Dual parameter flow cytometric analysis of DNA content, activation antigen expression, and T cell subset proliferation in the human mixed lymphocyte reaction

This study provides direct correlation via dual parameter flow cytometry (simultaneous assessment of immunofluorescence and DNA content) between mixed lymphocyte reaction (MLR) responder cell entry into the S/G2/M phases of the cell cycle with the kinetics of expression of two activation-associated cell surface proteins, Tac (IL 2 receptor) and 4F2 (unknown metabolic function). A small population of activated cells was identifiable by expression of both Tac and 4F2 antigens before peak DNA synthesis in the MLR. This population of activation antigen-positive cells expanded linearly in size from days 3 to 7 of culture. Treatment of immature MLR cultures with anti-4F2 Mab and complement (C) before DNA synthesis (treatment on day 3, peak DNA synthesis on days 5 to 6) resulted in blunted proliferation and activation antigen expression when the same culture was analyzed after maturation on day 6, indicating that the activated population had been previously detected and removed by anti-4F2 Mab + C. The 4F2 antigen was expressed on a greater percentage of cells in the MLR at all times (days 3 to 9) than was Tac, was present on virtually all S/G2/M phase responder cells, and a large fraction of cells remained intensely 4F2+ subsequent to peak DNA synthesis. In contrast, after initially preceding responder cell entry into the S phase of the cell cycle, the kinetics of Tac antigen expression closely paralleled the kinetics of responder cell proliferation. A subpopulation of cycling responder cells was noted in all MLR cultures studied that expressed Tac antigen weakly or not at all. Cells within both T4 and T8 cell subsets proliferate with similar kinetics in response to alloantigen. The possibility that activation antigens can be utilized to study effector cell generation in the MLR and that this flow cytometric technique may be utilized to analyze the response to various alloantigens is discussed.     

An evaluation of the importance of sulfate reduction and temperature to P fluxes from aerobic-surfaced, lacustrine sediments

We examined the influence of temperature and sulfate reduction rates onP-release from aerobic-surfaced, littoral sediments in a coolingreservoir. Annually, significant differences in P release from sediments at twosites (thermal effluent and non-effluent) were related to differenttemperature regimes, with higher rates of P release and decomposition at highertemperatures. Site-specific differences in solute fluxes were mostpronounced in late summer, when water temperatures reached40°Cat the effluent site and solute fluxes increased dramatically. The thermaleffluent site retained 65% of its annual P load, while thenon-effluent site retained 92%. Relative P release(P-release normalized to inorganic carbon fluxes; RPR) data indicatedthat lake sediments retained P selectively (relative to C) throughout the year,except at the effluent site during late summer when stored P was released inexcess of supply rates.Sulfate reduction rates were often typical of those measured in otherfreshwater lakes, but unusually high rates were measured at thethermal-effluent site especially in early fall and suggested higher DICfluxes than we measured. These high rates suggest that sulfate reduction rateswere overestimated and/or that most sulfide was recycled within the sediments.In any case, the highest sulfate reduction rates did not coincide with thehighest P release rates. Furthermore, the total reduced inorganic sulfurcontentof surficial sediments did not significantly correlate to RPR, althoughconcentrations varied widely throughout the year. Temperature was the onlyvariable examined that significantly correlated to RPR (R² =0.53, P-value = 0.017). Coupling between temperature and sedimentP release was likely mediated through temperature effects on bioturbation andmicrobial metabolic rates.     

Habitat occupancy by riparian muskrats reveals tolerance to urbanization and invasive vegetation

Wildlife communities are being altered by rapid environmental change including habitat loss and fragmentation, urbanization, and spread of invasive species. To predict consequences of these anthropogenic changes to landscapes, it is necessary to identify not only species that are negatively affected, but also species that are unaffected or even thrive. We used occupancy modeling to examine the spatial distribution of muskrats (Ondatra zibethicus) in riparian habitat within an agricultural region of east-central Illinois from 2007 to 2008. We examined whether site occupancy was related to local habitat conditions and anthropogenic landscape alterations including urbanization and dominance of invasive reed canary grass (Phalaris arundinacea). We sampled 90 study sites (200-m stream segments) for occupancy by muskrats based on presence of tracks, scat, and feeding sign. Per-survey detection probability was 0.79 (SE = 0.04) in 2007 and 0.76 (SE = 0.04) in 2008. Detection was related positively to Julian date and negatively to abundance of woody debris and emergent rocks. Site occupancy by muskrats was 0.59 (SE = 0.09) in 2007 and 0.69 (SE = 0.06) in 2008, a year with above-average precipitation. Occupancy was related positively to urban land cover surrounding sites, which could reflect higher baseflows and reduced risk from predation and trapping in urban areas. Occupancy was unrelated to site dominance by invasive reed canary grass, but muskrats occurred more often at larger, deeper streams and those with greater bank heights and less sandy bank soils. Turnover between years was driven by stream size and water availability. Muskrats exhibited tolerance to key aspects of environmental change, and muskrats might even be urban adapters when occupying riparian habitat that remains adequately connected in urbanizing landscapes. © 2011 The Wildlife Society.     

Unassembled HLA-DR beta monomers are degraded rapidly by a nonlysosomal mechanism.

We previously observed that in a mutant B lymphoblastoid cell line which has a homozygous HLA-DR alpha deletion, DR beta-chains appeared to be unstable. In the present study, we have studied the pathway that leads to degradation of unassembled DR beta-chains. Unassembled DR beta-chains are degraded rapidly in the DR alpha deletion mutant cells, compared with the assembled DR heterodimers present in non-mutant cells. Accelerated DR beta turnover in 9.22.3 cells is specific; class I molecules in these DR alpha-deficient cells turned over slowly. DR beta-chains assemble with Ii in the DR alpha deficient cell line, but this did not protect DR beta-chains from degradation. The maturation of unassembled beta-chains is arrested before their reaching the medial Golgi compartment, and this degradation proceeds by a nonlysosomal, nonendosomal pathway. Degradation of DR beta-chains is blocked when cells are cultured at 16 degrees C, a temperature known to prevent vesicular transport between the endoplasmic reticulum (ER) and the Golgi apparatus. Degradation is also inhibited by carbonyl cyanide m-chlorophenylhydrazone, a drug that is also known to inhibit protein transport from the ER. The results, taken together, suggest that degradation of unassembled DR beta-chains occurs by a nonlysosomal, nonendosomal pathway which involves transport of DR beta-chains out of the ER.     

Educational Malpractice: The Impact of Including Creationism in High School Biology Courses

College students whose recollections of their high school biology courses included creationism were significantly more likely to invoke creationism-based answers on questions derived from the Material Acceptance of the Theory of Evolution (MATE) instrument than were students whose recollections of their high school biology courses included evolution but not creationism. On average, students who were taught neither evolution nor creationism in their high school biology courses exhibited intermediary responses on the MATE instrument. These results suggest that (1) high school teachers’ treatments of evolution and creationism have a lasting impact and (2) the inclusion of creationism in high school biology courses increases the probability that students accept creationism and reject evolution when they arrive at college. These results are discussed relative to the impact of high school biology courses on students’ subsequent acceptance of evolution and creationism.     

Temperature Affects Stoichiometry and Biochemical Composition of Escherichia coli

Temperature is a master variable controlling biochemical processes in organisms, and its effects are manifested on many organizational levels in organisms and ecosystems. We examined the effects of temperature on the biochemical composition and stoichiometry of a model heterotrophic bacterium, Escherichia coli K-12, held at constant growth rate in chemostats. Increasing temperature led to increased cellular organic carbon (C) and organic nitrogen (N) with decreased phosphorus (P) content, leading to increased C/P and N/P biomass ratios. P content was related to cellular RNA, which is P-rich (9–10% by weight) and nonnucleic acid P (presumably composed of mostly phospholipids, intracellular phosphate, and polyphosphate). These results indicate that E. coli allocates an increased proportion of its P cell quota toward assembly (ribosomes) at low temperatures and an increasing proportion toward resource acquisition machinery (membranes) at higher temperatures. If these results are relevant to the behavior of prokaryotic heterotrophs in natural settings (the gut, soils, lakes, oceans, etc.), it suggests greater nutrient regeneration and less microbial nutrient retention as temperatures increase.