Identification and localization of polyketide synthase genes from cultures of diarrheic shellfish poisoning toxin producing dinoflagellates of the genus Prorocentrum

Aquatic toxins are responsible for a number of acute and chronic diseases in humans. Okadaic acid (OA) and other dinoflagellate derived polyketide toxins pose serious health risks on a global scale. Ingestion of OA contaminated shellfish causes diarrheic shellfish poisoning (DSP). Some evidence also suggests tumor promotion in the liver by OA. Microcystin-LR (MC-LR) is produced by cyanobacteria and is believed to be the most common freshwater toxin in the US. Humans may be exposed to this acute hepatotoxin through drinking or recreational use of contaminated waters. ^ OA producing dinoflagellates have not been cultured axenically. The presence of associated bacteria raises questions about the ultimate source of OA. Identification of the toxin-producing organism(s) is the first step in identifying the biosynthetic pathways involved in toxin production. Polyketide synthase (PKS) genes of toxic and non-toxic species were surveyed by construction of clonal libraries from PCR amplicons of various toxic and non-toxic species of Prorocentrum in an effort to identify genes, which may be part of the biosynthetic pathway of OA. Analysis of the PKS sequences revealed that toxic species shared identical PKS genes not present in non-toxic species. Interestingly, the same PKS genes were identified in a library constructed from associated bacteria. ^ Subsequent bacterial small subunit RNA (16S) clonal libraries identified several common bacterial species. The most frequent 16S sequences found were identified as species of the genus Roseobacter which has previously been implicated in the production of OA. Attempts to culture commonly occurring bacteria resulted in the isolation of Oceanicaulis alexandrii , a novel marine bacterium previously isolated from the dinoflagellate Alexandrium tamarense, from both P. lima, and P. hoffmanianum. ^ Metabolic studies of microcystin-LR, were conducted to probe the activity of the major human liver cytochromes (CYP) towards the toxin. CYPs may provide alternate routes of detoxification of toxins when the usual routes have been inhibited. For example, some research indicates that cyanobacterial xenobiotics, in particular, lipopolysaccharides may inhibit glutathione S-transferases allowing the toxin to persist long enough to be acted upon by other enzymes. These studies found that at least one human liver CYP was capable of metabolizing the toxin. ^

The influence of study and travel abroad on the personal and professional development of students in architecture design programs

International travel has significant implications on the study of architecture. This study analyzed ways in which undergraduate and graduate students benefited from the experience of international travel and study abroad. Taken from the perspective of 15 individuals who were currently or had been architecture students at the University of Miami and Florida International University or who were alumni of the University of Florida and Syracuse University, the research explored how international travel and study abroad enhanced their awareness and understanding of architecture, and how it complemented their architecture curricula. This study also addressed a more personal aspect of international travel in order to learn how the experience and exposure to foreign cultures had positively influenced the personal and professional development of the participants.^ Participants’ individual and two-person semi-structured interviews about study abroad experiences were electronically recorded and transcribed for analysis. A second interview was conducted with five of the participants to obtain feedback concerning the accuracy of the transcripts and the interpretation of the data. Sketch journals and design projects were also analyzed from five participants and used as data for the purposes of better understanding what these individuals learned and experienced as part of their study abroad.^ Findings indicated that study abroad experiences helped to broaden student understanding about architecture and urban development. These experiences also opened the possibilities of creative and professional expression. For many, this was the most important aspect of their education as architects because it heightened their interest in architecture. These individuals talked about how they had the opportunity to experience contemporary and ancient buildings that they had learned about in their history and design classes on their home campuses. In terms of personal and professional development, many of the participants remarked that they became more independent and self-reliant because of their study abroad experiences. They also displayed a sense of global awareness and were interested in the cultures of their host nations. The study abroad experiences also had a lasting influence on their professional development.^

A preliminary characterization of mercury uptake by the sulfate-reducing bacteria Desulfovibrio desulfuricans

The focus of this research is to determine if a relationship exists between the stability constant and the initial uptake rate of a mercury species by bacteria. Cultures of the sulfate-reducing bacteria (SRB) strain Desulfovibrio desulfuricans G20 were washed with a bicarbonate buffer solution containing either lactate and sulfate or pyruvate and fumarate. The washed cell solutions were then spiked with either mercury bound to natural organic matter (Hg-NOM) or neutral mercury chloride (HgCl2), followed by sampling over time to provide kinetic data. Despite the significantly different stability constants for Hg-NOM and HgCl2, the calculated initial rate constants for mercury uptake for these two types of complexes appeared to be comparable. Uptake of mercury sulfide species was inconclusive due to possible formation of cinnabar. A simple model that is based on assumptions of passive diffusion and facilitated uptake of mercury by bacteria was evaluated for its potential to simulate the uptake. The model results only agreed with experimental data for HgCl2 uptake.