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Abstract Background As the field of implementation science continues to grow, its key concepts are being transferred into new contexts globally, such as Low and Middle Income Countries (LMICs), and its use is constantly being reexamined and expanded. Theoretical and methodological positions commonly used in implementation research and practice have great utility in our work but in many cases are at odds with LMIC contexts. As a team of implementation scientists based in Zambia, we offer this commentary as a critical self-reflection on what has worked and what could limit us from fully utilizing the field’s promise for addressing health problems with contextual understanding. Main body We used a ‘premortem,’ an approach used to generate potential alternatives from failed assumptions about a particular phenomenon, as a way to reflect on our experiences conducting implementation research and practice. By utilizing prospectively imagined hindsights, we were able to reflect on the past, present and possible future of the field in Zambia. Six key challenges identified were: (i) epistemic injustices; (ii) simplified conceptualizations of evidence-informed interventions; (iii) limited theorization of the complexity of low-resource contexts and it impacts on implementation; (iv) persistent lags in transforming research into practice; (v) limited focus on strategic dissemination of implementation science knowledge and (vi) existing training and capacity building initiatives’ failure to engage a broad range of actors including practitioners through diverse learning models. Conclusion Implementation science offers great promise in addressing many health problems in Zambia. Through this commentary, we hope to spur discussions on how implementation scientists can reimagine the future of the field by contemplating on lessons from our experiences in LMIC settings.

Aims: Limited data exist on outcomes in very elderly ICD recipients. We describe outcomes in new ICD and Cardiac Resynchronisation Therapy with Defibrillator (CRT-D) implants in octogenarians at our institution. Methods: Patients aged 80 years and above who underwent de novo ICD or CRT-D implantation from January 2006 to July 2012 were identified. Clinical data were collected from the procedural record, medical and ICD notes. Baseline characteristics were compared using independent sample t test for continuous variables and Fisher’s exact test for categorical variables. Kaplan-Meier curves were constructed. Results: Ten per cent of all new ICD/CRT-D implants were aged 80 years and over. Median age was 83.0 years. Median follow-up was 29 months. Death occurred in 17 (34%). Median time to death was 23 months. Three deaths (6%) occurred within 12 months of ICD implantation. Appropriate therapy (ATP or shock) occurred in 19 (38%). Inappropriate therapy occurred in 6 (12%). Rates of appropriate shocks and inappropriate therapy (shocks and ATP) and significant valvular incompetence were higher amongst deceased patients (P=0.03 OR 5.9 95% CI 1.3-27) and (P=0.02 OR 12 95% CI 1.3-112). Univariate analysis identified diuretic use (P=0.008 95% C.I. 0.05 to 0.63) and appropriate shock (P= 0.025 95% C.I. 1.25 to 26.3) as predictors of mortality. Conclusion: Octogenarians make up a small but increasing number of ICD recipients. This study highlights high survival rates at one year with acceptable rates of appropriate and inappropriate device therapy. Ongoing debate regarding the appropriateness of ICD in very elderly patients is warranted.

Background: Control of antimicrobial resistance (AMR) relies on local knowledge and local intervention implementation. Effective antibiotic stewardship requires locally-suitable prescribing guidelines. We aimed to use a novel digital tool (the ZARIApp) and a participatory approach to help develop locally-relevant empiric antibiotic prescribing guidelines for two hospitals in Lusaka, Zambia. Methods: We produced an AMR report using samples collected locally and routinely from adults within the prior two years (April 2020 – April 2022). We developed the ZARIApp, which provides prescribing recommendations based on local resistance data and antibiotic prescribing practices. We used qualitative evaluation of focus group discussions among healthcare professionals to assess the feasibility and acceptability of using the ZARIApp and identify the barriers to and enablers of this stewardship approach. Results: Resistance prevalence was high for many key pathogens: for example, 73% of 41 Escherichia coli isolates were resistant to ceftriaxone. We identified that high resistance rates were likely due to low levels of requesting and processing of microbiology samples from patients leading to insufficient and unrepresentative microbiology data. This emerged as the major barrier to generating locally-relevant guidelines. Through active stakeholder engagement, we modified the ZARIApp to better support users to generate empirical antibiotic guidelines within this context of unrepresentative microbiology data. Qualitative evaluation of focus group discussions suggested that the resulting ZARIApp was useful and easy to use. New antibiotic guidelines for key syndromes are now in place in the two study hospitals, but these have substantial residual uncertainty. Conclusions: Tools such as the free online ZARIApp can empower local settings to better understand and optimise how sampling and prescribing can help to improve patient care and reduce future AMR. However, the usability of the ZARIApp is severely limited by unrepresentative microbiology data; improved routine microbiology surveillance is vitally needed.

Clarifying multifactorial musculoskeletal disorder etiologies supports risk analysis, development of targeted prevention, and treatment modalities. Deep learning enables comprehensive risk factor identification through systematic analyses of disease data sets but does not provide sufficient context for mechanistic understanding, limiting clinical applicability for etiological investigations. Conversely, multiscale biomechanical modeling can evaluate mechanistic etiology within the relevant biomechanical and physiological context. We propose a hybrid approach combining 3D explainable deep learning and multiscale biomechanical modeling; we applied this approach to investigate temporomandibular joint (TMJ) disorder etiology by systematically identifying risk factors and elucidating mechanistic relationships between risk factors and TMJ biomechanics and mechanobiology. Our 3D convolutional neural network recognized TMJ disorder patients through participant-specific morphological features in condylar, ramus, and chin. Driven by deep learning model outputs, biomechanical modeling revealed that small mandibular size and flat condylar shape were associated with increased TMJ disorder risk through increased joint force, decreased tissue nutrient availability and cell ATP production, and increased TMJ disc strain energy density. Combining explainable deep learning and multiscale biomechanical modeling addresses the "mechanism unknown" limitation undermining translational confidence in clinical applications of deep learning and increases methodological accessibility for smaller clinical data sets by providing the crucial biomechanical context.

Background and Aim: Filarial infections affect over 150 million people in the tropics. One of the major forms of filarial pathologies is lymphedema; a condition where the immune response is significantly altered, resulting in changes in the normal flora. Staphylococcus hominis , a human skin commensal, can also be pathogenic in immunocompromised individuals. Therefore, there is the possibility that S. hominis could assume a different behavior in filarial lymphedema patients. To this end, we investigated the levels of antibiotic resistance and extent of mec A gene carriage in S. hominis among individuals presenting with filarial lymphedema in rural Ghana., Method: We recruited 160 individuals with stages I-VII lymphedema, in a cross-sectional study in the Ahanta West District of the Western Region of Ghana. Swabs from lymphedematous limb ulcers, pus, and cutaneous surfaces were cultured using standard culture-based techniques. The culture isolates were subjected to Matrix-Assisted Laser Desorption/Ionization Time of Flight (MALDI-TOF) mass spectrometry for bacterial identification. Antimicrobial susceptibility testing (AST) was performed using the Kirby-Bauer method. mec A genes were targeted by polymerase chain reaction for strains that were cefoxitin resistant., Results: In all, 112 S . hominis were isolated. The AST results showed resistance to chloramphenicol (87.5%), tetracycline (83.3%), penicillin (79.2%), and trimethoprim/sulphamethoxazole (45.8%). Of the 112 strains of S. hominis , 51 (45.5%) were resistant to cefoxitin, and 37 (72.5%) of the cefoxitin-resistant S. hominis haboured the mec A gene., Conclusion: This study indicates a heightened level of methicillin-resistant S. hominis isolated among filarial lymphedema patients. As a result, opportunistic infections of S. hominis among the already burdened filarial lymphedema patients in rural Ghana may have reduced treatment success with antibiotics., Competing Interests: The authors declare no conflicts of interest., (© 2023 The Authors. Health Science Reports published by Wiley Periodicals LLC.)

The human temporomandibular joint (TMJ) lateral capsule ligament (LCL) complex is debated as a fibrous capsule with distinct ligaments or ligamentous thickening, necessitating further evaluation of the complex and its role in TMJ anatomy and mechanics. This study explores the ultrastructural arrangement, biomechanical tensile properties, and biochemical composition of the human LCL complex including region-specific differences to explore the presence of a distinct temporomandibular ligament and sex-specific differences to inform evaluations of potential etiological mechanisms. LCL complex ultrastructural arrangement, biomechanical properties, and biochemical composition were determined using cadaveric samples. Statistical modeling assessed sex- and region-specific effects on LCL complex tissue properties. Collagen fiber coherency, collagen fiber bundle size, and elastin fiber count did not differ between sexes, but females trended higher in elastin fiber count. LCL complex water and sGAG content did not differ between sexes or regions, but collagen content was higher in the anterior region (311.0 ± 185.6 μg/mg) compared to the posterior region (221.0 ± 124.9 μg/mg) (p = 0.045) across sexes and in males (339.6 ± 170.6 μg/mg) compared to females (204.5 ± 130.7 μg/mg) (p = 0.006) across regions. Anterior failure stress (1.1 ± 0.7 MPa) was larger than posterior failure stress (0.6 ± 0.4 MPa) (p = 0.024). Regional differences confirm the presence of a mechanically and compositionally distinct temporomandibular ligament. Baseline sex-specific differences are critical for etiological investigations of sex disparities in TMJ disorders. These results have important biomechanical and clinical ramifications, providing critical baseline tissue material properties, informing the development of TMJ musculoskeletal models, and identifying new areas for etiologic investigations for temporomandibular disorders., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

White-nose syndrome (WNS) is a devastating disease of hibernating bats caused by the fungus Pseudogymnoascus destructans . We obtained 383 fungal and bacterial isolates from the Soudan Iron Mine, an important bat hibernaculum in Minnesota, then screened this library for antifungal activity to develop biological control treatments for WNS. An extract from the fungus Oidiodendron truncatum was subjected to bioassay-guided fractionation, which led to the isolation of 14 norditerpene and three anthraquinone metabolites. Ten of these compounds were previously described in the literature, and here we present the structures of seven new norditerpene analogues. Additionally, this is the first report of 4-chlorophyscion from a natural source, previously identified as a semisynthetic product. The compounds PR 1388 and LL-Z1271α were the only inhibitors of P. destructans (MIC = 7.5 and 15 μg/mL, respectively). Compounds were tested for cytotoxicity against fibroblast cell cultures obtained from Myotis septentrionalis (northern long eared bat) and M. grisescens (gray bat) using a standard MTT viability assay. The most active antifungal compound, PR 1388, was nontoxic toward cells from both bat species (IC 50 > 100 μM). We discuss the implications of these results in the context of the challenges and logistics of developing a substrate treatment or prophylactic for WNS.

Peritonsillar abscess (PTA) is a difficult diagnosis to make clinically, with clinical examination of even otolaryngologists showing poor sensitivity and specificity. Machine learning is a form of artificial intelligence that "learns" from data to make predictions. We developed a machine learning classifier to predict the diagnosis of PTA based on patient symptoms. We retrospectively collected clinical data and symptomatology from 916 patients who underwent attempted needle aspiration for PTA. Machine learning classifiers were trained on a subset of the data to predict the presence or absence of purulence on attempted aspiration. The performance of the model was evaluated on a holdout set. The accuracy of the top-performing algorithm, the artificial neural network, was 72.3%. Artificial neural networks can use patient symptoms to exceed human ability to predict PTA in patients with clinical suspicion for PTA. Similar models can assist medical decision making for clinicians who have suspicion of PTA.

White-nose syndrome (WNS) is a devastating fungal disease that has been causing the mass mortality of hibernating bats in North America since 2006 and is caused by the psychrophilic dermatophyte Pseudogymnoascus destructans. Infected bats shed conidia into hibernaculum sediments and surfaces, but it is unknown if P. destructans can form stable, reproductive populations outside its bat hosts. Previous studies have found non-pathogenic Pseudogymnoascus in bat hibernacula, and these fungi may provide insight into the natural history of P. destructans. We compared the relatedness, resource capture, and competitive ability of non-pathogenic Pseudogymnoascus isolates with P. destructans to determine if they have similar adaptations for survival in hibernacula sediment. All non-pathogenic Pseudogymnoascus isolates grew faster, utilized a broader range of substrates with higher efficiency, and were generally more resistant to antifungals compared to P. destructans. All isolates also showed the ability to displace P. destructans in co-culture assays, but only some produced extractible antifungal metabolites. These results suggest that P. destructans would perform poorly in the same environmental niche as non-pathogenic Pseudogymnoascus, and must have an alternative saprophytic survival strategy if it establishes active populations in hibernaculum sediment and non-host surfaces.

Honey bees, Apis mellifera, collect antimicrobial plant resins from the environment and deposit them in their nests as propolis. This behavior is of practical concern to beekeepers since the presence of propolis in the hive has a variety of benefits, including the suppression of disease symptoms. To connect the benefits that bees derive from propolis with particular resinous plants, we determined the identity and botanical origin of propolis compounds active against bee pathogens using bioassay-guided fractionation against the bacterium Paenibacillus larvae, the causative agent of American foulbrood. Eleven dihydroflavonols were isolated from propolis collected in Fallon, NV, including pinobanksin-3-octanoate. This hitherto unknown derivative and five other 3-acyl-dihydroflavonols showed inhibitory activity against both P. larvae (IC 50  = 17-68 μM) and Ascosphaera apis (IC 50  = 8-23 μM), the fungal agent of chalkbrood. A structure-activity relationship between acyl group size and antimicrobial activity was found, with longer acyl groups increasing activity against P. larvae and shorter acyl groups increasing activity against A. apis. Finally, it was determined that the isolated 3-acyl-dihydroflavonols originated from Populus fremontii, and further analysis showed these compounds can also be found in other North American Populus spp., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objective: To explore the feasibility of ecological momentary assessments (EMAs) as a tool to more accurately assess the level of bother from tinnitus., Study Design: Longitudinal observational study., Setting: Washington University Department of Otolaryngology-Head and Neck Surgery faculty practice plan., Subjects and Methods: Twenty participants with moderately to severely bothersome tinnitus were enrolled. All participants owned a smartphone device, and all communications were conducted via email, phone, and text messaging. Participants received 4 EMAs per day for 2 weeks via text message and a final survey on the 15th day. In each survey, participants recorded their level of tinnitus bother, their location at the time of response, their stress level, how they were feeling, and what they were doing. Response rates as a proxy for the feasibility of the program., Results: There were a total of 1120 surveys sent to 20 participants (56 surveys per participant), and 889 (79.4%) of the surveys were completed and returned. The median time to response from the moment of receiving the text message was 7 minutes. The distribution of responses to the EMA question, "In the last 5 minutes, how bothered have you been by your tinnitus?" displayed both high between- and within-subject variability. At the end of 2 weeks, the median score on the Tinnitus Handicap Inventory was 37, with a range of 10 to 82 points; the median Tinnitus Functional Index score was 43, with a range of 10 to 82 points., Conclusion: This study suggests bothered tinnitus patients will use smartphones as part of EMA., (© American Academy of Otolaryngology-Head and Neck Surgery Foundation 2015.)

Propolis is a substance derived from antimicrobial plant resins that honey bees use in the construction of their nests. Propolis use in the hive is an important component of honey bee social immunity and confers a number of positive physiological benefits to bees. The benefits that bees derive from resins are mostly due to their antimicrobial properties, but it is unknown how the diversity of antimicrobial activities among resins might impact bee health. In our previous work, we found that resins from different North American Populus spp. differed in their ability to inhibit in vitro growth of the bee bacterial pathogen Paenibacillus larvae. The goal of our current work was to characterize the antimicrobial activity of propolis from 12 climatically diverse regions across the US against the bee pathogens P. larvae and Ascosphaera apis and compare the metabolite profiles among those samples using LC-MS-based metabolomic methods. Samples differed greatly in their ability to inhibit both bacterial and fungal growth in vitro, but propolis from Nevada, Texas, and California displayed high activity against both pathogens. Interestingly, propolis from Georgia, New York, Louisiana, and Minnesota were active against A. apis, but not very active against P. larvae. Metabolomic analysis of regional propolis samples revealed that each sample was compositionally distinct, and LC-FTMS profiles from each sample contained a unique number of shared and exclusive peaks. Propolis from Aspen, CO, Tuscon, AZ, and Raleigh, NC, contained relatively large numbers of exclusive peaks, which may indicate that these samples originated from relatively unique botanical sources. This is the first study to characterize how the diversity of bee preferred resinous plants in the US may affect bee health, and could guide future studies on the therapeutic potential of propolis for bees., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Programmed-temperature gas chromatographic (GC) retention information is difficult to share because it depends on so many experimental factors that vary among laboratories. Though linear retention indexing cannot properly account for experimental differences, retention times can be accurately calculated, or "projected", from shared isothermal retention vs. temperature (T) relationships, but only if the temperature program and hold-up time vs. T profile produced by a GC is known with great precision. The effort required to measure these profiles were previously impractical, but we recently showed that they can be easily back-calculated from the programmed-temperature retention times of a set of 25 n-alkanes using open-source software at www.retentionprediction.org/gc. In a multi-lab study, the approach was shown to account for both intentional and unintentional differences in the temperature programs, flow rates, and inlet pressures produced by the GCs. Here, we tested 16 other experimental factors and found that only 5 could reduce accuracy in retention projections: injection history, exposure to very high levels of oxygen at high temperature, a very low transfer line temperature, an overloaded column, and a very short column (≤15m). We find that the retention projection methodology acts as a hybrid of conventional retention projection and retention indexing, drawing on the advantages of both; it properly accounts for a wide range of experimental conditions while accommodating the effects of experimental factors not properly taken into account in the calculations. Finally, we developed a four-step protocol to efficiently troubleshoot a GC system after it is found to be yielding inaccurate retention projections., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Dental school admissions committees continue to grapple with challenges associated with recruiting, admitting, and enrolling students who best meet the mission of the particular institution. For many students, particularly underrepresented minority students and those from non-traditional backgrounds, standardized tests such as the Dental Admission Test (DAT) are poor predictors of their potential for success. Noncognitive assessment methods can be used in conjunction with traditional cognitive measures in making admissions decisions. These methods have been employed by hundreds of higher education institutions, foundations, and scholarship programs, but are relatively new in dental education. This article describes a prevalent assessment model and discusses one dental school's implementation of the model, with the outcome of enrolling students from diverse backgrounds whose career goals parallel the mission of the school.

East Carolina University School of Dental Medicine is responding to the changes in today's health care system by implementing an innovative model of community-based dental education that prepares tomorrow's dentists to meet North Carolina's future oral health challenges while also providing much-needed care in many underserved areas.

Gas chromatography/mass spectrometry (GC/MS) is a primary tool used to identify compounds in complex samples. Both mass spectra and GC retention times are matched to those of standards; however, it is often impractical to have standards on hand for every compound of interest, so we must rely on shared databases of MS data and GC retention information. Unfortunately, retention databases (e.g., linear retention index libraries) are experimentally restrictive, notoriously unreliable, and strongly instrument dependent, relegating GC retention information to a minor, often negligible role in compound identification despite its potential power. A new methodology called "retention projection" has great potential to overcome the limitations of shared chromatographic databases. In this work, we tested the reliability of the methodology in five independent laboratories. We found that, even when each lab ran nominally the same method, the methodology was 3-fold more accurate than retention indexing because it properly accounted for unintentional differences between the GC/MS systems. When the laboratories used different methods of their own choosing, retention projections were 4- to 165-fold more accurate. More importantly, the distribution of error in the retention projections was predictable across different methods and laboratories, thus enabling automatic calculation of retention time tolerance windows. Tolerance windows at 99% confidence were generally narrower than those widely used even when physical standards are on hand to measure their retention. With its high accuracy and reliability, the new retention projection methodology makes GC retention a reliable, precise tool for compound identification, even when standards are not available to the user.

The deposition of antimicrobial plant resins in honey bee, Apis mellifera, nests has important physiological benefits. Resin foraging is difficult to approach experimentally because resin composition is highly variable among and between plant families, the environmental and plant-genotypic effects on resins are unknown, and resin foragers are relatively rare and often forage in unobservable tree canopies. Subsequently, little is known about the botanical origins of resins in many regions or the benefits of specific resins to bees. We used metabolomic methods as a type of environmental forensics to track individual resin forager behavior through comparisons of global resin metabolite patterns. The resin from the corbiculae of a single bee was sufficient to identify that resin's botanical source without prior knowledge of resin composition. Bees from our apiary discriminately foraged for resin from eastern cottonwood (Populus deltoides), and balsam poplar (P. balsamifera) among many available, even closely related, resinous plants. Cottonwood and balsam poplar resin composition did not show significant seasonal or regional changes in composition. Metabolomic analysis of resin from 6 North American Populus spp. and 5 hybrids revealed peaks characteristic to taxonomic nodes within Populus, while antimicrobial analysis revealed that resin from different species varied in inhibition of the bee bacterial pathogen, Paenibacillus larvae. We conclude that honey bees make discrete choices among many resinous plant species, even among closely related species. Bees also maintained fidelity to a single source during a foraging trip. Furthermore, the differential inhibition of P. larvae by Populus spp., thought to be preferential for resin collection in temperate regions, suggests that resins from closely related plant species many have different benefits to bees.

Introduction: We have previously identified a rare subpopulation of variant human mammary epithelial cells (vHMEC) with repressed p16INK4A that exist in disease-free women yet display premalignant properties, suggesting that they have engaged the process of malignant transformation. In order to gain insight into the molecular alterations required for vHMEC to progress to malignancy, and to characterize the epigenetic events associated with early progression, we examined the effect of oncogenic stress on the behavior of these cells., Methods: HMEC that express p16INK4A and vHMEC that do not, were transduced with constitutively active Ha-rasV12 and subsequently exposed to serum to determine whether signals from the cellular microenvironment could cooperate with ras to promote the malignant transformation of vHMEC. Epigenetic alterations were assessed using methylation-specific polymerase chain reaction (PCR)., Results: vHMEC expressing Ha-rasV12 (vHMEC-ras) bypassed the classic proliferative arrest that has been previously documented in normal fibroblasts following oncogenic stress, and that we also observe here in normal HMEC. Moreover, vHMEC-ras cells exhibited many additional alterations that are observed during progression to malignancy such as the generation of chromosomal abnormalities, upregulation of telomerase activity, immortalization following exposure to serum, and anchorage-independent growth, but they did not form tumors following orthotopic injection in vivo. Associated with their early progression to malignancy was an increase in the number of genes methylated, two of which (RASSF1A and SFRP1) were also methylated in other immortalized mammary cell lines as well as in breast cancer cells and tissues., Conclusions: We have characterized a mammary progression model that recapitulates molecular and methylation alterations observed in many breast cancers. Our data suggest that concomitant methylation of RASSF1A and SFRP1 marks an early event in mammary transformation and may thus have prognostic potential.

The active acquisition of epigenetic changes is a poorly understood but important process in development, differentiation, and disease. Our work has shown that repression of the p16/pRb pathway in human epithelial cells, a condition common to stem cells and many tumor cells, induces dynamic epigenetic remodeling resulting in the targeted methylation of a selected group of CpG islands. We hypothesized that cells in this epigenetically plastic state could be programmed by the microenvironment to acquire epigenetic changes associated with tumorigenesis. Here, we describe an in vitro model system where epigenetically plastic cells were placed in an environment that induced epithelial to mesenchymal transition (EMT) and led to a program of acquired de novo DNA methylation at targeted sites. In this model, we found that repression of E-cadherin transcription preceded the subsequent acquisition of methylated CpG sites. Furthermore, the induction of EMT was accompanied by de novo methylation of several other gene promoters, including those of the estrogen receptor and Twist. These data demonstrate that signals from the microenvironment can induce phenotypic and gene expression changes associated with targeted de novo epigenetic alterations important in tumor progression, and that these alterations occur through a deterministic, rather than stochastic, mechanism. Given the dynamic epigenetic reprogramming that occurs in these cells, DNA methylation profiles observed in human tumors may reflect the history of environmental exposures during the genesis of a tumor.

Bcr-Abl is the oncogenic protein-tyrosine kinase responsible for chronic myelogenous leukemia. Recently, we observed that inhibition of myeloid Src family kinase activity (e.g. Hck, Lyn, and Fyn) induces growth arrest and apoptosis in Bcr-Abl-transformed cells, suggesting that cell transformation by Bcr-Abl involves Src family kinases (Wilson, M. B., Schreiner, S. J., Choi, H. J., Kamens, J., and Smithgall, T. E. (2002) Oncogene 21, 8075-8088). Here, we report the unexpected observation that Hck, Lyn, and Fyn strongly phosphorylate the SH3-SH2 region of Bcr-Abl. Seven phosphorylation sites were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry: Tyr89 and Tyr134 in the Abl-derived SH3 domain; Tyr147 in the SH3-SH2 connector; and Tyr158, Tyr191, Tyr204, and Tyr234 in the SH2 domain. SH3 domain Tyr89, the most prominent phosphorylation site in vitro, was strongly phosphorylated in chronic myelogenous leukemia cells in a Src family kinase-dependent manner. Substitution of the SH3-SH2 tyrosine phosphorylation sites with phenylalanine substantially reduced Bcr-Abl-mediated transformation of TF-1 myeloid cells to cytokine independence. The positions of these tyrosines in the crystal structure of the c-Abl core and the transformation defect of the corresponding Bcr-Abl mutants together suggest that phosphorylation of the SH3-SH2 region by Src family kinases impacts Bcr-Abl protein conformation and signaling.

Alterations in DNA methylation are important in cancer, but the acquisition of these alterations is poorly understood. Using an unbiased global screen for CpG island methylation events, we have identified a non-random pattern of DNA hypermethylation acquired in p16-repressed cells. Interestingly, this pattern included loci located upstream of a number of homeobox genes. Upon removal of p16(INK4A) activity in primary human mammary epithelial cells, polycomb repressors, EZH2 and SUZ12, are up-regulated and recruited to HOXA9, a locus expressed during normal breast development and epigenetically silenced in breast cancer. We demonstrate that at this targeted locus, the up-regulation of polycomb repressors is accompanied by the recruitment of DNA methyltransferases and the hypermethylation of DNA, an endpoint, which we show to be dependent on SUZ12 expression. These results demonstrate a causal role of p16(INK4A) disruption in modulating DNA hypermethylation, and identify a dynamic and active process whereby epigenetic modulation of gene expression is activated as an early event in breast tumor progression.

This review finds that college GPA and DAT scores provide dental schools in the United States and Canada with defensible methods for selecting students. College GPA seems the best predictor of academic performance in dental school. The academic average (AA) of the DAT is a better predictor than is the perceptual ability test (PAT), but dental educators who believe that evidence of manual dexterity or perceptual ability must be a part of the admissions decision can find enough supporting evidence to justify doing so. When added to college GPA and the AA, information from the PAT may in fact enhance predictability. There is also evidence, however, that manual skills can be learned during routine dental curricular experiences. Overall, conventional admissions criteria at best account for about 40 percent of the variance in dental school performance, and most of this variance occurs during the early years of the curriculum. Studies are lacking for evaluating criteria that may predict success in admitting students for preferentially addressing current challenges, including achieving diversity of the workforce, ensuring access to care for all, interprofessional health care, ethics and professionalism, filling faculty positions, and conducting needed research. Schools should periodically validate all of their admissions criteria against expected performances and make corresponding adjustments.

Chronic myelogenous leukemia (CML) is defined by the presence of the Philadelphia (Ph) chromosome, which results in the expression of the 210 kDa Bcr-Abl tyrosine kinase. Bcr-Abl constitutively activates several signaling proteins important for the proliferation and survival of myeloid progenitors, including the Src family kinases Hck and Lyn, the Stat5 transcription factor and upstream components of the Ras/Erk pathway. Recently, we found that kinase-defective Hck blocks Bcr-Abl-induced transformation of DAGM myeloid leukemia cells to cytokine independence, suggesting that activation of the Src kinase family may be essential to oncogenic signaling by Bcr-Abl. To investigate the contribution of Src kinases to Bcr-Abl signaling in vivo, we used the pyrrolo-pyrimidine Src kinase inhibitors PP2 and A-419259. Treatment of the Ph+ CML cell lines K-562 and Meg-01 with either compound resulted in growth arrest and induction of apoptosis, while the Ph- leukemia cell lines TF-1 and HEL were unaffected over the same concentration ranges. Suppression of Ph+ cell growth by PP2 and A-419259 correlated with a decrease in Src kinase autophosphorylation. Both inhibitors blocked Stat5 and Erk activation, consistent with the suppressive effects of the compounds on survival and proliferation. In contrast, the phosphotyrosine content of Bcr-Abl and its endogenous substrate CrkL was unchanged at inhibitor concentrations that induced apoptosis, blocked oncogenic signaling and inhibited Src kinases. These data implicate the Src kinase family in Stat5 and Erk activation downstream of Bcr-Abl, and identify myeloid-specific Src kinases as potential drug targets in CML.

We have previously reported that the Jak2 tyrosine kinase but not Jak1 is tyrosine phosphorylated in the absence of IL-3 in Bcr-Abl positive M3.16 cells, which are rendered IL-3 independent by BCR-ABL gene expression. We have explored the involvement of Jak2 tyrosine phosphorylation in Bcr-Abl oncogenic effects. Our results indicate that Jak2 became tyrosine-phosphorylated in a number of cell lines expressing Bcr-Abl, when maintained in medium lacking IL-3, whereas Bcr-Abl negative cells lacked Jak2 tyrosine phosphorylation. Jak2 was poorly tyrosine-phosphorylated in cells expressing the SH2 deletion mutant of Bcr-Abl compared to either wild-type Bcr-Abl or its SH3 deletion mutant. Moreover, tyrosine phosphorylation of Jak2 by Bcr-Abl was inhibited by the Abl tyrosine kinase inhibitor, STI 571, in a dose-dependent manner. This inhibition of Bcr-Abl kinase by the drug did not interfere with the ability of Jak2 and Bcr-Abl to form a complex. Studies with deletion mutants of Bcr-Abl indicated that the C-terminal domain of Abl within Bcr-Abl was involved in complex formation with Jak2. Similarly, GST-Abl pull-down assays confirmed the strong binding to Jak2 by the C-terminus of Abl. Jak2 peptide substrate studies indicated that the Bcr-Abl and Abl tyrosine kinases specifically phosphorylated Y1007 of Jak2 but only poorly phosphorylated Y1008. Phosphorylation of Y1007 of Jak2 is known to be critical for its tyrosine kinase activation. Tyrosine residue 1007 of Jak2 was phosphorylated in 32Dp210 cells as measured by Western blotting with a phosphotyrosine 1007 sequence-specific antibody. A kinase-inactive Jak2 mutant blocked the colony forming ability of K562 cells. Tumor formation of K562 cells in nude mice was similarly inhibited by this kinase-inactive Jak2 mutant. This inhibition was independent of Stat5 tyrosine phosphorylation. Furthermore, tyrosine-phosphorylated Jak2 was detected in blood cells from CML patients in blast crisis but not in a normal marrow sample. In summary, these findings provide strong evidence that the Jak2 tyrosine kinase is a critical factor in Bcr-Abl malignant transformation.

Bcr-Abl is the constitutively active protein-tyrosine kinase expressed as a result of the Philadelphia translocation in chronic myelogenous leukemia. Bcr-Abl is coupled to many of the same signaling pathways normally regulated by hematopoietic cytokines. Recent work shows that Hck, a member of the Src tyrosine kinase family with myeloid-restricted expression, associates with and is activated by Bcr-Abl. Here we investigated the mechanism of Hck interaction with Bcr-Abl and the requirement for Hck activation in Bcr-Abl transformation signaling. Binding studies demonstrated that the Hck SH3 and SH2 domains are sufficient for interaction with Bcr-Abl in vitro. Hck binding localizes to the Abl SH2, SH3, and kinase domains as well as the distal portion of the C-terminal tail. To address the requirement for endogenous Src family kinase activation in Bcr-Abl signaling, a kinase-defective mutant of Hck was stably expressed in the cytokine-dependent myeloid leukemia cell line DAGM. Kinase-defective Hck dramatically suppressed Bcr-Abl-induced outgrowth of these cells in the absence of cytokine compared with a control cell line expressing beta-galactosidase. In contrast, kinase-defective Hck did not affect cell proliferation in response to interleukin-3, suggesting that the effect is specific for Bcr-Abl. These data show that Hck interacts with Bcr-Abl through a complex mechanism involving kinase-dependent and -independent components and that interaction with Hck or other Src family members is essential for transformation signaling by Bcr-Abl.

Hematopoiesis involves a complex array of growth factors that regulate the survival and proliferation of immature progenitors, influence differentiation commitment, and modulate end-stage cell functions. This mini-review is focused on the role of Stat activation in the development of myeloid cells in response to hematopoietic cytokines. Much of the evidence implicating Stats in these cellular processes comes from studies of mutant cytokine receptors selectively uncoupled from Stat activation, dominant-inhibitory Stat mutants, and mice with targeted disruptions of Stat genes. Together these approaches provide strong evidence that Stat activation, particularly of Stat3 and Stat5, plays an important role in myeloid differentiation and survival. Oncogene (2000).

BACKGROUND: Arterial tonometry is a noninvasive technique for monitoring the arterial blood pressure in a continuous manner. Real-time arterial blood pressure measurements are advantageous in many clinical settings, but arterial tonometric devices must be validated prior to clinical acceptance. Guidelines for accuracy for noninvasive blood pressure monitors have been established by the Association for the Advancement of Medical Instrumentation. OBJECTIVE: To test the Colin Pilot 9200 configured with an arterial tonometry module with 20 patients and to compare tonometric blood pressure measurements with intra-arterial blood pressure measurements. METHODS: All of the patients in the study were aged over 14 years and weighed over 35 kg; testing was performed in the operating room or in the intensive care unit. Data from each patient consisted of multiple simultaneous recordings of tonometric and intra-arterial blood pressure values. The data were then compared; the mean and SD of the difference between the two measurement devices were then calculated. RESULTS: Tonometric values were slightly less than the intra-arterial pressure measurements; the mean difference for systolic blood pressure was 2.24 +/- 8.7 mmHg and for diastolic pressure was 0.26 +/- 8.88 mmHg. CONCLUSION: The arterial tonometry module incorporated into the Colin Pilot 9200 was investigated for use with selected adult and pediatric populations. With our patients, it generated accurate data throughout a wide blood pressure range. It satisfied Association for the Advancement of Medical Instrumentation standards for mean systolic and diastolic blood pressure measurements and only minimally exceeded the allowable SD. This technology should prove to be a valuable tool for noninvasive blood pressure monitoring of various patient populations.

Thirty-six cases of hepatic toxicity associated with felbamate therapy have been collected by the Food and Drug Administration. Five patients died. We describe a case of massive acute hepatic necrosis and death within 40 days of initiation of felbamate therapy for a generalized tonic-clonic seizure disorder. We describe the clinical and histopathologic features.

Methods: Twelve adult patients with bilateral facial telangiectasias were treated with two yellow light lasers. A section of each involved area was treated with a copper vapor laser and a similar section in the same patient was treated with the flashlamp pumped dye laser., Results: Both lasers provided satisfactory clearance at 2 and 6 weeks. Treatment was not painless with either laser; however, no patient needed local or general anesthesia. The time taken to treat equivalent areas was similar for both lasers. Postoperative swelling was greater with the flashlamp pumped dye laser than with the copper vapor laser and the time required for healing was longer with the flash-lamp pumped dye laser., Conclusion: The larger purpuric macules produced post-operatively by the flashlamp pumped dye laser were less cosmetically acceptable to patients when compared with the thin linear crusting produced by the copper vapor laser. Scarring or textural changes were not seen with either laser.

Background: We compared the relative frequency of pathogens isolated from 1985 to 1989 (N = 4358) with those isolated from 1980 to 1984 (N = 5290) in a university hospital to determine trends in the relative importance of pathogens causing nosocomial infection., Methods: Our study was based on surveillance data prospectively obtained between 1980 and 1989 from a 600-bed university hospital. Statistically significant trends occurring from 1980 to 1984 to 1985 to 1989 were determined by chi 2 tests with Bonferroni corrections (i.e., p less than [0.05/17])., Results: Overall an increased frequency of isolation occurred for Candida and other yeasts and for Haemophilus species. A decreased frequency was noted for Proteus species, non-Bacteroides anaerobes, and Serratia species. Comparison of 1985 to 1989 with 1980 to 1984 revealed that the most significant change in nosocomial pathogens was the marked increase in infections with yeast, principally Candida species. Candida and other yeast infections increased 40%, from 7.6% (rank, 5) to 10.6% (rank, 3) of all pathogens isolated. Increases, which occurred in urine, blood, and wound isolates, were especially marked among surgical patients. In addition, a significant increase was noted among blood isolates in the isolation of yeast other than Candida albicans., Conclusions: We conclude that Candida and other yeasts are being isolated increasingly as causative agents of nosocomial infection.

Solid and papillary epithelial neoplasm of the pancreas (SPENP) is a rare lesion characteristically occurring in young women. By contrast with pancreatic ductal adenocarcinomas, SPENP is a slow-growing tumor that rarely metastasizes or is fatal. The current report describes light and electron microscopic and histochemical findings with DNA flow cytometric analyses of two cases of SPENP. The first patient was a 24-year-old woman; the second, a 72-year-old man. Although SPENP is rare in older men, both patients had characteristic radiographic and light microscopic features of SPENP. Ultrastructural evidence of acinar differentiation was seen in the first patient; the second patient had focal neuroendocrine differentiation. Flow cytometric analysis of the first tumor demonstrated diploid-range DNA content with a 5.8% S-phase fraction (SPF). The DNA cytometric analysis of a biopsy specimen from the second tumor revealed diploid-range DNA content with a 6.1% SPF, although subsequent sampling of the resected tumor showed an aneuploid population with a DNA index of 1.8 and SPF of 2.1%.

As part of a prospective cohort study initiated in 1983, the human immunodeficiency virus type 1 (HIV-1) status has been periodically determined for patients with clotting disorders (hemophilia A or B, von Willebrand's disease, miscellaneous). The University of North Carolina Hospitals has conducted comprehensive surveillance for nosocomial infections (NI) using modified Centers for Disease Control criteria since 1980 and entered this information in a computerized data base. Cross-matching of our NI data base and hemophiliac/HIV-1 study data base for the time period 1980-1989 revealed that 13 NI occurred in 11 patients during 659 hospitalizations (5,723 hospital days). NI rates per 100 admissions (per 1,000 hospital days) by HIV-1 status were as follows: HIV-1 negative = 0.91 (1.18), HIV-1 positive pre-AIDS = 1.65 (1.84), and AIDS = 6.67 (6.48). NI occurred with a similar frequency in HIV-1 positive pre-AIDS hemophiliacs and HIV-1 negative hemophiliacs (Fisher's exact test, p greater than 0.10). However, NI occurred more frequently in hemophiliacs with AIDS versus HIV-1 positive or negative hemophiliacs (Fisher's exact test, p less than 0.05). We conclude that HIV-1 infection does not appreciably alter the risk of developing a NI, but that patients who have progressed to AIDS are at significantly increased risk of developing a NI per hospital day or per hospitalization.

A group of 14 intensive care unit (ICU) patients were studied to determine if manual ventilation bags (MVB) could serve as a source of bacterial or fungal pathogens that could colonize the respiratory tract of intubated patients. A total of 51 cultures were simultaneously obtained of patient's sputum, the exterior MVB surface, MVB port, and MVB interior (postexhalation valve). Pathogens colonizing or infecting the respiratory tract of intubated ICU patients were frequently simultaneously present on the exterior surface of the MVB and inside the MVB port used to connect the MVB with the endotracheal tube. In addition, coagulase-negative staphylococci and yeast were frequently present on the exterior surface of the MVB. The interior of the MVB was usually sterile. In three instances pathogens were isolated from the MVB before isolation from the patient's sputum. MVB may serve as a source for colonizing the respiratory tract of intubated ICU patients and/or the hands of medical personnel. The exterior surface and port of MVB should be cleaned of visible debris and disinfected at least once a day.

Two cases of congenital or infantile fibrosarcoma are described that were incompletely excised at the time of primary excision and have not recurred or metastasized after 3 years. The tumors were composed of densely cellular spindle cells with a high mitotic index. Immunohistochemical stains were positive for vimentin but negative for desmin and S-100. The tumor cells were grown in vitro, and a karyotype was obtained. Both tumors had normal diploid modal karyotypes. In addition, fragments of the primary tumor from both cases were injected subcutaneously into nude mice; neither tumor could be heterotransplanted. The clinical course and biologic features of these two tumors suggest that congenital or infantile sarcoma does not have the properties of a malignant neoplasm, and thus the designation of these tumors as a sarcoma may be a misnomer.

Plaster of Paris and nonsterile cast padding have previously been implicated in wound infections. The observation of three pin site infections with Bacillus cereus following pin placement and plaster fixation of an open forearm fracture led to an epidemiologic investigation. Two hundred sixty-one bacterial cultures were taken from materials in the Steinmann pin application and plaster fixation procedures. B. cereus with a similar antibiogram and biotype as was identified in the pin site cultures was recovered from three of the 22 (14%) plaster-impregnated gauze rolls and six of seven (81%) tapwater samples. The Bacillus sp contamination rate of plaster when dry or wetted with sterile water or tapwater was 58%, 25% and 40% respectively. All cultures of plaster samples were negative after steam or gas sterilization. These results suggest that the patient may have acquired the infection from the plaster-impregnated gauze wetted in tapwater. In this case, the Steinmann pins may have facilitated infection by guiding the plaster-associated bacteria to the pin insertion site. The use of gas sterilized plaster materials should be considered in certain circumstances.

Proteins were covalently coupled in good yield to periodate-oxidized Sephadex. The reaction was studied in detail with respect to Sephadex type, sodium m-periodate concentration, protein (albumin or immunoglobulin) concentration, time course, temperature dependence, and pH optimum. The complexes were stable and were used successfully for the immunoadsorption of sheep anti-rabbit immunoglobulin antibodies.

An increase in the recovery of Aspergillus flavus from respiratory specimens occurred among hospitalized patients during a several-month period in 1977. Notably, 86% of the patients with positive culture results were located in an older hospital section adjacent to a building construction site that created great amounts of dust in the hospital's vicinity. Airborne contamination was suspected and results of air sampling showed A. flavus in 80% of old wing patient rooms (average, 8 A. flavus/positive room) compared with only 23% of the contiguous newer wing patient rooms (average, 1 A. flavus/positive room). Inspection of the main mechanical ventilation units in the two hospital sections disclosed numerous defects in the unit in the old wing and a properly functioning unit in the new wing. After repairing the defective unit, a significant reduction in the recovery of A. flavus from clinical and environmental specimens was noted.

Subjects in two different clinical trials who had been advised to stop smoking were asked if they had done so. Some 22% of subjects (11 out of 51) in the first trial and 40% (33/82) in the second trial who said they had stopped smoking were found to have raised carboxyhaemoglobin concentrations. Deception appears to be common in people trying to stop smoking.

There are important reasons for considering nitrite inhalation as a factor in the development of AIDS-related KS in young male homosexuals. These are (1) the pharmacologic properties of amyl, butyl, and isobutyl nitrites, which are toxic; (2) the mutagenic, teratogenic, and carcinogenic products resulting from metabolism of N-nitroso compounds; (3) the potent carcinogenicity of N-nitroso compounds in 39 different animal species; and (4) the deleterious effects of volatile nitrites on human lymphocytes both in vitro and in vivo. Specifically related to this epidemic, there are additional reasons for pursuing the connection between nitrite inhalation and development of KS. These include: (1) the timing of the production and sales of volatile nitrites for use as recreational drugs and the subsequent outbreak of the AIDS epidemic (7 to 10 years); (2) the extensive use of nitrites among male homosexuals; (3) the virtual universal history of nitrite use by young male homosexuals in whom KS has developed during the past 3 years; and (4) the age group in which KS is developing is consistent with a cohort initially exposed 7 to 10 years ago.

The aim of this study was to define the post-traumatic changes in body fluid compartments and to evaluate the effect of plasma colloid osmotic pressure (COP) on the partitioning of body fluid between these compartments. Forty-two measurements of plasma volume (green dye), extracellular volume (bromine), and total body water (deuterium) were done in ten traumatized patients (mean Injury Severity Score, ISS, = 34) and 23 similar control studies were done in eight healthy volunteers who were in stable fluid balance. Interstitial volume, intracellular volume, and blood volume were calculated from measured fluid spaces and hematocrit; COP was directly measured. Studies in volunteers on consecutive days indicated good reproducibility, with coefficients of variation equal to 3.5% for COP, 6.3% for plasma volume, 4.5% for extracellular volume, and 4.9% for total body water. COP values extended over the entire range seen clinically, from 10 to 30 mmHg. Interstitial volume was increased by 55% in patients, but intracellular volume was decreased by 10%. We conclude (1) that posttraumatic peripheral edema resulting from hemodilution is located in the interstitial compartment, with no intracellular space expansion; and (2) that interstitial volume, but not intracellular volume, is closely related to plasma COP.