Microrheology of bacterial biofilms in vitro: Staphylococcus aureus and Pseudomonas aeruginosa.

The rheology of bacterial biofilms at the micron scale is an important step to understanding the communal lifecycles of bacteria that adhere to solid surfaces, as it measures how they mutually adhere and desorb. Improvements in particle-tracking software and imaging hardware have allowed us to successfully employ particle-tracking microrheology to measuring single-species bacterial biofilms, based on Staphlococcus aureus and Pseudomonas aeruginosa. By tracking displacements of the cells at a range of timescales, we separate active and thermal contributions to the cell motion. The S. aureus biofilms in particular show power-law rheology, in common with other dense colloidal suspensions. By calculating the mean compliance of S. aureus biofilms, we observe them becoming less compliant during growth, and more compliant during starvation. The biofilms are rheologically inhomogeneous on the micron scale, as a result of the strength of initial adhesion to the flow cell surface, the arrangement of individual bacteria, and larger-scale structures such as flocs of P. aeruginosa. Our S. aureus biofilms became homogeneous as a function of height as they matured: the rheological environment experienced by a bacterium became independent of how far it lived from the flow cell surface. Particle-tracking microrheology provides a quantitative measure of the "strength" of a biofilm. It may therefore prove useful in identifying drug targets and characterizing the effect of specific molecular changes on the micron-scale rheology of biofilms.

Investigating the permanent electric dipole moment of beta-lactoglobulin fibrils, using transient electric birefringence.

Amyloid fibrils, which are polymeric assemblies of protein molecules, have been intensively studied on a structural level, yet due to complications such as the disorder within the molecules, several aspects of their structure remain mysterious. Similarly, the kinetics of assembly are not well understood. Here we investigate the electric dipole moment of beta-lactoglobulin fibrils, a model amyloid fibril system, by applying the technique of transient electric birefringence. This moment appears to be large, and comparable to the total moment of the constituent protein monomers if they were joined in a chain, head-to-tail, without changing conformation, suggesting an ordered joining of monomers in the fibril. Such an ordered assembly may have implications for the assembly mechanism of beta-lactoglobulin fibrils in particular, and amyloid fibrils in general.

Electric birefringence study of an amyloid fibril system: the short end of the length distribution.

In this article, a system of amyloid fibrils, based on the protein beta-lactoglobulin, is studied by transient electric birefringence. Single pulses of an electric field were applied to the solution, and the initial rise and subsequent decay of birefringence analysed. The decay takes place on a range of relaxation times, and therefore contains information about the length distribution of fibrils in the system. The information can be extracted using theories of the electric polarisability of polyelectrolyte rods, since the fibrils are an example of these. Despite the long-standing complications of such theories, useful quantitative information about the system can still be obtained. Using the Fixman model of polyelectrolyte polarisability, we obtain a measurement of the short end of the length distribution which shows the fibril concentration as a function of length rising linearly from 0.02-2 microm. The short end of the length distribution was unobtainable in our previous study using rheo-optics (S.S. Rogers et al., Macromolecules 38, 2948 (2005)), but reasonable agreement between the two techniques shows they are complementary.

The mechanism of amyloid spherulite formation by bovine insulin.

The formation of amyloid-containing spherulite-like structures has been observed in some instances of amyloid diseases, as well as in amyloid fibril-containing solutions in vitro. In this article we describe the structure and kinetics of bovine insulin amyloid fibril spherulites formed in the presence and absence of different salts and at different salt concentrations. The general spherulite structure consists of radially oriented amyloid fibrils, as shown by optical microscopy and environmental scanning electron microscopy. In the center of each spherulite, a "core" of less regularly oriented material is observed, whose size decreases when the spherulites are formed in the presence of increasing concentrations of NaCl. Similarly, amyloid fibrils form faster in the presence of NaCl than in its absence. A smaller enhancement of the rate of formation with salt concentration is observed for spherulites. These data suggest that both amyloid fibril formation and random aggregation occur concurrently under the conditions tested. Changes in their relative rates result in the different-sized cores observed in the spherulites. This mechanism can be likened to that leading to the formation of spherulites of polyethylene, in agreement with observations that polypeptide chains under partially denaturing conditions can exhibit behavior not dissimilar to that of synthetic polymers.

Spine-sparing postmastectomy irradiation.

Postoperative irradiation of the chest wall and regional lymphatics following mastectomy for operable breast carcinoma is an effective means of reducing the chance of postoperative locoregional recurrence. The majority of patients who fail will ultimately develop bony metastases, especially in the thoracic and lumbar spinal regions. Our technique of postoperative irradiation avoids exit irradiation of the spine. This is important for patients who may later develop bony metastases or epidural spinal cord compression and require spinal irradiation. The details of the technique are presented.

Extended-field isocentric irradiation for Hodgkin's disease.

Extended-field therapeutic irradiation is the treatment of choice for the majority of patients diagnosed with pathologic stages I and II Hodgkin's disease, and total nodal irradiation can be effectively used to treat selected stage III Hodgkin's patients. Standard 100-cm source-to-axis distance extended-field isocentric technique and results are presented.

Survival of patients with unresectable non-small cell lung cancer following 6,000 rad megavoltage radiotherapy.

One hundred and fifty-seven patients with histologically proven, locally advanced, unresectable or inoperable non-small cell lung cancer were treated in a uniform fashion with external-beam megavoltage radiotherapy. Patients received a continuous course of 6,000 rad in 7½ weeks (four fractions of 200 rad per fraction each week). The comparatively high, uncorrected one-year survival rate of 48.7 percent, and uncorrected two-year survival rate of 25.7 percent, would be expected following uniform treatment with a relatively high external-beam tumor dose. The five-year uncorrected actuarial survival rate of 2.7 percent demonstrates that although the median survival can be increased by the use of higher radiation doses and better delivery technique, the ultimate cure rate and prognosis for inoperable and unresectable non-small cell lung cancer remains poor with current equipment and methods of radiotherapy.