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Biologists who use electron microscopy (EM) images to build nanoscale 3D models of whole cells and their organelles have historically been limited to small numbers of cells and cellular features due to constraints in imaging and analysis. This has been a major factor limiting insight into the complex variability of cellular environments. Modern EM can produce gigavoxel image volumes containing large numbers of cells, but accurate manual segmentation of image features is slow and limits the creation of cell models. Segmentation algorithms based on convolutional neural networks can process large volumes quickly, but achieving EM task accuracy goals often challenges current techniques. Here, we define dense cellular segmentation as a multiclass semantic segmentation task for modeling cells and large numbers of their organelles, and give an example in human blood platelets. We present an algorithm using novel hybrid 2D-3D segmentation networks to produce dense cellular segmentations with accuracy levels that outperform baseline methods and approach those of human annotators. To our knowledge, this work represents the first published approach to automating the creation of cell models with this level of structural detail.

Biogeographical transition zones are important areas to investigate evolutionary ecological questions, but long-term population monitoring is needed to better understand ecological processes that govern population variations in such edge environments. The southernmost Brazilian rocky reefs are the southern limit of distribution for 96% of the tropical ichthyofauna of the western Atlantic. The Arvoredo Marine Biological Reserve is the only nearshore no-take marine-protected area (MPA) located in this transition zone. The main aim was to investigate how the populations of rocky reef fish species vary in density and biomass in space and over time, inside and outside the Arvoredo MPA. This study presents results based on a 9 year (2008-2017) underwater visual census monitoring study to evaluate the density and biomass of key fish species. Variations in density and biomass were detected for most species. Factors and mechanisms that may have influenced spatial variation are habitat structural complexity and protection from fisheries. Temporal variations, otherwise, may have been influenced by species proximity to their distributional limit, in synergy with density-dependent mechanisms and stochastic winter temperature oscillations. The MPAs harbour higher density and biomass for most species. Nonetheless, a prominent temporal decline in the recruitment of Epinephelus marginatus calls into question the continuous effectiveness of the MPA., (© 2020 The Fisheries Society of the British Isles.)

Introduction: The relationship between volume and outcome of total knee arthroplasties is a concern in both the civilian and military patient populations. We sought to compare surgeons and hospital procedure volumes performed on military service members and define factors leading to increased civilian referrals., Materials and Methods: The Military Health System Data Repository (MDR) contains patient information on all healthcare beneficiary encounters, including care provided both in Military Health System (MHS) facilities and in civilian network facilities. The Military Analysis and Reporting Tool (M2) queried the MDR for all patients between 2011 and 2015 with a CPT code for hip or knee arthroplasty associated with a provider HIPAA taxonomy code for orthopedic surgery. M2 enrollee encounters were used to calculate the total number of arthroplasty procedures performed by both military and civilian orthopedic surgeons on MHS enrollees as well as the incidence rate of arthroplasty procedures. Logistic regression was used to predict which cases were more likely to have been treated at military treatment facilities using patient gender, sponsor service branch, age, and beneficiary category., Results: During the study period, a total of 12,627 military facility arthroplasty cases and a total of 142,637 civilian facility arthroplasty cases were performed on TRICARE enrolled patients. The total number of military surgeons performing arthroplasty on TRICARE enrolled patients was 323, while the total number of civilian surgeons performing arthroplasty was 10,245 during the same time period; the number of military surgeons performing arthroplasty on active duty patients was 176, and the total number of civilian surgeons performing arthroplasty on military patients was 1045. Overall, including retirees and activity duty service members, more procedures are performed by civilian network surgeons than military surgeons in all states. In an adjusted model, male patients were slightly more likely to receive care at an military treatment facilitie than female patients (OR = 1.47, 95% CI: 1.41-1.53). Furthermore, with respect to service, patients with Air Force (OR: 1.08, 95% CI: 1.02-1.15) and Navy sponsors (OR: 1.61, 95% CI: 1.51-1.71) were more likely to receive military care than patients with Army sponsors., Conclusions: Based on our findings, we recommend the MHS focus attention to recapturing the Army active duty male patients who are more likely to receive care outside of the military healthcare network. Further analysis of the many factors including, but not limited to, referral process for total joint arthroplasty, time to procedure, and facility resources is required, in addition to assessing patient outcomes following the procedures., (© The Author(s) 2020. Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Human-mediated species invasions are recognized as a leading cause of global biotic homogenization and extinction. Studies on colonization events since early stages, establishment of new populations and range extension are scarce because of their rarity, difficult detection and monitoring. Chromis limbata is a reef-associated and non-migratory marine fish from the family Pomacentridae found in depths ranging between 3 and 45 m. The original distribution of the species encompassed exclusively the eastern Atlantic, including the Azores, Madeira and the Canary Islands. It is also commonly reported from West Africa between Senegal and Pointe Noire, Congo. In 2008, vagrant individuals of C. limbata were recorded off the east coast of Santa Catarina Island, South Brazil (27° 41' 44″ S, 48° 27' 53″ W). This study evaluated the increasing densities of C. limbata populations in Santa Catarina State shoreline. Two recent expansions, northwards to São Paulo State and southwards to Rio Grande do Sul State, are discussed, and a niche model of maximum entropy (MaxEnt) was performed to evaluate suitable C. limbata habitats. Brazilian populations are established and significantly increasing in most sites where the species has been detected. The distributional boundaries predicted by the model are clearly wider than their known range of occurrence, evidencing environmental suitability in both hemispheres from areas where the species still does not occur. Ecological processes such as competition, predation and specially habitat selectivity may regulate their populations and overall distribution range. A long-term monitoring programme and population genetics studies are necessary for a better understanding of this invasion and its consequences to natural communities., (© 2020 The Fisheries Society of the British Isles.)

There remains a dearth of research on and general knowledge with regard to materials used for wound closure and soft-tissue repair and approximation. Critical suture properties include physical configuration, fluid absorption and capillarity, caliber or diameter, tensile strength, torsion, absorbability, elasticity, plasticity, memory, coefficient of friction, and knot security. The optimal ranges of each of these characteristics remain undefined for most sutures and indications. Needle types and basic design characteristics affect suture passage and require further consideration with regard to specific suture-needle selection. Suture must perform its intended purpose with a minimum of undesirable reaction and infectious potential, adequate duration of efficacy, and adequate strength. However, stronger, or high tensile strength, suture is not always better because of the requisite increase in suture caliber as well as the potential for inadvertent tissue strangulation, possibly increasing inflammatory reactivity. Sometimes, we seek stable, watertight fascial closure; occasionally, strong and durable tendon repair; and other times, gentle, cosmetically friendly, skin eversion and opposition. A variety of common suture types differ in these critical characteristics and may be optimally utilized for contrasting, but sometimes overlapping, indications.

Background: Machine-learning methods such as the Bayesian belief network, random forest, gradient boosting machine, and decision trees have been used to develop decision-support tools in other clinical settings. Opioid abuse is a problem among civilians and military service members, and it is difficult to anticipate which patients are at risk for prolonged opioid use., Questions/purposes: (1) To build a cross-validated model that predicts risk of prolonged opioid use after a specific orthopaedic procedure (ACL reconstruction), (2) To describe the relationships between prognostic and outcome variables, and (3) To determine the clinical utility of a predictive model using a decision curve analysis (as measured by our predictive system's ability to effectively identify high-risk patients and allow for preventative measures to be taken to ensure a successful procedure process)., Methods: We used the Military Analysis and Reporting Tool (M2) to search the Military Health System Data Repository for all patients undergoing arthroscopically assisted ACL reconstruction (Current Procedure Terminology code 29888) from January 2012 through December 2015 with a minimum of 90 days postoperative follow-up. In total, 10,919 patients met the inclusion criteria, most of whom were young men on active duty. We obtained complete opioid prescription filling histories from the Military Health System Data Repository's pharmacy records. We extracted data including patient demographics, military characteristics, and pharmacy data. A total of 3.3% of the data was missing. To curate and impute all missing variables, we used a random forest algorithm. We shuffled and split the data into 80% training and 20% hold-out sets, balanced by outcome variable (Outcome90Days). Next, the training set was further split into training and validation sets. Each model was built on the training data set, tuned with the validation set as applicable, and finally tested on the separate hold-out dataset. We chose four predictive models to develop, at the end choosing the best-fit model for implementation. Logistic regression, random forest, Bayesian belief network, and gradient boosting machine models were the four chosen models based on type of analysis (classification). Each were trained to estimate the likelihood of prolonged opioid use, defined as any opioid prescription filled more than 90 days after anterior cruciate reconstruction. After this, we tested the models on our holdout set and performed an area under the curve analysis concordance statistic, calculated the Brier score, and performed a decision curve analysis for validation. Then, we chose the method that produced the most suitable analysis results and, consequently, predictive power across the three calculations. Based on the calculations, the gradient boosting machine model was selected for future implementation. We systematically selected features and tuned the gradient boosting machine to produce a working predictive model. We performed area under the curve, Brier, and decision curve analysis calculations for the final model to test its viability and gain an understanding of whether it is possible to predict prolonged opioid use., Results: Four predictive models were successfully developed using gradient boosting machine, logistic regression, Bayesian belief network, and random forest methods. After applying the Boruta algorithm for feature selection based on a 100-tree random forest algorithm, features were narrowed to a final seven features. The most influential features with a positive association with prolonged opioid use are preoperative morphine equivalents (yes), particular pharmacy ordering sites locations, shorter deployment time, and younger age. Those observed to have a negative association with prolonged opioid use are particular pharmacy ordering sites locations, preoperative morphine equivalents (no), longer deployment, race (American Indian or Alaskan native) and rank (junior enlisted).On internal validation, the models showed accuracy for predicting prolonged opioid use with AUC greater than our benchmark cutoff 0.70; random forest were 0.76 (95% confidence interval 0.73 to 0.79), 0.76 (95% CI 0.73 to 0.78), 0.73 (95% CI 0.71 to 0.76), and 0.72 (95% CI 0.69 to 0.75), respectively. Although the results from logistic regression and gradient boosting machines were very similar, only one model can be used in implementation. Based on our calculation of the Brier score, area under the curve, and decision curve analysis, we chose the gradient boosting machine as the final model. After selecting features and tuning the chosen gradient boosting machine, we saw an incremental improvement in our implementation model; the final model is accurate, with a Brier score of 0.10 (95% CI 0.09 to 0.11) and area under the curve of 0.77 (95% CI 0.75 to 0.80). It also shows the best clinical utility in a decision curve analysis., Conclusions: These scores support our claim that it is possible to predict which patients are at risk of prolonged opioid use, as seen by the appropriate range of hold-out analysis calculations. Current opioid guidelines recommend preoperative identification of at-risk patients, but available tools for this purpose are crude, largely focusing on identifying the presence (but not relative contributions) of various risk factors and screening for depression. The power of this model is that it will permit the development of a true clinical decision-support tool, which risk-stratifies individual patients with a single numerical score that is easily understandable to both patient and surgeon. Probabilistic models provide insight into how clinical factors are conditionally related. Not only will this gradient boosting machine be used to help understand factors contributing to opiate misuse after ACL reconstruction, but also it will allow orthopaedic surgeons to identify at-risk patients before surgery and offer increased support and monitoring to prevent opioid abuse and dependency., Level of Evidence: Level III, therapeutic study.

Background: Limited data are available regarding excessive opioid prescribing in the perioperative period after routine orthopaedic procedures in US military personnel., Purpose: To examine the demographic profile of the patients receiving these medications and to identify potential risk factors for prolonged opioid use after anterior cruciate ligament reconstruction (ACLR) in the active duty military population., Study Design: Case-control study; Level of evidence, 3., Methods: The Military Analysis and Reporting Tool (M2) was used to search the Military Health System Data Repository (MDR) for patients undergoing ACLR from 2012 through 2015 and specifically for active duty personnel with an arthroscopically assisted ACLR (Current Procedural Terminology [CPT] code 29888). Complete opioid prescription filling history was also obtained. This study had 2 primary outcomes: (1) use of opiate analgesics more than 90 days after surgery, representing prolonged opiate prescriptions, and (2) high levels of postoperative opiate use, defined as having filled prescriptions accounting for greater than the 95th percentile of morphine equivalents for patients in the study cohort. Data were analyzed via multivariate regression analysis to identify potential associations with the primary outcomes., Results: A total of 9474 patients met the inclusion criteria. Median patient age was 27 years, and the sample included 1316 (14%) female and 8158 (86%) male patients. A total of 66 (0.7%) patients had a preoperative diagnosis for substance abuse; 2656 (28%) patients continued to receive opioid prescriptions more than 90 days after surgery, and 502 (5%) patients were in the top 95th percentile of all opioid users within the study cohort. Total preoperative morphine equivalents per day and total perioperative morphine equivalents per day were highly important risk factors for both outcomes, although other demographic factors such as race, sex, and age may play minor roles., Conclusion: We identified total preoperative morphine equivalents, total perioperative morphine equivalents, sex, and race as potential predictors of prolonged opioid use after ACLR. This information may prove useful in developing a predictive model to identify at-risk patients before surgery. This could help mitigate future misuse or abuse and improve preoperative patient counseling regarding pain management expectations., Competing Interests: One or more of the authors has declared the following potential conflict of interest or source of funding: G.C.B. has received grant support from Arthrex and educational support from Arthrex, Smith & Nephew, and Supreme Orthopedic Systems. B.K.P. has received hospitality payments from Zimmer Biomet. J.A.F. has received educational support from Zimmer Biomet and consulting fees from The Solsidan Group. J.F.D. has received educational support from Supreme Orthopedic Systems. AOSSM checks author disclosures against the Open Payments Database (OPD). AOSSM has not conducted an independent investigation on the OPD and disclaims any liability or responsibility relating thereto., (© The Author(s) 2020.)

Smokers who undergo total joint arthroplasty (TJA) face increased rates of medical and surgical complications that can be reduced by preoperative smoking cessation. We investigated the long-term durability of preoperative smoking cessation among TJA patients. Twenty-seven TJA patients who were identified as having an active history of smoking at the preoperative appointment before TJA consented to telephone survey about their perioperative and current smoking status. Average time from operation to survey was 3.7 years. Of the 27 patients, 21 (77.8%) were identified as having quit smoking prior to surgery. Of these 21 patients, 10 (47.6%) self-reported continued abstinence from smoking at the time of survey. Our cessation rate was significantly lower than reported long-term smoking cessation rates with standard therapies (p < 0.001). Our results suggest that preoperative counseling and a requirement for smoking-cessation prior to elective TJA may have long-term durability that exceeds that of popular reported methods. (Journal of Surgical Orthopaedic Advances 29(2):103-105, 2020).

Background: PATHFx is a clinical decision-support tool based on machine learning capable of estimating the likelihood of survival after surgery for patients with skeletal metastases. The applicability of any machine-learning tool depends not only on successful external validation in unique patient populations but also on remaining relevant as more effective systemic treatments are introduced. With advancements in the treatment of metastatic disease, it is our responsibility to patients to ensure clinical support tools remain contemporary and accurate., Question/purposes: Therefore, we sought to (1) generate updated PATHFx models using recent data from patients treated at one large, urban tertiary referral center and (2) externally validate the models using two contemporary patient populations treated either surgically or nonsurgically with external-beam radiotherapy alone for symptomatic skeletal metastases for symptomatic lesions., Methods: After obtaining institutional review board approval, we collected data on 208 patients undergoing surgical treatment for pathologic fractures at Memorial Sloan Kettering Cancer Center between 2015 and 2018. These data were combined with the original PATHFx training set (n = 189) to create the final training set (n = 397). We then created six Bayesian belief networks designed to estimate the likelihood of 1-month, 3-month, 6-month, 12-month, 18-month, and 24-month survival after treatment. Bayesian belief analysis is a statistical method that allows data-driven learning to arise from conditional probabilities by exploring relationships between variables to estimate the likelihood of an outcome using observed data. For external validation, we extracted the records of patients treated between 2016 and 2018 from the International Bone Metastasis Registry and records of patients treated nonoperatively with external-beam radiation therapy for symptomatic skeletal metastases from 2012 to 2016 using the Military Health System Data Repository (radiotherapy-only group). From each record, we collected the date of treatment, laboratory values at the time of treatment initiation, demographic data, details of diagnosis, and the date of death. All records reported sufficient follow-up to establish survival (yes/no) at 24-months after treatment. For external validation, we applied the data from each record to the new PATHFx models. We assessed calibration (calibration plots), accuracy (Brier score), discriminatory ability (area under the receiver operating characteristic curve [AUC])., Results: The updated PATHFx version 3.0 models successfully classified survival at each time interval in both external validation sets and demonstrated appropriate discriminatory ability and model calibration. The Bayesian models were reasonably calibrated to the Memorial Sloan Kettering Cancer Center training set. External validation with 197 records from the International Bone Metastasis Registry and 192 records from the Military Health System Data Repository for analysis found Brier scores that were all less than 0.20, with upper bounds of the 95% confidence intervals all less than 0.25, both for the radiotherapy-only and International Bone Metastasis Registry groups. Additionally, AUC estimates were all greater than 0.70, with lower bounds of the 95% CI all greater than 0.68, except for the 1-month radiotherapy-only group. To complete external validation, decision curve analysis demonstrated clinical utility. This means it was better to use the PATHFx models when compared to the default assumption that all or no patients would survive at all time periods except for the 1-month models. We believe the favorable Brier scores (< 0.20) as well as DCA indicate these models are suitable for clinical use., Conclusions: We successfully updated PATHFx using contemporary data from patients undergoing either surgical or nonsurgical treatment for symptomatic skeletal metastases. These models have been incorporated for clinical use on PATHFx version 3.0 (https://www.pathfx.org). Clinically, external validation suggests it is better to use PATHFx version 3.0 for all time periods except when deciding whether to give radiotherapy to patients with the life expectancy of less than 1 month. This is partly because most patients survived 1-month after treatment. With the advancement of medical technology in treatment and diagnosis for patients with metastatic bone disease, part of our fiduciary responsibility is to the main current clinical support tools., Level of Evidence: Level III, therapeutic study.

Introduction: Acute pain and chronic pain are significant burdens in the Department of Defense, compounded by the ongoing opioid crisis. Given the ubiquity of (leftover) opioid prescriptions following orthopedic surgery, it is essential to identify feasible and acceptable avenues of opioid risk mitigation efforts. The present quality improvement project builds on recent studies by evaluating factors related to opioid prescribing decisions in a sample of orthopedic surgery providers., Materials and Methods: This quality improvement project received a Determination of Not Research and was conducted through a collaboration between the Department of Orthopaedic Surgery and the Department of Anesthesiology and Pain Management at Walter Reed National Military Medical Center. Providers in the Department of Orthopaedic Surgery completed an anonymous online survey assessing opioid prescribing education, factors influencing prescribing practices, opioid-safety practices, and perspectives on potential opioid safety initiatives., Results: In total, 39 respondents completed surveys. There was variability in exposure to different types of opioid prescribing education, with some variation between attendings/physician assistants and residents. Patients' acute postsurgical pain, using a standardized amount for most patients, and prescription histories were the three most influential factors. Concern of patients running out and fear of patient dissatisfaction were the least influential factors. Respondents commonly reported engagement in promoting nonpharmacological pain management, as well as coordinating with chronic pain providers when applicable, but did not commonly report educating patients on leftover opioid disposal. Respondents indicated that a barrier to opioid risk mitigation was the difficulty of accessing appropriate electronic health record data to inform decisions. Lastly, they reported openness to proposed opioid safety initiatives., Conclusion: The results of this quality improvement project identified several target areas for future initiatives focused on improving opioid prescribing practices. This included a provider training program, improved patient education system, increased awareness and use of opioid tracking databases, and development of a standardized (but adaptable per patient characteristics and history) recommended dose for common orthopedic surgeries. Future projects will target tailored development, implementation, and evaluation of such efforts., (Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States 2019. This work is written by (a) US Government employee(s) and is in the public domain in the US.)

Environmental conditions shape the occurrence and abundance of habitat-building organisms at global scales. Rhodolith beds structure important hard substrate habitats for a large number of marine benthic organisms. These organisms can benefit local biodiversity levels, but also compete with rhodoliths for essential resources. Therefore, understanding the factors shaping the distribution of rhodoliths and their associated communities along entire distributional ranges is of much relevance for conservational biology, particularly in the scope of future environmental changes. Here we predict suitable habitat areas and identify the main environmental drivers of rhodoliths' variability and of associated epiphytes along a large-scale latitudinal gradient. Occurrence and abundance data were collected throughout the South-western Atlantic coast (SWA) and modelled against high resolution environmental predictors extracted from Bio-Oracle. The main drivers for rhodolith occurrence were light availability and temperature at the bottom of the ocean, while abundance was explained by nitrate, temperature and current velocity. Tropical regions showed the highest abundance of rhodoliths. No latitudinal pattern was detected in the variability of epiphytes abundance. However, significant differences were found between sampled sites regarding the composition of predominant taxa. The predictors influencing such differences were temperature and nitrate. The Tropical region is abundant in species with warm-water affinities, decreasing toward warm temperate region. The expressive occurrence of tropical species not referred before for warm temperate beds indicate a plausible tropicalization event., Competing Interests: Declaration of competing interest None., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Previous studies have shown relatively high rates of inappropriate referrals to pediatric orthopedic surgery. The purpose of this study was to determine the rate of inappropriate referrals in an egalitarian healthcare system. We reviewed all 400 referrals to our pediatric orthopedic surgery practice over a 2-year period and assessed the appropriateness of each based on the American Academy of Pediatrics guidelines. A total 206 (52%) referrals were deemed appropriate. There is a high rate of inappropriate referral to pediatric orthopedic surgery even within a closed, egalitarian healthcare system. Targeted education of primary care providers may help reduce this rate.

Blood flow restriction (BFR) has been shown to produce beneficial adaptations to skeletal muscle. These adaptations have been documented in the civilian and military populations. BFR therapy may provide patients a safe method to begin strength training at earlier stages of rehabilitation to allow for earlier and more effective return to activity and improved military readiness. The purpose was to review BFR therapy physiology, complications, side effects, standardized treatment algorithms, and long-term patient outcomes.

Introduction: Women account for approximately 15% of the active duty US Army, and studies show that women may be at an increased risk of musculoskeletal injury during sport and military training. Nationally, the field of orthopedic surgery comprises 14% women, lagging behind other surgical fields. Demographics for US Military orthopedic surgeons are not readily available. Similarly, demographic data of graduating medical students entering Military Medicine are not reported. We hypothesize that a gender disparity within military orthopedics will be apparent. We will compare the demographic profile of providers to our patients and hypothesize that the two groups are dissimilar. Secondarily, we examine the demographics of military medical students potentially entering orthopedics from the Uniformed Services University of the Health Sciences (USUHS) or the Health Professions Scholarship Program., Methods: A census was formed of all US Army active duty orthopedic surgeons to include staff surgeons and residents, as well as US Army medical student graduates and orthopedic patients., Results: There are 252 Army orthopedic surgeons and trainees; 26 (10.3%) are women and 226 (89.7%) are men. There were no significant demographic differences between residents and staff. Between 2014 and 2017, the 672 members of the USUHS graduating classes included 246 Army graduates. Of those, 62 (25%) were female. Army Health Professions Scholarship Program graduated 1,072 medical students, with women comprising 300 (28%) of the group. No statistical trends were seen over the 4 yr at USUHS or in Health Professions Scholarship Program. In total, 2,993 orthopedic clinic visits during the study period were by Army service members, 23.6% were women., Conclusion: There exists a gender disparity among US Army orthopedic surgeons, similar to that seen in civilian orthopedics. Gender equity is also lacking among medical students who feed into Army graduate medical education programs. The gender profile of our patient population is not reflected by that of providers. Because patients prefer providers of the same gender, this is a limitation to patient satisfaction and access to care for musculoskeletal injuries. Further study is underway to identify perceptions and potential causes of these disparities, including the critical perspective of our patients. In addition to the inherent benefits offered by diversity (e.g., expanding the talent pool and more perspectives for decision-making), ultimately it affords a greater ability to maintain a fit and ready force., (Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States 2017. This work is written by (a) US Government employee(s) and is in the public domain in the US.)

Inadequate pain management after total knee arthroplasty (TKA) can interfere with participation in and progression of physical rehabilitation, and thereby prolong hospital stay, and increase costs and overall dissatisfaction with the procedure. At our institution, TKA traditionally has been performed with femoral nerve catheters (FNCs) for postoperative pain control. We conducted a retrospective, longitudinal, repeated -measures study to compare FNC and long-acting liposomal bupivacaine (LALB) with respect to pain control, range of motion, ability to ambulate, and hospital length of stay. Twenty-three patients underwent separately staged bilateral TKAs, the first with FNC and the second with periarticular injection of LALB. Statistically significant differences favoring LALB over FNC were found for hospital length of stay (P < .01), per-attempt walking distance during hospitalization (P < .01), total range of motion (extension plus flexion) at 3-week follow-up (P = .02), and total morphine-equivalent dose during hospitalization (P = .02). Our results showed that, compared with patients who received FNC, patients who received LALB had comparable pain control, improved knee range of motion, and shorter hospital stays. Additional clinical studies are needed to better determine the efficacy and cost-effectiveness of LALB and other long-acting local anesthetic formulations., Competing Interests: Authors’ Disclosure Statement: The authors report no actual or potential conflict of interest in relation to this article.

The damselfish Chromis limbata is native to the Macaronesian Archipelagos (Azores, Madeira and Canaries) and the western coast of Africa between Senegal and Angola. During the austral summers of 2008 and 2009 the species was recorded for the first time in the south-western Atlantic Ocean around Campeche and Xavier Islands, in Florianópolis, Santa Catarina State, Brazil. Here, the progression of C. limbata in southern Brazilian waters is described using visual counts and genetic surveys and changes in the density of the native congener Chromis multilineata were also investigated. Underwater visual censuses of both Chromis species were carried out from 2009 to 2014. Chromis limbata tissue samples were collected and the mtDNA control region was sequenced and compared with mtDNA haplotypes from the natural range to confirm species identity, compare genetic diversity and to infer connectivity between newly established Brazilian populations. The Brazilian population of C. limbata increased significantly over the past 5 years and the effect on C. multilineata is still an open question, longer time-series data will be necessary to clarify possible interactions. The molecular analyses confirmed species identity, revealed strong haplotype connectivity among Brazilian study sites and showed a low genetic diversity in Brazil when compared with the native populations, suggesting few individuals started the invasion. Four hypotheses could explain this colonizing event: C. limbata was released by aquarium fish keepers; larvae or juveniles were transported via ship ballast water; the species has rafted alongside oil rigs; they crossed the Atlantic Ocean through normal larval dispersal or naturally rafting alongside drifting objects. The rafting hypotheses are favoured, but all four possibilities are plausible and could have happened in combination., (© 2017 The Fisheries Society of the British Isles.)

Introduction: Provoked vestibulodynia (PVD) is a common vulvovaginal pain condition that negatively impacts women's psychological and sexual well-being. Controlled studies have found that women with PVD report greater negative and less positive cognitions about penetration; however, associations between these types of cognitions and women's pain and sexual well-being remain unknown. Further, researchers have yet to examine how interpersonal variables such as sexual communication may impact the association between women's penetration cognitions and PVD outcomes., Aim: We examined associations between vaginal penetration cognitions and sexual satisfaction, sexual function, and pain in women with PVD, as well as the moderating role of sexual communication., Methods: Seventy-seven women (M age = 28.32, SD = 6.19) diagnosed with PVD completed the catastrophic and pain cognitions and positive cognitions subscales of the Vaginal Penetration Cognition Questionnaire, as well as the Dyadic Sexual Communication Scale. Participants also completed measures of sexual satisfaction, sexual function, and pain., Main Outcome Measures: Dependent measures were the (i) Global Measure of Sexual Satisfaction Scale; (ii) Female Sexual Function Index; and (iii) Present Pain Intensity scale of the McGill Pain Questionnaire, with reference to pain during vaginal intercourse., Results: Women's lower catastrophic and pain cognitions, higher positive cognitions, and higher sexual communication were each uniquely associated with higher sexual satisfaction and sexual function. Lower catastrophic and pain cognitions also were associated with women's lower pain. For women who reported higher sexual communication, as positive cognitions increased, there was a significantly greater decrease in pain intensity during intercourse compared to women who reported lower levels of sexual communication., Conclusion: Findings may inform cognitive-behavioral interventions aimed at improving the pain and sexual well-being of women with PVD. Targeting the couple's sexual communication and women's penetration cognitions may improve women's sexual adjustment and reduce pain., (Copyright © 2016 International Society for Sexual Medicine. Published by Elsevier Inc. All rights reserved.)

It has been shown recently that when reactants and products are well modeled within a comprehensive self-consistent theory for the electrochemical interface, accurate predictions are possible for reversible potentials, Urev, in acid electrolyte for reactions such as reduction of H(+)(aq) to form under potential deposited H(ads) and oxidation of an OH bond of H2O(ads) to deposit OH(ads). Predictions are based on calculated Gibbs energies for the reactant and product being equal at the reversible potential, which is the potential at the crossing point for reaction and product Gibbs energies, plotted as functions of electrode potential. In this Letter, it is demonstrated that the same capability holds for these reactions in basic electrolyte. This demonstration opens up the opportunity for predictions of reversible potentials and mechanisms for other electrocatalytic reactions in base.

Distraction from erotic cues during sexual encounters is a major contributor to sexual difficulties in men and women. Being able to assess distraction in studies of sexual arousal will help clarify underlying contributions to sexual problems. The current study aimed to identify the most accurate assessment of distraction from erotic cues in healthy men (n = 29) and women (n = 38). Participants were assigned to a no distraction, low distraction, or high distraction condition. Distraction was induced using an auditory distraction task presented during the viewing of an erotic video. Attention to erotic cues was assessed using three methods: a written quiz, a visual quiz, and a self-reported distraction measure. Genital and psychological sexual responses were also measured. Self-reported distraction and written quiz scores most accurately represented the level of distraction present, while self-reported distraction also corresponded with a decrease in genital arousal. Findings support the usefulness of self-report measures in conjunction with a brief quiz on the erotic material as the most accurate and sensitive ways to simply measure experimentally-induced distraction. Insight into distraction assessment techniques will enable evaluation of naturally occurring distraction in patients suffering from sexual problems.

The effects of military training activities on the land condition of Army installations vary spatially and temporally. Training activities observably degrade land condition while also increasing biodiversity and stabilizing ecosystems. Moreover, other anthropogenic activities regularly occur on military lands such as prescribed burns and agricultural haying-adding to the dynamics of land condition. Thus, spatially and temporally assessing the impacts of military training, prescribed burning, agricultural haying, and their interactions is critical to the management of military lands. In this study, the spatial distributions and patterns of military training-induced disturbance frequency were derived using plot observation and point observation-based method, at Fort Riley, Kansas from 1989 to 2001. Moreover, spatial and variance analysis of cumulative impacts due to military training, burning, haying, and their interactions on the land condition of Fort Riley were conducted. The results showed that: (1) low disturbance intensity dominated the majority of the study area with exception of concentrated training within centralized areas; (2) high and low values of disturbance frequency were spatially clustered and had spatial patterns that differed significantly from a random distribution; and (3) interactions between prescribed burning and agricultural haying were not significant in terms of either soil erosion or disturbance intensity although their means and variances differed significantly between the burned and non-burned areas and between the hayed and non-hayed areas.

Quantum chemical theory is used to identify the reasons for platinum's limitations as an electrocatalyst for oxidizing methanol at fuel cell anodes. The linear Gibbs energy relation (LGER) method is employed to predict reversible potentials for reaction steps for intermediates on the electrode surface. In this procedure, standard reversible potentials are calculated for the reactions in bulk solution phase and then they are perturbed using calculated adsorption bond strengths to the electrode surface, yielding the equilibrium potentials for each electron transfer step for adsorbed intermediates. Adsorption properties of ideal electrocatalysts for the methanol oxidation are found by imposing the condition that the reversible potential of each electron transfer step equals that for the overall reaction. The adsorption bond strengths that provide the ideal properties also apply to formic acid oxidation and carbon dioxide reduction. It is instructive to think of the ideal electrocatalyst as a lens that focusses the reversible potentials for the n individual electron transfer steps to the reversible potential for the n-electron process. It is found that the ideal catalyst will adsorb many intermediates, including HOOC, CO, OCH, HOC, HOCH, HOCH2, and OCH3 more weakly than platinum, and OOCH and OH more strongly. For example, for one possible pathway it is necessary to weaken adsorption bond strengths for HOCH2, HOCH, OCH, HOOC by about 0.5 eV, weaken adsorption CO by about 1.1 eV and strengthen OH adsorption by about 0.6 eV. These results imply a need for developing new multi-component catalysts.

This article aspires to an embodiment of dynamic living versus mere survival. The term cancer survivor, including a survivor who is in remission, has been legitimated (Berger and Luckmann, The social construction of reality, p. 94 1967) by language which creates knowledge of what a cancer survivor is and does. Because we act under descriptions (Hacking, The social construction of what?, p. 103 1999), those of us who have passed through illnesses such as cancer not only have been given the name and the idea of survivor, we have assumed and conform to some or most of the characteristics assigned to it; examples of some of those characteristics are discussed throughout this project. Whether or not we choose to enact all that falls under the grammar of the classification of survivor, we still live with, create, and experience ourselves and others as legitimated by such a classification. The term survivor operates through a number of institutions (medical, capitalism, and media) resulting in individuals' awareness of such classifications about themselves and others. Many, if not most, who are aware of being classified as survivors may wish to modify or resist the constraining aspects of those classifications and their descriptions. Through layered accounts of interviews and prose, I interact with this term as one who is both caught in and wants to go against the stream of classification and description. I want to transcend what I know, yet I am aware that whatever story I make and tell is a part of the whole-my story is part of two other survivor's stories which I include in the following telling of my own. All of our stories matter. Still, I want to look beyond what is in front of me, move beyond it, dream. I do so with a desire to tell my story as part of other survivors' stories.

When standard reversible potentials for bulk solution reactions, U(0), are known, the reversible potentials when the reactant and product are adsorbed on an electrocatalyst surface, U(surf)(rev), are given in terms of these potentials and the adsorption Gibbs energy bond strengths: U(surf)(rev) = U0 + D(ads)G (Ox)/F-Δ(ads)G (R)/F (i). When the Δ(ads)G (Ox) and Δ(ads)G (Red) values are known at potential U, this equation is exact. When the overpotential for a multi-electron transfer reaction is minimal, each electron transfer takes place at the standard reversible potential for the overall reaction. In the case of O(2) reduction to water via the intermediate step OOH(aq) → O(aq) + OH(aq), or via O(2)(g) → 2O(aq), the respective endergonic O-O dissociation Gibbs energies are shown to be 2.52 eV and 4.76 eV. When the oxygen product and water reactant adsorb weakly, as on platinum, the adsorption Gibbs energies, Δ(ads)G, for O, OH, and OOH intermediates can be uniquely predicted using these data. All of the above depend exclusively on experimentally determined data. Reversible potentials have been calculated for oxygen reduction steps on the platinum electrocatalyst surface using Interface 1.0, a comprehensive computational code for the potential dependence of the electrochemical interface. Using these results as benchmarks, is found to be accurate to around 0.1 V when the Δ(ads)G are values calculated for the potentials of zero charge, instead of 1.229 V, which is a significant simplification. The variation in Δ(ads)G values between the calculated potentials of zero charge and 1.229 V are found to be 0.2 eV V(-1) or less. Prior work, using internal adsorption energies calculated at the potential of zero charge in place of Gibbs energies in was found to be accurate to within about 0.2 V. On platinum Δ(ads)G of the reaction OOH(ads) → O(ads) + OH(ads) is calculated at the potential of zero charge for the reactant and product to be about 1.2 eV exergonic under Langmuir conditions, and this Gibbs energy loss reduces the 1.229 V four-electron reversible potential on the platinum surface to an effective reversible potential of about 0.93 V for this mechanism on platinum. The effective reversible potential is a consequence of efficiency loss, not kinetics. Based on these values, the onset potential for four-electron oxygen reduction will be less than or equal to the effective reversible potential and on pure Pt(111) it appears to be equal to it.

Advances in our knowledge of the molecular mechanisms involved in cancer biology have contributed to an increase in novel target-specific oncology therapeutics. Unfortunately, clinical development of new drugs is an expensive and slow process, and the patient and financial resources needed to study the vast number of potential therapies are limited, requiring novel approaches to clinical trial design and patient recruitment. In addition, traditional efficacy endpoints may not be adequate to fully determine the therapeutic worth of the new classes of targeted agents. In this new era of drug development, it has become increasingly clear that new clinical trial design paradigms that examine nontraditional endpoints have become necessary to assist in prioritizing the development of the most promising agents. It is also vital that individual patient management be considered, and the subpopulations of patients most likely to derive benefit or experience harm from a new therapy be identified as early as possible. Phase I and II clinical trials allow investigators doing clinical research the opportunity to define these critical endpoints and subpopulations early on, before conducting large-scale randomized phase III clinical trials, which require an abundance of financial and patient resources., (©2010 AACR.)

Vehicle use during military training activities results in soil disturbance and vegetation loss. The capacity of lands to sustain training is a function of the sensitivity of lands to vehicle use and the pattern of land use. The sensitivity of land to vehicle use has been extensively studied. Less well understood are the spatial patterns of vehicle disturbance. Since disturbance from off-road vehicular traffic moving through complex landscapes varies spatially, a spatially explicit nonlinear regression model (disturbance model) was used to predict the pattern of vehicle disturbance across a training facility. An uncertainty analysis of the model predictions assessed the spatial distribution of prediction uncertainty and the contribution of different error sources to that uncertainty. For the most part, this analysis showed that mapping and modeling process errors contributed more than 95% of the total uncertainty of predicted disturbance, while satellite imagery error contributed less than 5% of the uncertainty. When the total uncertainty was larger than a threshold, modeling error contributed 60% to 90% of the prediction uncertainty. Otherwise, mapping error contributed about 10% to 50% of the total uncertainty. These uncertainty sources were further partitioned spatially based on other sources of uncertainties associated with vehicle moment, landscape characterization, satellite imagery, etc., (2009 Elsevier Ltd. All rights reserved.)

Preclinical evaluation of vaginal microbicides includes screening against lactobacilli. However, there is no consensus regarding the species to be tested. This study was carried out to determine if results with one species would apply to other species, and to evaluate the utility of turbidometry as a screening tool. One current (PPCM; previously designated sulfuric acid-modified mandelic acid, SAMMA) and two former (cellulose sulfate, CS; and polystyrene sulfonate, PSS) candidate microbicides were evaluated. Bacterial growth was measured turbidometrically and by direct cell count. No microbicide affected Lact. gasseri, measured by either method. Apparent inhibition of Lact. jensenii by CS, PSS, and PPCM, and of Lact. crispatus by CS, occurred with turbidometric measurement. This was not substantiated with direct cell count. PSS and PPCM inhibited Lact. crispatus and Lact. acidophilus with both methods. These findings agree with results from vaginal isolates, which included Lact. gasseri, jensenii, acidophillus, crispatus, rhamnosis, casei, and paracasei. We conclude that sensitivities of similar lactobacilli to at least three microbicides are different. A single species is inadequate for screening vaginal products. Turbidometric evaluation is a sensitive, but not specific, screening method. We recommend that this method be used to screen candidate microbicides against several species of prevalent Lactobacillus species as an initial measure of microbicide safety evaluation.

Charge transfer between diamond and an electrochemical redox couple in an adsorbed water film has recently been shown to pin the Fermi level in hydrogen-terminated diamond. Here we show that this effect is a more general phenomenon and influences the properties of other semiconductors when the band lineup between the ambient and electronic states in the semiconductor is appropriate. We find that the luminescent intensities from GaN and ZnO change in different, but predictable, ways when exposed to HCl and NH3 vapors in humid air. The effect is reversible and has been observed on single crystals, nanowires, flakes, and powders. These observations are explained by electron exchange between the oxygen electrochemical redox couple in an adsorbed water film and electronic states in the semiconductor. This effect can take place in parallel with other processes such as defect formation, chemisorption, and surface reconstruction and may play an important, but previously unrecognized, role when electronic and optical measurements are made in air.

Undoped, high-quality diamond is, under almost all circumstances, one of the best insulators known. However, diamond covered with chemically bound hydrogen shows a pronounced conductivity when exposed to air. This conductivity arises from positive-charge carriers (holes) and is confined to a narrow near-surface region. Although several explanations have been proposed, none has received wide acceptance, and the mechanism remains controversial. Here, we report the interactions of hydrogen-terminated, macroscopic diamonds and diamond powders with aqueous solutions of controlled pH and oxygen concentration. We show that electrons transfer between the diamond and an electrochemical reduction/oxidation couple involving oxygen. This charge transfer is responsible for the surface conductivity and also influences contact angles and zeta potentials. The effect is not confined to diamond and may play a previously unrecognized role in other disparate systems.

Off-road vehicles increase soil erosion by reducing vegetation cover and other types of ground cover, and by changing the structure of soil. The investigation of the relationship between disturbance from off-road vehicles and the intensity of the activities that involve use of vehicles is essential for water and soil conservation and facility management. Models have been developed in a previous study to predict disturbance caused by off-road vehicles. However, the effect of data on model quality and model performance, and the appropriate structure of models have not been previously investigated. In order to improve the quality and performance of disturbance models, this study was designed to investigate the effects of model structure and data. The experiment considered and tested: (1) two measures of disturbance based on the Vegetation Cover Factor (C Factor) of the Revised Universal Soil Loss Equation (RUSLE) and Disturbance Intensity; (2) model structure using two modeling approaches; and (3) three subsets of data. The adjusted R-square and residuals from validation data are used to represent model quality and performance, respectively. Analysis of variance (ANOVA) is used to identify factors which have significant effects on model quality and performance. The results of the ANOVA show that subsets of data have significant effects on both model quality and performance for both measures of disturbance. The ANOVA also detected that the C Factor models have higher quality and performance than the Disturbance models. Although modeling approaches are not a significant factor based on the ANOVA tests, models containing interaction terms can increase the adjusted R-squares for nearly all tested conditions and the maximum improvement can reach 31%.

Cost-efficient sample designs for collection of ground data and accurate mapping of variables are required to monitor natural resources and environmental and ecological systems. In this study, a sample design and mapping method was developed by integrating stratification, model updating, and cokriging with Landsat Thematic Mapper (TM) imagery. This method is based on the spatial autocorrelation of variables and the spatial cross-correlation among them. It can lead to sample designs with variable grid spacing, where sampling distances between plots vary depending on spatial variability of the variables from location to location. This has potential cost-efficiencies in terms of sample design and mapping. This method is also applicable for mapping in the case in which no ground data can be collected in some parts of a study area because of the high cost. The method was validated in a case study in which a ground and vegetation cover factor was sampled and mapped for monitoring soil erosion. The results showed that when the sample obtained with three strata using the developed method was used for sampling and mapping the cover factor, the sampling cost was greatly decreased, although the error of the map was slightly increased compared to that without stratification; that is, the sample cost-efficiency quantified by the product of cost and error was greatly increased. The increase of cost-efficiency was more obvious when the cover factor values of the plots within the no-significant-change stratum were updated by a model developed using the previous observations instead of remeasuring them in the field.

A biocompatible, sustained-release subretinal drug-delivery platform was developed to overcome the therapeutic accessibility limitations of current retinal disease treatments. The prototype implants were fabricated by coating nitinol, poly(methyl methacrylate) or chromic gut core filaments, with a drug-eluting polymer matrix. The polymer coatings are manufactured and coated by SurModics. The coating is a mixture of poly(butyl methacrylate) and poly(ethylene-co-vinyl acetate). The drug is either triamcinolone acetonide or sirolimus. The rods were successfully implanted into the subretinal space of 20/24 rabbits. Four rabbits were lost to early surgery from a dysfunctional infusion line and hemorrhage. No serious complications were observed during the 4-week follow-up period. Slight conjunctival redness was reported in all rabbits by 1-day follow-up, but the redness had subsided by the following week. Intraocular lens touch occurred in six rabbits during the implantations; of these, four had a lensectomy at the time of surgery, and the remaining two developed cataract. Corneal edema developed in three rabbits by 1-week follow-up, but subsided within 2 weeks. Initial observations of the implantation and elution characteristics revealed that the implants are well tolerated by the retinal tissue and that the implant can elute triamcinolone acetonide for a period of at least 4 weeks without eliciting an inflammatory response or complications. There were adverse clinical indications with the sirolimus-loaded implants at the delivered dose. Device retrieval required an uncomplicated surgical procedure, and revealed no associated or adherent tissue. Implant drug content analysis and opacity changes to the polymer matrix coating following retrieval demonstrated the sustained elution of the drug.

An experimental and theoretical study of electroreduction of oxygen to hydrogen peroxide is presented. The experimental measurements of nitrided Ketjenblack indicated an onset potential for reduction of approximately 0.5 V (SHE) compared to the onset potential of 0.2 V observed for untreated carbon. Quantum calculations on cluster models of nitrided and un-nitrided graphite sheets show that carbon radical sites formed adjacent to substitutional N in graphite are active for O2 electroreduction to H2O2 via and adsorbed OOH intermediate. The weak catalytic effect of untreated carbon is attributed to weaker bonding of OOH to the H atom-terminated graphite edges. Substitutional N atoms that are far from graphite sheet edges will be active, and those that are close to the edges will be less active. Interference from electrochemical reduction of H atoms on the reactive sites is considered, and it is shown that in the potential range of H2O2 formation the reactive sites are not blocked by adsorbed H atoms.

The slab band quantum computational approach in the Vienna ab initio simulation package (VASP) is used to calculate the adsorption energies of reactants, reaction intermediates, and products in O2 reduction and in water oxidation in acid on three crystallographic surfaces of pentlandite structure Co9S8. Reversible potentials for the reaction steps involving electron and proton transfer are determined by using the energies in a linear Gibbs free energy relationship. On the basis of these results, we find that the partially OH-covered (202) surface is active toward O2 reduction and should have overpotential behavior similar to that observed for platinum electrodes. One structure in the predicted four-electron reduction mechanism is novel: S2- provides an adsorption site for O following O-O bond scission, which, unlike the case of platinum electrodes, takes place prior to the first reduction step.

Determining a remeasurement frequency of variables over time is required in monitoring environmental systems. This article demonstrates methods based on regression modeling and spatio-temporal variability to determine the time interval to remeasure the ground and vegetation cover factor on permanent plots for monitoring a soil erosion system. The spatio-temporal variability methods include use of historical data to predict semivariograms, modeling average temporal variability, and temporal interpolation by two-step kriging. The results show that for the cover factor, the relative errors of the prediction increase with an increased length of time interval between remeasurements when using the regression and semivariogram models. Given precision or accuracy requirements, appropriate time intervals can be determined. However, the remeasurement frequency also varies depending on the prediction interval time. As an alternative method, the range parameter of a semivariogram model can be used to quantify average temporal variability that approximates the maximum time interval between remeasurements. This method is simpler than regression and semivariogram modeling, but it requires a long-term dataset based on permanent plots. In addition, the temporal interpolation by two-step kriging is also used to determine the time interval. This method is applicable when remeasurements in time are not sufficient. If spatial and temporal remeasurements are sufficient, it can be expanded and applied to design spatial and temporal sampling simultaneously.

Two redox reactions on platinum electrodes in base, the formation of underpotential deposited hydrogen, forming a Pt-H bond, and the electro-oxidation of water, forming a Pt-OH bond, were studied by two methods. The first applies a linear relationship between reaction energy in solution and standard reversible potential, an approach recently used in this lab to predict the formation potential of the surface-bonded species. This method depends on the availability of accurate surface adsorption bond strengths from measurement or theory and can be applied in two formats, the empirical model and the linear correlation model. The second method treats the reaction within the so-called double-layer model where reactants and products on the surface are well defined and are experiencing the influence of the electrolyte. When this approach is used, two coordination shells of hydrogen bonded water molecules are found necessary to sufficiently stabilize the hydroxide ion in this model, unlike acid for which past work showed only one shell around the hydronium ion is needed. The calculated reversible potentials for both reactions by the empirical and linear correlation models are in good agreement with the experimental onset potentials observed in cyclic voltammetry measurements for Pt(111) surface electrodes when empirical or accurately calculated H, OH, and H(2)O adsorption energies are used. The double layer models for these reactions also yield satisfactory results, and it is concluded that the models should be useful for studying electron-transfer reactions in base, as has already been done for forming Pt-H and Pt-OH in acid solution.

A combined theoretical and experimental analysis of the electrode potential dependencies of activation energies is presented for the first step in oxygen reduction over platinum and platinum alloy catalysts in both polycrystalline and carbon supported form. Tafel data for several of the catalysts are used to predict potential-dependent activation energies for oxygen reduction over the 0.6-0.9 V range in strong and weak acid. Comparisons with the theoretical curve show good agreement above 0.8 V, suggesting a fairly constant preexponential factor. Arrhenius determinations of activation energies over the 0.7-0.9 V range yield little trend for weak acid, possibly because of the larger uncertainties in the Arrhenius fits, but the strong acid results have smaller uncertainties and for them the measured activation energies trend up with potential.

Employing combination coatings, whether multiple coatings on the same surface or different coatings on different parts of the same device offer medical devices a number of benefits. Examples of what can be achieved are reported here.

Escherichia coli has four gluconate transporters, GntP, GntU, GntT, and IdnT, which are members of the major facilitator superfamily. The physiological function of GntP was previously unknown and is the subject of this study. GntP is not induced by gluconate, and despite being located adjacent to genes involved in glucuronate catabolism, gntP does not encode a glucuronate transporter. Here we identify gntP as the gene which encodes the fructuronate transporter. We show that gntP is induced by fructuronate and is a new member of the UxuR regulon: gntP is derepressed in an uxuR strain, UxuR binds in vitro specifically to an operator site that overlaps the gntP promoter, and UxuR binding is eliminated by fructuronate. Transcription of gntP requires activation by cyclic AMP (cAMP)-cAMP receptor protein. A gntP mutant cannot grow on fructuronate but grows normally on glucuronate and gluconate. Thus, the UxuR regulon is a module of sugar acid catabolism whose physiological role is for growth on fructuronate. Glucuronate, because it proceeds through a fructuronate intermediate, must induce the UxuR regulon and must also induce the ExuR regulon, which encodes the glucuronate transporter, ExuT, and the first step in its catabolism, UxaC. Thus, hexuronate catabolism in E. coli requires both the ExuR and UxuR regulons, while fructuronate catabolism requires only the UxuR regulon.

Micellar electrokinetic capillary chromatography with laser-induced fluorescence detection was used to detect the differences in doxorubicin metabolite accumulation in four subcellular fractions isolated from the CCRF-CEM and the CEM/C2 human leukemia cell lines. Five fluorescent metabolites and doxorubicin make up the metabolite profile of these cell lines upon treatment with 10 microM doxorubicin for 12 h, cell lysis, and fractionation by differential centrifugation. Based on the relative electrophoretic mobility of synthetic standards, we tentatively identify one metabolite as 7-deoxydoxorubicinone and suggest that doxorubicinone is not among those metabolites detected. Although the obvious difference between the derived cell line (CEM/C2) and the parent cell line (CCRF-CEM) is the decreased topoisomerase I activity in the former, the results presented here indicate that each cell line has a unique distribution of metabolites in each one of four subcellular fractions: nuclear-enriched, heavy-organelle-enriched, light-organelle-enriched, and cytoplasmic fractions.

We report the doxorubicin content in individual organelles following their capillary electrophoretic separation and illustrate that chemical accumulation at the subcellular level is highly heterogeneous. In individual mitochondria from cultured human leukemia cells DOX amount is around 50 zmol, 2 orders of magnitude higher than expected from diffusion during drug treatment, and spans 2 orders of magnitude.

Whole-genome expression profiling revealed Escherichia coli MG1655 genes induced by growth on mucus, conditions designed to mimic nutrient availability in the mammalian intestine. Most were nutritional genes corresponding to catabolic pathways for nutrients found in mucus. We knocked out several pathways and tested the relative fitness of the mutants for colonization of the mouse intestine in competition with their wild-type parent. We found that only mutations in sugar pathways affected colonization, not phospholipid and amino acid catabolism, not gluconeogenesis, not the tricarboxylic acid cycle, and not the pentose phosphate pathway. Gluconate appeared to be a major carbon source used by E. coli MG1655 to colonize, having an impact on both the initiation and maintenance stages. N-acetylglucosamine and N-acetylneuraminic acid appeared to be involved in initiation, but not maintenance. Glucuronate, mannose, fucose, and ribose appeared to be involved in maintenance, but not initiation. The in vitro order of preference for these seven sugars paralleled the relative impact of the corresponding metabolic lesions on colonization: gluconate > N-acetylglucosamine > N-acetylneuraminic acid = glucuronate > mannose > fucose > ribose. The results of this systematic analysis of nutrients used by E. coli MG1655 to colonize the mouse intestine are intriguing in light of the nutrient-niche hypothesis, which states that the ecological niches within the intestine are defined by nutrient availability. Because humans are presumably colonized with different commensal strains, differences in nutrient availability may provide an open niche for infecting E. coli pathogens in some individuals and a barrier to infection in others.

Studies on the interactions of tissue plasminogen activator (tPA) and plasminogen with polyurethane surfaces containing epsilon-lysine moieties (epsilon-amino group free) are reported. These surfaces are considered to have the potential to dissolve nascent clots that may be formed on them. For adsorption from both single protein solutions and plasma, the surfaces were found to have a high capacity for tPA as well as plasminogen. A significant fraction of preadsorbed tPA was displaced from the epsilon-lysine surfaces upon contact with plasma. These surfaces, when preadsorbed with tPA and then incubated with plasma, were able to dissolve incipient clots formed around them. However, the clot-dissolving capacity diminished as the time of plasma incubation increased, presumably due to loss of tPA. It was also shown that in plasma, preadsorbed tPA is displaced from these surfaces largely by plasminogen, which thus appears to have a greater binding affinity than tPA for the epsilon-lysine moieties. Finally, it was found that in plasma, the epsilon-lysine surfaces interact with plasminogen in a dynamic manner, and that about 70% of the bound plasminogen is exchanging continuously with plasminogen in the plasma., (Copyright 2003 Wiley Periodicals, Inc.)

The targeted, controlled local delivery of a drug by an implant is the optimum method of treating a range of conditions. This review of product developments discusses potential future device applications and progress with novel techniques employing drug delivery matrices and coating methods.

Doxorubicin (DOX) treatment of NS-1 mouse hybridoma cells results in the formation of zeptomole amounts of metabolites per cell that are difficult to determine by confocal microscopy or HPLC. The native fluorescence of DOX and its metabolites together with laser-induced fluorescence detection (HF) has previously been used to detect a maximum of four components. In this study, we use capillary electrophoresis with postcolumn LIF (CE-LIF) to separate and detect 12 components attributed to DOX metabolism, resulting from treatment of NS-1 cells with 25 microM DOX for 8 h. The so-called metabolites 8 and 10 have been identified as doxorubicinone (DOXone) and 7-deoxydoxorubicinone (7-deoxyDOXone), respectively, by comigration with the corresponding synthetic standard. Due to comigration of DOX with doxorubicinol (DOXone), the presence of DOXone had to be determined separately by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The rest of the metabolites remain unidentified and are referred to by their number assignment. In comparison with the whole cell lysate, fractionation by differential centrifugation results in a better separation resolution of metabolites due to reduced amounts of metabolites in each fraction. This approach was chosen to compare the distribution of 13 metabolites in three subcellular fractions that form a pellet at < 1,400 g, 1,400-14,000 g, and > 14, 000 g and that generically are enriched in nuclei, organelles (mitochondria and lysosomes), and cytosolic components, respectively. The most abundant metabolite, DOXone, was estimated to be 90 +/- 15, 18 +/- 2, and 60 +/- 12 amol/cell (n = 5) in the nuclear-enriched, organelle-enriched, and cytosole-enriched fractions, respectively. In contrast, the total amount of other metabolites in a given fraction varied from 0 to 1,300 zmol. 7-DeoxyDOXone is the only metabolite that was present at similar levels in the three fractions. Other salient observations are metabolites 3, 7, and 11 are not detectable in the nuclear-enriched, organelle-enriched, and cytosole-enriched fractions, respectively; metabolite 9 and DOXone are more abundant in the nuclear-enriched fraction than in the other two fractions. The observations presented here suggest that subcellular fractionation followed by CE-LIF could be a powerful diagnostic for monitoring drug distribution, which is highly relevant to DOX cytoxicity studies.

Capillary electrophoresis with laser-induced fluorescence detection was used to separate and detect doxorubicin and at least five metabolites from NS-1 cells that were treated with 25 microM doxorubicin for 8 h. Using 10 mM borate, 10 mM sodium dodecyl sulfate (pH 9.3) as separation buffer, the 488-nm argon-ion laser line for fluorescence excitation, and a 635 +/- 27.5 nm bandpass filter for detection, the limit of detection (S/N=3) for doxorubicin is 61 +/- 13 zmol. This low limit of detection allows for the detection of a larger number of metabolites than previously reported. Two extraction procedures were performed: a bulk liquid-liquid extraction and an in-capillary single-cell lysis. While in the bulk liquid-liquid extraction procedure, recovery for doxorubicin range from 50 to 99%, in single cell analysis the recovery is expected to be complete. Furthermore performing lysis of a single cell inside the separation capillary prevents doxorubicin or metabolite loss or degradation during handling. Based on the bulk method the calculated metabolite abundance is in the sub-amol per cell range while it varies from 0.1 to 1.1 fmol per cell in single cell analysis confirming metabolite loss during handling. Each metabolite was found at a level less than 0.1% of the doxorubicin content in either method, suggesting a slow metabolism in the NS-1 cell system or effective removal of metabolites by the cell.

The Land Condition Trend Analysis (LCTA) program is the US Army's standard for land inventory and monitoring, employing standardized methods of natural resources data collection, analyses, and reporting designed to meet multiple goals and objectives. Critical to using LCTA data in natural resources management decisions is the ability of the LCTA protocols to detect changes in natural resources. To quantify the ability of LCTA protocols to detect resource changes, power analysis techniques were used to estimate minimum detectable effect sizes (MDES) for selected primary and secondary management variables for three Army installations. MDES for a subset of primary variables were estimated using data from 27 installation LCTA programs. MDES for primary and secondary variables varied widely. However, LCTA programs implemented at larger installations with lower sampling intensities detected changes in installation resources as well as programs implemented at smaller more intensively sampled installations. As a national monitoring program that is implemented at individual installations, LCTA protocols provide relatively consistent monitoring data to detect changes in resources despite diverse resource characteristics and implementation constraints.

Heparin-based coatings substantially improve the performance of a variety of blood-contacting medical devices. Recently, it has also been shown that nonheparin-based, synthetic coatings are promising for medical device applications where biological molecules are not desired or appropriate. This article reports on recent work.

Prentice and Sheppard (1995, Biometrika 82, 113-125) proposed a method for estimating relative risks associated with poorly measured exposures using disease rates from multiple populations and exposure and confounding factor data from sample surveys of persons in each population. The method involved an assumption of independence of disease rates across populations, conditional on exposures and confounding factors. Here, this assumption is relaxed by allowing dependencies among the disease rates within a partition of the populations, perhaps defined on the basis of geographic proximity or cultural similarity. Such dependencies could, for example, derive from unmeasured risk factors that are shared among neighboring populations. Dependencies within an element of the partition are modeled by allowing a common correlation between the disease rates of contiguous populations and by inducing correlations between the rates for noncontiguous populations using a multivariate lognormal assumption. Estimating equations are proposed for relative risk parameter estimation, and robustness and efficiency properties are assessed through simulations. The relaxed estimation procedure is shown to yield useful efficiency gains when moderate or strong disease rate dependencies are present, especially when disease rate variances are large relative to binomial variance.

Since 1971, most states have been subject to lawsuits seeking to reform their education funding systems. These cases are litigated on the basis of state (not federal) constitutional language and generally seek either greater equity in funding among school districts or a guaranteed level of adequate funding for education. State supreme courts have found the finance systems unconstitutional in 16 states, and many states are still actively involved in litigation. Even where litigation has not occurred or has not succeeded, the prospect of litigation has prompted revisions of state funding policies. Despite the predominant role equity and adequacy play in litigation, there are no universally accepted definitions for either of these words in education funding. Most commonly, equity is measured in terms of the variation in per-pupil revenues among school districts in a single state. By this measure, some states have greater funding equity than others, and in most states wealthy districts have significantly higher per-pupil expenditures than do poor districts. Equity is likely to be greater when the residents of poor districts pay higher taxes. (In some states, residents in poorer areas pay twice as much of their income in local taxes as do residents of wealthier communities.) Equity is also greater in those states where the state's share of the education budget is higher and where the state consistently targets its contributions to lower-income districts. Much of current litigation and legislative activity in education funding seeks to assure "adequacy," that is, a sufficient level of funding to deliver an adequate education to every student in the state. Most states have not explicitly addressed the questions of how much education is "adequate" or how educational standards can be converted to a finance formula. Several approaches to calculating the cost of an adequate education are described.