Specific binding of the mammalian high mobility group protein AT-hook 2 to the minor groove of AT -rich DNAs: Thermodynamic and specificity studies

The mammalian high mobility group protein AT-hook 2 (HMGA2) is a small transcriptional factor involved in cell development and oncogenesis. It contains three "AT-hook" DNA binding domains, which specifically recognize the minor groove of AT-rich DNA sequences. It also has an acidic C-terminal motif. Previous studies showed that HMGA2 mediates all its biological effects through interactions with AT-rich DNA sequences in the promoter regions. In this dissertation, I used a variety of biochemical and biophysical methods to examine the physical properties of HMGA2 and to further investigate HMGA2's interactions with AT-rich DNA sequences. The following are three avenues perused in this study: (1) due to the asymmetrical charge distribution of HMGA2, I have developed a rapid procedure to purify HMGA2 in the milligram range. Preparation of large amounts of HMGA2 makes biophysical studies possible; (2) Since HMGA2 binds to different AT-rich sequences in the promoter regions, I used a combination of isothermal titration calorimetry (ITC) and DNA UV melting experiment to characterize interactions of HMGA2 with poly(dA-dT) 2 and poly(dA)poly(dT). My results demonstrated that (i) each HMGA2 molecule binds to 15 AT bp; (ii) HMGA2 binds to both AT DNAs with very high affinity. However, the binding reaction of HMGA2 to poly(dA-dT) 2 is enthalpy-driven and the binding reaction of HMGA2 with poly(dA)poly(dT) is entropy-driven; (iii) the binding reactions are strongly depended on salt concentrations; (3) Previous studies showed that HMGA2 may have sequence specificity. In this study, I used a PCR-based SELEX procedure to examine the DNA binding specificity of HMGA2. Two consensus sequences for HMGA2 have been identified: 5'-ATATTCGCGAWWATT-3' and 5'-ATATTGCGCAWWATT-3', where W represents A or T. These consensus sequences have a unique feature: the first five base pairs are AT-rich, the middle four to five base pairs are GC-rich, and the last five to six base pairs are AT-rich. All three segments are critical for high affinity binding. Replacing either one of the AT-rich sequences to a non-AT-rich sequence causes at least 100-fold decrease in the binding affinity. Intriguingly, if the GC-segment is substituted by an AT-rich segment, the binding affinity of HMGA2 is reduced approximately 5-fold. Identification of the consensus sequences for HMGA2 represents an important step towards finding its binding sites within the genome.

The Effect of Nutrient-Rich Effluents from Shrimp Farming on Mangrove Soil Carbon Storage and Geochemistry Under Semi-Arid Climate Conditions in Northern Brazil

A semi-arid mangrove estuary system in the northeast Brazilian coast (Ceará state) was selected for this study to (i) evaluate the impact of shrimp farm nutrient-rich wastewater effluents on the soil geochemistry and organic carbon (OC) storage and (ii) estimate the total amount of OC stored in mangrove soils (0–40 cm). Wastewater-affected mangrove forests were referred to as WAM and undisturbed areas as Non-WAM. Redox conditions and OC content were statistically correlated (P < 0.05) with seasonality and type of land use (WAM vs. Non-WAM). Eh values were from anoxic to oxic conditions in the wet season (from − 5 to 68 mV in WAM and from < 40 to > 400 mV in Non-WAM soils) and significantly higher (from 66 to 411 mV) in the dry season (P < 0.01). OC contents (0–40 cm soil depth) were significantly higher (P < 0.01) in the wet season than the dry season, and higher in Non-WAM soils than in WAM soils (values of 8.1 and 6.7 kg m− 2 in the wet and dry seasons, respectively, for Non-WAM, and values of 3.8 and 2.9 kg m− 2 in the wet and dry seasons, respectively, for WAM soils; P < 0.01). Iron partitioning was significantly dependent (P < 0.05) on type of land use, with a smaller degree of pyritization and lower Fe-pyrite presence in WAM soils compared to Non-WAM soils. Basal respiration of soil sediments was significantly influenced (P < 0.01) by type of land use with highest CO2 flux rates measured in the WAM soils (mean values of 0.20 mg CO2 h− 1–g− 1 C vs. 0.04 mg CO2 h− 1–g− 1 C). The OC storage reduction in WAM soils was potentially caused (i) by an increase in microbial activity induced by loading of nutrient-rich effluents and (ii) by an increase of strong electron acceptors [e.g., NO3−] that promote a decrease in pyrite concentration and hence a reduction in soil OC burial. The current estimated OC stored in mangrove soils (0–40 cm) in the state of Ceará is approximately 1 million t.

The Development of an Optimized System of Narcotic and Explosive Contraband Mimics for Calibration and Training of Biological Detectors

Current commercially available mimics contain varying amounts of either the actual explosive/drug or the chemical compound of suspected interest by biological detectors. As a result, there is significant interest in determining the dominant chemical odor signatures of the mimics, often referred to as pseudos, particularly when compared to the genuine contraband material. This dissertation discusses results obtained from the analysis of drug and explosive headspace related to the odor profiles as recognized by trained detection canines. Analysis was performed through the use of headspace solid phase microextraction in conjunction with gas chromatography mass spectrometry (HS-SPME-GC-MS). Upon determination of specific odors, field trials were held using a combination of the target odors with COMPS. Piperonal was shown to be a dominant odor compound in the headspace of some ecstasy samples and a recognizable odor mimic by trained detection canines. It was also shown that detection canines could be imprinted on piperonal COMPS and correctly identify ecstasy samples at a threshold level of approximately 100ng/s. Isosafrole and/or MDP-2-POH show potential as training aid mimics for non-piperonal based MDMA. Acetic acid was shown to be dominant in the headspace of heroin samples and verified as a dominant odor in commercial vinegar samples; however, no common, secondary compound was detected in the headspace of either. Because of the similarities detected within respective explosive classes, several compounds were chosen for explosive mimics. A single based smokeless powder with a detectable level of 2,4-dinitrotoluene, a double based smokeless powder with a detectable level of nitroglycerine, 2-ethyl-1-hexanol, DMNB, ethyl centralite and diphenylamine were shown to be accurate mimics for TNT-based explosives, NG-based explosives, plastic explosives, tagged explosives, and smokeless powders, respectively. The combination of these six odors represents a comprehensive explosive odor kit with positive results for imprint on detection canines. As a proof of concept, the chemical compound PFTBA showed promise as a possible universal, non-target odor compound for comparison and calibration of detection canines and instrumentation. In a comparison study of shape versus vibration odor theory, the detection of d-methyl benzoate and methyl benzoate was explored using canine detectors. While results did not overwhelmingly substantiate either theory, shape odor theory provides a better explanation of the canine and human subject responses.

Environmental analysis of polar herbicides in complex organic-rich matrices by high performance liquid chromatography atmospheric pressure ionization mass spectrometry (HPLC-API-MS)

A comprehensive forensic investigation of sensitive ecosystems in the Everglades Area is presented. Assessing the background levels of contamination in these ecosystems represents a vital resource to build up forensic evidence required to enforce future environmental crimes within the studied areas. This investigation presents the development and validation of a fractionation and isolation method for two families of herbicides commonly applied in the vicinity of the study area, including phenoxy acids like 2,4-D, MCPA, and silvex; as well as the most common triazine-based herbicides like atrazine, prometyne, simazine and related metabolites like DIA and DEA. Accelerated solvent extraction (ASE) and solid phase extraction (SPE) were used to isolate the analytes from abiotic matrices containing large amounts of organic material. Atmospheric-pressure ionization (API) with electrospray ionization in negative mode (ESP-), and Chemical Ionization in the positive mode (APCI+) were used to perform the characterization of the herbicides of interest.

Inventory of thermo-erosional valleys and streams in three ice-rich permafrost lowlands adjacent to the Laptev Sea, with links to maps in ArcGIS format

This dataset provides an inventory of thermo-erosional landforms and streams in three lowland areas underlain by ice-rich permafrost of the Yedoma-type Ice Complex at the Siberian Laptev Sea coast. It consists of two shapefiles per study region: one shapefile for the digitized thermo-erosional landforms and streams, one for the study area extent. Thermo-erosional landforms were manually digitized from topographic maps and satellite data as line features and subsequently analyzed in a Geographic Information System (GIS) using ArcGIS 10.0. The mapping included in particular thermo-erosional gullies and valleys as well as streams and rivers, since development of all of these features potentially involved thermo-erosional processes. For the Cape Mamontov Klyk site, data from Grosse et al. [2006], which had been digitized from 1:100000 topographic map sheets, were clipped to the Ice Complex extent of Cape Mamontov Klyk, which excludes the hill range in the southwest with outcropping bedrock and rocky slope debris, coastal barrens, and a large sandy floodplain area in the southeast. The mapped features (streams, intermittent streams) were then visually compared with panchromatic Landsat-7 ETM+ satellite data (4 August 2000, 15 m spatial resolution) and panchromatic Hexagon data (14 July 1975, 10 m spatial resolution). Smaller valleys and gullies not captured in the maps were subsequently digitized from the satellite data. The criterion for the mapping of linear features as thermo-erosional valleys and gullies was their clear incision into the surface with visible slopes. Thermo-erosional features of the Lena Delta site were mapped on the basis of a Landsat-7 ETM+ image mosaic (2000 and 2001, 30 m ground resolution) [Schneider et al., 2009] and a Hexagon satellite image mosaic (1975, 10 m ground resolution) [G. Grosse, unpublished data] of the Lena River Delta within the extent of the Lena Delta Ice Complex [Morgenstern et al., 2011]. For the Buor Khaya Peninsula, data from Arcos [2012], which had been digitized based on RapidEye satellite data (8 August 2010, 6.5 m ground resolution), were completed for smaller thermo-erosional features using the same RapidEye scene as a mapping basis. The spatial resolution, acquisition date, time of the day, and viewing geometry of the satellite data used may have influenced the identification of thermo-erosional landforms in the images. For Cape Mamontov Klyk and the Lena Delta, thermo-erosional features were digitized using both Hexagon and Landsat data; Hexagon provided higher resolution and Landsat provided the modern extent of features. Allowance of up to decameters was made for the lateral expansion of features between Hexagon and Landsat acquisitions (between 1975 and 2000).

Enhanced microbial activity in carbon-rich pillow lavas, Ordovician, Great Britain and Ireland

Date of acceptance: 09/07/2015

Enhanced microbial activity in carbon-rich pillow lavas, Ordovician, Great Britain and Ireland

Date of acceptance: 09/07/2015

Transplanting the leafy liverwort Herbertus hutchinsiae : A suitable conservation tool to maintain oceanic-montane liverwort-rich heath?

Peer reviewed

Availability and dose response of phytophenols from a wheat bran-rich cereal product in healthy human volunteers

Funding: Kellogg’s and the Scottish Government Food Land and People Programme.

Enhanced microbial activity in carbon-rich pillow lavas, Ordovician, Great Britain and Ireland

Date of acceptance: 09/07/2015

Poet Adrienne Rich

General note: Title and date provided by Bettye Lane.

Catharine Stimpson, Barbara Deming, Adrienne Rich, and Gloria Orenstein attending The Woman's Salon.

Inscription: Verso: The Woman's Salon, New York. Left to right: Catharine Stimpson, Barbara Deming, Adrienne Rich, and Gloria Orenstein.

Left to right: Adrienne Rich, Barbara Demin, Erika Duncan, Glora Orenstein at the Woman's Salon

Inscriptions: Verso: [stamped] Photograph by Freda Leinwand. [463 West Street, Studio 229G, New York, NY 10014].

Barbara Deming, Erika Duncan, Adrienne Rich, and Gloria Orenstein at The Woman's Salon

Inscription: Verso: The Woman's Salon, New York. Seated on platform: Barbara Deming, Erika Duncan, Adrienne Rich, and Gloria Orenstein.

(Table 1, page 126) Concentration of elements in Fe and Mn rich pisoliths (nodules) from Lake Storsjoen, South Norway

A detailed description of the ores of Lake Storsjoen was given by Vogt J. H. L., 1915 who pointed out that the ores may be divided into two principal types; first, iron ore with 2% or less of manganese (ex: Ertemalm), and, second, ores with manganese contents of up to 30% (ex: Korinter). The iron-rich ore sometimes occurs as a conglomerate embedded in manganese-rich ores, clearly demonstrating that two distinctly different precipitates are involved. In the iron-rich ore, a concentric structure is common of which light brown layers of loose, almost dusty material alternate with hard and brittle black layers, the thickness of each being 0.5 mm or less. The analyses presented in this paper seem to demonstrate that the composition of the sedimentary ores of Lake Storsjden could result from fluctuations in the composition of ground waters feeding the lake.