Monitoring bacterially induced calcite precipitation in porous media using magnetic resonance imaging and flow measurements.

A range of nuclear magnetic resonance (NMR) techniques are employed to provide novel, non-invasive measurements of both the structure and transport properties of porous media following a biologically mediated calcite precipitation reaction. Both a model glass bead pack and a sandstone rock core were considered. Structure was probed using magnetic resonance imaging (MRI) via a combination of quantitative one-dimensional profiles and three-dimensional images, applied before and after the formation of calcite in order to characterise the spatial distribution of the precipitate. It was shown through modification and variations of the calcite precipitation treatment that differences in the calcite fill would occur but all methods were successful in partially blocking the different porous media. Precipitation was seen to occur predominantly at the inlet of the bead pack, whereas precipitation occurred almost uniformly along the sandstone core. Transport properties are quantified using pulse field gradient (PFG) NMR measurements which provide probability distributions of molecular displacement over a set observation time (propagators), supplementing conventional permeability measurements. Propagators quantify the local effect of calcite formation on system hydrodynamics and the extent of stagnant region formation. Collectively, the combination of NMR measurements utilised here provides a toolkit for determining the efficacy of a biological-precipitation reaction for partially blocking porous materials.

Chorda tympani nerve function after myringoplasty.

This is a cross-sectional study design aimed to determine the prevalence of Chorda Tympani Nerve (CTN) injury and related symptoms following myringoplasty. Thirty patients were included in this study. The methods used were measuring taste thresholds using electrogustometer to map taste threshold on the anterior two-third of the tongue on the operated side with the non operated side as the control. Reading is taken when the subject experiences sour/metallic taste. All corresponding threshold values and findings were recorded and compared to control. Results showed 50% of patients had elevated threshold levels suggestive of CTN injury. However, none of the patients reported subjective taste loss. This study concludes that the prevelance rate of CTN injury in post myringoplasty patients is about 50% but this is not associated with altered taste sensation.

Inverse treatment planning by physically constrained minimization of a biological objective function.

In the current state-of-the art of clinical inverse planning, the design of clinically acceptable IMRT plans is predominantly based on the optimization of physical rather than biological objective functions. A major impetus for this trend is the unproven predictive power of radiobiological models, which is largely due to the scarcity of data sets for an accurate evaluation of the model parameters. On the other hand, these models do capture the currently known dose-volume effects in tissue dose-response, which should be accounted for in the process of optimization. In order to incorporate radiobiological information in clinical treatment planning optimization, we propose a hybrid physico-biological approach to inverse treatment planning based on the application of a continuous penalty function method to the constrained minimization of a biological objective. The objective is defined as the weighted sum of normal tissue complication probabilities evaluated with the Lyman normal-tissue complication probability model. Physical constraints specify the admissible minimum and maximum target dose. The continuous penalty function method is then used to find an approximate solution of the resulting large-scale constrained minimization problem. Plans generated by our approach are compared to ones produced by a commercial planning system incorporating physical optimization. The comparisons show clinically negligible differences, with the advantage that the hybrid technique does not require specifications of any dose-volume constraints to the normal tissues. This indicates that the proposed hybrid physico-biological method can be used for the generation of clinically acceptable plans.

On the implementation of dose-volume objectives in gradient algorithms for inverse treatment planning.

A method that allows a straightforward implementation of dose-volume constraints in gradient algorithms for inverse treatment planning is presented. The method is consistent with the penalty function approach, which requires the formulation of an objective function with penalty terms proportional to the magnitudes of constraint violations. Dose constraints with respect to minimum and maximum target dose levels are incorporated in quadratic, dose-penalty terms. Analogously, quadratic volume-penalty terms in the objective function reflect the violation of dose-volume constraints imposing limits on the fractions of healthy organ volumes that can be irradiated above specified dose levels. It has been demonstrated that within the framework of this formulation neither modified objective functions nor finite difference gradient calculations are necessary for the incorporation of gradient minimization algorithms. As an example, a simple steepest descent algorithm is presented along with its application to illustrate prostate and lung cases.

Cell kinetics and repopulation parameters of irradiated xenograft tumours in SCID mice: comparison of two dose-fractionation regimens.

The extent and mechanism(s) of repopulation were assessed in SiHa (human cervical squamous cell carcinoma) xenografts in SCID mice for two fractionated irradiation regimens. Mice in one arm of the study received 50 Gy in 20 fractions over 23 days with a 14 day split between 10 fraction, 5 day courses. The other tumours were treated with 50 Gy in 20 fractions over 10 consecutive days. Cell kinetics and tumour regrowth parameters were monitored during and after treatment by measuring tumour volume and analysing cellular DNA content and proliferation parameters with flow cytometry. Repopulation occurred rapidly, beginning during irradiation and largely attributable to an increased growth fraction and decreased potential doubling time, apparently triggered by increased cell loss. Cell cycle time, in contrast, remained relatively constant throughout. Extrapolation of these results to humans suggests that treatment times should be minimised whenever possible, since regrowth rates exceeded those predicted from pretreatment Tpot measurements.

Repopulation characteristics and cell kinetic parameters resulting from multi-fraction irradiation of xenograft tumors in SCID mice.

PURPOSE: Cell kinetics and repopulation rates during multifraction irradiation have previously been measured in SiHa human cervical carcinoma cells grown as spheroids. The current study applied similar techniques to SiHa tumor xenografts with the ultimate goal of assessing the clinical prognostic value of in situ cell kinetics. METHODS AND MATERIALS: SiHa (human squamous cell cervical tumor) cells were inoculated subcutaneously in the flank or back of SCID mice. When tumors reached a size of 200-300 mg, they received 25 Gy in 10 fractions over 5 days. Tumor regrowth and cell kinetics parameters were followed during treatment, and for 10 days after completion by measuring tumor volume and analyzing cellular BrdUrd and IdUrd incorporation with flow cytometry. RESULTS: Tumor volume was of limited use in assessing response to irradiation. The fraction of proliferating cells increased early during irradiation as did the labeling index; potential doubling time (Tpot) decreased during treatment and returned to the pre-irradiation value after treatment. Cell cycle time remained relatively constant throughout the experiments. CONCLUSION: These results confirm the feasibility of evaluating cell cycle kinetics and repopulation parameters in a murine tumor model undergoing a fractionated course of irradiation. Repopulation of clonogenic tumor cells occurred more rapidly than predicted by pretreatment measurements, primarily due to an increased growth fraction and consequent decrease in Tpot.

The lifetime of hypoxic human tumor cells.

PURPOSE: For hypoxic and anoxic cells in solid tumors to be a therapeutic problem, they must live long enough to be therapeutically relevant, or else be rapidly recruited into the proliferating compartment during therapy. We have, therefore, estimated lifetime and recruitment rate of hypoxic human tumor cells in multicell spheroids in vitro, or in xenografted tumors in SCID mice. MATERIALS AND METHODS: Cell turnover was followed by flow cytometry techniques, using antibodies directed at incorporated halogenated pyrimidines. The disappearance of labeled cells was quantified, and verified to be cell loss rather than label dilution. Repopulation was studied in SiHa tumor xenografts during twice-daily 2.5-Gy radiation exposures. RESULTS: The longevity of hypoxic human tumor cells in spheroids or xenografts exceeded that of rodent cell lines, and cell turnover was slower in xenografts than under static growth as spheroids. Human tumor cells remained viable in the hypoxic regions of xenografts for 4-10 days, compared to 3-5 days in spheroids, and 1-3 days for most rodent cells in spheroids. Repopulation was observed within the first few radiation treatments for the SiHa xenografts and, with accumulated doses of more than 10 Gy, virtually all recovered cells had progressed through at least one S-phase. CONCLUSION: Our results suggest an important difference in the ability of human vs. rodent tumor cells to withstand hypoxia, and raise questions concerning the increased longevity seen in vivo relative to the steady-state spheroid system.

Cell kinetics and repopulation mechanisms during multifraction irradiation of spheroids.

BACKGROUND AND PURPOSE: The objective of the present study was to evaluate the predictive potential of cell kinetic parameters and repopulation rates determined by flow cytometry during multifraction irradiation of spheroids, a system in which the fate of all cells can be determined with high precision. Ultimately, similar analytical techniques should provide a reproducible and prognostically significant clinical predictive assay. MATERIALS AND METHODS: Multicellular spheroids of Chinese hamster V79 lung cells were irradiated with 2.5 Gy of 250 kVp X-rays twice daily to a total dose of 25 Gy. Repopulation parameters and cell kinetic parameters were followed throughout the irradiation period and for 5 days after completion of exposure. RESULTS: (1) Regrowth (RG) took place early during multifraction irradiation. (2) Potential doubling time (Tpot) decreased steadily from the early part of treatment, remaining of short duration until the spheroids almost attained the pre-treatment number of clonogenic cells. (3) Accelerated repopulation was mainly due to a decreased cell loss factor (phi) and increased growth fraction (GF), although a modest decrease in cell cycle time (tc) was suggested. (4) Phi decreased during exponential RG. (5) Other parameters such as observed doubling time (td) and labelling index (LI) paralleled these findings. CONCLUSIONS: Clonogen repopulation that began early in the irradiation scheme and accelerated rapidly is not consistent with the prevailing view that accelerated repopulation begins several weeks into clinical protocols. Also, pre-treatment Tpot did not adequately estimate the repopulation speed in the spheroids. Equivalent studies in animal tumour systems, and then in the clinic, are consequently indicated and of some urgency.