Phase I Cultural Resources Survey Of The Proposed Texas Eastern Transmission, Lp, Dot 2015 - Mexi-Stfe Project, In Hidalgo County, Texas

This document describes the results of Phase I cultural resources survey of the proposed Texas Eastern Transmission, LP, DOT 2015 – MEXI-STFE Project, in Hidalgo County, Texas (Figures 1.1 and 1.2). The project entailed the examination of a 275 m (2378.6 ft) long pipeline replacement, as well as 2.1 ha (5.2 ac) of additional temporary workspace and 826.0 m (2710 ft) of project access roads. Together these project items totaled 4.4 ha (10.9 ac) in area. This investigation was completed on behalf of Texas Eastern Transmission, LP, by R. Christopher Goodwin & Associates, Inc. in May of 2015. The goal of fieldwork was to identify and to evaluate all historic properties, archeological sites, cultural resources loci, standing structures, and/or cemeteries that may be impacted adversely by the proposed project. All work was performed in accordance with the procedures outlined in the National Historic Preservation Act of 1966, as amended; the Archaeological and Historic Preservation Act of 1974; the Archaeological Resources Protection Act of 1979, as amended; and Title 36 of the Code of Federal Regulations, Parts 60-66 and 800, as appropriate. Additionally, this survey effort abided by the standards set forth in the Federal Energy Regulatory Commission, Office of Pipeline Regulation’s Guidelines for Reporting on Cultural Resources Investigations (1994:11-13), the Archeology and Historic Preservation: The Secretary of the Interior’s Guidelines, and with guidelines contained within the Texas Historical Commission’s Preserving Our Heritage: A Statewide Plan for Texas. Information provided by the Council of Texas Archeologists, the Texas Archaeological Research Laboratory, the Antiquities Code of Texas, and the Texas Historical Commission’s Rules of Practice and Procedure for the Antiquities Code of Texas, also was utilized.. All of the proposed project items fell within the bounds of the Louisiana-Rio Grande Canal Company Irrigation System National Register District. The historic property consists of two pumping stations and a network of canals, irrigation pipes, and drainage ditches that extend approximately 500 miles total in length (Meyers and Weitze 1995). The two pumping stations are located outside the project area, but the project area is in the vicinity of drainage ditches likely part of the former Louisiana – Rio Grande Canal Company system, now under the jurisdiction of the Hidalgo County Irrigation District No. 2. Despite the proximity of the district, the proposed projects, currently defined as replacement of an existing pipeline and hydrostatic testing, will not pose an adverse effect to, or have any additional visual impacts on, the district. If the project scope changes, further investigation of this national register property may be warranted. Furthermore, no cultural resources were identified within the limits of the current survey areas. No additional investigation or recordation of the project items associated with the Texas Eastern Transmission, LP, Project Name Project is recommended

Transcriptome Analysis of Single Cells

Many gene expression analysis techniques rely on material isolated from heterogeneous populations of cells from tissue homogenates or cells in culture.1,2,3 In the case of the brain, regions such as the hippocampus contain a complex arrangement of different cell types, each with distinct mRNA profiles. The ability to harvest single cells allows for a more in depth investigation into the molecular differences between and within cell populations. We describe a simple and rapid method for harvesting cells for further processing. Pipettes often used in electrophysiology are utilized to isolate (using aspiration) a cell of interest and conveniently deposit it into an Eppendorf tube for further processing with any number of molecular biology techniques. Our protocol can be modified for the harvest of dendrites from cell culture or even individual cells from acute slices

The Iddm14 gene is Tcrbv-13S1A1: Prevention of Autoimmune Diabetes in the Rat with an Allele-Specific Depleting Antibody That Recognizes the Vβ13a T Cell Receptor Beta Chain

To identify new intervention strategies for autoimmune type 1 diabetes (T1D), we investigated several rat models of the disorder. We dissected the powerful Iddm14 diabetes susceptibility locus in eight T1D susceptible vs. resistant rat strains by single nucleotide polymorphism (SNP) haplotyping. We identified an allele of a T cell receptor (TCR) beta chain gene, Tcrb-V13S1A1 (encoding V13βa) as a candidate gene. In three separate trials, treating LEW.1WR1 rats, which are susceptible to T1D, with a depleting anti-Vβ13 monoclonal antibody reduced diabetes frequency from 75% (N=50) to 17% (N=30, p\u3c0.001. Anti-Vβ13 monoclonal antibody also prevented T1D in spontaneously diabetic BBDP rats. We then analyzed the phenotype of infiltrating T cells recovered from the cultured islets of LEW.1WR1 rats exposed to a diabetogenic trigger. Within 5 days, up to 22% of CD4+ T cells recovered from islets were V13β+, most of these CD25+FoxP3-. We also recovered Vβ13 transcripts from pre-diabetic islets and observed a limited number of Jβ variant transcripts, indicating an oligoclonal TCR response to pancreatic beta cells. These data indicate that, in susceptible rats, V13βa on diabetogenic T cells is required to recognize a critical T1D autoantigen. Interestingly, the diabetogenic and non-diabetogenic alleles of Vβ13 have non-conservative sequence differences in both CRR1 and CDR2. The data suggest that it is possible to prevent T1D in the rat with a very narrowly targeted deletional therapy. Preliminary data suggest that a specific alpha chain may preferentially pair with Vβ13a. We are currently generating rat T cell hybridoma clones with which to analyze the interaction of putative autoantigens with a diabetogenic TCR

In situ reverse transcription: the magic of strength and anonymity

In this study, we describe an approach that enables a highly specific, effective and fast detection of polyadenylated RNA sequences in situ at the light and electron microscopy levels. The method developed is based on the incorporation of 5-bromo-2′-deoxyuridine into the growing cDNA strand by means of the reverse transcriptase. We have shown that unlike the previously used deoxyuridine tagged with biotin or digoxigenin, 5-bromo-2′-deoxyuridine is ‘invisible’ in the DNA–DNA duplex but easily detectable in the DNA–RNA duplex. This feature is an important pre-requisite for the correct interpretation of the data obtained, as our results strongly indicate that reverse transcriptase uses DNA breaks as primers efficiently. We have also shown that the replacement of deoxythymidine by 5-bromo-2′-deoxyuridine considerably stabilizes the growing DNA–RNA duplex, thus enabling the one-step detection of polyadenylated RNA in structurally well-preserved cells. The method developed provides a highly specific signal with the signal/noise ratio higher than 130 for permeabilized cells and 25 for conventional acrylic resin sections under the conditions used. When the high pressure freezing technique followed by the freeze substitution is employed for the cell's preparation, the ratio is higher than 80

The Missing Heritability in T1D and Potential New Targets for Prevention

Type 1 diabetes (T1D) is a T cell-mediated disease. It is strongly associated with susceptibility haplotypes within the major histocompatibility complex, but this association accounts for an estimated 50% of susceptibility. Other studies have identified as many as 50 additional susceptibility loci, but the effect of most is very modest (odds ratio (OR) 5) and that deletion of V beta 13+ T cells prevents diabetes. A role for the TCR is also suspected in NOD mice, but TCR regions have not been associated with human T1D. To investigate this disparity, we tested the hypothesis in silico that previous studies of human T1D genetics were underpowered to detect MHC-contingent TCR susceptibility. We show that stratifying by MHC markedly increases statistical power to detect potential TCR susceptibility alleles. We suggest that the TCR regions are viable candidates for T1D susceptibility genes, could account for missing heritability, and could be targets for prevention