The pedestrian watchmaker: Genetic clocks from engineered oscillators

The crucial role of time-keeping has required organisms to develop sophisticated regulatory networks to ensure the reliable propagation of periodic behavior. These biological clocks have long been a focus of research; however, a clear understanding of how they maintain oscillations in the face of unpredictable environments and the inherent noise of biological systems remains elusive. Here, we review the current understanding of circadian oscillations using Drosophila melanogaster as a typical example and discuss the utility of an alternative synthetic biology approach to studying these highly intricate systems.     

Natural atmospheric microbial conditions in a typical suburban area

Ambient outdoor concentrations and size distributions of airborne microbial particles were measured approximately weekly for 2 years in a Washington, D.C., suburban area. The study objective was to characterize microbial air quality in the vicinity of a proposed sewage sludge composting facility. During the study, 379 samples were taken at 17 stations, using Andersen microbial samplers. Concentration ranges (in viable particles per cubic meter) were as follows: airborne mesophilic fungi, 0 to 7,220 with a geometric mean of 273; thermophilic fungi, 0 to 193 with a median of 2.1; Aspergillus fumigatus, 0 to 71 with a median of 1.0; aerobic bacteria, 4.2 to 1,640 with a geometric mean of 79; and fecal streptococci, 0 to 5.7 with a median of 0. No fecal coliforms were recovered. The potentially respirable fraction (less than 8 microns) averaged 34% for total bacteria, 56% for mesophilic fungi, 91% for thermophilic fungi, and 95% for A. fumigatus. The specific sampling location was not a major factor affecting microbial particle concentrations or size distributions. Conversely, the time of year was an important determinant of viable particle concentrations for all groups of microorganisms studied. The highest concentrations were observed in summer and fall, with significantly lower levels detected in winter. In general, the microbial data did not correlate with other variables, including weather conditions, measured in this study.     

Elevation of oil body integrity and emulsion stability by polyoleosins,multiple oleosin units joined in tandem head‐to‐tail fusions

We have successfully created polyoleosins by joining multiple oleosin units in tandem head‐to‐tail fusions. Constructs encoding recombinant proteins of 1, 3 and 6 oleosin repeats were purposely expressed both in planta and in Escherichia coli. Recombinant polyoleosins accumulated in the seed oil bodies of transgenic plants and in the inclusion bodies of E. coli. Although polyoleosin was estimated to only accumulate to <2% of the total oil body protein in planta, their presence increased the freezing tolerance of imbibed seeds as well as emulsion stability and structural integrity of purified oil bodies; these increases were greater with increasing oleosin repeat number. Interestingly, the hexameric form of polyoleosin also led to an observable delay in germination which could be overcome with the addition of external sucrose. Prokaryotically produced polyoleosin was purified and used to generate artificial oil bodies and the increase in structural integrity of artificial oil bodies‐containing polyoleosin was found to mimic those produced in planta. We describe here the construction of polyoleosins, their purification from E. coli, and properties imparted on seeds as well as native and artificial oil bodies. A putative mechanism to account for these properties is also proposed.     

Degradation artefacts during sample preparation for sodium dodecyl sulphate polyacrylamide gel electrophoresis

Preparation of samples for sodium dodecyl sulphate polyacrylamide gel electrophoresis routinely involves heating the protein in solution containing detergent and reducing agent for at least two minutes. Here we show that this treatment causes fragmentation of the protein glycogen phosphorylase, whether purified or as a component of a skeletal muscle preparation. The fragments are detected as minor bands on western blots and represent the products of discrete breakage point in the peptide sequence. Protease inhibitors cannot suppress the fragmentation.Such small amounts of immunoreactive fragments may be incorrectly identified on western blots as contaminants that were originally present in the antigen preparation. They may also be a source of ambiguity in studies that search for degradation intermediates during proteolysis.     

Renal function in the choline deficient rat.

Abnormalties in renal concentrating ability and free water reabsorption, and a diminished sodium excretion, glomerular filtration rate, and effective renal plasma flow were observed in adolescent rats which ingested a lipotrope deficient diet for 10 months.