Insights from an ancient gymnosperm lineage: ambient temperature and light and the timing of thermogenesis in cycad cones.

PREMISE: Although maintaining the appropriate mid-day timing of the diel thermogenic events of cones of the dioecious cycads Macrozamia lucida and M. macleayi is central to the survival of both plant and pollinator in this obligate pollination mutualism, the nature of the underlying mechanism remains obscure. We investigated whether it is under circadian control. Circadian mechanisms control the timing of many ecologically important processes in angiosperms, yet only a few gymnosperms have been studied in this regard. METHODS: We subjected cones to different ambient temperature and lighting regimens (constant temperature and darkness; stepwise cool/warm ambient temperatures in constant darkness; stepwise dark/light exposures at constant temperature) to determine whether the resulting timing of their thermogenic events was consistent with circadian control. RESULTS: Cones exposed to constant ambient temperature and darkness generated multiple temperature peaks endogenously, with an average interpeak-temperature period of 20.7 (±0.20) h that is temperature-compensated (Q10 = 1.02). Exposure to 24-h ambient temperature cycles (12 h cool/12 h warm, constant darkness) yielded an interpeak-temperature period of 24.0 (±0.05) h, accurately and precisely replicating the ambient temperature period. Exposure to 24-h photo-cycles (12 h light/12 h dark, constant ambient temperature) yielded a shorter, more variable interpeak-temperature period of 23 (±0.23) h. CONCLUSIONS: Our results indicate that cycad cone thermogenesis is under circadian clock control and differentially affected by ambient temperature and light cycles. Our data from cycads (an ancient gymnosperm lineage) adds to what little is known about circadian timing in gymnosperms, which have rarely been studied from the circadian perspective.

Unique chemistry associated with diversification in a tightly coupled cycad-thrips obligate pollination mutualism.

Cycad cone thermogenesis and its associated volatiles are intimately involved in mediating the behavior of their obligate specialist pollinators. In eastern Australia, thrips in the Cycadothrips chadwicki species complex are the sole pollinators of many Macrozamia cycads. Further, they feed and reproduce entirely in the pollen cones. M. miquelii, found only in the northern range of this genus, is pollinated only by a C. chadwicki cryptic species that is the most distantly related to others in the complex. We examined the volatile profile from M. miquelii pollen and ovulate (receptive and non-receptive) cones to determine how this mediates pollination mechanistically, using GC-MS (gas chromatography-mass spectrometry) and behavioral tests. Monoterpenes comprise the bulk of M. miquelii volatile emissions, as in other Macrozamia species, but we also identified compounds not reported previously in any cycad, including three aliphatic esters (prenyl acetate and two of uncertain identity) and two aliphatic alcohols. The two unknown esters were confirmed as prenyl (3-methylbut-2-enyl) esters of butyric and crotonic ((E))-but-2-enoic) acids after chemical synthesis. Prenyl crotonate is a major component in emissions from pollen and receptive ovulate cones, is essentially absent from non-receptive cones, and has not been reported from any other natural source. In field bioassays, Cycadothrips were attracted only to those volatile treatments containing prenyl crotonate. We discuss M. miquelii cone odorants relative to those of other cycads, especially with respect to prenyl crotonate being a species-specific signal to this northern C. chadwicki cryptic species, and how this system may have diversified.

Thermogenic respiratory processes drive the exponential increase of volatile organic compound emissions in Macrozamia cycad cones.

An important outcome of plant thermogenesis is increased emissions of volatiles that mediate pollinator behaviour. We investigated whether the large increase in emissions, mainly the monoterpene ß-myrcene (>90%), during daily thermogenic events of Macrozamia macleayi and lucida cycad cones are due solely to the influence of high cone temperatures or are, instead, a result of increased respiratory rates during thermogenesis. We concurrently measured temperature, oxygen consumption and ß-myrcene emission profiles during thermogenesis of pollen cones under typical environmental temperatures and during experimental manipulations of cone temperatures and aerobic conditions, all in the dark. The exponential rise in ß-myrcene emissions never occurred without a prior, large increase in respiration, whereas an increase in cone temperature alone did not increase emissions. When respiration during thermogenesis was interrupted by anoxic conditions, ß-myrcene emissions decreased. The increased emission rates are not a result of increased cone temperature per se (through increased enzyme activity or volatilization of stored volatiles) but are dependent on biosynthetic pathways associated with increased respiration during thermogenesis that provide the carbon, energy (ATP) and reducing compounds (NADPH) required for ß-myrcene production through the methylerythritol phosphate (MEP) pathway. These findings establish the significant contribution of respiration to volatile production during thermogenesis.

Adaptive model-predictive controller for magnetic resonance guided focused ultrasound therapy.

PURPOSE: Minimising treatment time and protecting healthy tissues are conflicting goals that play major roles in making magnetic resonance image-guided focused ultrasound (MRgFUS) therapies clinically practical. We have developed and tested in vivo an adaptive model-predictive controller (AMPC) that reduces treatment time, ensures safety and efficacy, and provides flexibility in treatment set-up. MATERIALS AND METHODS: The controller realises time savings by modelling the heated treatment cell's future temperatures and thermal dose accumulation in order to anticipate the optimal time to switch to the next cell. Selected tissues are safeguarded by a configurable temperature constraint. Simulations quantified the time savings realised by each controller feature as well as the trade-offs between competing safety and treatment time parameters. In vivo experiments in rabbit thighs established the controller's effectiveness and reliability. RESULTS: In all in vivo experiments the target thermal dose of at least 240 CEM43 was delivered everywhere in the treatment volume. The controller's temperature safety limit reliably activated and constrained all protected tissues to <9 CEM43. Simulations demonstrated the path independence of the controller, and that a path which successively proceeds to the hottest untreated neighbouring cell leads to significant time savings, e.g. when compared to a concentric spiral path. Use of the AMPC produced a compounding time-saving effect; reducing the treatment cells' heating times concurrently reduced heating of normal tissues, which eliminated cooling periods. CONCLUSIONS: Adaptive model-predictive control can automatically deliver safe, effective MRgFUS treatments while significantly reducing treatment times.

The accuracy and precision of two non-invasive, magnetic resonance-guided focused ultrasound-based thermal diffusivity estimation methods.

PURPOSE: The use of correct tissue thermal diffusivity values is necessary for making accurate thermal modelling predictions during magnetic resonance-guided focused ultrasound (MRgFUS) treatment planning. This study evaluates the accuracy and precision of two non-invasive thermal diffusivity estimation methods, a Gaussian temperature method and a Gaussian specific absorption rate (SAR) method. MATERIALS AND METHODS: Both methods utilise MRgFUS temperature data obtained during cooling following a short (<25 s) heating pulse. The Gaussian SAR method can also use temperatures obtained during heating. Experiments were performed at low heating levels (ΔT∼10 °C) in ex vivo pork muscle and in vivo rabbit back muscle. The non-invasive MRgFUS thermal diffusivity estimates were compared with measurements from two standard invasive methods. RESULTS: Both non-invasive methods accurately estimated thermal diffusivity when using MR temperature cooling data (overall ex vivo error <6%, in vivo <12%). Including heating data in the Gaussian SAR method further reduced errors (ex vivo error <2%, in vivo <3%). The significantly lower standard deviation values (p < 0.03) of the Gaussian SAR method indicated that it had better precision than the Gaussian temperature method. CONCLUSIONS: With repeated sonications, either MR-based method could provide accurate thermal diffusivity values for MRgFUS therapies. Fitting to more data simultaneously likely made the Gaussian SAR method less susceptible to noise, and using heating data helped it converge more consistently to the FUS fitting parameters and thermal diffusivity. These effects led to the improved precision of the Gaussian SAR method.

Cone thermogenesis and its limits in the tropical Cycas micronesica (Cycadaceae): association with cone growth, dehiscence, and post-dehiscence phases.

PREMISE OF THE STUDY: Thermogenesis is a prominent pollination-related feature of cycad cones and is generally assumed to play a role in pollination. Although typically studied just before, during, and immediately after the cones' pollination phase, thermogenesis may be present in other cone developmental phases. • METHODS: We assayed thermogenesis in Cycas micronesica, Guam's endangered cycad, over successive cone developmental phases by measuring temperatures in shaded and unshaded in situ cones for up to 7 wk. We also studied the effect of ambient conditions on cone thermogenesis in laboratory experiments and estimated the cones' metabolic heating rates. • KEY RESULTS: Pollen cones exhibit a continuous, but small, metabolically generated thermogenesis for multiple weeks, including a single thermogenic peak temperature greater than peak ambient each day. The magnitudes of those daily peak temperature elevations above ambient reach maxima twice during cone development: a few days before dehiscence and approximately 1 wk post-dehiscence. Excised cones in dark, fixed temperature environments generated multiple thermogenic events (∼24 h period) over ∼10 d. Cones appear to initiate a protective temperature regulatory response at temperatures ≥∼38°C. • CONCLUSIONS: Cycas micronesica pollen cones exhibit several thermogenic attributes not reported in other cycads, including continuous thermogenesis for many weeks. These cones grow in a hot tropical environment that likely confines their metabolically generated temperature increases to a small thermogenic window beyond which they encounter heat stress. These findings suggest the presence of thermogenic functions not strictly related to pollination and a potential vulnerability to warming climates.

Isolated kidney phantom for development of biothermal vascular models with application to high intensity focused ultrasound therapy.

A methodology using magnetic resonance angiography (MRA) is presented for identifying thermally significant blood vessels in isolated kidneys, specifically for use in biothermal model development with application to high intensity focused ultrasound (HIFU). A combination of a proven preservation technique, newly developed MR-compatible experimental procedures and the refinement of MR pulse sequence parameters was used to determine vascular characteristics using high-resolution three-dimensional time-of-flight MRA image of flow through isolated kidneys. Results presented are twofold. First, improved vessel visibility was attained through decreasing the magnetic resonance imaging bandwidth from 150 to 30 Hz/pixel while simultaneously increasing the echo time, repetition time, and flip angle; vascular center line extraction showed an 18% improvement in the number of vessel segments detected and a 23% increase in length of the terminal segments over a base line technique without improvements. Second, the overall system was shown to be practical to determine vascular flow effects during HIFU heating; testing results from heating the kidney with HIFU are presented, showing a decrease of average kidney temperature with an increase of flow rate through the kidney with localized cooling demonstrated surrounding known vessel locations.

Unstable, self-limiting thermochemical temperature oscillations in Macrozamia cycads.

Field measurements and laboratory experiments on the Australian cycads Macrozamia lucida and Macrozamia macleayi demonstrate that their cones' diel peak thermogenic temperature increase varies systematically with cone stage, with single thermogenic temperature peaks occurring daily for up to 2 weeks and reaching 12 degrees C above ambient at midstage. The initiation, magnitude and timing of those peaks are strongly modulated by ambient temperature; the period between successive thermogenic temperature peaks is not circadian, and light is neither necessary nor sufficient to initiate a thermogenic event. A mathematical analysis is developed that provides a unified explanation of the experimental results. It describes these unstable, self-limiting thermogenic events in terms of conservation of energy and a first-order chemical reaction rate model that includes an Arrhenius equation dependence of the cone's metabolic heating rate on the cone temperature.

A comparative analysis of three self-balancing wheelchair balancing mechanisms.

In the last 20 years, three different basic, dynamic balancing designs have been proposed for a self-balancing wheelchair (SBW) that allows the wheelchair user to transition from driving on all four wheels to driving while balanced on the two large rear wheels. The dynamic performance of these three SBW designs, the hanging pendulum counterweight (HPC), the single inverted pendulum (SIP), and the double inverted pendulum (DIP), are compared when controlled by a common state space controller. The four dynamic performance considerations of stability, driver dynamic stress, maneuverability and technical requirements were used to compare these designs while performing the following five tests: 1) transition from four-wheel to two-wheel, balancing mode; 2) stationary, self-balancing stability when subjected to an impact disturbance; 3) movement initiation, and stopping while balancing; 4) response to impact disturbances while moving; and 5) stability on low traction surfaces. In addition, the movement initiation and stopping test was repeated with increased chair mass and inertia to investigate the sensitivity of model performance to changes in model parameters. After comparing the three models it was determined that the HPC mechanism is the best choice for further development based on the criteria of stability, driver dynamic stress, maneuverability, and technical requirements. The HPC ranked equal or better compared to the SIP and DIP on 15 of 29 stability and performance factors. It was also the only design that was stable for all normally expected driving conditions.

Readdressing the issue of thermally significant blood vessels using a countercurrent vessel network.

A physiologically realistic arterio-venous countercurrent vessel network model consisting of ten branching vessel generations, where the diameter of each generation of vessels is smaller than the previous ones, has been created and used to determine the thermal significance of different vessel generations by investigating their ability to exchange thermal energy with the tissue. The temperature distribution in the 3D network (8178 vessels; diameters from 10 to 1000 microm) is obtained by solving the conduction equation in the tissue and the convective energy equation with a specified Nusselt number in the vessels. The sensitivity of the exchange of energy between the vessels and the tissue to changes in the network parameters is studied for two cases; a high temperature thermal therapy case when tissue is heated by a uniformly distributed source term and the network cools the tissue, and a hypothermia related case, when tissue is cooled from the surface and the blood heats the tissue. Results show that first, the relative roles of vessels of different diameters are strongly determined by the inlet temperatures to those vessels (e.g., as affected by changing mass flow rates), and the surrounding tissue temperature, but not by their diameter. Second, changes in the following do not significantly affect the heat transfer rates between tissue and vessels; (a) the ratio of arterial to venous vessel diameter, (b) the diameter reduction coefficient (the ratio of diameters of successive vessel generations), and (c) the Nusselt number. Third, both arteries and veins play significant roles in the exchange of energy between tissue and vessels, with arteries playing a more significant role. These results suggest that the determination of which diameter vessels are thermally important should be performed on a case-by-case, problem dependent basis. And, that in the development of site-specific vessel network models, reasonable predictions of the relative roles of different vessel diameters can be obtained by using any physiologically realistic values of Nusselt number and the diameter reduction coefficient.

Control of thermal therapies with moving power deposition field.

A thermal therapy feedback control approach to control thermal dose using a moving power deposition field is developed and evaluated using simulations. A normal tissue safety objective is incorporated in the controller design by imposing constraints on temperature elevations at selected normal tissue locations. The proposed control technique consists of two stages. The first stage uses a model-based sliding mode controller that dynamically generates an 'ideal' power deposition profile which is generally unrealizable with available heating modalities. Subsequently, in order to approximately realize this spatially distributed idealized power deposition, a constrained quadratic optimizer is implemented to compute intensities and dwell times for a set of pre-selected power deposition fields created by a scanned focused transducer. The dwell times for various power deposition profiles are dynamically generated online as opposed to the commonly employed a priori-decided heating strategies. Dynamic intensity and trajectory generation safeguards the treatment outcome against modelling uncertainties and unknown disturbances. The controller is designed to enforce simultaneous activation of multiple normal tissue temperature constraints by rapidly switching between various power deposition profiles. The hypothesis behind the controller design is that the simultaneous activation of multiple constraints substantially reduces treatment time without compromising normal tissue safety. The controller performance and robustness with respect to parameter uncertainties is evaluated using simulations. The results demonstrate that the proposed controller can successfully deliver the desired thermal dose to the target while maintaining the temperatures at the user-specified normal tissue locations at or below the maximum allowable values. Although demonstrated for the case of a scanned focused ultrasound transducer, the developed approach can be extended to other heating modalities with moving deposition fields, such as external and interstitial ultrasound phased arrays, multiple radiofrequency needle applicators and microwave antennae.

MR thermometry-based feedback control of efficacy and safety in minimum-time thermal therapies: phantom and in-vivo evaluations.

The experimental validation of a model-based, thermal therapy control system which automatically and simultaneously achieves the specified efficacy and safety objectives of the treatment is reported. MR-thermometry measurements are used in real-time to control the power of a stationary, focused ultrasound transducer in order to achieve the desired treatment outcome in minimum time without violating the imposed safety constraints. Treatment efficacy is quantified in terms of the thermal dose delivered to the target. Normal tissue safety is ensured by automatically maintaining normal tissue temperature below the imposed limit in the user-specified locations. To reflect hardware limitations, constraints on the maximum applied power are also imposed. At the pretreatment stage, MR imaging and thermometry are used to localize the treatment target and identify thermal and actuation models. The results of phantom and canine experiments demonstrate that spatially-distributed, real-time MR temperature measurements enhance one's ability to robustly achieve the desired treatment outcome in minimum time without violating safety constraints. Post-treatment evaluation of the outcome using T2-weighted images of canine muscle showed good spatial correlation between the sonicated area and thermally damaged tissue.

An analytical study of 'Poisson conduction shape factors' for two thermally significant vessels in a finite, heated tissue.

To conveniently and properly account for the vessel to vessel and vessel to tissue heat transfer rates to predict in vivo tissue temperature distributions, this paper analyses two different types of Poisson conduction shape factors (PCSFs) for unheated and/or uniformly heated, non-insulated, finite tissue domains. One is related to the heat transfer rate from one vessel to another (vessel-vessel PCSF (VVPCSF)) and the other is related to the vessel to tissue heat transfer rates (vessel-tissue PCSF (VTPCSF)). Two alternative formulations for the VTPCSFs are studied; one is based on the difference between the vessel wall and tissue boundary temperatures, and the other on the difference between the vessel wall and the average tissue temperatures. The effects of a uniform source term and of the diameters and locations of the two vessels on the PCSFs are studied for two different cases: one, when the vessel wall temperatures are lower than the tissue boundary temperature, i.e., the vessels cool the tissue, and vice versa. Results show that, first, the VVPCSFs are only geometry dependent and they do not depend on the applied source term and the vessel wall and tissue boundary temperatures. Conversely, the VTPCSFs are strong functions of the source term and of the temperatures of the vessel walls and tissue boundary. These results suggest that to account for the vessel to vessel heat transfer rates, the VVPCSFs can be evaluated solely based on the vessel network geometry. However, to account for the vessel to tissue heat transfer rates, the VTPCSFs should be used iteratively while solving for the tissue temperature distributions. Second, unlike the tissue boundary temperature-based VTPCSFs which may become singular only in heated tissues, the average tissue temperature-based VTPCSFs have the potential to become singular in both unheated and heated tissues. These results suggest that caution should be exercised in the use of the VTPCSFs since they may approach singularity by virtue of their definition and thus may introduce large errors in the evaluation of tissue temperature distribution. Presented results are new and complementary to the previous shape factor results since these include the effect of (1) source term and (2) unequal vessel-tissue heat transfer rates from the two vessels to the tissue.

Direct thermal dose control of constrained focused ultrasound treatments: phantom and in vivo evaluation.

The first treatment control system that explicitly and automatically balances the efficacy and safety goals of noninvasive thermal therapies is described, and its performance is evaluated in phantoms and in vivo using ultrasound heating with a fixed, focused transducer. The treatment efficacy is quantified in terms of thermal dose delivered to the target. The developed feedback thermal dose controller has a cascade structure with the main nonlinear dose controller continuously generating the reference temperature trajectory for the secondary, constrained, model predictive temperature controller. The control system ensures thermal safety of the normal tissue by automatically complying with user-specified constraints on the maximum allowable normal tissue temperatures. To reflect hardware limitations and to prevent cavitation, constraints on the maximum transducer power can also be imposed. It is shown that the developed controller can be used to achieve the minimum-time delivery of the desired thermal dose to the target without violating safety constraints, which is a novel and clinically desirable feature. The developed controller is model based, and requires patient- and site-specific models for its operation. These models were obtained during pre-treatment identification experiments. In our implementation, predictive models, internally used by the automatic treatment controller, are dynamically updated each time new temperature measurements become available. The adaptability of internal models safeguards against adverse effects of modelling errors, and ensures robust performance of the control system in the presence of a priori unknown treatment disturbances. The successful validation with two experimental models of considerably different thermal and ultrasound properties suggests the applicability of the developed treatment control system to different anatomical sites.

Blood perfusion and thermal conduction effects in Gaussian beam, minimum time single-pulse thermal therapies.

A previous analytical study has shown that the minimum obtainable treatment time for a single pulse that delivers a given thermal dose to a specified point at a specified time occurs when the temperature at that point is rapidly raised to its maximum allowable value. The present study extends that result by investigating the spatial distribution of thermal effects of a single Gaussian shaped focal zone pulse that reaches that maximum allowable temperature at the center point of the focal zone. Analytical solutions are obtained that separately include the effects of perfusion and conduction. This situation is analyzed for a conservative treatment strategy in which the desired thermal dose is delivered when the tumor cools down to basal conditions. The results show that for a specified thermal dose delivered by a spherical Gaussian beam with focal widths below approximately 4 mm, the maximum allowable temperature, the minimum obtainable treatment time, and the size of the treatment zone (as a percentage of the size of the Gaussian beam) are all independent of the tissue blood perfusion, and are only functions of the focal zone size. Conversely, for focal widths above approximately 20 cm, these results are independent of the focal width and are only functions of blood perfusion. Between these two sizes (where most practical treatments will occur, since single pulses with widths of <4 mm and >20 cm will be uncommon in practice) a transition zone exists in which both perfusion and conduction effects are important. Thus while it is possible to implement a truly perfusion-independent, single pulse thermal treatment by using focal widths of <4 mm, in practice many such pulses will be needed to treat most tumors. This is especially true since the nonlinear temperature/thermal dose relationship causes the width of the delivered dose distribution to be only approximately 25%-30% of the width of the focal zone. However, shorter overall treatment times can be obtained when multiple pulses are linked together by using larger focal zone sizes, but this gain in treatment time is accompanied by increased effects of perfusion, illustrating the conflict between attaining both perfusion-independence and minimal treatment time for multiple-pulse thermal treatments.

Minimum-time thermal dose control of thermal therapies.

The problem of controlling noninvasive thermal therapies is formulated as the problem of directly controlling thermal dose of the target. To limit the damage to the surrounding normal tissue, the constraints on the peak allowable temperatures in the selected spacial locations are imposed. The developed controller has a cascade structure with a linear, constrained, model predictive temperature controller in the secondary loop. The temperature controller manipulates the intensity of the ultrasound transducer with saturation constraints, which noninvasively heats the spatially distributed target. The main nonlinear thermal dose controller dynamically generates the reference temperature trajectories for the temperature controller. The thermal dose controller is designed to force the treatment progression at either the actuation or temperature constraints, which is required to minimize the treatment time. The developed controller is applicable to high and low-intensity treatments, such as thermal ablation and thermoradiotherapy. The developed approach is tested using computer simulations for a one-dimensional model of a tumor with constraints on the maximum allowable temperature in the normal tissue and a constrained power output of the ultrasound transducer. The simulation results demonstrate that the proposed approach is effective at delivering the desired thermal dose in a near minimum time without violating constraints on the maximum allowable temperature in healthy tissue, despite significant plant-model mismatch introduced during numerical simulation. The results of in vitro and in vivo validation are reported elsewhere.

Model-predictive control of hyperthermia treatments.

A model-predictive controller (MPC) of the thermal dose in hyperthermia cancer treatments has been developed and evaluated using simulations with one-point and one-dimensional models of a tumor. The developed controller is the first effort in: 1) the application of feedback control to pulsed, high-temperature hyperthermia treatments; 2) the direct control of the treatment thermal dose rather than the treatment temperatures; and 3) the application of MPC to hyperthermia treatments. Simulations were performed with different blood flow rates in the tumor and constraints on temperatures in normal tissues. The results demonstrate that 1) thermal dose can be controlled in the presence of plant-model mismatch and 2) constraints on the maximum allowable temperatures in normal tissue and/or the pulsed power magnitude can be directly incorporated into MPC and met while delivering the desired thermal dose to the tumor. For relatively high blood flow rates and low transducer surface intensities--factors that limit the range of temperature variations in the tumor, the linear MPC, obtained by piece-wise linearization of the dose-temperature relationship, provides an adequate performance. For large temperature variations, the development of nonlinear MPC is necessary.