Detection of bacteria in suspension by using a superconducting quantum interference device.

We demonstrate a technique for detecting magnetically labeled Listeria monocytogenes and for measuring the binding rate between antibody-linked magnetic particles and bacteria. This sensitive assay quantifies specific bacteria in a sample without the need to immobilize them or wash away unbound magnetic particles. In the measurement, we add 50-nm-diameter superparamagnetic magnetite particles, coated with antibodies, to an aqueous sample containing L. monocytogenes. We apply a pulsed magnetic field to align the magnetic dipole moments and use a high-transition temperature superconducting quantum interference device, an extremely sensitive detector of magnetic flux, to measure the magnetic relaxation signal when the field is turned off. Unbound particles randomize direction by Brownian rotation too quickly to be detected. In contrast, particles bound to L. monocytogenes are effectively immobilized and relax in about 1 s by rotation of the internal dipole moment. This Néel relaxation process is detected by the superconducting quantum interference device. The measurements indicate a detection limit of (5.6 +/- 1.1) x 10(6) L. monocytogenes in our sample volume of 20 microl. If the sample volume were reduced to 1 nl, we estimate that the detection limit could be improved to 230 +/- 40 L. monocytogenes cells. Time-resolved measurements yield the binding rate between the particles and bacteria.

Gastric damage in the rat with nitrogen-containing bisphosphonates depends on pH.

BACKGROUND: The use of nitrogen-containing bisphosphonates (N-BPs) has been reported to be associated with gastrointestinal intolerance. The fasted, indomethacin-treated rat provides a model for assessing the gastrointestinal effects of these compounds. AIMS: The aims of this study were to elucidate the effect of pH on N-BP-induced gastric damage, and to evaluate the structure-activity relationship between N-BP anti-resorptive and gastric effects. METHODS: Fasted rats were dosed concomitantly with indomethacin (40 mg/kg, subcutaneously) and an N-BP (pamidronate, alendronate, or risedronate at 150 or 300 mg/kg, orally), with the N-BP dosing solutions adjusted to pH 2, 4 or 7. The aminopentane and aminohexane N-BPs (150, 225 or 300 mg/kg, orally) were only tested at pH 4 only. RESULTS: Nitrogen-containing bisphosphonate-induced gastric damage was pH-dependent, with increased damage at increasing pH. CONCLUSIONS: Gastric damage potential did not correlate with bone anti-resorptive effects, and the more potent anti-resorptive N-BPs were not necessarily more damaging to the stomach.

Evaluation of changes in trabecular bone architecture and mechanical properties of minipig vertebrae by three-dimensional magnetic resonance microimaging and finite element modeling.

The study objective was to analyze the three-dimensional (3D) trabecular architecture and mechanical properties in vertebral specimens of young and mature Sinclair minipigs to assess the relative contribution of architecture to bone strength. We used 3D magnetic resonance microimaging (MRmicroI) and direct image analysis to evaluate a set of standard structural measurements and new architectural descriptors of trabecular bone in biopsy specimens from L2, L3, and L4 vertebrae (n = 16 in each group) from young (mean age, 1.2 years) and mature (mean age, 4.8 years) minipigs. The measurements included bone volume/tissue volume (BV/TV), marrow star volume (Ma.St.V), connectivity density (ConnD), and two new parameters, percent platelike trabeculae (% plate) and percent bone in the load direction (% boneLD). The % plate, calculated from surface curvature, allowed the delineation of plates from rods. The % boneLD quantified the percentage of bone oriented along the long axis of the vertebral body. We showed that 3D MRmicroI can detect the subtle changes in trabecular architecture between the two age groups. ConnD, star volume, % plate, % boneLD, and BV/TV were found to be more effective than the model-based, derived indices (trabecular thickness [Tb.Th], trabecular separation [Tb.Sp], and trabecular number [Tb.N]) in differentiating the structural changes. BV/TV, % plate, and % boneLD significantly increased (p < 0.05) in all three vertebral sites of the mature minipigs. The significant decrease in ConnD and star volume in the mature vertebra was consistent with the concurrent increase of platelike trabecular bone (p < 0.05). Overall, ConnD, star volume, % plate, and % boneLD provided a coherent picture of the architectural changes between the two age groups. Apparent modulus and maximum stress were determined experimentally on biopsy specimens from L2 vertebrae (n = 16). When apparent modulus was predicted using 3D MRmicroI data sets as input for finite element modeling (FEM), the results were similar to the experimentally determined apparent modulus (p = 0.12). Both methods were then used to compare the young and the mature animals; the experimental and predicted apparent modulus were significantly higher for the mature group (p = 0.003 and 0.012, respectively). The experimental maximum stress in the vertebra of the mature animals was twice as high as that for the young animals (p = 0.006). Bone quantity (BV/TV or bone mineral content [BMC]) alone could explain approximately 74-85% of the total variability in stress and modulus. The inclusion of either ConnD or % boneLD with BV/TV in a multiple regression analysis significantly improved the predictability of maximum stress, indicating that architecture makes additional contributions to compressive strength in normal minipig vertebra.

Application and maintenance habits do make a difference in adhesion of Alora transdermal systems.

OBJECTIVES: To explore and evaluate Alora placebo patch application and maintenance habits of women in order to identify the factors that influence adhesion success. METHODS: This single-center, open-label, placebo, randomized, multiple-application, parallel-group study involved 99 healthy naïve users of transdermal patches. Participants applied and wore an Alora placebo patch for ten consecutive applications of approximately 3.5 days each and evaluated adhesion of the patches twice-daily. Three subgroups comprising participants achieving low, moderate or high adhesion success took part in focus groups to discuss their wear habits, practices and attitudes regarding transdermal patches. RESULTS: There was a significant behavioral component involved in patch adhesion. The habits, practices and attitudes of high achievers were clearly different from the other two subgroups. The three most important issues identified to improve adhesion were: mastering the removal of the patch liner, identifying the best site of application, and developing and implementing techniques to maintain patch adhesion. The Alora placebo patch was well tolerated throughout the study. CONCLUSION: The data showed that there is a learning curve involved in achieving maximal adhesion with a transdermal patch. During the study, a novel patch application method ('press, fold and slide') was demonstrated for the participants. This method was very well received by all participants and was more easily executed than the previous method. An adaptation of this method was incorporated into the Alora patient information leaflet, together with several other changes to help improve adhesion success.

Optical coherence microscopy. A technology for rapid, in vivo, non-destructive visualization of plants and plant cells.

We describe the development and utilization of a new imaging technology for plant biology, optical coherence microscopy (OCM), which allows true in vivo visualization of plants and plant cells. This novel technology allows the direct, in situ (e.g. plants in soil), three-dimensional visualization of cells and events in shoot tissues without causing damage. With OCM we can image cells or groups of cells that are up to 1 mm deep in living tissues, resolving structures less than 5 microm in size, with a typical collection time of 5 to 6 min. OCM measures the inherent light-scattering properties of biological tissues and cells. These optical properties vary and provide endogenous developmental markers. Singly scattered photons from small (e.g. 5 x 5 x 10 microm) volume elements (voxels) are collected, assembled, and quantitatively false-colored to form a three-dimensional image. These images can be cropped or sliced in any plane. Adjusting the colors and opacities assigned to voxels allows us to enhance different features within the tissues and cells. We show that light-scattering properties are the greatest in regions of the Arabidopsis shoot undergoing developmental processes. In large cells, high light scattering is produced from nuclei, intermediate light scatter is produced from cytoplasm, and little if any light scattering originates from the vacuole and cell wall. OCM allows the rapid, repetitive, non-destructive collection of quantitative data about inherent properties of cells, so it provides a means of continuously monitoring plants and plant cells during development and in response to exogenous stimuli.

Improvement of pagetic bone lesions with risedronate treatment: a radiologic study.

Risedronate is a potent pyridinyl bisphosphonate in clinical development for treatment and prevention of osteoporosis, and has been recently approved for treatment of Paget's disease in the United States. An open-label study was conducted to determine the effect of risedronate treatment on pagetic bone lesions in patients with moderate to severe Paget's disease (mean serum alkaline phosphatase levels [ALP] approximately seven times the upper limit of normal). Patients were treated with 30 mg/day oral risedronate for 84 days followed by a 112-day nontreatment period. This 196-day cycle was repeated once in patients whose ALP did not normalize or who experienced relapse, defined as a > or =25% increase in ALP from the lowest value measured. Radiographs of affected anatomical sites in 26 patients were collected at baseline, 6 months, and/or 12 months. Eleven patients received one course and 15 patients received two courses of treatment. Radiographs were examined by a skeletal radiologist who was blinded to their time sequence. Changes in pagetic lesions were categorized as "improved," "deteriorated," or "no change." Between baseline and 6 months, 16 patients improved and 3 deteriorated; at 12 months, 11 patients improved and 2 deteriorated. Most lesions remained unchanged between 6 and 12 months. Improvements were noted in all skeletal sites (tibia, femur, humerus, forearm, pelvis, spine, and skull), but were most pronounced in weight-bearing long bones. In weight-bearing bones, nine lesions had osteolytic fronts. Of these, seven improved and two remained unchanged at 6 months; at 12 months, all but one lesion (which improved) remained unchanged. This radiographic assessment demonstrates that oral risedronate, 30 mg/day in one or two 3-month courses, is highly effective for improving bone lesions in patients with Paget's disease. Risedronate treatment had no deleterious effect on osteolytic lesions in weight-bearing bones; indeed, the majority of lesions with osteolytic fronts were improved after 6 months of risedronate treatment.

Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part III--Validation.

This article, the last in a series of three, describes the validation phase of a study conducted to test the correlation of respirator fit factors to the subject's actual exposure using biological sampling. The study consisted of three phases: protocol development, method comparison testing, and validation. Six quantitative fit-test methods were evaluated in the method comparison testing phase. The two fit methods with the highest correlation with the wearers' measured exposure were a corn oil method (R2 = 0.81) and an ambient aerosol method (R2 = 0.78). Because the ambient aerosol method is more commonly used in the workplace, it was selected for further analysis. In this validation phase, the fit factors measured during the ambient aerosol fit-test were used to calculate the exposures to Freon-113 by using the model determined in the method comparison testing phase of the study. The actual Freon-113 exposures were then measured and compared with the predicted exposures. The results verified that the ambient aerosol method fit factors are highly correlated to the total Freon-113 exposure dose and thus that the model had a predictive ability.

Pain after periodontal scaling and root planing.

BACKGROUND: Although periodontal scaling and root planing, or SRP, is one of the most common procedures used in dental practice, there is little information available about the degree of postprocedural pain associated with it. The authors undertook this study to document the intensity and duration of pain after SRP with a view toward helping practitioners and their patients manage postprocedural discomfort. METHODS: Using the Heft-Parker self-assessment pain scale, 52 adults with moderate periodontitis evaluated their pain before and after SRP conducted with local anesthetic. RESULTS: After SRP, 28 percent of all patients reported faint-to-weak pain, 18 percent experienced weak-to-mild pain, 28 percent experienced mild-to-moderate pain, 8 percent had moderate-to-strong pain and 8 percent reported strong-to-intense pain. The average time to onset of maximum pain was approximately three hours after SRP, and the average duration of mild or greater pain was about six hours. Upon awakening the morning after SRP, subjects found that pain had returned to pre-SRP levels. Overall, 23 percent of all patients reported self-medicating with analgesics to relieve postprocedural pain. Women self-medicated earlier (P < .05) and more often than men (43 percent vs. 10 percent; P < .05). CONCLUSIONS: Patients experienced significant duration and magnitude of pain after SRP. This pain peaked between two and eight hours after SRP, lasted about six hours, and returned to pre-SRP levels by the morning after the procedure. Almost 25 percent of all patients self-medicated to relieve pain after SRP, and women took analgesic medication earlier and more often than men. CLINICAL IMPLICATIONS: Practitioners should consider using appropriate analgesic drugs to alleviate mild-to-moderate pain after SRP. On the basis of this study, it would appear that an analgesic that has a peak effect two to eight hours after the completion of SRP would be the most appropriate medication. Moreover, it is unlikely that analgesic medication would be needed by most patients beyond the day on which SRP was performed.

Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part I--Protocol development.

Quantitative fit tests (QNFT) have been assumed to be predictive of the protection respirators would provide to a wearer in the workplace. Workplace studies have consistently found no correlation between quantitative fit factors and workplace protection factors. This article is the first in a series of three describing a study designed to compare the fit factors from six QNFT methods against the actual dose of 1,1,2 trichloro-1,2,2 trifluoroethane (Freon-113) received under the same laboratory conditions. Five preliminary studies conducted to develop the protocol to assess the respirator wearer's dose through end-exhaled air analysis are described in this article: (1) chamber characterization, (2) end-exhaled air sampling, (3) skin absorption testing, (4) pharmacokinetic modeling, and (5) subject characterization. It was established that the concentration of corn oil aerosol and Freon-113 could be generated simultaneously in the chamber. It was ascertained that the optimum time to sample the exhaled breath was 30 minutes after the subject exited the chamber. It was also found that in a chamber concentration of 500 ppm, without any respiratory exposure, Freon-113 was still present in the end-exhaled air. This was attributed to skin absorption. The end-exhaled air of subjects exposed to 0.5, 3, 5, 25, 50, and 100 ppm (30 minute time-weighted average) of Freon-113 was evaluated at 30 minutes postexposure. This characterization was then used to predict the actual dose of Freon-113 received during the method comparison and validation testing to be described in subsequent articles.

Comparison of six respirator fit-test methods with an actual measurement of exposure in a simulated health care environment: Part II--Method comparison testing.

This article, the second in a series of three, describes the method comparison testing portion of a study conducted to compare the fit factors from six quantitative fit-tests (QNFT) with a measure of a respirator wearer's actual exposure assessed by end-exhaled air analysis for 1,1,2-trichloro-1,2,2-trifluoroethane (Freon-113) under the same conditions. The six QNFT methods were (1) continuous low flow, flush probe; (2) continuous high flow, deep probe (CHD); (3) exhalation valve discharge (EVD); (4) controlled negative pressure; (5) 10-minute Ambient Aerosol 1 (AA1); and (6) 30-minute Ambient Aerosol 2. The first three methods utilized corn oil and a forward light scattering photometer. The last two methods used the TSI Portacount. Respirators used in the study were both disposable and elastomeric organic vapor/high efficiency half-masks. The characterization equations from the preliminary research (described previously) were used to determine the actual exposure to Freon-113 during the method comparison testing. The fit factors resulting from the QNFT methods were then individually correlated with the Freon-113 exposures using the coefficient of determination, R2. The lowest R2 value, 0.20, was found with the EVD method. The highest R2 values, 0.81 and 0.78, were associated, respectively, with the CHD and AA1 methods. This study suggests that some QNFT methods may be used to estimate actual respirator performance under laboratory conditions.

Field performance measurements of half-facepiece respirators: steel mill operations.

Ambient and in-facepiece samples to evaluate the protection provided by negative-pressure, half-facepiece respirators were collected on workers in different areas of a steel mill including a sinter plant and a basic oxygen process shop. Protection was assessed by workplace protection factors (WPF). All the in-facepiece concentrations were dramatically less than the corresponding ambient concentration levels or permissible exposure limits. The geometric mean (GM) ambient and in-facepiece concentrations of iron were found to vary among tasks. Significant differences were also found to occur between the GM ambient exposure levels in which some of the respirators were used. Significant differences in respirator performance as measured by WPF or in-facepiece iron concentration were observed among different brands of respirators. For all job classifications and at all levels of airborne exposure, the fifth percentile estimates for the WPF distributions for each brand of respirator were all greater than 20.

Field performance measurements of half-facepiece respirators: developing probability estimates to evaluate the adequacy of an APF of 10.

To evaluate the protection provided by negative-pressure, half-facepiece respirators, ambient and in-facepiece samples were collected on workers in foundry, aircraft-painting, and steel-manufacturing operations. Protection was assessed by workplace protection factors (WPF). The appropriateness of the assigned protection factor (APF) for half-facepiece respirators was evaluated with a new approach using the WPF data from these and other studies previously published. The new approach utilizes binomial statistics based on the number of successes (no overexposure) and failures (overexposure) and is illustrated with a graphical representation of WPF data. With this consideration of the data, the probability of overexposure occurring during a wearing period for workers wearing the half-facepiece respirators represented by the studies referenced here was 0.5%, with a 95% confidence interval of 0.01 to 2.7%. If 50% in-facepiece sampling errors are considered, the probability of overexposure was 2.9%, with a 95% confidence interval of 1.1 to 6.3%. The authors believe the current APF of 10 for half-facepiece respirators is appropriate.

Clinical and clinicopathological assessment of serial phlebotomy in the Sprague Dawley rat.

Two studies, designed to mimic a single-dose, cross-over pharmacokinetic protocol, were conducted to gain a better understanding of the rat's response to multiple, frequent blood sampling. Parameters evaluated included body weight, clinical signs of disease, hematologic and serum biochemical analytes, organ weights, and histopathologic features. Study groups consisted of either 6 or 8 male, viral antibody-free, Sprague Dawley rats. These included controls and blood-collection groups that represented withdrawal of 10, 15, 20, 30, and 40% of estimated total blood volume. Volume of blood collected per time point was based on the total volume to be withdrawn divided by the 13 samples that were collected over 24 h. This regimen was repeated 2 weeks later. Samples were taken for clinical pathologic evaluation on the days subsequent to blood collection for both studies as follows: 0, 1, 2, and 3 days; 7, 8, or 9 days; and either 13 or 14 days. In Study 1, samples were also taken on either days 15 or 16, and on 17 or 18 after the second blood collection. Approximately 2 weeks after the second blood collection regimen, animals were euthanized. Animals in one study were necropsied, and selected tissues were taken for histologic examination. Analysis of variance, based on changes from baseline, was used to assess group differences. Results indicate that the rate of body-weight gain for the bled groups was not significantly different from that of the controls. Group differences in multiple hematologic parameters were significant. Changes were typical of acute blood-loss anemia, with positive or negative trends relating to the volume of blood removed. In addition, these changes were characterized by recovery to control values within approximately 14 days. Few statistically significant group differences were detected in serum biochemical values, and those detected were not biologically relevant. Although organ weights of bled groups were similar to those of controls, minimal to mild splenic hematopoiesis was present in all bled groups, compared with controls. These data indicate that removal of up to 40% of a rat's total blood volume over a 24-h period, and repeated 2 weeks later, caused no gross ill effects.

Nonclinical model for assessing gastric effects of bisphosphonates.

Gastrointestinal intolerance has been associated with amino bisphosphonate therapy in the clinic. The objective of this study was to develop a model for assessing bisphosphonate-induced gastric damage that may aid in the development of future bisphosphonate therapies. Rats were dosed concomitantly with indomethacin (40 mg/kg, subcutaneously) and an amino or pyridinyl bisphosphonate (orally at. 150, 225 or 300 mg/kg). The bisphosphonates studied were pamidronate and alendronate (primary amino bisphosphonates) and risedronate and NE-97221 (pyridinyl bisphosphonates). Macroscopically, alendronate induced significantly (P < 0.05) more antral damage (both lesion length and number) than pamidronate and risedronate at 225 and 300 mg/kg, and more than NE-97221 at 300 mg/kg. NE-97221 induced significantly more antral damage (lesion length) than risedronate at 225 mg/kg and a greater number of lesions compared to pamidronate and risedronate at 225 and 300 mg/kg. The model was validated histologically, and macroscopic findings correlated with histologic evidence of antral mucosal necrosis and inflammatory infiltration of the lamina propria. The calcium chelators EGTA and EDTA did not induce gastric damage in this model when dosed according to the same protocol as the nitrogen-containing bisphosphonates. This suggests that calcium chelation does not account for the gastric effects in this model. The fasted, indomethacin-treated rat provides a novel nonclinical model to assess gastric effects of bisphosphonates, which may aid in the development of future bisphosphonate therapies. These data suggest that when expressed on an actual or anticipated clinical dose basis for osteoporosis (pamidronate, 150 mg; alendronate, 5-10 mg; risedronate and NE-97221, 5 mg), primary amino bisphosphonates may have a greater potential for inducing gastric damage than do pyridinyl bisphosphonates.

Two-stage designs for the logistic regression model in single-agent bioassays.

In this paper we focus on the use of a two-stage procedure for logistic regression that emphasizes predicting response through the use of the Q-optimality criterion. The use of D-optimality in the first stage is primarily to allow best possible parameter estimates as one enters the second stage. However, it is important to understand that there are many ways to formulate the two-stage procedure. It may involve any optimality criterion in either stage. In fact, theoretically, one need not stop at two stages. It was our intention in this paper to demonstrate the potential in the two-stage procedure in cases in which good initial parameter estimates are not available. Those investigators who are interested in the software for the two-stage procedure described here should contact Dr. William R. Myers.

Field performance measurements of half-facepiece respirators--foundry operations.

A series of field studies was conducted to measure workplace protection factors (WPF) provided by elastomeric and disposable half-facepiece respirators against different particulate contaminants. The research protocol developed for the study has been described in a previous article. This article presents results from dust and fume exposures at three foundries. The major components of the airborne exposures in these foundries were zinc, lead, and silicon. The major components of the in-facepiece samples were zinc, chlorine, and lead. Significant differences were observed in ambient zinc and lead concentration levels among foundries; however, no significant difference was observed in the in-facepiece concentrations of these elements among foundries. Respirator performance varied within each foundry, but there was no difference in performance when pooling all foundry data. The 5th, 10th and 50th percentile estimates for the pooled foundry and respirator WPF data were about 9, 16, and 114 respectively. The infacepiece concentration data clearly indicate that dust-fume-mist (DFM) class half-facepiece respirators, when conscientiously used, worn, and maintained, in conjunction with other existing controls in these foundries, provided effective worker protection.

The effects of respirator use on workers' productivity in a mentally stressing task.

This article quantitatively determines the effect of wearing half-facepiece respirators, the most widely used type in industry, on worker productivity and efficiency in a mentally stressing task. An experiment investigated the performance of 10 subjects in a task simulating inspection of printed circuit boards. Accuracy and mean reaction time were tested automatically. Paced and unpaced task simulations were used to evaluate the effect of wearing the respirator on response times and the accuracy of subjects' responses for different product complexities and quality levels. The results indicated that wearing the respirator, in general, did not affect the speed or accuracy of subjects performing simulated inspection tasks. Pacing the task resulted in longer response times but did not affect accuracy. More complex products resulted in longer response times but also did not affect accuracy. High product quality resulted in longer response times and a higher miss rate. In terms of accuracy and reaction times a statistically significant difference was found in the performance of subjects.

Evaluation of new in-facepiece sampling procedures for full and half facepieces.

Precision and bias were determined on five different methods of conducting in-facepiece sampling. In-board penetration occurred through fixed, circular, leak geometries positioned at different areas on the face-seal of full and half facepiece negative-pressure respirators. The sampling procedures evaluated in the study were: (1) a continuous, low sampling rate, flush on the respirator, mid nose-mouth probing (CLF) procedure; (2) a continuous, high sampling rate, deep front-of-mouth probing (CHD) procedure; (3) a pulsed exhalation, deep front-of-mouth probing (PED) procedure; (4) an exhalation value discharge (EVD) procedure; and (5) a pulsed inhalation, deep front-of-mouth, probing (PID) procedure. The CLF procedure represents a recommended in-facepiece sampling procedure in the United States. Evaluations were done on populations of nine full facepiece respirators and five half facepieces. The full facepieces were not equipped with nose cups. The average sampling biases on the full facepieces were: (1) CLF procedure, -21%; (2) CHD procedure, -3%; (3) PED procedure, 0.7%; (4) EVD procedure, -14%; and (5) PID procedure, -12.3%. On the five half facepiece respirators the average sampling biases were: (1) CLF procedure, -26%; (2) CHD procedure, -13%; (3) PED procedure, -4%; (4) EVD procedure, -2%; and (5) PID procedure, -24%. The bias observed with each method was found to be affected, to some extent, by the location of the face-seal penetration.

Review of respirator performance testing in the workplace: issues and concerns.

Performance capability of respirators has traditionally been evaluated by testing components of the respirator (e.g., filter efficiency), facepiece fit, total inward leakage, or some other measure of performance evaluated under laboratory conditions. In recent years, increased emphasis has been placed on development of test methods suitable for evaluating respirator performance in the workplace. The goal of such testing is to evaluate the level of protection provided by respirators in the work environment. The AIHA Respiratory Protection Committee believes that workplace testing of respirators has the potential to be an excellent tool for increasing knowledge about the effectiveness of respiratory protection. However, a number of technical issues remain to be addressed before optimal test protocols and data analysis methods can be defined. The progress made to date in workplace testing will be reviewed, and broader discussion about key elements that must be considered when developing guidelines for testing respirators in the workplace will be initiated.