Rational design of a bacterial import system for new-to-nature molecules.

Integration of novel compounds into biological processes holds significant potential for modifying or expanding existing cellular functions. However, the cellular uptake of these compounds is often hindered by selectively permeable membranes. We present a novel bacterial transport system that has been rationally designed to address this challenge.

Controlled E. coli Aggregation Mediated by DNA and XNA Hybridization.

Chemical cell surface modification is a fast-growing field of research, due to its enormous potential in tissue engineering, cell-based immunotherapy, and regenerative medicine. However, engineering of bacterial tissues by chemical cell surface modification has been vastly underexplored and the identification of suitable molecular handles is in dire need. We present here, an orthogonal nucleic acid-protein conjugation strategy to promote artificial bacterial aggregation.

Projected mid-century rainfall erosivity under climate change over the southeastern United States.

Recent observations and climate change projections indicate that changes in rainfall energy, intensity, duration, and frequency, which determine the erosive power of rainfall, will amplify erosion rates around the world. However, the magnitude and scope of these future changes in erosive power of rainfall remain largely unknown, particularly at finer-resolutions and local scales. Due to a lack of available projected future sub-hourly climate data, previous studies relied on aggregates (hourly, daily) rainfall data.

Temporal disaggregation of hourly precipitation under changing climate over the Southeast United States.

Climate change impacts on precipitation characteristics will alter the hydrologic characteristics, such as peak flows, time to peak, and erosion potential of watersheds. However, many of the currently available climate change datasets are provided at temporal and spatial resolutions that are inadequate to quantify projected changes in hydrologic characteristics of a watershed. Therefore, it is critical to temporally disaggregate coarse-resolution precipitation data to finer resolutions for studies sensitive to precipitation characteristics.

Improved forest dynamics leads to better hydrological predictions in watershed modeling.

This study explored how the characterization of forest processes in hydrologic models affects watershed hydrological responses. To that end, we applied the widely used Soil and Water Assessment Tool (SWAT) model to two forested watersheds in the southeastern United States. Although forests can cover a large portion of watersheds, tree attributes such as leaf area index (LAI), biomass accumulation, and processes such as evapotranspiration (ET) are rarely calibrated in hydrological modeling studies.

Improving the representation of forests in hydrological models.

Forests play a critical role in the hydrologic cycle, impacting the surface and groundwater dynamics of watersheds through transpiration, interception, shading, and modification of the atmospheric boundary layer. It is therefore critical that forest dynamics are adequately represented in watershed models, such as the widely applied Soil and Water Assessment Tool (SWAT). SWAT's default parameterization generally produces unrealistic forest growth predictions, which we address here through an improved representation of forest dynamics using species-specific re-parameterizations.

Improved agricultural Water management in data-scarce semi-arid watersheds: Value of integrating remotely sensed leaf area index in hydrological modeling.

In watersheds located in semi-arid regions, vegetation dynamics, evapotranspiration (ET), and associated water and energy balances collectively play a major role in controlling hydrological regimes and crop yield. As such, it is challenging to predict the complex hydrological processes and biophysical dynamics. This challenge increases in areas with limited data availability.

-Glycosylated 8-Azapurine and Methylated Purine Nucleobases: Synthesis and Study of Base Pairing Properties.

In this report, we present the synthesis of -glycosylated 8-aza-2-methylhypoxanthine and 8-aza-6-thiohypoxanthine 2'-deoxynucleosides as well as methylated 2'-deoxynebularine derivatives. In vitro base pairing properties between each modified and canonical nucleobase were studied. As demonstrated by , incorporation of the modified bases in DNA resulted, with few exceptions, in low stability of duplexes.

Mutant Variants of the Substrate-Binding Protein DppA from Escherichia coli Enhance Growth on Nonstandard γ-Glutamyl Amide-Containing Peptides.

The import of nonnatural molecules is a recurring problem in fundamental and applied aspects of microbiology. The dipeptide permease (Dpp) of is an ABC-type multicomponent transporter system located in the cytoplasmic membrane, which is capable of transporting a wide range of di- and tripeptides with structurally and chemically diverse amino acid side chains into the cell. Given this low degree of specificity, Dpp was previously used as an entry gate to deliver natural and nonnatural cargo molecules into the cell by attaching them to amino acid side chains of peptides, in particular, the γ-carboxyl group of glutamate residues.

Synthesis and antiviral evaluation of cyclopentyl nucleoside phosphonates.

The synthesis of both 2'-hydroxy-3'-deoxy and 2'-deoxy-3'-hydroxy cyclopentyl nucleoside phosphonates with the natural nucleobases adenine, thymine, cytosine and guanine from a single precursor has been performed. The guanine containing analogues showed antiviral activity. Especially the 3'-deoxy congener 23 was active, displaying an EC of 5.

Overcoming the membrane barrier: Recruitment of γ-glutamyl transferase for intracellular release of metabolic cargo from peptide vectors.

Semipermeable membranes of cells frequently pose an obstacle in metabolic engineering by limiting uptake of substrates, intermediates, or xenobiotics. Previous attempts to overcome this barrier relied on the promiscuous nature of peptide transport systems, but often suffered from low versatility or chemical instability. Here, we present an alternative strategy to transport cargo molecules across the inner membrane of Escherichia coli based on chemical synthesis of a stable cargo-peptide vector construct, transport through the peptide import system, and efficient intracellular release of the cargo by the promiscuous enzyme γ-glutamyl transferase (GGT).

Incorporation of Amino Acids with Long-Chain Terminal Olefins into Proteins.

The increasing need for site-specific protein decorations that mimic natural posttranslational modifications requires access to a variety of noncanonical amino acids with moieties enabling bioorthogonal conjugation chemistry. Here we present the incorporation of long-chain olefinic amino acids into model proteins with rational variants of pyrrolysyl-tRNA synthetase (PylRS). Nε-heptenoyl lysine was incorporated for the first time using the known promiscuous variant PylRS(Y306A/Y384F), and Nε-pentenoyl lysine was incorporated in significant yields with the novel variant PylRS(C348A/Y384F).

Design and synthesis of nucleolipids as possible activated precursors for oligomer formation via intramolecular catalysis: stability study and supramolecular organization.

Background: Fatty acid vesicles are an important part of protocell models currently studied. As protocells can be considered as pre-biological precursors of cells, the models try to contribute to a better understanding of the (cellular) origin of life and emphasize on 2 major aspects: compartmentalization and replication. It has been demonstrated that lipid-based membranes are amenable to growth and division (shell replication).

Nutrient loss in leachate and surface runoff from surface-broadcast and subsurface-banded broiler litter.

Subsurface band application of poultry litter has been shown to reduce the transport of nutrients from fields in surface runoff compared with conventional surface broadcast application. Little research has been conducted to determine the effects of surface broadcast application and subsurface banding of litter on nutrients in leachate. Therefore, a field experiment was conducted to determine the effects of subsurface band application and surface broadcast application of poultry litter on nutrient losses in leachate.

Watershed-level comparison of predictability and sensitivity of two phosphorus models.

Buildup of phosphorus (P) in agricultural soils and transport of P to nearby surface waters due to excessive, long-term application of poultry litter is an environmental concern in many poultry-producing states. Watershed models are often used to quantify soil and water quality impacts of poultry litter applications. However, depending on how P transport is simulated in watershed models, the anticipated impact could be quite different.

Effects of chronic bhang (cannabis) administration on the reproductive system of male mice.

Background: The purpose of this study was to investigate the effect of chronic uptake of bhang, prepared from the Cannabis sativa, on male reproductive physiology in adult male Parkes strain (P) mice. An attempt was also made to investigate the presence of cannabinoid 1 (CB1) and cannabinoid 2 (CB2) receptors, and fatty acid amide hydrolase (FAAH) in the testis and to evaluate any changes in it resulting from chronic intake of bhang in mice.

Methods And Results: Adult male mice were given bhang (3 or 6 mg/kg body weight/day) orally for 36 consecutive days.

An ecologically-sustainable surface water withdrawal framework for cropland irrigation: a case study in Alabama.

Agricultural production in the state of Alabama, USA, is mostly rain-fed, because of which it is vulnerable to drought during growing season. Since Alabama receives a significant portion of its annual precipitation during winter months, the goal of this study was to evaluate the feasibility of water withdrawal from streams during winter months for irrigation in the growing season. The Soil and Water Assessment Tool (SWAT) was used to estimate the quantity of water that can be sustainably withdrawn from streams during winter high flow periods.

A large-scale chemical modification screen identifies design rules to generate siRNAs with high activity, high stability and low toxicity.

The use of chemically synthesized short interfering RNAs (siRNAs) is currently the method of choice to manipulate gene expression in mammalian cell culture, yet improvements of siRNA design is expectably required for successful application in vivo. Several studies have aimed at improving siRNA performance through the introduction of chemical modifications but a direct comparison of these results is difficult. We have directly compared the effect of 21 types of chemical modifications on siRNA activity and toxicity in a total of 2160 siRNA duplexes.

Assessment of economic and water quality impacts of land use change using a simple bioeconomic model.

The objective of this study is to assess the economic and water quality impact of land use change in a small watershed in the Wiregrass region of Alabama. The study compares changes in water quality and revenue from agricultural and timber production due to changes in land use between years 1992 and 2001. The study was completed in two stages.

Five- and six-membered conformationally locked 2',4'-carbocyclic ribo-thymidines: synthesis, structure, and biochemical studies.

Two unusual reactions involving the 5-hexenyl or the 6-heptenyl radical cyclization of a distant double bond at C4' and the radical center at C2' of the ribofuranose ring of thymidine have been used as key steps to synthesize North-type conformationally constrained cis-fused bicyclic five-membered and six-membered carbocyclic analogues of LNA (carbocyclic-LNA-T) and ENA (carbocyclic-ENA-T) in high yields. Their structures have been confirmed unambiguously by long range 1H-13C NMR correlation (HMBC), TOCSY, COSY, and NOE experiments. The carbocyclic-LNA-T and carbocyclic-ENA-T were subsequently incorporated into the antisense oligonucleotides (AONs) to show that they enhance the Tm of the modified AON/RNA heteroduplexes by 3.