Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry Characterization of Primary Amine End-Functionalized Polystyrene and Poly(methyl methacrylate) Synthesized by Living Anionic Polymerization Techniques

The reaction of living anionic polymers with 2,2,5,5-tetramethyl-1-(3-bromopropyl)-1-aza-2,5- disilacyclopentane (1) was investigated using coupled thin layer chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Structures of byproducts as well as the major product were determined. The anionic initiator having a protected primary amine functional group, 2,2,5,5-tetramethyl- 1-(3-lithiopropyl)-1-aza-2,5-disilacyclopentane (2), was synthesized using all-glass high-vacuum techniques, which allows the long-term stability of this initiator to be maintained. The use of 2 in the preparation of well-defined aliphatic primary amine R-end-functionalized polystyrene and poly(methyl methacrylate) was investigated. Primary amino R-end-functionalized poly(methyl methacrylate) can be obtained near-quantitatively by reacting 2 with 1,1-diphenylethylene in tetrahydrofuran at room temperature prior to polymerizing methyl methacrylate at -78 °C. When 2 is used to initiate styrene at room temperature in benzene, an additive such as N,N,N',N'- tetramethylethylenediamine is necessary to activate the polymerization. However, although the resulting polymers have narrow molecular weight distributions and well-controlled molecular weights, our mass spectra data suggest that the yield of primary amine α-end-functionalized polystyrene from these syntheses is very low. The majority of the products are methyl α-end-functionalized polystyrene.

Evaluation of an In Vitro Blood-Based Assay to Detect Production of Interferon-γ By Mycobacterium Bovis–Infected White-Tailed Deer (Odocoileus Virginianus)

Tuberculosis due to Mycobacterium bovis in captive Cervidae was identified as an important disease in the United States in 1990 and prompted the addition of captive Cervidae to the USDA Uniform Methods and Rules for eradication of bovine tuberculosis. As well, M. bovis infection was identified in free-ranging white-tailed deer in northeast Michigan in 1995. Tuberculosis in both captive and free-ranging Cervidae represents a serious challenge to the eradication of M. bovis infection from the United States. Currently, the only approved antemortem tests for tuberculosis in Cervidae are the intradermal tuberculin skin test and the blood tuberculosis test (BTB). At present, the BTB is not available in North America. Tuberculin skin testing of Cervidae is time-consuming and involves repeated animal handling and risk of injury to animals and humans. This study evaluated the potential of a new blood-based assay for tuberculosis in Cervidae that would decrease animal handling, stress, and losses due to injury. In addition, a blood-based assay could provide a more rapid diagnosis. Twenty 6–9-month-old white-tailed deer, male and female, were experimentally inoculated by instillation of 300 colony-forming units of M. bovis in the tonsillar crypts. Seven, age-matched uninfected deer served as controls. Blood was collected on days 90, 126, 158, 180, 210, 238, 263, and 307 after inoculation and was analyzed for the production of interferon-γ (IFN-γ) in response to incubation with M. bovis purified protein derivative (PPDb), M. avium PPDa, pokeweed mitogen (PWM), or media alone. Production of IFN-g in response to PPDb was significantly greater (P < 0.05) at all time points in samples from M. bovis–infected deer as compared with uninfected control deer, whereas IFN-γ production to PWM did not differ significantly between infected and control deer. Measurement of IFN-γ production to PPDb may serve as a useful assay for the antemortem diagnosis of tuberculosis in Cervidae.

A Pilot Study of the Relationship of Calcium Intake and Frequency of Injuries In High School Athletes

Little information is available related to adolescent calcium intake and relationships with injuries they might suffer from sport participation. To determine calcium intake of high school athletes, to assess their self reported injury rates, and to examine the relationship between the two over a 12 month period of time. Participants received a questionnaire at their school and completed it anywhere they found convenient. Adolescent athletes in the Lincoln Public School system (n=43) that participated in at least one sport in the past year. Four age groups participated in the study with sixteen year olds having a significantly higher calcium intake at 1297 mg that of fourteen year olds. A variety of sports were represented with largest number of respondents participating in baseball/or softball at (55%). The next most played sport was basketball at (18%). Median total diet calcium was 1144.5 mg with a mean of 1182 mg + 567 mg. For the frequency of injuries that caused a missed practice or game in the past year, ankle injuries were the most common (25%). Knee injuries were the second most common (17%), followed closely by hand injuries (8%). Mean total diet calcium of athletes with five or more injuries that caused a missed practice or game was significantly higher at 1966 mg (P<.05) than athletes mean diet calcium with zero, one, two, and three injuries. Total milk calcium of those who reported three injuries that resulted in broken or fractured bones or dislocated joints was significantly higher (P<.05) at 1286 mg of total milk calcium than those who reported having zero, one, or two breaks or fractures. Athletes with higher calcium intakes have a higher number of reported injuries. This may be the result of increased vigorous activity which leads to increased calorie and calcium consumption. More importantly, this increased activity leads to an increased chance of injury. The greater calcium intake correlated with greater number of injuries may also be because of third parties advising the athletes who get injured to drink more milk and get more calcium in their diets because they have been injuries already.