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Description
Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR.

Zoonotic pathogens that cause leprosy (Mycobacterium leprae) and tuberculosis (Mycobacterium tuberculosis complex, MTBC) continue to impact modern human populations. Therefore, methods able to survey mycobacterial infection in potential animal hosts are necessary for proper evaluation of human exposure threats. Here we tested for mycobacterial-specific single- and multi-copy loci using qPCR. In a trial study in which armadillos were artificially infected with M. leprae, these techniques were specific and sensitive to pathogen detection, while more traditional ELISAs were only specific. These assays were then employed in a case study to detect M. leprae as well as MTBC in wild marmosets. All marmosets were negative for M. leprae DNA, but 14 were positive for the mycobacterial rpoB gene assay. Targeted capture and sequencing of rpoB and other MTBC genes validated the presence of mycobacterial DNA in these samples and revealed that qPCR is useful for identifying mycobacterial-infected animal hosts.
Created2015-11-16
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Description

Nutrient availability and ratios can play an important role in shaping microbial communities of freshwater ecosystems. The Cuatro Ciénegas Basin (CCB) in Mexico is a desert oasis where, perhaps paradoxically, high microbial diversity coincides with extreme oligotrophy. To better understand the effects of nutrients on microbial communities in CCB, a

Nutrient availability and ratios can play an important role in shaping microbial communities of freshwater ecosystems. The Cuatro Ciénegas Basin (CCB) in Mexico is a desert oasis where, perhaps paradoxically, high microbial diversity coincides with extreme oligotrophy. To better understand the effects of nutrients on microbial communities in CCB, a mesocosm experiment was implemented in a stoichiometrically imbalanced pond, Lagunita, which has an average TN:TP ratio of 122 (atomic). The experiment had four treatments, each with five spatial replicates – unamended controls and three fertilization treatments with different nitrogen:phosphorus (N:P) regimes (P only, N:P = 16 and N:P = 75 by atoms). In the water column, quantitative PCR of the 16S rRNA gene indicated that P enrichment alone favored proliferation of bacterial taxa with high rRNA gene copy number, consistent with a previously hypothesized but untested connection between rRNA gene copy number and P requirement. Bacterial and microbial eukaryotic community structure was investigated by pyrosequencing of 16S and 18S rRNA genes from the planktonic and surficial sediment samples. Nutrient enrichment shifted the composition of the planktonic community in a treatment-specific manner and promoted the growth of previously rare bacterial taxa at the expense of the more abundant, potentially endemic, taxa. The eukaryotic community was highly enriched with phototrophic populations in the fertilized treatment. The sediment microbial community exhibited high beta diversity among replicates within treatments, which obscured any changes due to fertilization. Overall, these results showed that nutrient stoichiometry can be an important factor in shaping microbial community structure.

ContributorsLee, Zarraz (Author) / Poret-Peterson, Amisha (Author) / Siefert, Janet L. (Author) / Kaul, Drishti (Author) / Moustafa, Ahmed (Author) / Allen, Andrew E. (Author) / Dupont, Chris L. (Author) / Eguiarte, Luis E. (Author) / Souza, Valeria (Author) / Elser, James (Author) / College of Liberal Arts and Sciences (Contributor) / School of Life Sciences (Contributor) / School of Earth and Space Exploration (Contributor)
Created2017-05-30
Description

Background:
Many pharmaceutical drugs are known to be ineffective or have negative side effects in a substantial proportion of patients. Genomic advances are revealing that some non-synonymous single nucleotide variants (nsSNVs) may cause differences in drug efficacy and side effects. Therefore, it is desirable to evaluate nsSNVs of interest in their

Background:
Many pharmaceutical drugs are known to be ineffective or have negative side effects in a substantial proportion of patients. Genomic advances are revealing that some non-synonymous single nucleotide variants (nsSNVs) may cause differences in drug efficacy and side effects. Therefore, it is desirable to evaluate nsSNVs of interest in their ability to modulate the drug response.

Results:
We found that the available data on the link between drug response and nsSNV is rather modest. There were only 31 distinct drug response-altering (DR-altering) and 43 distinct drug response-neutral (DR-neutral) nsSNVs in the whole Pharmacogenomics Knowledge Base (PharmGKB). However, even with this modest dataset, it was clear that existing bioinformatics tools have difficulties in correctly predicting the known DR-altering and DR-neutral nsSNVs. They exhibited an overall accuracy of less than 50%, which was not better than random diagnosis. We found that the underlying problem is the markedly different evolutionary properties between positions harboring nsSNVs linked to drug responses and those observed for inherited diseases. To solve this problem, we developed a new diagnosis method, Drug-EvoD, which was trained on the evolutionary properties of nsSNVs associated with drug responses in a sparse learning framework. Drug-EvoD achieves a TPR of 84% and a TNR of 53%, with a balanced accuracy of 69%, which improves upon other methods significantly.

Conclusions:
The new tool will enable researchers to computationally identify nsSNVs that may affect drug responses. However, much larger training and testing datasets are needed to develop more reliable and accurate tools.

ContributorsGerek, Nevin Z. (Author) / Liu, Li (Author) / Gerold, Kristyn (Author) / Biparva, Pegah (Author) / Thomas, Eric D. (Author) / Kumar, Sudhir (Author) / Biodesign Institute (Contributor) / Center for Evolution and Medicine (Contributor)
Created2015-01-15
Description
Microalgae-derived lipids are good sources of biofuel, but extracting them involves high cost, energy
expenditure, and environmental risk. Surfactant treatment to disrupt Scenedesmus biomass was evaluated
as a means to make solvent extraction more efficient. Surfactant treatment increased the recovery of fatty
acid methyl ester (FAME) by as much as 16-fold vs. untreated

Microalgae-derived lipids are good sources of biofuel, but extracting them involves high cost, energy
expenditure, and environmental risk. Surfactant treatment to disrupt Scenedesmus biomass was evaluated
as a means to make solvent extraction more efficient. Surfactant treatment increased the recovery of fatty
acid methyl ester (FAME) by as much as 16-fold vs. untreated biomass using isopropanol extraction, and
nearly 100% FAME recovery was possible without any Folch solvent, which is toxic and expensive. Surfactant
treatment caused cell disruption and morphological changes to the cell membrane, as documented by
transmission electron microscopy and flow cytometry. Surfactant treatment made it possible to extract wet
biomass at room temperature, which avoids the expense and energy cost associated with heating
and drying of biomass during the extraction process. The best FAME recovery was obtained from highlipid
biomass treated with Myristyltrimethylammonium bromide (MTAB)- and 3-(decyldimethylammonio)-
propanesulfonate inner salt (3_DAPS)-surfactants using a mixed solvent (hexane : isopropanol = 1 : 1, v/v)
vortexed for just 1 min; this was as much as 160-fold higher than untreated biomass. The critical micelle
concentration of the surfactants played a major role in dictating extraction performance, but the growth
stage of the biomass had an even larger impact on how well the surfactants disrupted the cells and
improved lipid extraction. Surfactant treatment had minimal impact on extracted-FAME profiles and,
consequently, fuel-feedstock quality. This work shows that surfactant treatment is a promising strategy for
more efficient, sustainable, and economical extraction of fuel feedstock from microalgae.
Created2015-10-20
Description
Using a CH[subscript 4]-based membrane biofilm reactor (MBfR), we studied perchlorate (ClO[subscript 4]–) reduction by a biofilm performing anaerobic methane oxidation coupled to denitrification (ANMO-D). We focused on the effects of nitrate (NO[subscript 3]–) and nitrite (NO[subscript 2]–) surface loadings on ClO[subscript 4]– reduction and on the biofilm community’s mechanism

Using a CH[subscript 4]-based membrane biofilm reactor (MBfR), we studied perchlorate (ClO[subscript 4]–) reduction by a biofilm performing anaerobic methane oxidation coupled to denitrification (ANMO-D). We focused on the effects of nitrate (NO[subscript 3]–) and nitrite (NO[subscript 2]–) surface loadings on ClO[subscript 4]– reduction and on the biofilm community’s mechanism for ClO[subscript 4]– reduction. The ANMO-D biofilm reduced up to 5 mg/L of ClO[subscript 4]– to a nondetectable level using CH[subscript 4] as the only electron donor and carbon source when CH[subscript 4] delivery was not limiting; NO[subscript 3]– was completely reduced as well when its surface loading was ≤0.32 g N/m[superscript 2]-d. When CH[subscript 4] delivery was limiting, NO[subscript 3]– inhibited ClO[subscript 4]– reduction by competing for the scarce electron donor. NO[subscript 2]– inhibited ClO[subscript 4]– reduction when its surface loading was ≥0.10 g N/m[superscript 2]-d, probably because of cellular toxicity. Although Archaea were present through all stages, Bacteria dominated the ClO[subscript 4]–-reducing ANMO-D biofilm, and gene copies of the particulate methane mono-oxygenase (pMMO) correlated to the increase of respiratory gene copies. These pieces of evidence support that ClO[subscript 4]– reduction by the MBfR biofilm involved chlorite (ClO[subscript 2]–) dismutation to generate the O[subscript 2] needed as a cosubstrate for the mono-oxygenation of CH[subscript 4].
ContributorsLuo, Yi-Hao (Author) / Chen, Ran (Author) / Wen, Li-Lian (Author) / Meng, Fan (Author) / Zhang, Yin (Author) / Lai, Chun-Yu (Author) / Rittmann, Bruce (Author) / Zhao, He-Ping (Author) / Zheng, Ping (Author) / Biodesign Institute (Contributor) / Swette Center for Environmental Biotechnology (Contributor)
Created2015-02-17
Description
Herbivores face various nutritional challenges in their life cycles, challenges that may become increasingly acute under ongoing environmental changes.
Here, focusing on calcium, phosphorus and nitrogen, we used nutritional geometry to analyse individual-based data on foraging and extraction efficiencies, and combined these with data on reproduction and migratory behaviour to understand

Herbivores face various nutritional challenges in their life cycles, challenges that may become increasingly acute under ongoing environmental changes.
Here, focusing on calcium, phosphorus and nitrogen, we used nutritional geometry to analyse individual-based data on foraging and extraction efficiencies, and combined these with data on reproduction and migratory behaviour to understand how a large herbivorous carnivore can complete its life cycle on a narrow and seemingly low quality bamboo diet.
Behavioural results showed that pandas during the year switched between four main food categories involving the leaves and shoots of two bamboo species available. Nutritional analysis suggests that these diet shifts are related to the concentrations and balances of calcium, phosphorus and nitrogen. Notably, successive shifts in range use and food type corresponded with a transition to higher concentrations and/or a more balanced intake of these multiple key constituents.
Our study suggests that pandas obligatorily synchronize their seasonal migration and reproduction with the disjunct nutritional phenologies of two bamboo species. This finding has potentially important implications for habitat conservation for this species and, more generally, draws attention to the need for understanding the nutritional basis of food selection in devising management plans for endangered species.
ContributorsNie, Yonggang (Author) / Zhang, Zejun (Author) / Raubenheimer, David (Author) / Elser, James (Author) / Wei, Wei (Author) / Wei, Fuwen (Author) / College of Liberal Arts and Sciences (Contributor) / School of Life Sciences (Contributor)
Created2015-01-01
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Description

I present the case for a fire-centric scholarship, and suggest the transition between burning living landscapes and lithic ones (in the form of fossil fuels) would make a good demonstration of what such scholarship might do and what its value could be.

ContributorsPyne, Stephen (Author) / College of Liberal Arts and Sciences (Contributor) / School of Life Sciences (Contributor)
Created2017-10-23
Description
To achieve nitrite accumulation for shortcut biological nitrogen removal (SBNR) in a biofilm process, we explored the simultaneous effects of oxygen limitation and free ammonia (FA) and free nitrous acid (FNA) inhibition in the nitrifying biofilm. We used the multi-species nitrifying biofilm model (MSNBM) to identify conditions that should or

To achieve nitrite accumulation for shortcut biological nitrogen removal (SBNR) in a biofilm process, we explored the simultaneous effects of oxygen limitation and free ammonia (FA) and free nitrous acid (FNA) inhibition in the nitrifying biofilm. We used the multi-species nitrifying biofilm model (MSNBM) to identify conditions that should or should not lead to nitrite accumulation, and evaluated the effectiveness of those conditions with experiments in continuous flow biofilm reactors (CFBRs). CFBR experiments were organized into four sets with these expected outcomes based on the MSNBM as follows: (i) Control, giving full nitrification; (ii) oxygen limitation, giving modest long-term nitrite build up; (iii) FA inhibition, giving no long-term nitrite accumulation; and (iv) FA inhibition plus oxygen limitation, giving major long-term nitrite accumulation. Consistent with MSNBM predictions, the experimental results showed that nitrite accumulated in sets 2–4 in the short term, but long-term nitrite accumulation was maintained only in sets 2 and 4, which involved oxygen limitation. Furthermore, nitrite accumulation was substantially greater in set 4, which also included FA inhibition. However, FA inhibition (and accompanying FNA inhibition) alone in set 3 did not maintained long-term nitrite accumulation. Nitrite-oxidizing bacteria (NOB) activity batch tests confirmed that little NOB or only a small fraction of NOB were present in the biofilms for sets 4 and 2, respectively. The experimental data supported the previous modeling results that nitrite accumulation could be achieved with a lower ammonium concentration than had been required for a suspended-growth process. Additional findings were that the biofilm exposed to low dissolved oxygen (DO) limitation and FA inhibition was substantially denser and probably had a lower detachment rate.
ContributorsPark, Seongjun (Author) / Chung, Jinwook (Author) / Rittmann, Bruce (Author) / Bae, Wookeun (Author) / Biodesign Institute (Contributor) / Swette Center for Environmental Biotechnology (Contributor)
Created2015-01-01
Description
This work presents a spectroscopic study of the thermally enhanced photoinduced electron emission from nitrogen-doped diamond films prepared on p-type silicon substrates. It has been shown that photon-enhanced thermionic emission (PETE) can substantially enhance thermionic emission intensity from a p-type semiconductor. An n-type diamond/p-type silicon structure was illuminated with 400–450

This work presents a spectroscopic study of the thermally enhanced photoinduced electron emission from nitrogen-doped diamond films prepared on p-type silicon substrates. It has been shown that photon-enhanced thermionic emission (PETE) can substantially enhance thermionic emission intensity from a p-type semiconductor. An n-type diamond/p-type silicon structure was illuminated with 400–450 nm light, and the spectra of the emitted electrons showed a work function less than 2 eV and nearly an order of magnitude increase in emission intensity as the temperature was increased from ambient to ∼400 °C. Thermionic emission was negligible in this temperature range. The results are modeled in terms of contributions from PETE and direct photoelectron emission, and the large increase is consistent with a PETE component. The results indicate possible application in combined solar/thermal energy conversion devices.
ContributorsSun, Tianyin (Author) / Koeck, Franz (Author) / Rezikyan, Aram (Author) / Treacy, Michael (Author) / Nemanich, Robert (Author) / College of Liberal Arts and Sciences (Contributor) / Department of Physics (Contributor)
Created2014-09-15
Description
Heterochromatin is a repressive chromatin compartment essential for maintaining genomic integrity. A hallmark of heterochromatin is the presence of specialized nonhistone proteins that alter chromatin structure to inhibit transcription and recombination. It is generally assumed that heterochromatin is highly condensed. However, surprisingly little is known about the structure of heterochromatin

Heterochromatin is a repressive chromatin compartment essential for maintaining genomic integrity. A hallmark of heterochromatin is the presence of specialized nonhistone proteins that alter chromatin structure to inhibit transcription and recombination. It is generally assumed that heterochromatin is highly condensed. However, surprisingly little is known about the structure of heterochromatin or its dynamics in solution. In budding yeast, formation of heterochromatin at telomeres and the homothallic silent mating type loci require the Sir3 protein. Here, we use a combination of sedimentation velocity, atomic force microscopy and nucleosomal array capture to characterize the stoichiometry and conformation of Sir3 nucleosomal arrays. The results indicate that Sir3 interacts with nucleosomal arrays with a stoichiometry of two Sir3 monomers per nucleosome. We also find that Sir3 fibres are less compact than canonical magnesium-induced 30 nm fibres. We suggest that heterochromatin proteins promote silencing by ‘coating’ nucleosomal arrays, stabilizing interactions between nucleosomal histones and DNA.
ContributorsSwygert, Sarah G. (Author) / Manning, Benjamin J. (Author) / Senapati, Subhadip (Author) / Kaur, Parminder (Author) / Lindsay, Stuart (Author) / Demeler, Borries (Author) / Peterson, Craig L. (Author) / Biodesign Institute (Contributor) / Single Molecule Biophysics (Contributor)
Created2014-08-01