Heather A. Bullen

Surface Complexation of Catechol to Metal Oxides: An ATR-FTIR, Adsorption, and Dissolution Study

The interaction of catechol with chromium(III) oxide (Cr(2)O(3)), manganese dioxide (MnO(2)), iron(III) oxide (Fe(2)O(3)), and titanium dioxide (TiO(2)) was evaluated as a function of pH conditions (pH 3-10) and ionic strength using a combined approach of bulk adsorption, attenuated total reflectance Fourier-transform infrared spectroscopy (ATR-FTIR), and dissolution analysis. Adsorption of catechol showed a strong pH-dependent behavior with the metal oxides, remaining constant under acidic-neutral pH (3-7) and increasing under more basic conditions. In situ ATR-FTIR measurements indicate that catechol binds predominately as an outer-sphere complex on MnO(2) and as an inner-sphere complex on Fe(2)O(3), TiO(2), and Cr(2)O(3) substrates. Catechol complexation on Fe(2)O(3), TiO(2), and Cr(2)O(3) promotes dissolution at pH >5, whereas MnO(2) dissolution occurs under acidic and basic conditions (pH 3-10).

Bacterial Calcium Carbonate Precipitation in Cave Environments: A Function of Calcium Homeostasis

To determine if microbial species play an active role in the development of calcium carbonate (CaCO 3 ) deposits (speleothems) in cave environments, we isolated 51 culturable bacteria from a coralloid speleothem and tested their ability to dissolve and precipitate CaCO 3 . The majority of these isolates could precipitate CaCO 3 minerals; scanning electron microscopy and X-ray diffractrometry demonstrated that aragonite, calcite and vaterite were produced in this process. Due to the inability of dead cells to precipitate these minerals, this suggested that calcification requires metabolic activity. Given growth of these species on calcium acetate, but the toxicity of Ca 2+ ions to bacteria, we created a loss-of-function gene knock-out in the Ca 2+ ion efflux protein ChaA. The loss of this protein inhibited growth on media containing calcium, suggesting that the need to remove Ca 2+ ions from the cell may drive calcification. With no carbonate in the media used in the calcification studies, we used stable isotope probing with C 13 O 2 to determine whether atmospheric CO 2 could be the source of these ions. The resultant crystals were significantly enriched in this heavy isotope, suggesting that extracellular CO 2 does indeed contribute to the mineral structure. The physiological adaptation of removing toxic Ca 2+ ions by calcification, while useful in numerous environments, would be particularly beneficial to bacteria in Ca 2+ -rich cave environments. Such activity may also create the initial crystal nucleation sites that contribute to the formation of secondary CaCO 3 deposits within caves.

Use of Attenuated Total Reflectance Fourier Transform Infrared Spectroscopy To Identify Microbial Metabolic Products on Carbonate Mineral Surfaces

ABSTRACT This paper demonstrates the use of attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy to detect microbial metabolic products on carbonate mineral surfaces. By creating an ATR-FTIR spectral database for specific organic acids using ATR-FTIR spectroscopy we were able to distinguish metabolic acids on calcite surfaces following Escherichia coli growth. The production of these acids by E. coli was verified using high-performance liquid chromatography with refractive index detection. The development of this technique has allowed us to identify microbial metabolic products on carbonate surfaces in nutrient-limited cave environments.

Analysis of biotinylated generation 4 poly(amidoamine) (PAMAM) dendrimer distribution in the rat brain and toxicity in a cellular model of the blood-brain barrier.

Dendrimers are highly customizable nanopolymers with qualities that make them ideal for drug delivery. The high binding affinity of biotin/avidin provides a useful approach to fluorescently label synthesized dendrimer-conjugates in cells and tissues. In addition, biotin may facilitate delivery of dendrimers through the blood-brain barrier (BBB) via carrier-mediated endocytosis. The purpose of this research was to: (1) measure toxicity using lactate dehydrogenase (LDH) assays of generation (G)4 biotinylated and non-biotinylated poly(amidoamine) (PAMAM) dendrimers in a co-culture model of the BBB, (2) determine distribution of dendrimers in the rat brain, kidney, and liver following systemic administration of dendrimers, and (3) conduct atomic force microscopy (AFM) on rat brain sections following systemic administration of dendrimers. LDH measurements showed that biotinylated dendrimers were toxic to cell co-culture after 48 h of treatment. Distribution studies showed evidence of biotinylated and non-biotinylated PAMAM dendrimers in brain. AFM studies showed evidence of dendrimers only in brain tissue of treated rats. These results indicate that biotinylation does not decrease toxicity associated with PAMAM dendrimers and that biotinylated PAMAM dendrimers distribute in the brain. Furthermore, this article provides evidence of nanoparticles in brain tissue following systemic administration of nanoparticles supported by both fluorescence microscopy and AFM.

Improved methodology for monitoring poly(amidoamine) dendrimers surface transformations and product quality by ultra performance liquid chromatography

Ultra performance liquid chromatography (UPLC) analysis was utilized for the first time as a methodology for monitoring poly(amidoamine) (PAMAM) dendrimer surface transformations and product quality. Results were compared to high-performance liquid chromatography (HPLC) and were found to provide a vastly improved analytical method for the characterization of dendrimer polydispersity and variance in a typical surface modification. The application of UPLC increased the average number of theoretical plates by a factor of 7 and reduced retention times of analytes by 36%, while improving the resolution capability to discriminate surface variances in dendrimers. The new UPLC procedures were used to monitor surface modification of [core: ethylenediamine]; (G=4); dendri-poly(amidoamine)-(NH2)64 (i.e., [EDA]; (G4); dendri-PAMAM-(NH2)64) to produce biotinylated dendrimer conjugates. The enhanced sensitivity and efficiency of the UPLC analyses allowed resolution of biotin substituent levels and a better characterization of the targeted dendrimer conjugates compared to traditional HPLC methodology.

Bioweathering Potential of Cultivable Fungi Associated with Semi-Arid Surface Microhabitats of Mayan Buildings

Soil and rock surfaces support microbial communities involved in mineral weathering processes. Using selective isolation, fungi were obtained from limestone surfaces of Mayan monuments in the semi-arid climate at Yucatan, Mexico. A total of 101 isolates representing 53 different taxa were studied. Common fungi such as Fusarium, Pestalotiopsis, Trichoderma, and Penicillium were associated with surfaces and were, probably derived from airborne spores. In contrast, unusual fungi such as Rosellinia, Annulohypoxylon, and Xylaria were predominantly identified from mycelium particles of biofilm biomass. Simulating oligotrophic conditions, agar amended with CaCO3 was inoculated with fungi to test for carbonate activity. A substantial proportion of fungi, in particular those isolated from mycelium (59%), were capable of solubilizing calcium by means of organic acid release, notably oxalic acid as evidenced by ion chromatography. Contrary to our hypothesis, nutrient level was not a variable influencing the CaCO3 solubilization ability among isolates. Particularly active fungi (Annulohypoxylon stygium, Penicillium oxalicum, and Rosellinia sp.) were selected as models for bioweathering experiments with limestone-containing mesocosms to identify if other mineral phases, in addition to oxalates, were linked to bioweathering processes. Fungal biofilms were seen heavily covering the stone surface, while a biomineralized front was also observed at the stone-biofilm interface, where network of hyphae and mycogenic crystals was observed. X-ray diffraction analysis (XRD) identified calcite as the main phase, along with whewellite and wedellite. In addition, lower levels of citrate were detected by Attenuated Total Reflectance-Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Overall, our results suggest that a diverse fungal community is associated with limestone surfaces insemi-arid climates. A subset of this community is geochemically active, excreting organic acids under quasi-oligotrophic conditions, suggesting that the high metabolic cost of exuding organic acids beneficial under nutrient limitation. Oxalic acid release may deteriorate or stabilize limestone surfaces, depending on microclimatic dynamics.

Retention of STEM majors using early undergraduate researchexperiences

An early undergraduate research program for rising sophomores and juniors at risk of leaving STEM degree programs is described. Students are paid a stipend to work part-time, at a maximum of twenty hours per week, as part of a research team. Faculty researchers are not financially compensated for working with students. The program successfully brings together STEM departments to target students who are at risk of leaving their major. Initial results demonstrate a positive influence of undergraduate research in retaining STEM majors and improvements in Student Assessment of Learning Gains. Future work, including institutionalization of the project, is discussed.

Low-Level Detection of Poly(amidoamine) PAMAM Dendrimers Using Immunoimaging Scanning Probe Microscopy

Immunoimaging scanning probe microscopy was utilized for the low-level detection and quantification of biotinylated G4 poly(amidoamine) PAMAM dendrimers. Results were compared to those of high-performance liquid chromatography (HPLC) and found to provide a vastly improved analytical method for the low-level detection of dendrimers, improving the limit of detection by a factor of 1000 (LOD = 2.5 × 10−13 moles). The biorecognition method is reproducible and shows high specificity and good accuracy. In addition, the capture assay platform shows a promising approach to patterning dendrimers for nanotechnology applications.

Student-driven independent research projects: developing a framework for success in analytical chemistry

As educators, we are always looking for new ways to engage our students within the classroom and laboratory setting. Students today want a learning environment that prepares them for their future careers. It is not only important that they see a connection between what they are learning and their world, but also that they see themselves within the discipline. Real-life applications are one way to achieve this. However, creating an environment that provides this can be challenging. Analytical chemistry courses are still often taught in a traditional format that generally introduces students to a wide variety of analysis methods in a series of short experiments. Efforts are being made to provide guided inquiry and problem-based approaches that allow students to become problem solvers in a collaborative environment [1–4]. These activities can greatly enhance the learning experience and allow students to conduct investigations that approximate how practicing scientists do science. However, instructors may still struggle with how to incorporate this type of learning into their curriculum, and/ or feel they are limited to the methods they can explore in the time frame of a course using this approach. Presented here is the development of an instrumental analysis course that utilizes a problem-based learning approach to integrate student-driven research projects into the curriculum. Students have a wide variety of “problems” that interest them. Allowing students to explore and develop questions of their own, rather than providing problems to solve, engaged students in a creative environment where they seized ownership of their learning, gaining important reasoning and critical thinking skills. To effectively implement this course design, independent projects were preceded by a series of introductory experiments designed to provide students with a necessary foundation.