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. ^

An analysis of student and instructor perceptions of students' writing skills at the university level

This study examined university students' writing skills as perceived by university students and their English instructors. The goal of the study was to provide English instructors with objective, quantified information about writing perceptions from both the students' and instructors' viewpoints. ^ A survey instrument was developed based on a survey instrument created by Newkirk, Cameron, and Selfe (1977) to identify instructors' perceived knowledge of student writing skills. The present study used a descriptive statistical design. It examined five writing skill areas: attitude, content, grammar and mechanics, literary considerations, and the writing process through a questionnaire completed by a convenience sample of summer and fall admitted freshmen who were enrolled in Essay Writing and Freshman Composition courses and English Department instructors at a large South Florida public university. ^ The study consisted of five phases. The first phase was modifying of the Newkirk, Cameron, and Selfe (1977) questionnaire. Two versions of the revised survey were developed - one for instructors and one for students. The second phase was pilot testing the questionnaire for evaluation of administration and scoring. The third phase was administering the questionnaire to 1,280 students and 48 instructors. The fourth phase was analyzing the data. The study found a significant difference in the perceptions of students and instructors in all areas of writing skills examined by the survey. Responses to 29 of 30 questions showed that students felt they had better attitudes toward writing and better writing skills than instructors thought. ^ The final phase was developing recommendations for practice. Based on findings and theory and empirical evidence drawn from the fields of adult education and composition research, learner-centered, self-directed curriculum guidelines are offered. ^ By objectively quantifying student and instructor perceptions of students' writing skills, this study contributes to a growing body of literature that: (a) encourages instructors to acknowledge the perception disparities between instructors and students; (b) gives instructors a better understanding of how to communicate with students; and (c) recommends the development of new curriculum, placement tests, and courses that meet the needs of students and enables English instructors to provide meaningful instruction. ^

The use of computer technology to compare and analyze community college dissertations

A purpose of this research study was to demonstrate the practical linguistic study and evaluation of dissertations by using two examples of the latest technology, the microcomputer and optical scanner. That involved developing efficient methods for data entry plus creating computer algorithms appropriate for personal, linguistic studies. The goal was to develop a prototype investigation which demonstrated practical solutions for maximizing the linguistic potential of the dissertation data base. The mode of text entry was from a Dest PC Scan 1000 Optical Scanner. The function of the optical scanner was to copy the complete stack of educational dissertations from the Florida Atlantic University Library into an I.B.M. XT microcomputer. The optical scanner demonstrated its practical value by copying 15,900 pages of dissertation text directly into the microcomputer. A total of 199 dissertations or 72% of the entire stack of education dissertations (277) were successfully copied into the microcomputer's word processor where each dissertation was analyzed for a variety of syntax frequencies. The results of the study demonstrated the practical use of the optical scanner for data entry, the microcomputer for data and statistical analysis, and the availability of the college library as a natural setting for text studies. A supplemental benefit was the establishment of a computerized dissertation corpus which could be used for future research and study. The final step was to build a linguistic model of the differences in dissertation writing styles by creating 7 factors from 55 dependent variables through principal components factor analysis. The 7 factors (textual components) were then named and described on a hypothetical construct defined as a continuum from a conversational, interactional style to a formal, academic writing style. The 7 factors were then grouped through discriminant analysis to create discriminant functions for each of the 7 independent variables. The results indicated that a conversational, interactional writing style was associated with more recent dissertations (1972-1987), an increase in author's age, females, and the department of Curriculum and Instruction. A formal, academic writing style was associated with older dissertations (1972-1987), younger authors, males, and the department of Administration and Supervision. It was concluded that there were no significant differences in writing style due to subject matter (community college studies) compared to other subject matter. It was also concluded that there were no significant differences in writing style due to the location of dissertation origin (Florida Atlantic University, University of Central Florida, Florida International University).