VERIFICATION OF DNA PREDICTED PROTEIN SEQUENCES BY ENZYME HYDROLYSIS AND MASS SPECTROMETRIC ANALYSIS

The focus of this thesis lies in the development of a sensitive method for the analysis of protein primary structure which can be easily used to confirm the DNA sequence of a protein's gene and determine the modifications which are made after translation. This technique involves the use of dipeptidyl aminopeptidase (DAP) and dipeptidyl carboxypeptidase (DCP) to hydrolyze the protein and the mass spectrometric analysis of the dipeptide products.^ Dipeptidyl carboxypeptidase was purified from human lung tissue and characterized with respect to its proteolytic activity. The results showed that the enzyme has a relatively unrestricted specificity, making it useful for the analysis of the C-terminal of proteins. Most of the dipeptide products were identified using gas chromatography/mass spectrometry (GC/MS). In order to analyze the peptides not hydrolyzed by DCP and DAP, as well as the dipeptides not identified by GC/MS, a FAB ion source was installed on a quadrupole mass spectrometer and its performance evaluated with a variety of compounds.^ Using these techniques, the sequences of the N-terminal and C-terminal regions and seven fragments of bacteriophage P22 tail protein have been verified. All of the dipeptides identified in these analysis were in the same DNA reading frame, thus ruling out the possibility of a single base being inserted or deleted from the DNA sequence. The verification of small sequences throughout the protein sequence also indicates that no large portions of the protein have been removed after translation. ^

Is body mass index associated with Barrett esophagus: A systematic literature review

Background. Risk factors underlying the development of Barrett's esophagus (BE) are poorly understood. Recent studies have examined the association between elevated body mass index (BMI) and BE with conflicting results. A systematic review of literature was performed to study this association.^ Methods. Cross-sectional, case control and cohort studies published through April 2007 meeting strict inclusion and exclusion criteria were included. A thorough data abstraction, including that of reported crude or adjusted odds ratios or mean BMI, was performed. Crude odds ratios were estimated from available information in 3 studies.^ Results. Of 630 publications identified by our search items, 59 were reviewed in detail and 12 included in the final analyses. 3 studies showed a statistically significant association between obesity and BE (30-32) while 2 studies found a statistically significant association between overweight and BE (31, 32). Two studies that reported BMI as a continuous variable found BMI in cases to be significantly higher than that in the comparison group (30, 32). Other studies failed to show an significant association between elevated BMI and BE.^ Conclusions. There is conflicting data regarding the association between elevated BMI and BE. It is important to identify other risk factors that in combination with elevated BMI may lead to BE. Further studies are needed to evaluate if the presence of reflux symptoms or any particular pattern of obesity, are independently associated with BE.^ Key words. Barrett's esophagus, obesity, Body Mass Index, gastroesophageal reflux disease, meta-analysis^

Characterization of the East Houston particulate matter <2.5 atmosphere; a novel method for desorbing organic constituents from airborne particulates

I have developed a novel approach to test for toxic organic substances adsorbed onto ultra fine particulate particles present in the ambient air in Northeast Houston, Texas. These particles are predominantly carbon soot with an aerodynamic diameter (AD) of <2.5 μm. If present in the ambient air, many of the organic substances will be absorbed to the surface of the particles (which act just like a charcoal air filter), and may be adducted into the respiratory system. Once imbedded into the lungs these particles may release the adsorbed toxic organic substances with serious health consequences. I used a Airmetrics portable Minivol air sampler time drawing the ambient air through collection filters samples from 6 separate sites in Northeast Houston, an area known for high ambient PM 2.5 released from chemical plants and other sources (e.g. vehicle emissions).(1) In practice, the mass of the collected particles were much less than the mass of the filters. My technique was designed to release the adsorbed organic substances on the fine carbon particles by heating the filter samples that included the PM 2.5 particles prior to identification by gas chromatography/mass spectrometry (GCMS). The results showed negligible amounts of target chemicals from the collection filters. However, the filters alone released organic substances and GCMS could not distinguish between the organic substances released from the soot particles from those released from the heated filter fabric. However, an efficacy tests of my method using two wax burning candles that released soot revealed high levels of benzene. This suggests that my method has the potential to reveal the organic substances adsorbed onto the PM 2.5 for analysis. In order to achieve this goal, I must refine the particle collection process which would be independent of the filters; the filters upon heating also release organic substances obscuring the contribution from the soot particles. To obtain pure soot particles I will have to filter more air so that the soot particles can be shaken off the filters and then analyzed by my new technique. ^