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PubMed2004

Production of Carbopol 974P and Carbopol 971P pellets by extrusion-spheronization: optimization of the processing parameters and water content.

Pellets obtained by extrusion-spheronization represent multiparticulate dosage forms whose interest in intestinal drug delivery can be potentiated and targeted through bioadhesive properties. However, adhesion itself makes the process difficult or even impossible. The problem of tackiness encountered with bioadhesive wet masses was previously eliminated by the use of electrolytes such as CaCl2. This approach is known to reduce the viscosity of polyacrylic acids by disturbing the interactions between carboxylate groups on adjacent polymer molecules, thereby decreasing their bioadhesive properties. The present study aimed at producing pellets containing carbomers without addition of electrolytes in order to maintain their bioadhesive potentiality at its maximum. Carbopol 974P (10%, 15% and 20%) and Carbopol 971P (10%) were used in combination with Avicel PH101. The extrusion speed (30, 45, 60, 90, and 150 rpm), spheronizer speed (350, 700, 960, 1000, and 1300 rpm), spheronization time (5, 10, 15, and 20 minutes) and amount of water (45%, 50%, 54%, and 58%) were optimized in order to obtain the highest yield of spherical pellets ranging 710-1000 microm in diameter. For pellets containing 10%, 15% Carbopol 974P or 10% Carbopol 971P and 45% water content, 30 rpm extrusion speed, 960 rpm, and 10 minutes spheronization speed and time led to the highest yields and sphericities, respectively, 72% and 0.91, 67% and 0.78, and 76% and 0.80. Production of pellets with 20% Carbopol 974P could be achieved through the increase of the water content up to 58% and implementation of 30 rpm extrusion speed, 1300 rpm, and 10 minutes spheronization speed and time. The yield and sphericity were 42% and 0.78 respectively.

Acrylates↗

PubMed2004

Effects of lactose as an inducer on expression of Helicobacter pylori rUreB and rHpaA, and Escherichia coli rLTKA63 and rLTB.

AIM: To demonstrate the effect of lactose as an inducer on expression of the recombinant proteins encoded by Helicobacter pylori ureB and hpaA, and Escherichia coli LTB and LTKA63 genes and to determine the optimal expression parameters. METHODS: By using SDS-PAGE and BIO-RAD gel image analysis system, the outputs of the target recombinant proteins expressed by pET32a-ureB-E.coliBL21, pET32a-hpaA-E.coliBL21, pET32a-LTKA63-E.coliBL21 and pET32a-LTB-E.coliBL21 were measured when using lactose as inducer at different dosages, original bacterial concentrations, various inducing temperatures and times. The results of the target protein expression induced by lactose were compared to those by isopropyl-beta-D-thiogalactoside (IPTG). The proteins were expressed in E.coli. RESULTS: Lactose showed higher efficiency of inducing the expression of rHpaA, rUreB, rLTB and rLTKA63 than IPTG. The expression outputs of the target recombinant proteins induced at 37 degrees were remarkably higher than those at 28 degrees. Other optimal expression parameters for the original bacterial concentrations, dosages of lactose and inducing time were 0.8, 50 g/L and 4 h for rHpaA; 0.8, 100 g/L and 4 h for rLTKA63; 1.2, 100 g/L and 5 h for both rUreB and rLTB, respectively. CONCLUSION: Lactose, a sugar with non-toxicity and low cost, is able to induce the recombinant genes to express the target proteins with higher efficiency than IPTG. The results in this study establish a beneficial foundation for industrial production of H pylori genetic engineering vaccine.

Adhesins, Bacterial↗

PubMedDate not supplied

3D virtual endoscopy of the upper airway: optimization of the scan parameters in a cadaver phantom and clinical assessment.

Our goal was to evaluate the influence of the scan parameters on the 3D virtual endoscopy of the larynx and trachea and the clinical assessment. Helical CT (HiSpeed Advantage; GE, Milwaukee, WI, U.S.A.) of a cadaver phantom was performed with increased collimation (1-10 mm) and pitch (0.5-3). Seventy-two patients with complaints of the upper airways were investigated by virtual endoscopy and their results were compared with the findings of regular endoscopy. Best correlation between virtual endoscopy and anatomical findings, diagnostic quality of the axial slices, and useful longitudinal coverage of the examination were obtained with a collimation of 3 mm and a pitch of 1.5. Space-occupying tumors and stenosis were detected correctly, but the mucous membrane could not be visualized. 3D virtual endoscopy proved to be a valuable method for displaying anatomical structures. For an optimal protocol, a collimation of 3 mm with a pitch of 1.5 is recommended.

Endoscopy↗

PubMed2006

Optimization of peak power of doubly Q-switched lasers with both an acousto-optic modulator and a Cr4+-doped saturable absorber.

A doubly Q-switched laser can obtain a shorter pulse with a stable repetition rate and high pulse peak power, which has been experimentally proved. By taking into account the Gaussian spatial distributions of the intracavity photon density and the initial population-inversion density as well as the influence of the acousto-optic (AO) Q switch, we introduce the coupled rate equations for a doubly Q-switched laser with both an AO modulator and a Cr(4+)-doped saturable absorber. These coupled rate equations are solved numerically. The key parameters of an optimally coupled doubly Q-switched laser are determined based on maximizing the peak power, which include the optimal normalized coupling parameter, the optimal normalized saturable absorber parameters, and the normalized parameters of the AO Q switch. The optimal normalized peak power, the corresponding normalized energy, and the normalized pulse width are also given, and a group of general curves are generated for the first time to our knowledge. The curves can give us a good understanding of the dependence of the optimal key parameters on the parameters of the gain medium, the saturable absorber, the AO Q switch, the resonator, and the spatial distributions of the intracavity photon density. The optimal calculations for a diode-pumped Nd(3+):YVO(4) laser with both an AO modulator and a Cr(4+):YAG saturable absorber are presented to demonstrate the use of the curves and the related formulas.

Journal Article↗

PubMed2004

A parameter study to determine the optimal source neutron energy in boron neutron capture therapy of brain tumours.

The values of the parameters used in boron neutron capture therapy (BNCT) to calculate a given dose to human tissue vary with patients due to different physical, biological and/or medical circumstances. Parameters include the tissue dimensions, the 10B concentration and the relative biological effectiveness (RBE) factors for the different dose components associated with BNCT. Because there is still no worldwide agreement on RBE values, more often than not, average values for these parameters are used. It turns out that the RBE-problem can be circumvented by taking into account all imaginable parameter values. Approaching this quest from another angle: the outcome will also provide the parameters (and values) which influence the optimal source neutron energy. For brain tumours it turns out that the 10B concentration, the RBE factors for 10B as well as fast neutrons, together with the dose limit set for healthy tissue, affect the optimal BNCT source neutron energy. By using source neutrons of a few keV together with neutrons of a few eV, it ensures that, under all imaginable circumstances, a maximum of alpha (and lithium) particles can be delivered in the tumour.

Algorithms↗

PubMed2001

[Cavitation bubble formation during Erbium:YAG laser vitrectomy].

BACKGROUND: Clinical and experimental studies demonstrated the potential advantages of Erbium:YAG laser vitrectomy for posterior segment surgery. However, a detailed knowledge on the laser-tissue interaction is needed for an optimization of new ophthalmic applications. The aim of this experimental work was to investigate the cavitation bubble formation during Erbium:YAG laser vitrectomy and to find optimized laser parameters and the best geometry of the aspiration port of the microsurgical probe. MATERIALS AND METHODS: We investigated the formation of cavitation bubbles in water by high-speed photography. The output energy at the quartz tip reached up to 50 mJ and the laser pulse duration ranged from 50 to 300 microseconds. Various commercially available microsurgery probes were investigated regarding the extent of the cavitation bubbles. RESULTS: The threshold for laser-induced cavitation bubble formation was found to be 0.32 +/- 0.1 mJ for a laser pulse duration of 130 microseconds and a core diameter of 320 microns of the quartz fiber tip. The length of the cavitation bubbles increases with the laser pulse energy up to a length of 1.6 mm at a pulse energy of 10 mJ. In contrast, the size of the vapor bubbles decreases with an increase of the laser pulse duration. A slit-shaped aspiration port led to a 50% smaller volume of the cavitation bubble exiting the port compared with a circular aspiration port. CONCLUSIONS: The optimized laser parameters and microsurgery probe geometry may significantly decrease the risk of intraoperative ocular damages during Erbium:YAG laser vitrectomy.

Animals↗

PubMed1992

MR imaging of joints: analytic optimization of GRE techniques at 1.5 T.

To clarify the choice of imaging parameters for optimal gradient-recalled echo MR scanning of joints, we analyzed the behavior of contrast-to-noise and signal-to-noise ratios for spoiled (i.e., fast low-angle shot [FLASH] or spoiled GRASS) and steady-state (i.e., gradient-recalled acquisition in the steady state [GRASS] or fast imaging with steady precession) techniques at 1.5 T. The analysis is based on tissue characteristics derived from spin-echo measurements of hyaline cartilage and synovial fluid signal in the patellofemoral joints of 11 volunteers. Separate analysis of contrast-to-noise and signal-to-noise ratios for multiplanar (long TR) acquisitions shows that these parameters are each improved compared with single-slice methods. At TRs greater than 250 msec, there is no significant difference in the contrast behavior of FLASH and GRASS. For optimal contrast-to-noise ratio (synovial fluid-cartilage), the best multiplanar sequence (for TE less than 23 msec) is with a short TE and a large flip angle (e.g., 400/9/73 degrees [TR/TE/flip angle]). If a single-scan or three-dimensional technique is desired, than a GRASS sequence at minimal TR and TE and intermediate flip angle (18/9/32 degrees) is best. For optimal signal-to-noise ratio (for both synovial fluid and hyaline cartilage), the best multiplanar sequence uses a short TE and an intermediate flip angle (e.g., 400/9/30 degrees). If a short TR, high signal-to-noise technique is desired, then GRASS (18/9/13 degrees) is superior to FLASH.

Cartilage, Articular↗

PubMed1995

Neodymium: yttrium-aluminum-garnet laser dosimetry for the prolase II side-firing delivery system in the human prostate.

OBJECTIVES: The objective of this study was to define optimal treatment parameters and quantitative dosimetry for neodymium:yttrium-aluminum-garnet (Nd:YAG) laser coagulation prostatectomy utilizing the ProLase II side-firing laser delivery system. METHODS: Transurethral Nd:YAG laser application was performed with the ProLase II side-firing laser fiber in adult human prostates prior to planned radical prostatectomy. Depth and volume of prostatic tissue coagulation for single, continuous laser applications were measured at variable power settings from 40 to 80 W, while holding total energy delivery constant. Holding the power setting constant, the extent of tissue coagulation was similarly measured for variable treatment times from 45 to 90 seconds. RESULTS: Peak tissue coagulation for the ProLase II was observed at 60 W--up to a maximum of 13 mm tissue penetration and 3.92 cc volume coagulation following a single spot laser application. The mean depth of tissue coagulation at 60 W power setting was 12.7 mm, with a mean volume of tissue coagulation of 3.07 cc. Holding the power setting constant at 60 W, and increasing treatment time from 45 to 60 seconds, the extent to tissue coagulation increased significantly. However, beyond 60 seconds continuous laser application at 60 W, a plateau in tissue effects was observed, with minimal increase in tissue coagulation between 60 and 90 seconds. CONCLUSIONS: Using the ProLase II side-firing laser fiber, 60-second Nd:YAG spot laser applications at 60 W power setting represent optimal treatment parameters. The quantitative dosimetry curves produced for the ProLase II fiber are compared with prior dosimetry studies with the Urolase delivery system.

Aged↗

PubMedDate not supplied

Production and optimization studies of cephalosporin C by solid state fermentation.

Production of cephalosporin C employing a strain, Cephalosporium sp. NCIM 1039 under solid state fermentation was optimized. Different substrates like wheat bran, wheat grains, rice grains, barley and rice bran were studied to optimize the best substrate. Wheat bran showed the highest antibiotic yield. The physical and chemical parameters were optimized. The maximum productivity of cephalosporin C (750 U/g) was achieved by employing wheat bran and with optimized nutritional and process parameters such as potato starch as additive 1% w/w, urea as additive 1% w/w, incubation period of 7 days, incubation temperature at 30 degrees C, inoculum level 10% w/v, moisture content of solid substrate 80% and pH 7.0.

Acremonium↗

PubMedDate not supplied

Technique for bilateral intracranial implantation of cells in monkeys using an automated delivery system.

Intracerebral grafting combined with gene transfer may provide a powerful technique for local delivery of therapeutic agents into the CNS. The present study was undertaken to: (i) develop a reliable and reproducible automated cell implantation system, (ii) determine optimal implantation parameters of cells into the striatum, (iii) determine upper safe limits of cellular implantation into the neostriatum of monkeys. Autologous fibroblasts were infused into six sites of the striatum in nonhuman primates (Macaca mulatta, n = 11). Twenty-six-gauge cannulae were inserted vertically through cortical entry sites into the striatum (two sites in the caudate nucleus and four sites in the putamen) at predefined coordinates based on magnetic resonance imaging (MRI). The cannulae were guided by an electronically operated, hydraulic micropositioner and withdrawn at controlled rates, while cells (5, 10, 20, 40, or 80 microl/site) were infused simultaneously. Varying infusion rates and cell concentrations were also evaluated. Visualization and evaluation of graft placement were performed using contrast MRI at 3-5 days postsurgery. Animals were monitored for signs of clinical complications and sacrificed 2 weeks following surgery. Postimplantation MRI revealed a tissue mass effect of the implant with shifting of midline, edema, and infiltration of the white tracts at 40 and 80 microl/site. In addition, these animals developed transient hemiparesis contralateral to the implant site. MRI of animals grafted with 20 microl/site exhibited columnar-shaped implants and evidence of infiltration into white matter tracts possibly due to a volume effect. No clinical side effects were seen in this group. At 14 days postsurgery, MRI scans showed consistent columnar grafts (measuring approximately 5 mm in height) throughout the striatum in animals implanted with 5 or 10 microl/site. No signs of clinical side effects were associated with these volumes and postmortem histological examination confirmed MRI observations. Optimal surgical parameters for delivery of cells into the striatum consist of a graft volume of 10 microl/site, an infusion rate of 1.6 microl/min, a cell concentration of 2.0 x 10(5) cells/microl, and a cannula withdrawal rate of 0.75 mm/min. These results show that infusion of cells into the striatum can be done in a safe and routine manner.

Animals↗

PubMed2005

[Determination of SARS-CoV by simplified nested fluorescent RT-PCR].

OBJECTIVE: To establish a simple rapid and sensitive nested RT-PCR method for detection of SARS coronavirus RNA by designing the specific primers for SARS and optimizing the parameters for PCR. METHODS: Primers and fluorescent probes were designed according to the sequences of SARS coronavirus genes available from GenBank. The optimization of the parameters for PCR was performed in PE 7700 thermal cycle. The 36 serum samples and 40 mouthwash of SARS patients and 80 samples of healthy people were tested. RESULTS: The positive rate of patient serum and mouthwash was 33.6%, (12/36) and 67.5%, (27/40), respectively, while the positive rate of healthy people was zero (0/160). CONCLUSION: The simple nested RT-PCR method was a rapid, efficient and sensitive one for SARS early diagnosis.

Bodily Secretions↗

PubMed2006

Endoscopic ablation of intestinal metaplasia containing high-grade dysplasia in esophagectomy patients using a balloon-based ablation system.

BACKGROUND: This study aimed to determine the optimal treatment parameters for the ablation of intestinal metaplasia (IM) containing high-grade dysplasia (HGD) using a balloon-based ablation system for patients undergoing esophagectomy. METHODS: Immediately before esophagectomy, patients underwent ablation of circumferential segments of the esophagus containing IM-HGD using the HALO360 system. The treatment settings were randomized to 10, 12, or 14 J/cm2 for two, three, or four applications. After esophagectomy, multiple sections from ablation zones were microscopically evaluated. Histologic end points included maximum ablation depth (histologic layer) and complete ablation of all IM-HGD (yes/no). RESULTS: Eight men with a mean age of 57 years (range, 45-71 years) were treated, and 10 treatment zones were created. There were no device-related adverse events. At resection, there was no evidence of a transmural thermal effect. Grossly, ablation zones were clearly demarcated sections of ablated epithelium. The maximum ablation depth was the lamina propria or muscularis mucosae. The highest energy (14 J/cm2, 4 applications) incurred edema in the superficial submucosa, but no submucosa ablation. Complete ablation of IM and HGD occurred in 9 of 10 ablation zones (90%), defined as complete removal of the epithelium with only small foci of "ghost cells" representing nonviable, ablated IM-HGD and demonstrating loss of nuclei and cytoarchitectural derangement. One focal area of viable IM-HGD remained at the margin of one ablation zone (12 J/cm2, 2 applications) because of incomplete overlap. CONCLUSION: Complete ablation of IM-HGD without ablation of submucosa is possible using the HALO360 system. Ablation depth is dose related and limited to the muscularis mucosae. In one patient, small residual foci of IM-HGD at the edge of the ablation zone were attributable to incomplete overlap, which can be avoided. This study, together with nonesophagectomy IM-HGD trials currently underway, will identify the optimal treatment parameters for IM-HGD patients who would otherwise undergo esophagectomy or photodynamic therapy.

Aged↗

PubMedDate not supplied

[An analysis of using the graphic monitoring of ventilation for an optimal choice of respiratory management parameters in patients with acute respiratory distress syndrome].

The potentialities of graphic ventilation monitoring (graphic monitor "Servo Screen 390", Siemens Elema, Sweden) were analyzed for optimizing the respiratory management parameters in 48 obstetric and gynecology patients with acute respiratory distress syndrome (ARDS). The ventilation loops and curves, ALV parameters, mechanical lung properties, gas blood composition and gas indices were dynamically evaluated during examination stages. The graphic ventilation monitoring, when used for respiratory management in patients with ARDS, provides for optimizing, in the real time mode, the PEEP and Vt levels, which is in line with the AVL "safety" concept.

Blood Gas Analysis↗

PubMed1992

Experimental tests of a superposition hypothesis to explain the relationship between the vestibuloocular reflex and smooth pursuit during horizontal combined eye-head tracking in humans.

1. We used a modeling approach to test the hypothesis that, in humans, the smooth pursuit (SP) system provides the primary signal for cancelling the vestibuloocular reflex (VOR) during combined eye-head tracking (CEHT) of a target moving smoothly in the horizontal plane. Separate models for SP and the VOR were developed. The optimal values of parameters of the two models were calculated using measured responses of four subjects to trials of SP and the visually enhanced VOR. After optimal parameter values were specified, each model generated waveforms that accurately reflected the subjects' responses to SP and vestibular stimuli. The models were then combined into a CEHT model wherein the final eye movement command signal was generated as the linear summation of the signals from the SP and VOR pathways. 2. The SP-VOR superposition hypothesis was tested using two types of CEHT stimuli, both of which involved passive rotation of subjects in a vestibular chair. The first stimulus consisted of a "chair brake" or sudden stop of the subject's head during CEHT; the visual target continued to move. The second stimulus consisted of a sudden change from the visually enhanced VOR to CEHT ("delayed target onset" paradigm); as the vestibular chair rotated past the angular position of the stationary visual stimulus, the latter started to move in synchrony with the chair. Data collected during experiments that employed these stimuli were compared quantitatively with predictions made by the CEHT model. 3. During CEHT, when the chair was suddenly and unexpectedly stopped, the eye promptly began to move in the orbit to track the moving target. Initially, gaze velocity did not completely match target velocity, however; this finally occurred approximately 100 ms after the brake onset. The model did predict the prompt onset of eye-in-orbit motion after the brake, but it did not predict that gaze velocity would initially be only approximately 70% of target velocity. One possible explanation for this discrepancy is that VOR gain can be dynamically modulated and, during sustained CEHT, it may assume a lower value. Consequently, during CEHT, a smaller-amplitude SP signal would be needed to cancel the lower-gain VOR. This reduction of the SP signal could account for the attenuated tracking response observed immediately after the brake. We found evidence for the dynamic modulation of VOR gain by noting differences in responses to the onset and offset of head rotation in trials of the visually enhanced VOR.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

PubMed2005

Sampling plans for fitting the psychometric function.

Research on estimation of a psychometric function psi has usually focused on comparing alternative algorithms to apply to the data, rarely addressing how best to gather the data themselves (i.e., what sampling plan best deploys the affordable number of trials). Simulation methods were used here to assess the performance of several sampling plans in yes-no and forced-choice tasks, including the QUEST method and several variants of up-down staircases and of the method of constant stimuli (MOCS). We also assessed the efficacy of four parameter estimation methods. Performance comparisons were based on analyses of usability (i.e., the percentage of times that a plan yields usable data for the estimation of all the parameters of psi) and of the resultant distributions of parameter estimates. Maximum likelihood turned out to be the best parameter estimation method. As for sampling plans, QUEST never exceeded 80% usability even when 1000 trials were administered and rendered accurate estimates of threshold but misestimated the remaining parameters. MOCS and up-down staircases yielded similar and acceptable usability (above 95% with 400-500 trials) and, although neither type of plan allowed estimating all parameters with optimal precision, each type appeared well suited to estimating a distinct subset of parameters. An analysis of the causes of this differential suitability allowed designing alternative sampling plans (all based on up-down staircases) for yes-no and forced-choice tasks. These alternative plans rendered near optimal distributions of estimates for all parameters. The results just described apply when the fitted psi has the same mathematical form as the actual psi generating the data; in case of form mismatch, all parameters except threshold were generally misestimated but the relative performance of all the sampling plans remained identical. Detailed practical recommendations are given.

Algorithms↗

PubMed1998

Application of the post-source pulse-focusing technique in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry: optimization of the experimental parameters and their influence on the capability of the method.

This paper describes experiments demonstrating the performance of a linear time-of-flight mass spectrometer equipped with the post-source pulse-focusing (PSPF) technique. The influence of various experimental parameters like the shape of the PSPF pulse on the performance of the instrument is discussed. The application of an ion lens in the acceleration region allows the focusing of high mass ions on the detector with a high yield. The observed mass resolution for matrix-assisted laser desorption/ionization (MALDI) generated ions is about 7000 (FWHM) and the mass accuracy using external standards for calibration is about 0.05% or better, in this work demonstrated for molecules with a mass up to 5700 Da. The percentage of the complete mass spectrum which can be detected with increased resolution and accuracy depends on the geometry of a particular setup. In order to cover a broad mass range several parameter sets have to be applied one after the other. Instead of the commonly employed microchannel plates, microsphere plates were used for detection of the ions in these experiments in order to test their applicability in combination with MALDI.

Amino Acid Sequence↗

PubMed2006

On the influence of oxygen and cell concentration in an SFPR whole cell biocatalytic Baeyer-Villiger oxidation process.

Efficient whole cell biotransformations, in particular microbial whole cell Baeyer-Villiger oxidation with molecular oxygen, demand comprehension and optimization of the process details involved. Optimal provision of oxygen and control of bioprocess parameters are pivotal for their success. The interrelation of cell density and oxygen supply in an in situ substrate feeding and product removal (SFPR) whole cell Baeyer-Villiger oxidation process was investigated in detail. Both parameters were optimized with respect to practical considerations. The outcome of this study supports a schematic process model, allows estimation of optimum process conditions and exploration of its limits.

Bioreactors↗

PubMed1993

Optimization of electric field parameters for the control of bone remodeling: exploitation of an indigenous mechanism for the prevention of osteopenia.

The discovery of piezoelectric potentials in loaded bone was instrumental in developing a plausible mechanism by which functional activity could intrinsically influence the tissue's cellular environment and thus affect skeletal mass and morphology. Using an in vivo model of osteopenia, we have demonstrated that the bone resorption that normally parallels disuse can be prevented or even reversed by the exogenous induction of electric fields. Importantly, the manner of the response (i.e., formation, turnover, resorption) is exceedingly sensitive to subtle changes in electric field parameters. Fields below 10 microV/cm, when induced at frequencies between 50 and 150 Hz for 1 h/day, were sufficient to maintain bone mass even in the absence of function. Reducing the frequency to 15 Hz made the field extremely osteogenic. Indeed, this frequency-specific sinusoidal field initiated more new bone formation than a more complex pulsed electromagnetic field (PEMF), though inducing only 0.1% of the electrical energy of the PEMF. The frequencies and field intensities most effective in the exogenous stimulation of bone formation are similar to those produced by normal functional activity. This lends strong support to the hypothesis that endogenous electric fields serve as a critical regulatory factor in both bone modeling and remodeling processes. Delineation of the field parameters most effective in retaining or promoting bone mass will accelerate the development of electricity as a unique and site-specific prophylaxis for osteopenia. Because fields of these frequencies and intensities are indigenous to bone tissue, it further suggests that such exogenous treatment can promote bone quantity and quality with minimal risk or consequence.

Animals↗