An evaluation of M2+ interference correction approaches associated with As and Se in ICP-MS using a multi-day dataset along with ICP-MS/MS/HR-ICP-MS based analysis and hierarchical modeling as a means of assessing bias in fortified drinking waters and single component matrices.

Three 1 2 mass oriented rare earth element (REE) M2+ correction approaches (fixed factor, a dual internal standard, and an in-sample) are evaluated for use in an ICP-MS environmental method update. The multi-variant-based evaluation includes analyzing the same 19 REE-fortified matrices on eight different days over a two-month period using two instrument tunes. These REE-fortified matrices were also analyzed using HR-ICP-MS and ICP-MS/MS to estimate the reference value for use in the principal component analysis (PCA) and hierarchical modeling evaluation. A fixed factor is unable to compensate for matrix and mass dependent drift and because of this it generates the largest across matrix, tune, and day 95th percent confidence bounds for the REE corrections on both As (1.1 ppb) and Se (23 ppb) using samples fortified with 100 ppb Nd, Sm & Gd. The PCA analysis indicated that M2+ ions cluster together across matrix, tune and day better than M1+ and these tighter correlations are reflected in reduced 95th percentile confidence bounds for dual M2+ internal standards (M2+; As = 0.3 ppb; Se = 5.4 ppb; n = 704) relative to M1+ internal standards (M1+; As = 0.6 ppb; Se = 12.0 ppb; n = 1056). The use of an in-sample M2+ correction produced comparable 95th percent confidence bounds (As = 0.2 ppb; Se = 3.4 ppb; n = 352) relative to the M2+ internal standard approaches. Finally, the hierarchical modeling indicated M2+ ions as internal standards tend to minimize the across day variability induced by cone changes and the daily reoccurring matrix shifts in the M2+/M1+ ratio associated with 250 ppm matrices of Na, Ca, and Mg. This internal standard driven reduction in variability can be beneficial in compliance monitoring methods.

Geospatial Patterns of Antimicrobial Resistance Genes in the US EPA National Rivers and Streams Assessment Survey.

Antimicrobial resistance (AR) is a serious global problem due to the overuse of antimicrobials in human, animal, and agriculture sectors. There is intense research to control the dissemination of AR, but little is known regarding the environmental drivers influencing its spread. Although AR genes (ARGs) are detected in many different environments, the risk associated with the spread of these genes to microbial pathogens is unknown. Recreational microbial exposure risks are likely to be greater in water bodies receiving discharge from human and animal waste in comparison to less disturbed aquatic environments. Given this scenario, research practitioners are encouraged to consider an ecological context to assess the effect of environmental ARGs on public health. Here, we use a stratified, probabilistic survey of nearly 2000 sites to determine national patterns of the anthropogenic indicator class I integron Integrase gene (intI1) and several ARGs in 1.2 million kilometers of United States (US) rivers and streams. Gene concentrations were greater in eastern than in western regions and in rivers and streams in poor condition. These first of their kind findings on the national distribution of intI1 and ARGs provide new information to aid risk assessment and implement mitigation strategies to protect public health.

Empirically-based modeling and mapping to consider the co-occurrence of ecological receptors and stressors.

Part of the ecological risk assessment process involves examining the potential for environmental stressors and ecological receptors to co-occur across a landscape. In this study, we introduce a Bayesian joint modeling framework for use in evaluating and mapping the co-occurrence of stressors and receptors using empirical data, open-source statistical software, and Geographic Information Systems tools and data. To illustrate the approach, we apply the framework to bioassessment data on stream fishes and nutrients collected from a watershed in southwestern Ohio. The results highlighted the joint model's ability to parse and exploit statistical dependencies in order to provide empirical insight into the potential environmental and ecotoxicological interactions influencing co-occurrence. We also demonstrate how probabilistic predictions can be generated and mapped to visualize spatial patterns in co-occurrences. For practitioners, we believe that this data-driven approach to modeling and mapping co-occurrence can lead to more quantitatively transparent and robust assessments of ecological risk.

Effects of Temperature and Spatial Scale on Rio Grande Cutthroat Trout Growth and Abundance.

Diversity in habitat and life-history strategies promote a species' long-term persistence. However, life-history strategies are most commonly studied at broad spatial and temporal scales. We applied longevity growth models and closed N-mixture models to examine within- versus between stream variability in life-history characteristics of Rio Grande Cutthroat Trout in northern New Mexico streams. We developed a von Bertalanffy growth model and a closed N-mixture model in a hierarchical Bayesian framework to examine the importance of fine-scale variability in temperature and density-dependence on growth and abundance. The model indicated that accumulation of degree days likely positively influenced instantaneous growth rates and, to a lesser extent, negatively affected asymptotic body length. A nonlinear response of abundance to temperature was also observed, suggesting that Cutthroat Trout productivity along the temperature continuum was affected by physiological limitations (e.g., optimal growth temperatures). Parameter variability was greatest at the segment level for asymptotic size and abundance, but greatest at the stream level for the rate at which asymptotic size is reached. In total, the results suggest that fine-scale habitat heterogeneity (i.e., temperature) may play important roles in the continued persistence of Rio Grande Cutthroat Trout. Management actions should, therefore, consider the role of fine-scale processes for improving the likelihood of future population persistence.

Hemostasis and sealing of air leaks in the lung using high-intensity focused ultrasound.

BACKGROUND: Operative management of parenchymal lung injury can be complicated by persistent hemorrhage and air leak, which might require resection. Techniques that preserve parenchyma are associated with improved survival. High-intensity focused ultrasound (HIFU) has been demonstrated as a useful method for hemostasis in experimental solid organ injuries. We wished to investigate whether this could be applied to lung injuries. METHODS: An intraoperative HIFU device (frequency of 5.7 MHz, acoustic power of 65 W), equipped with a titanium coupler, was used. Incisions (average length of 2.5 cm, and depth of 5 mm) were made in the lungs of 11 pigs, which created both parenchymal hemorrhage and air leakage. In treatment experiments, 70 incisions were sealed with HIFU. The HIFU application started within 10 seconds of inducing the injury. Hemostasis was assessed by visual observation of sealed incisions. The possible air leakage was determined by submersing the sealed incision under the layer of water and observing for air bubble formation. In control experiments, five incisions were left untreated to monitor air leaks and bleeding for 2 minutes. RESULTS: Hemostasis and pneumostasis (sealing of air leaks) of the treated incisions were achieved in 51 +/- 37 seconds (mean +/- SD) (range of 10-210 seconds) of HIFU application time. Over 95% of incisions were hemostatic within 2 minutes of HIFU application. The treatment time was not dependent on the incision length or depth. In control experiments, the air leaking and bleeding were still present at 2 minutes after the injury. CONCLUSION: Intraoperative HIFU might provide an effective method of hemostasis and control of air leaks from lacerations caused by trauma.

Hemorrhage control in arteries using high-intensity focused ultrasound: a survival study.

High-intensity focused ultrasound (HIFU) has been shown to provide an effective method for hemorrhage control of blood vessels in acute animal studies. The objective of the current study was to investigate the long-term effects of HIFU-induced hemostasis in punctured arteries. The femoral arteries ( approximately 2mm in diameter) of 25 adult anesthetized rabbits were surgically exposed, and either punctured and treated with HIFU (n=15), served as control (no puncture and no HIFU application: n=7), or were punctured and left untreated (n=3). Treated animals were allowed to recover, and examined and/or sacrificed on days 0, 1, 3, 7, 14, 28, and 60 after treatment to obtain ultrasound images and samples of blood and tissue. Hemostasis (arrest of bleeding) was achieved in all 15 of the HIFU-treated arteries. Eleven of the arteries were patent after HIFU treatment, and four arteries were occluded, as determined by Doppler ultrasound. The median HIFU application time to achieve hemostasis was 20s (range 7-55 s) for the patent arteries and 110 s (range 50-134 s) for the occluded arteries. In untreated animals, bleeding had not stopped after 120 s. One of the occluded arteries had reopened by day 14. No immediate or delayed re-bleeding was observed after HIFU treatment. Maximal blood flow velocities were similar in HIFU-treated patent vessels and control vessels. No significant difference in hematocrits was found between HIFU-treated and control groups at different time points after the procedure. Light microscopy observations of the HIFU-treated arteries showed disorganization of adventitia, and coagulation and thinning of the tunica media. The general organization of the adventitia and tunica media recovered to normal appearance within 28 days, with some thinning of the tunica media observed up to day 60. Neointimal hyperplasia was observed on days 14 and 28. The results show that HIFU can produce effective and long-term (up to 60 days) hemostasis of punctured femoral arteries while preserving normal blood flow and vessel wall structure in the majority of vessels.

Intra-operative acoustic hemostasis of liver: production of a homogenate for effective treatment.

OBJECTIVE: We have shown that High-Intensity Focused Ultrasound (HIFU) can effectively control bleeding from injuries to solid organs such as liver, spleen, and lung. Achievement of hemostasis was augmented when a homogenate of tissue and blood was formed. The objective of this study was to investigate quantitatively the effect of homogenate production on HIFU application time for hemostasis. Possible mechanisms involved in homogenate production were also studied. METHODS: Ten anesthetized rabbits had laparotomy and liver exposure. Liver incisions, 15-25 mm long and 3-4 mm deep, were made followed immediately by HIFU application. Two electrical powers of 80 and 100 W corresponding to focal acoustic intensities of 2264 and 2829 W/cm(2), respectively were used. Tissue and homogenate temperatures were measured. Smear and histological tissue sample analysis using light microscopy were performed. RESULTS: In treatments with homogenate formation, hemostasis was achieved in 76+/-1.3 s (Mean+/-Standard Error Mean: SEM) at 80 W. In treatments without homogenate formation (at 80 W), hemostasis was achieved in 106+/-0.87 s. At 100 W, hemostasis was achieved in 46+/-0.3 s. The time required for homogenate formation, at 80 and 100 W were 60+/-2.5 and 23+/-0.3 s, respectively. The homogenate temperature was 83 degrees C (SEM 0.6 degrees C), and the non-homogenate tissue temperature at the treatment site was 60 degrees C (SEM 0.4 degrees C). The smear and histological analysis showed significant blood components and cellular debris in the homogenate, with some intact cells. CONCLUSION: The HIFU-induced homogenate of blood and tissue resulted in a statistically significant shorter HIFU application time for hemostasis. The incisions with homogenate had higher temperatures as compared to incisions without homogenate. Further studies of the correlation between homogenate formation and temperature must be done, as well as studies on the long-term effects of homogenate in achieving hemostasis.

Dental dolorimetry for human pain research: methods and apparatus.

Electrical stimulation of human tooth pulp provides a means of safely producing human pain in the laboratory. This paper describes a dolorimetry and data collection system for stimulating volunteers, recording responses and analyzing data. The system allows multilevel stimulation in pseudorandom sequences and analysis of results using the methods of Sensory Decision Theory. It consists of modified commercial equipment, specially designed circuitry, an interface, and a programmable calculator. Fundamental problems and safety considerations for electrical dental stimulation are reviewed. Reliability of stimulation and response measurement is discussed.

Comparative effects of acupuncture in Japan and the United States on dental pain perception.

The effect of low frequency electrical acupunctural stimulation on the perception of induced dental pain were compared in two cultural settings. Twenty Japanese and 20 American subjects (consisting of 10 Caucasians and 10 second or third generation Japanese) were tested in two functionally identical laboratories, one at Tottori University in Yonago, Japan and the other at the University of Washington. Each subject underwent a control and an acupuncture session on separate days wtih subjects counterbalanced for carry-over order effects. Sensory decision theory (SDT) analysis demonstrated a significant reduction in perceptual capability and an increased bias against reporting stimuli as painful following the acupuncture as treatment which was performed bilaterally at traditional facial points. No significant differences between groups in alteration of perceptual capability, bias or pain threshold were demonstrated, indicating that the cultural and racial differences studied did not influence responses to acupuncture in a laboratory setting.

Three-dimensional echocardiographic measurement of left ventricular wall thickness: In vitro and in vivo validation.

INTRODUCTION: Three-dimensional (3D) echocardiography has been shown to accurately measure left ventricular (LV) volume and mass. This study evaluated the accuracy of 3D echocardiography and the CenterSurface method for measuring LV wall thickness in vitro and in vivo. METHOD: Three-dimensional echocardiography scans, obtained from 7 LV phantoms and subjects having healthy (n = 5) or diseased (n = 8) hearts, were digitized. Endocardial and epicardial borders were outlined and used in 3D LV reconstruction. In vitro wall thickness was compared with true micrometer measurements. Three-dimensional in vivo wall thickness was compared with 2-dimensional (2D) thickness measured by the centerline method. RESULTS: The in vitro 3D echocardiography measurements agreed closely with true wall thickness (P <.0001), as did in vivo measurements (P <.0001). CONCLUSION: Three-dimensional echocardiography reconstruction has previously been shown to provide accurate representation of LV shape in addition to volume and mass. This study demonstrates that the CenterSurface method provides accurate quantification of wall thickness.

An image-guided high intensity focused ultrasound device for uterine fibroids treatment.

A high intensity focused ultrasound (HIFU) device was developed for treating uterine fibroid tumors. This prototype device enables image-guided therapy by aligning a commercially available abdominal ultrasound image probe to a vaginal HIFU transducer so the HIFU focus is in the image plane. The device was designed based on anatomical constraints of the female pelvic structures. HIFU was generated using a 3.5 MHz PZT-8 crystal, 25.4 mm in diameter, bonded to an aluminum lens. Computer simulations were performed to ensure that effective focusing was achievable at a fixed focal depth of 40 mm. Transducer efficiency was empirically determined to be 58%, and the half pressure maximum focal dimensions were 11 mm in length and 1.2 mm in width. A water-filled latex condom surrounding the transducer provided acoustic coupling, a stand-off, and allowed water circulation for transducer cooling. In vitro experiments in a tissue-mimicking gel phantom and in turkey breast demonstrated ultrasound image-guided lesion formation, or tissue necrosis, at the focus due to HIFU induced thermal and cavitation effects. The HIFU treatment site appeared as a hyperechoic spot on the ultrasound image at intensities above 1250 W/cm2. The results of in vitro experiments and in vivo ergonomic testing in six human volunteers indicated that the device has the potential of providing a nonsurgical approach for uterine fibroid treatment. Future in vivo studies in large animal models and fibroids patients are planned.

Ultrasound-enhanced transcorneal drug delivery.

PURPOSE: Ultrasound has been shown to enhance, by up to 10 times, the corneal permeability to different compounds such as beta-blockers and fluorescein. Here, we report on our investigation of the mechanisms of ultrasound-enhanced drug delivery through the cornea using light and electron microscopy. METHODS: Enhancement of permeability for a hydrophilic compound, sodium fluorescein, in rabbit cornea in vitro was achieved using ultrasound at a frequency of 880 kHz and intensities of 0.19-0.56 W/cm2 with an exposure duration of 5 minutes. Light and electron microscopy (transmission and scanning) were used to observe ultrasound-induced structural changes in the cornea. RESULTS: The permeability increased by 2.1, 2.5, and 4.2 times when ultrasound was applied at 0.19, 0.34, and 0.56 W/cm2, respectively (P<0.05). The surface cells of corneal epithelium exposed to ultrasound appeared swollen and lighter in color (indications of membrane rupture) as compared with the control cells. Some of the surface epithelial cells were absent. The cells in the inner layers of the epithelium were occasionally lighter in color. Also, holes 3-10 microm in diameter were observed on the epithelial surface. No structural changes were observed in the stroma. CONCLUSION: Ultrasound enhancement of drug delivery through the cornea appears to result from minor structural alterations in the epithelium. Careful investigation of the recovery of cornea structure and barrier function after the ultrasound application, in vivo, is needed.

Ocular drug delivery using 20-kHz ultrasound.

The cornea is a major pathway for drug delivery to diseased eye structures. We have investigated the application of 1-s bursts of 20-kHz ultrasound, at I(SAPA) of 14 W/cm(2) (I(SATA) of 2 W/cm(2)), for enhancement of corneal permeability to glaucoma drugs of different lipophilicity (atenolol, carteolol, timolol and betaxolol). The permeability of rabbit cornea increased by 2.6 times for atenolol, 2.8 for carteolol, 1.9 for timolol and 4.4 times for betaxolol (all p-values < 0.05), after 60 min of ultrasound (US) exposure in vitro. The differences between the treatment and control experiments were statistically significant after 10 to 30 min of US exposure for all four drugs. US application appeared to produce epithelial disorganization and structural changes in the corneal stroma. Further studies are needed to determine the optimal US parameters for a safe and effective treatment.

Attenuation of porcine tissues in vivo after high-intensity ultrasound treatment.

The attenuation of ultrasound (US) waves in biologic tissues is an important determinant of energy absorption and wave propagation; thus, important in optimization of high-intensity focused US (HIFU) therapy. We measured attenuation of selected porcine tissues (liver, spleen and abdominal wall) in vivo in the frequency range of 1 to 5 MHz, using the pulse-transmission method, before and after HIFU treatment. In all tissues, an increase in attenuation was observed with increasing frequency. The attenuation coefficient was higher in HIFU-treated tissues than in the untreated tissues. The lowest attenuation was measured in the liver, both in normal and HIFU-treated cases. Mechanisms that may be responsible for the observed attenuation coefficient increase in HIFU-treated tissues include thermally induced change in the tissue macromolecular structure and presence of gas/vapor bubbles due to cavitation and/or boiling.

Polyacrylamide gel as an acoustic coupling medium for focused ultrasound therapy.

A hydrogel acoustic coupling medium was investigated as a practical alternative to water for clinical applications of focused ultrasound (US) therapy. Material characterization and functional testing of polyacrylamide gel couplers were performed. Acoustic, bulk and thermal properties were measured. Conical couplers were designed and fabricated to fit a 3.5-MHz, spherically concave transducer for functional tests, including Schlieren imaging, power efficiency measurements and in vivo hemostasis experiments. Polyacrylamide was shown to have favorable acoustic properties that varied linearly with acrylamide concentration from 10% to 20% weight in volume. Attenuation coefficient, sound speed and impedance ranged from 0.08 to 0.14 dB/cm at 1 MHz, 1546 to 1595 m/s and 1.58 to 1.68 Mrayl, respectively. An intraoperative in vivo hemostasis experiment in a sheep model demonstrated that the gel-coupled transducer was capable of inducing hemostasis in actively bleeding splenic and hepatic incisions. The results of this study show that polyacrylamide may be a promising coupling material for focused US therapy.

Water-cooled, high-intensity ultrasound surgical applicators with frequency tracking.

High-intensity, focused ultrasound (HIFU) applicators have been developed for arresting bleeding with the ultimate intent of use in surgery. The design uses a tapered titanium component for transmission coupling of the ultrasound energy from a spherically curved transducer to biological tissues. The nominal operating frequency is 5.5 MHz, in a highly resonant mode (quality factor of 327 with water load). Liquid cooling is used to remove energy loss important at net applied power greater than 18 W/cm2 at the surface of the piezoelectric element. A downward resonance frequency shift (>20 kHz) occurs, even with cooling, as the applicator warms with normal operation. A feedback technique is used for maintaining the excitation near optimum resonance. Standing wave ratios of the applied power of 1.6 or less are thus sustained. The system and applicators have been found to be highly robust, effective in achieving hemostasis in the hemorrhaging liver, spleen, lung, or blood vessels in rabbit and pig experiments. One unit has been operated for over 1.7 hours in treating organ hemorrhage in blunt trauma experiments with nine swine with electrical net power of up to 158 W (31 W/cm2 across the transducer) and intensity of 2560 W/cm2 at focus.

Quantitative investigation of acoustic streaming in blood.

Acoustic streaming may have practical utility in diagnostic medical ultrasound in distinguishing between stagnant blood and tissue as well as clotted and unclotted blood. This distinction can be difficult with conventional ultrasound but have high value in managing trauma patients with internal hemorrhage. Ultrasound energy applies a force to blood by momentum transfer, resulting in bulk streaming that is a function of the acoustic attenuation, sound speed, acoustic intensity, blood viscosity, and the boundary conditions posed by the geometry around the hematoma. A simple tubular model was studied analytically, by finite element simulation, and experimentally by in vitro measurement. The simulation agreed closely with measurements while the analytic solutions were found to be valid only for beam diameters approximating the diameter of the tubular channel. Experimentally, the acoustic streaming in blood decreased as the blood began to clot and the streaming flow was not detected in clotted blood. In contrast, the echogenicity of the same blood samples did not change appreciably from the unclotted to the clotted state for the stagnant blood studied. Streaming detection appears to offer a potential tool for improving hemorrhage diagnosis.

High-intensity ultrasound treatment of blunt abdominal solid organ injury: an animal model.

BACKGROUND: High-intensity focused ultrasound (HIFU) is effective in producing hemostasis in injuries from organ lacerations and punctures in animals but has not been evaluated in impact injuries. METHOD: High-energy blows were applied to 11 heparinized and anesthetized pigs, resulting in solid organ injury. HIFU was applied to injuries via laparotomy. The animals were closed, administered saline, observed under general anesthesia for 3.6 +/- 0.4 hours, reopened, and inspected, and abdominal free fluid was aspirated. RESULTS: Organ hemostasis was achieved (mean +/- SD) with 15 +/- 6 minutes of HIFU treatment and 54 +/- 3 minutes of operating time, and 18.8 +/- 13.1 mL/kg of blood was recovered from the abdomen. One animal died from an untreated occult injury to a large vein. HIFU-treated sites were hemostatic at relaparotomy, with 8.6 +/- 6.2 mL/kg abdominal serosanguinous fluid recovered. CONCLUSION: HIFU is effective in producing hemostasis by direct treatment of injured parenchyma in blunt trauma.

Treatment of uterine leiomyosarcoma in a xenograft nude mouse model using high-intensity focused ultrasound: a potential treatment modality for recurrent pelvic disease.

OBJECTIVE: The objective was to test the efficacy of high-intensity focused ultrasound (HIFU) for treatment of uterine leiomyosarcoma in a Xenograft nude mouse model. METHODS: A total of 65 athymic nude mice were inoculated subcutaneously with 5 to 7 x 10(6) ELT-5B cells, a uterine leiomyosarcoma cell line derived from the Eker rat. Thirty animals showed tumor growth. The tumor volume was measured transcutaneously once a week. Animals were randomly assigned to three groups: HIFU treatment (n = 17), sham treatment (n = 7), and control (n = 6). A HIFU device, operating at a frequency of 2.0 MHz and an intensity of 2000 W/cm(2), was used for treatment. RESULTS: Within 3 weeks of a single HIFU treatment, 100% reduction in tumor volume was observed in all animals, except one. A second HIFU treatment was applied to that animal, resulting in 100% reduction in tumor volume. The tumors in the sham-treated animals continued to grow at a similar rate to that of the control group to approximately 500% of the tumor volume at the time of treatment. All animals were monitored for a maximum of 3 months. No metastasis was observed in the HIFU-treated animals. Histological examination confirmed a complete tumor disappearance after HIFU treatment. CONCLUSION: We have shown that HIFU can effectively treat uterine leiomyosarcoma tumors inoculated in Xenograft nude mice, demonstrating HIFU's potential use for treatment of recurrent uterine leiomyosarcoma.

Three-dimensional echocardiographic measurement of left and right ventricular mass and volume: in vitro validation.

INTRODUCTION: Three-dimensional (3D) echocardiography has been shown to offer highly accurate measurements of left ventricular (LV) volume and mass. The present study evaluated the accuracy of 3D surface reconstruction by the piecewise smooth subdivision method in measuring volume and mass not only in the LV but also in the more complexly shaped right ventricle (RV). METHODS: 3D echo scans were obtained of in vitro LV's (n = 15) and RVs (n = 10). From digitized images, ventricular borders were traced and used in surface reconstructions. Mass and volume determined from the reconstructions were compared to true volume and mass determined prior to imaging. Additionally casts of two RVs were made and laser-scanned. Distances between the laser-identified points on the RV surface and the corresponding 3D echo reconstructions were measured. RESULTS: 3D LV volume agreed well with the true volume (y = 0.99x + 1.73, r = 0.99, SEE = 3.35 ml, p < 0.0001), as did 3D LV mass (y = 0.99x - 4.71, r = 0.99, SEE = 9.85 g, p < 0.0001). 3D RV volume overestimated true volume (y = 1.11x + 1.77, r = 0.99, SEE = 3.36 ml, p < 0.001) by 6.23+/-3.70 ml (p < 0.0001). 3D mass agreed well with RV mass (y = 0.78x + 17.32, r2 = 0.93, SEE = 3.54 g, p < 0.0001). 3D echo reconstructions matched the laser-scanned RV closely with residual distances of 1.1+/-0.9 and 1.4+/-1.2 mm, respectively. CONCLUSIONS: 3D echo using freehand scanning combined with surface reconstruction by the piecewise smooth subdivision surface method enables accurate determination of LV mass and volume, of RV mass and volume, and of the RV's complex shape.