-like solution (Menke et al., 2017, Front. Microbiol. 8) in a 1:10 sediment: RNAlater-like solution ratio as previously validated (Jensen et al. (2021, Micro. Spec. 2021, 9:2)
Project description:Purpose: The goal of this study is to compare endothelial small RNA transcriptome to identify the target of OASL under basal or stimulated conditions by utilizing miRNA-seq. Methods: Endothelial miRNA profilies of siCTL or siOASL transfected HUVECs were generated by illumina sequencing method, in duplicate. After sequencing, the raw sequence reads are filtered based on quality. The adapter sequences are also trimmed off the raw sequence reads. rRNA removed reads are sequentially aligned to reference genome (GRCh38) and miRNA prediction is performed by miRDeep2. Results: We identified known miRNA in species (miRDeep2) in the HUVECs transfected with siCTL or siOASL. The expression profile of mature miRNA is used to analyze differentially expressed miRNA(DE miRNA). Conclusions: Our study represents the first analysis of endothelial miRNA profiles affected by OASL knockdown with biologic replicates.
Project description:A cDNA library was constructed by Novogene (CA, USA) using a Small RNA Sample Pre Kit, and Illumina sequencing was conducted according to company workflow, using 20 million reads. Raw data were filtered for quality as determined by reads with a quality score > 5, reads containing N < 10%, no 5' primer contaminants, and reads with a 3' primer and insert tag. The 3' primer sequence was trimmed and reads with a poly A/T/G/C were removed
Project description:Whole exome sequencing of 5 HCLc tumor-germline pairs. Genomic DNA from HCLc tumor cells and T-cells for germline was used. Whole exome enrichment was performed with either Agilent SureSelect (50Mb, samples S3G/T, S5G/T, S9G/T) or Roche Nimblegen (44.1Mb, samples S4G/T and S6G/T). The resulting exome libraries were sequenced on the Illumina HiSeq platform with paired-end 100bp reads to an average depth of 120-134x. Bam files were generated using NovoalignMPI (v3.0) to align the raw fastq files to the reference genome sequence (hg19) and picard tools (v1.34) to flag duplicate reads (optical or pcr), unmapped reads, reads mapping to more than one location, and reads failing vendor QC.
Project description:Mitochondria are complex organelles containing 13 proteins encoded by mitochondrial DNA and over 1000 proteins encoded on nuclear DNA. Many mitochondrial proteins are associated with the inner or outer mitochondrial membranes, either peripherally or as integral membrane proteins, while others reside in either of the two soluble mitochondrial compartments, the mitochondrial matrix and the intermembrane space. The biogenesis of the five complexes of the oxidative phosphorylation system are exemplars of this complexity. These large multi-subunit complexes are built through the tightly controlled coalescence of more than 80 proteins with both membrane integral and peripheral associations, with progression between different assembly steps dependent on soluble, membrane integral and peripherally associated assembly factor proteins. Understanding the membrane association of subunits and assembly factors during each assembly step is critical to understanding OXPHOS complex biogenesis. Here we couple sodium carbonate extraction with quantitative mass spectrometry to track changes in the membrane association of the mitochondrial proteome across multiple human complex III knockout cell lines. In addition to identifying the membrane association status of over 840 human mitochondrial proteins, we identify a previously undetected intermediate step of complex III biogenesis. Sodium carbonate extraction with quantitative mass spectrometry (SCE-MS) is thus an easy to apply tool with general utility in the study of mitochondrial respiratory chain biogenesis.
Project description:We use nucleosome maps obtained by high-throughput sequencing to study sequence specificity of intrinsic histone-DNA interactions. In contrast with previous approaches, we employ an analogy between a classical one-dimensional fluid of finite-size particles in an arbitrary external potential and arrays of DNA-bound histone octamers. We derive an analytical solution to infer free energies of nucleosome formation directly from nucleosome occupancies measured in high-throughput experiments. The sequence-specific part of free energies is then captured by fitting them to a sum of energies assigned to individual nucleotide motifs. We have developed hierarchical models of increasing complexity and spatial resolution, establishing that nucleosome occupancies can be explained by systematic differences in mono- and dinucleotide content between nucleosomal and linker DNA sequences, with periodic dinucleotide distributions and longer sequence motifs playing a secondary role. Furthermore, similar sequence signatures are exhibited by control experiments in which genomic DNA is either sonicated or digested with micrococcal nuclease in the absence of nucleosomes, making it possible that current predictions based on highthroughput nucleosome positioning maps are biased by experimental artifacts. Included are raw (eland) and mapped (wig) reads. The mapped reads are provided in eland and wiggle formats, and the raw reads are included in the eland file. This series includes only Mnase control data. The sonicated control is part of this already published accession, as is a in vitro nucleosome map: http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE15188 We also studied data (in vitro and in vivo maps as well as a model) from http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE13622 and from: http://www.ncbi.nlm.nih.gov/sra/?term=SRA001023