Search

We are experiencing a revolution in cancer. Advances in screening, targeted and immune therapies, big data, computational methodologies, and significant new knowledge of cancer biology are transforming the ways in which we prevent, detect, diagnose, treat, and survive cancer. These advances are enabling durable progress in the goal to achieve personalized cancer care. Despite these gains, more work is needed to develop better tools and strategies to limit cancer as a major health concern. One persistent gap is the inconsistent coordination among researchers and caregivers to implement evidence-based programs that rely on a fuller understanding of the molecular, cellular, and systems biology mechanisms underpinning different types of cancer. Here, the authors integrate conversations with over 90 leading cancer experts to highlight current challenges, encourage a robust and diverse national research portfolio, and capture timely opportunities to advance evidence-based approaches for all patients with cancer and for all communities.

IntroductionPersonal digital devices that provide health information, such as the Apple Watch, have developed an increasing array of cardiopulmonary tracking features which have received regulatory clearance and are directly marketed to consumers. Despite their widespread and increasing use, data about the impact of personal digital device use on patient-reported outcomes and healthcare utilization are sparse. Among a population of patients with atrial fibrillation undergoing cardioversion, our primary aim is to determine the impact of the heart rate measurement, irregular rhythm notification, and electrocardiogram features of the Apple Watch on quality of life and healthcare utilization.Methods and analysisWe are conducting a prospective, open-label multicenter pragmatic randomized clinical trial, leveraging a unique patient-centered health data sharing platform for enrollment and follow-up. A total of 150 patients undergoing cardioversion for atrial fibrillation (or atrial flutter, if they have a history of atrial fibrillation) will be randomized 1:1 to receive the Apple Watch Series 6 or Withings Move at the time of cardioversion. The primary outcome is the difference in the Atrial Fibrillation Effect on QualiTy-of-life (AFEQT) global score at six months post-cardioversion. Secondary outcomes include inpatient and outpatient healthcare utilization. Additional secondary outcomes include a comparison of the Apple Watch ECG and pulse oximeter features with gold standard data obtained in routine clinical care settings.Ethics and disseminationThe Institutional Review Boards at Yale University, Mayo Clinic, and Duke University Health System have approved the trial protocol. This trial will provide important data to policymakers, clinicians, and patients about the impact of the heart rate, irregular rhythm notification, and electrocardiogram features of widely used personal digital devices on patient quality of life and healthcare utilization. Findings will be disseminated to study participants, at professional society meetings, and in peer-reviewed journals.Trial registration numberNCT04468321Strengths and limitations of this studyThis randomized clinical trial will use a patient-centered health data sharing platform for study enrollment and follow-up, leveraging patient ownership over health data and patient engagement with researchers.This is the first study to examine the impact of electrocardiogram and irregular rhythm notification features of personal digital technologies on patient-reported quality-of-life among patients with atrial fibrillation.Due to costs of personal digital devices, our study is powered to identify a difference of 8.8 on the primary outcome of Atrial Fibrillation Effect on QualiTy-of-life (AFEQT) global score, which is greater than the minimal clinically important difference of 5.Our study focuses on a subset of patients with atrial fibrillation, and the findings may not be generalizable to those who do not have atrial fibrillation.

Background More than 600 000 coronary stents are implanted during percutaneous coronary interventions ( PCI s) annually in the United States. Because no real‐world surveillance system exists to monitor their long‐term safety, claims data are often used for this purpose. The extent to which adverse events identified with claims data can be reasonably attributed to a specific medical device is uncertain. Methods and Results We used deterministic matching to link the NCDR (National Cardiovascular Data Registry) Cath PCI Registry to Medicare fee‐for‐service claims for patients aged ≥65 years who underwent PCI with drug‐eluting stents ( DESs ) between July 1, 2009 and December 31, 2013. We identified subsequent PCI s within 1 year of the index procedure in Medicare claims as potential safety events. We linked these subsequent PCI s back to the NCDR Cath PCI Registry to ascertain how often the revascularization could be reasonably attributed to the same coronary artery as the index PCI (ie, target vessel revascularization). Of 415 306 DES placements in 368 194 patients, 33 174 repeat PCI s were identified in Medicare claims within 1 year. Of these, 28 632 (86.3%) could be linked back to the NCDR Cath PCI Registry; 16 942 (51.1% of repeat PCI s) were target vessel revascularizations. Of these, 8544 (50.4%) were within a previously placed DES : 7652 for in‐stent restenosis and 1341 for stent thrombosis. Of 16 176 patients with a claim for acute myocardial infarction in the follow‐up period, 4446 (27.5%) were attributed to the same coronary artery in which the DES was implanted during the index PCI (ie, target vessel myocardial infarction). Of 24 288 patients whose death was identified in claims data, 278 (1.1%) were attributed to the same coronary artery in which the DES was implanted during the index PCI . Conclusions Most repeat PCI s following DES stent implantation identified in longitudinal claims data could be linked to real‐world registry data, but only half could be reasonably attributed to the same coronary artery as the index procedure. Attribution among those with acute myocardial infarction or who died was even less frequent. Safety signals identified using claims data alone will require more in‐depth examination to accurately assess stent safety.

INTRODUCTION Embedded care managers are increasingly implemented as part of the care team within primary care practices, yet previous studies have indicated variability in acceptance by physicians and staff. This study assesses the acceptability of care managers among staff and physicians within the Michigan Primary Care Transformation (MiPCT) demonstration. METHODS Care manager acceptance was measured using a web-based survey distributed to practices participating in the MiPCT demonstration. RESULTS Both physicians and staff reported high levels of care manager acceptance. Longer length of care manager employment at the practice, higher care manager FTE dedicated to care management, and care manager employed by practice were all significantly associated with care manager acceptance. DISCUSSION The MiPCT demonstration found high care manager acceptance across all care team members. The high level of acceptance may be due to the structures and processes developed by MiPCT to support implementation of care managers and the length of the intervention period. CONCLUSION The MiPCT demonstration confirms that following three years of implementation, embedded care managers are acceptable to both physicians and staff within primary care practices. Importantly, embeddedness, or the amount of time care managers are located within practices, is associated with increased acceptance.

OBJECTIVE To examine the effect of acceleration and deceleration distance (0, 1, 2 and 3 m) on the comfortable and maximum walking speeds in: (i) the 5-m walk test (5mWT); and (ii) the 10-m walk test (10mWT) in people with chronic stroke. DESIGN Cross-sectional study. SETTING University-based rehabilitation centre. SUBJECTS Thirty individuals with chronic stroke. METHODS Timed walking at comfortable and maximum walking speeds in the 5mWT and 10mWT with different acceleration and deceleration distances (0, 1, 2 and 3 m). RESULTS The comfortable walking speed in the 5mWT with 0 m acceleration and deceleration distance was significantly slower than that with 1, 2 or 3 m acceleration and deceleration distances (p

Bacterial antimicrobial resistance represents an important current and future problem in infectious disease public health. When reviewing longitudinal studies (research studies involving repeated observations of a location over a period of time), very little information was found regarding the proliferation and dissemination of foodborne bacteria in a new dairy farm environment. Iowa State University (ISU; Ames IA) was awarded 887 acres of land by a donor family. An academic teaching farm was later constructed, The Dairy/Animal Science Education Facility (or the ISU Dairy Farm). Prior to building the ISU Dairy Farm, the land was only used for recreational activities-no production livestock had ever been introduced. It was decided to study the bacteria microbiota Enterobacteriaceae (particularly Salmonella), Enterococcus and Campylobacter. Microbiological sampling was conducted using a variety of techniques-from phenotypic methods (e.g. detecting bacteria by using specialized media) to genotypic methods (e.g. Pulsed Field Gel Electrophoresis). Sampling took place prior to dairy cattle placement (20 October 2007 and 13 November 2007) and after placement (27 January 2008, 29 February 2008, 12 March 2008, 23 April 2008 and 28 May 2008). A literature review discussing the introduction of dairy cattle to the United States, antibiotics used in animal production, antibiotic resistance, and the “prudent” (or responsible) use of antibiotics is discussed in chapter 1. Data from the ISU Dairy Farm Studies are included in chapter 2 (the establishment and proliferation of antibiotic-resistant Campylobacter), chapter 3 (the establishment and proliferation of Enterococcus spp.) and chapter 4 (the establishment of Enterobacteriaceae and Salmonella london). Chapter 5 addresses general conclusions of this work and recommendations for future research.

Joseph S Ross,1–4 Jonathan Bates,4 Craig S Parzynski,4 Joseph G Akar,4,5 Jeptha P Curtis,4,5 Nihar R Desai,4,5 James V Freeman,4,5 Ginger M Gamble,4 Richard Kuntz,6 Shu-Xia Li,4 Danica Marinac-Dabic,7 Frederick A Masoudi,8 Sharon-Lise T Normand,9,10 Isuru Ranasinghe,11 Richard E Shaw,12 Harlan M Krumholz2–5 1Section of General Medicine, Department of Medicine, 2Robert Wood Johnson Foundation Clinical Scholars Program, Yale School of Medicine, 3Department of Health Policy and Management, Yale School of Public Health, 4Center for Outcomes Research and Evaluation, Yale–New Haven Hospital, 5Section of Cardiovascular Medicine, Department of Medicine, Yale School of Medicine, New Haven, CT, 6Medtronic Inc, Minneapolis, MN, 7Division of Epidemiology, Center for Devices and Radiological Health, US Food and Drug Administration, Silver Spring, MD, 8Division of Cardiology, Department of Medicine, University of Colorado, Aurora, CO, 9Department of Health Care Policy, Harvard Medical School, 10Department of Biostatistics, Harvard TH Chan School of Public Health, Boston, MA, USA; 11Discipline of Medicine, University of Adelaide, Adelaide, SA, Australia; 12Department of Clinical Informatics, California Pacific Medical Center, San Francisco, CA, USA Background: Machine learning methods may complement traditional analytic methods for medical device surveillance.Methods and results: Using data from the National Cardiovascular Data Registry for implantable cardioverter–defibrillators (ICDs) linked to Medicare administrative claims for longitudinal follow-up, we applied three statistical approaches to safety-signal detection for commonly used dual-chamber ICDs that used two propensity score (PS) models: one specified by subject-matter experts (PS-SME), and the other one by machine learning-based selection (PS-ML). The first approach used PS-SME and cumulative incidence (time-to-event), the second approach used PS-SME and cumulative risk (Data Extraction and Longitudinal Trend Analysis [DELTA]), and the third approach used PS-ML and cumulative risk (embedded feature selection). Safety-signal surveillance was conducted for eleven dual-chamber ICD models implanted at least 2,000 times over 3 years. Between 2006 and 2010, there were 71,948 Medicare fee-for-service beneficiaries who received dual-chamber ICDs. Cumulative device-specific unadjusted 3-year event rates varied for three surveyed safety signals: death from any cause, 12.8%–20.9%; nonfatal ICD-related adverse events, 19.3%–26.3%; and death from any cause or nonfatal ICD-related adverse event, 27.1%–37.6%. Agreement among safety signals detected/not detected between the time-to-event and DELTA approaches was 90.9% (360 of 396, k=0.068), between the time-to-event and embedded feature-selection approaches was 91.7% (363 of 396, k=–0.028), and between the DELTA and embedded feature selection approaches was 88.1% (349 of 396, k=–0.042).Conclusion: Three statistical approaches, including one machine learning method, identified important safety signals, but without exact agreement. Ensemble methods may be needed to detect all safety signals for further evaluation during medical device surveillance. Keywords: implanted cardioverter–defibrillator, methodology, surveillance

HIV-1's Rev protein forms a homo-oligomeric adaptor complex linking viral RNAs to the cellular CRM1/Ran-GTP nuclear export machinery through the activity of Rev's prototypical leucine-rich nuclear export signal (NES). In this study, we used a functional fluorescently tagged Rev fusion protein as a platform to study the effects of modulating Rev NES identity, number, position, or strength on Rev subcellular trafficking, viral RNA nuclear export, and infectious virion production. We found that Rev activity was remarkably tolerant of diverse NES sequences, including supraphysiological NES (SNES) peptides that otherwise arrest CRM1 transport complexes at nuclear pores. Rev's ability to tolerate a SNES was both position and multimerization dependent, an observation consistent with a model wherein Rev self-association acts to transiently mask the NES peptide(s), thereby biasing Rev's trafficking into the nucleus. Combined imaging and functional assays also indicated that NES masking underpins Rev's well-known tendency to accumulate at the nucleolus, as well as Rev's capacity to activate optimal levels of late viral gene expression. We propose that Rev multimerization and NES masking regulates Rev's trafficking to and retention within the nucleus even prior to RNA binding. IMPORTANCE HIV-1 infects more than 34 million people worldwide causing >1 million deaths per year. Infectious virion production is activated by the essential viral Rev protein that mediates nuclear export of intron-bearing late-stage viral mRNAs. Rev's shuttling into and out of the nucleus is regulated by the antagonistic activities of both a peptide-encoded N-terminal nuclear localization signal and C-terminal nuclear export signal (NES). How Rev and related viral proteins balance strong import and export activities in order to achieve optimal levels of viral gene expression is incompletely understood. We provide evidence that multimerization provides a mechanism by which Rev transiently masks its NES peptide, thereby biasing its trafficking to and retention within the nucleus. Targeted pharmacological disruption of Rev-Rev interactions should perturb multiple Rev activities, both Rev-RNA binding and Rev's trafficking to the nucleus in the first place.

This article includes current recommendations for screening breast ultrasonography, a review of the literature, a discussion of problems associated with the implementation of screening breast ultrasonography, and a brief discussion of the political and economic factors influencing the use of screening breast ultrasonography.

Purpose. To investigate fundamental mechanisms of regimes of laser induced damage to the retina and the morphological changes associated with the damage response.Methods. Varying grades of photothermal, photochemical, and photomechanical retinal laser damage were produced in eyes of eight cynomolgus monkeys. An adaptive optics confocal scanning laser ophthalmoscope and spectral domain optical coherence tomographer were combined to simultaneously collect complementaryin vivoimages of retinal laser damage during and following exposure. Baseline color fundus photography was performed to complement high-resolution imaging. Monkeys were perfused with 10% buffered formalin and eyes were enucleated for histological analysis.Results. Laser energies for visible retinal damage in this study were consistent with previously reported damage thresholds. Lesions were identified in OCT images that were not visible in direct ophthalmoscopic examination or fundus photos. Unique diagnostic characteristics, specific to each damage regime, were identified and associated with shape and localization of lesions to specific retinal layers. Previously undocumented retinal healing response to blue continuous wave laser exposure was recorded through a novel experimental methodology.Conclusion. This study revealed increased sensitivity of lesion detection and improved specificity to the laser of origin utilizing high-resolution imaging when compared to traditional ophthalmic imaging techniques in the retina.

Retroviruses encode cis-acting RNA nuclear export elements that override nuclear retention of intron-containing viral mRNAs including the full-length, unspliced genomic RNAs (gRNAs) packaged into assembling virions. The HIV-1 Rev-response element (RRE) recruits the cellular nuclear export receptor CRM1 (also known as exportin-1/XPO1) using the viral protein Rev, while simple retroviruses encode constitutive transport elements (CTEs) that directly recruit components of the NXF1(Tap)/NXT1(p15) mRNA nuclear export machinery. How gRNA nuclear export is linked to trafficking machineries in the cytoplasm upstream of virus particle assembly is unknown. Here we used long-term (>24 h), multicolor live cell imaging to directly visualize HIV-1 gRNA nuclear export, translation, cytoplasmic trafficking, and virus particle production in single cells. We show that the HIV-1 RRE regulates unique, en masse, Rev- and CRM1-dependent “burst-like” transitions of mRNAs from the nucleus to flood the cytoplasm in a non-localized fashion. By contrast, the CTE derived from Mason-Pfizer monkey virus (M-PMV) links gRNAs to microtubules in the cytoplasm, driving them to cluster markedly to the centrosome that forms the pericentriolar core of the microtubule-organizing center (MTOC). Adding each export element to selected heterologous mRNAs was sufficient to confer each distinct export behavior, as was directing Rev/CRM1 or NXF1/NXT1 transport modules to mRNAs using a site-specific RNA tethering strategy. Moreover, multiple CTEs per transcript enhanced MTOC targeting, suggesting that a cooperative mechanism links NXF1/NXT1 to microtubules. Combined, these results reveal striking, unexpected features of retroviral gRNA nucleocytoplasmic transport and demonstrate roles for mRNA export elements that extend beyond nuclear pores to impact gRNA distribution in the cytoplasm., Author Summary How cellular and viral mRNAs are trafficked inside the cell is poorly understood. In this study, we use fluorescence microscopy to visualize HIV-1 mRNA nuclear export, translation, trafficking, and virus particle assembly in single living cells, focusing on the full-length viral genomic RNAs (gRNAs) that are packaged into virus particles. We demonstrate that HIV-1 gRNAs exhibit dramatic “burst”-like transitions from the nucleus to fill the cytoplasm prior to the onset of virus particle assembly. This process is regulated by the viral Rev protein and the cellular nuclear export factor CRM1. By contrast, Mason-Pfizer monkey virus (M-PMV), a pathogenic non-human retrovirus that does not exploit CRM1, links gRNAs to the microtubule cytoskeleton in a mechanism regulated by an alternative cellular nuclear export factor, NXF1. Combined, our comparative studies reveal two unanticipated mRNA nucleocytoplasmic transport behaviors programmed by contrasting mRNA nuclear export factors. Further elucidation of the molecular details underpinning these unique trafficking behaviors may inform the development of antiviral strategies targeting viral gene expression and/or infectious virion assembly.

Enterococci are Gram-positive, facultative anaerobic cocci. They are found in many environments (including milk and dairy products, vegetables, plants, cereals, and meats). Enterococci are considered commensal organisms, but can also be opportunistic pathogens associated with morbidity and mortality of humans and animals. A longitudinal study of antibiotic resistance of Enterococcus to ampicillin, erythromycin, and tetracycline was conducted on an academic teaching farm. Environmental samples were collected by drag swabs at select locations prior to and after the introduction of livestock. All samples were initially processed and screened with specialized media, and then replica plated on tryptic soy agar containing a predetermined amount of antibiotic. There was some variation in the quantity of bacterial and antibiotic-resistant colonies; however, resistance to tetracycline was extremely high. The increases of too numerous to count populations were not time-dependent and appeared consistently after the placement of cows. There is little information on the prevalence and epidemiology of antibiotic resistance of Enterococci outside of the hospital setting, including on dairy farms. Longitudinal studies are important in providing insight into the dynamics of establishment and proliferation of bacteria and of antibiotic resistance.

Several sources of data suggest that the Yellow-billed Magpie (Pica nuttalli), a corvid endemic to California, is extremely susceptible to West Nile virus (WNV) and that its abundance has decreased since the establishment of WNV throughout California in 2004. From 2004 to 2006, 12,211 Yellow-billed Magpie carcasses were reported to the California Department of Health Services. Seventy-eight percent of the 1,007 Yellow-billed Magpie carcasses tested were WNV-positive, and this was the highest proportion of WNV-positive carcasses of all California bird species with reasonable sample sizes (>20). Assuming a starting population size of 180,000 in 2003, California Department of Health Services data suggest that Yellow-billed Magpie populations may have been reduced by 49% in just two years. Breeding Bird Survey (BBS) data show a 22% decline as of 2005, and Christmas Bird Count (CBC) data show a 42% decline as of 2006. Furthermore, flock size at three traditional urban communal roosts monitored in Sacr...

Dynamic NMR and molecular modeling techniques were employed to investigate the influence of structural characteristics on the ring inversion processes of inositol phosphates. The results indicate that inositol phosphates that contain the syn-1,3,5-triaxial trisphosphate arrangement show great proclivity to adopt the sterically hindered conformation. These results suggest that the ability to form chelation cages with counter-ions is a major factor in the ring inversion processes of inositol phosphates.

Recently Hunt, Holloway, & Scordalakes (1999) described a novel procedure for examining how social interactions with an intoxicated sibling can enhance periadolescent rats' voluntary intake of ethanol. In the present series of experiments we extend these findings to preweanlings. In Experiment 1, same-sex sibling 16-day-olds were assigned to be either (a) a demonstrator that was administered 1.5 g/kg ethanol or water control or (b) an observer that was tested for ethanol intake following a brief interaction with the demonstrator. Observers interacting with EtOH demonstrators exhibited increased intake of ethanol relative to observers interacting with water demonstrators. In Experiment 2, subjects were 8, 12, or 16 days of age and at all ages, ethanol intakes increased following exposure to an intoxicated sibling. In Experiment 3, repeated exposures to ethanol demonstrators on days 12, 14, and 16 was found to promote ethanol intake after weaning (on postnatal day 22). Collectively these data indicate that exposure to ethanol cues in the context of home/social cues can lead to modifications in ethanol acceptance, and that repeated exposures to such cues during infancy can impact ethanol ingestion after weaning.

The effects of treadmill exercise (20 m/min, five days/wk, one hr/day) on growth and tissue ascorbic acid (AA) concentrations were investigated in 40 young, male guinea pigs. Animals were fed ad libitium an AA-free diet which was supplemented with AA. AA was administered daily via gavage feeding. Animals were divided into two groups, exercise (E) and sedentary (S). Food consumption and body weight were recorded. After eight weeks, animals were sacrificed and blood taken via heart puncture. Liver, spleen, adrenals and kidneys were removed, weighed and frozen. Plasma and organ tissues were analyzed for AA. Results indicated that exercise had no significant effect on growth. Growth was within normal range. Exercised animals had significantly (p

The spatial and temporal activation of NF-kB (p65) was monitored in the retina of a transgenic mouse model (cis-NFkB-EGFP) in vivo after receiving varying grades of laser induced thermal injury in one eye. Baseline images of the retinas from 26 mice were collected prior to injury and up to five months post-exposure using a Heidelberg Spectralis HRA confocal scanning laser ophthalmoscope (cSLO) with a spectral domain optical coherence tomographer (SDOCT). Injured and control eyes were enucleated at discrete time points following laser exposure for cryosectioning to determine localization of NF-kB dependent enhanced green fluorescent protein (EGFP) reporter gene expression within the retina using fluorescence microscopy. In addition, EGFP basal expression in brain and retinal tissue from the cis-NFkB-EGFP was characterized using two-photon imaging. Regions of the retina exposed to threshold and supra-threshold laser damage evaluated using fluorescence cSLO showed increased EGFP fluorescence localized to the exposed region for a duration that was dependent upon the degree of injury. Fluorescence microscopy of threshold damage revealed EGFP localized to the outer nuclear region and retinal pigment epithelial layer. Basal expression of EGFP imaged using two-photon microscopy was heterogeneously distributed throughout brain tissue and confined to the inner retina. Results show cis-NF-kB-EGFP reporter mouse can be used for in vivo studies of light induced injury to the retina and possibly brain injury.

The first therapies to reliably eradicate Helicobacter pylori used combinations of bismuth, metronidazole, and tetracycline or amoxicillin. The increasing frequency of metronidazole resistance, however, has limited the effectiveness of the original combination therapies. As with the use of bismuth compounds, proton pump inhibitors can be used to kill H pylori but are themselves insufficient to eradicate the infection. Proton pump inhibitors markedly increase gastric pH. Because this action may make antimicrobials more effective, we evaluated omeprazole as an adjuvant to antimicrobial therapy. We tested omeprazole 20 mg TID for 7 days as an adjuvant to oral clarithromycin 500 mg TID or gentamicin 20 mg TID (each was given for the final 5 days of the 7-day treatment course). Eradication was defined as the absence of evidence of H pylori 4 or more weeks after cessation of therapy. Eleven patients with active peptic ulcer disease were treated with omeprazole/clarithromycin; the eradication rate was 64%. Six healthy H pylori -infected volunteers were treated with omeprazole/gentamicin; no eradication was achieved. Omeprazole appears to be a useful adjuvant to some therapies but the results cannot be predicted. Combination therapy with omeprazole and clarithromycin warrants further study.

Balb/c wild type mice were used to perform in vivo experiments of laser-induced thermal damage to the retina. A Heidelberg Spectralis HRA confocal scanning laser ophthalmoscope with a spectral domain optical coherence tomographer was used to obtain fundus and cross-sectional images of laser induced injury in the retina. Sub-threshold, threshold, and supra-threshold lesions were observed using optical coherence tomography (OCT), infrared reflectance, red-free reflectance, fluorescence angiography, and autofluorescence imaging modalities at different time points post-exposure. Lesions observed using all imaging modalities, except autofluorescence, were not visible immediately after exposure but did resolve within an hour and grew in size over a 24 hour period. There was a decrease in fundus autofluorescence at exposure sites immediately following exposure that developed into hyper-fluorescence 24-48 hours later. OCT images revealed threshold damage that was localized to the RPE but extended into the neural retina over a 24 hour period. Volumetric representations of the mouse retina were created to visualize the extent of damage within the retina over a 24 hour period. Multimodal imaging provides complementary information regarding damage mechanisms that may be used to quantify the extent of the damage as well as the effectiveness of treatments without need for histology.

We evaluated whether therapy designed to eradicate Helicobacter pylori infection resulted in a reduction in rebleeding in patients with peptic ulcer disease. Patients presenting because of major upper gastrointestinal hemorrhage from peptic ulcer and whose ulcers healed in a study in which they were randomized to receive ranitidine alone or triple therapy plus ranitidine were followed up regularly with endoscopy. No maintenance anti-ulcer therapy was given after ulcer healing.Patients received ranitidine, 300 mg, or ranitidine plus triple therapy. Triple therapy consisted of tetracycline, 2 g; metronidazole, 750 mg; and bismuth subsalicylate, 5 or 8 tablets (151 mg bismuth per tablet), and was administered for the first 2 weeks of treatment; ranitidine therapy was continued until the ulcer had healed or 16 weeks had elapsed. After ulcer healing, no maintenance antiulcer therapy was given. Development of ulcer recurrence with or without recurrent upper gastrointestinal bleeding was evaluated.Thirty-one patients with major upper gastrointestinal bleeding from peptic ulcer were studied; 17 received triple therapy and 14 ranitidine alone. Major rebleeding occurred significantly (p = 0.031) more often in those in the ranitidine group (28.6%), compared with none (0%) in the triple therapy group.Eradication of H. pylori infection reduces the rate of ulcer recurrence and rebleeding in complicated ulcer disease.

Salmonella spp. are important zoonotic pathogens in humans and animals. A longitudinal study was conducted at the Iowa State University's campus (at the Dairy/Animal Science Education and Discovery Facility) to observe change in Enterobacteriaceae (specifically Salmonella) before and after the placement of dairy livestock. To our knowledge, this is the first study that evaluated environmental changes of Gram-negative organisms in a new dairy farm environment. Environmental samples were taken using drag swabs and immediately processed in the laboratory using phenotypic methods (replica plating, the BBL Crystal Identification System for enteric/nonfermenter organisms™, and plating on specialized media/broths). Genotypic methods were also used (BAX PCR™ and pulsed-field gel electrophoresis). Organisms identified as Salmonella were sent to the National Veterinary Services Laboratory (Ames, IA) for confirmatory serotyping. Resistance to antibiotics (ampicillin, nalidixic acid, and tetracycline) was determined by replica plating of Enterobacteriaceae and Salmonella isolates using the guidelines of the National Antimicrobial Resistance Monitoring System and Clinical and Laboratory Standards Institute. The microflora of Enterobacteriaceae changed as cattle were introduced and as time progressed. Additionally, multidrug-resistant isolates began to appear immediately after cattle were introduced (multidrug-resistant isolates were rare prior to introduction of livestock). Variables such as temperature and humidity did not affect the proliferation of bacterial organisms. Seventeen Salmonella isolates were identified as Salmonella London and three isolates as Salmonella Montevideo. Based on pulsed-field gel electrophoresis-generated dendrograms, it is likely that 17 Salmonella London isolates and 3 Salmonella Montevideo isolates are clonal.

A study of retinal damage thresholds in non-human primates (NHP) in the near-infrared (NIR) wavelengths of 1110, 1130, 1150, and 1319 nm has recently been reported. The progression of damage in retinal areas that received exposures below, greater than, and at threshold values for each respective wavelength are compared. Subjects were imaged using an Adaptive Optics (AO) enhanced Spectral Domain Optical Coherence Tomographer (SD-OCT) a year post laser exposure to examine damage characteristics and localization. The subject's retinas within the study exhibited a delayed response to NIR exposures in that many of the lesions that were not visible at the 1-hour observation period continued to grow in size over the 24-hour period and or became visible. Thermal lensing is believed to play a significant role in the formation or retinal lesions in the NIR and may explain the delayed response.

Near threshold retinal lesions were created in the eyes of non-human primate (NHP) subjects in the near infrared (NIR) wavelength range of 1100 to 1319 nm, with 80 to 100 ms laser exposures. Two new in vivo imagining techniques, Adaptive Optic enhanced-Spectral Domain Optical Coherence Tomography (AO-SDOCT) and Adaptive Optic enhanced confocal Scanning Laser Ophthalmoscope imagery (AOcSLO) we re utilized to pinpoint areas of chronic damage within the retinal layers resulting from laser e xposure. Advantages and limitations of each technology with regard to the study of laser retinal tissue interaction are highlighted. Keywords: Adaptive Optics, MVL, NIR, laser threshold, in vivo imaging, confocal, scanning laser ophthalmoscope, SLO, Spectral Domain, OCT 1. INTRODUCTION High-fidelity imaging of lesions in the NHP retina is vital for determining damage thresholds for laser exposure. Retinal damage thresholds are traditionally determined from direct funduscopic observation with an ophthalmoscopic camera. The ability to discern a change from healthy to denatured retinal tissue in vivo is limited primarily by the optical transfer function (OTF) of the eye, with observable lesions of 25 um in extent (or larger) typically reported as being a minimally visible lesion (MVL).[1] However, damage widths which correspond to the extent of a single cell at the RPE and photoreceptor layers in the retina for ultrashort laser exposures have been reported with histological confirmation by Toth.[2] Recent observations of changes in autofluorescence in the retina also suggest that unresolved damage as a result of subthreshold exposure levels can exist without detection via traditional imaging methods in threshold studies. [3-5] These observations and the lack of data reported for similarly sized lesions resulting from photothermal exposures accentuate the possibility that damage which is not directly observable through standard funduscopic observation is present at exposure levels which are “subthreshold.” Given the 25-um size purported to be the smallest repeatedly observable lesion in thermal studies, the observation of small lesions in ultrashort laser studies and the subthreshold detection of changes in autofluorescence of RPE cells it is apparent that a method of observation which is both more sensitive (improved spatial resolution) and specific (able to isolate damaged tissue within the layers of the retina) is needed. Therefore, imaging techniques which overcome the li mitations of the OTF of the eye and which enable imaging at depth in turbid media are highly desired for use in retinal laser threshold studies. Two adaptive-optic-enabled retinal imaging systems were delivered to Brooks City-Base as the product of Air Force Research Laboratory sponsored Small Business Initiative for Re search (SBIR) technology de velopment contracts. The first system developed by PSI (Andover, MA) is a spectral domain optical coherence tomography (AO-SDOCT) system. Details of this system are published elsewhere.[6]. The second system, developed by IRIS Electro-Optics (Berkeley, CA), is a confocal scanning ophthalmoscope (AOcSLO). [7] A schematic for each system is shown in Figure 1. This paper details the initial use of these imaging systems to observe laser damage in the NHP retina which were produced by exposure to NIR laser wavelengths.

Recent studies have shown that the exaggerated meal-stimulated gastrin release in patients with duodenal ulcer abates after eradication of Helicobacter pylori infection. Bombesin-stimulated gastrin release was compared in 11 H. pylori -infected patients with chronic duodenal ulcer and 8 uninfected healthy volunteers both before and after therapy to eradicate H. pylori . Bombesin infusion significantly increased the gastrin release both in control subjects and in patients with duodenal ulcer. Antimicrobial therapy (bismuth, tetracycline, and metronidazole) to eradicate the H. pylori infection was associated with a significant reduction in bombesin-stimulated gastrin release in patients with duodenal ulcer (from 116.9 ± 19 pg/mL to 69.5 ± 7 pg/mL following 50 pmol · kg −1 · h −1 bombesin; and from 158 ± 29 to 83.4 ± 10 following 200 pmol · kg −1 · h −1 bombesin: P = 0.01 for each). Antimicrobial therapy had no effect on gastrin release in uninfected volunteers, thus excluding a nonspecific effect of antimicrobial therapy on antral G-cell function. Serum gastrin was also not increased by feeding 500 mg of urea to 5 H. pylori -infected volunteers. This suggests that access of hydrogen ion to the pH-sensitive sites governing gastrin release by mucosal ammonia produced by H. pylori urease is not a critical factor. These data suggest that exaggerated gastrin release present in patients with duodenal ulcer disease is secondary to H. pylori infection.

The reflectance and absorption of the skin plays a vital role in determining how much radiation will be absorbed by human tissue. Any substance covering the skin would change the way radiation is reflected and absorbed and thus the extent of thermal injury. Hairless guinea pigs ( cavia porcellus ) in vivo were used to evaluate how the minimum visible lesion threshold for single-pulse laser exposure is changed with a topical agent applied to the skin. The ED 50 for visible lesions due to an Er: glass laser at 1540-nm with a pulse width of 50-ns was determined, and the results were compared with model predictions using a skin thermal model. The ED50 is compared with the damage threshold of skin coated with a highly absorbing topical cream at 1540 nm to determine its effect on damage pathology and threshold. The ED 50 for the guinea pig was then compared to similar studies using Yucatan minipigs and Yorkshire pigs at 1540-nm and nanosecond pulse duration. 1,2 The damage threshold at 24-hours of a Yorkshire pig for a 2.5-3.5-mm diameter beam for 100 ns was 3.2 Jcm -2 ; very similar to our ED 50 of 3.00 Jcm -2 for the hairless guinea pigs.

Helicobacter pylori infection is associated with exaggerated gastrin release. We investigated whether this abnormality was due to the bacteria or the immune response. Fasting and meal-stimulated 'total' and amidated gastrin were measured in 10 H. pylori-infected volunteers before eradication therapy, after 2 and 14 days of therapy, and 4 weeks after completion of therapy. The exaggerated meal-stimulated gastrin concentration remained unchanged after 2 days of therapy, although the polymorphonuclear cell infiltrate and H. pylori bacteria were no longer evident. The expected fall in gastrin concentration after 14 days of therapy was associated with a reduction in the density of mucosal mononuclear cells, suggesting exaggerated gastrin release was related to chronic inflammation or to H. pylori or its products. The effect of H. pylori on normal progastrin processing was also assessed; 2 control groups were included: 10 H. pylori-uninfected volunteers and 13 patients with H. pylori peptic ulcers. There was a significant difference in the proportion of circulating gastrins that were biologically active amidated gastrins between ulcer patients and uninfected controls (56.7 +/- 4% versus 33.8 +/- 4%, p0.001). The proportion of amidated to total gastrins did not increase after successful eradication.

It has recently been recognized that Helicobacter pylori infection is associated with abnormalities in the regulation of gastrin secretion. We investigated whether there was a relationship between H. pylori infection and G-cell and D-cell numbers.The numbers of antral G cells and D cells were compared between 20 patients with duodenal ulcer and 24 volunteers, 12 with and 12 without H. pylori infection. The effect of eradication of H. pylori infection on G-cell number was also evaluated. Antral mucosal biopsy specimens were examined using immunohistochemical techniques specific for the presence of gastrin and somatostatin.The number of G cells was significantly (P0.02) less in patients with duodenal ulcer than in either infected or uninfected controls (3.7 +/- 0.3 vs. 6.2 +/- 0.6 and 5.3 +/- 0.5 G cells per gland for infected and uninfected controls, respectively). The ratio of G-cells to D-cells was similar in duodenal ulcer patients (2.2) and uninfected controls (2.0). It was found that, although eradication of the H. pylori infection results in a dramatic reduction in stimulated gastrin secretion, it is not associated with a change in the numbers of antral G cells or D cells in patients with duodenal ulcer.It is concluded that H. pylori infection-associated increase in gastrin secretion appear to be related to local factors regulating G-cell function.

Triple therapies using bismuth, metronidazole and tetracycline or amoxicillin were the first truly successful anti-H. pylori therapies. Metronidazole resistance has become an increasing problem that has severely limited the usefulness of the original triple therapy. Resistance to tetracycline or amoxicillin has not been reported and both are effective against H. pylori. We therefore tested a new triple therapy consisting of 500 mg tetracycline, 500 mg amoxicillin, and 2 tablets of bismuth subsalicylate each administered four times daily (with meals and at bedtime) for 14 days during treatment with ranitidine 300 mg daily. H. pylori eradication was defined as no evidence of H. pylori one or more months after stopping therapy. H. pylori status was evaluated by a combination of urea breath test and histology. Sixteen patients with H. pylori infection and active peptic ulcers were enrolled. The new triple therapy was successful in only 7 individuals (43%). Metronidazole appears to be critical for the effectiveness of the original triple therapy. An alternative to metronidazole will be required for a new successful triple therapy.

Eradication of Helicobacter pylori infections has proven to be difficult [2–4, 6–8, 13]. Therapy with a single or with two antimicrobials has proven ineffective as evidenced by eradication rates much below 50% [3]. Recently, combinations of three antimicrobial drugs have been shown to eradicate more than 50% of H. pylori infections. Despite these apparent successes, the results have been variable and the factors influencing effectiveness of therapy remain unclear [3, 7, 8]. In this study we evaluated the effectiveness of a combination of three antimicrobial drugs (triple therapy) for eradication of H. pylori infection with special emphasis on the factors that might predict good versus poor responses.

OBJECTIVE To determine the effect of treating Helicobacter pylori infection on the recurrence of gastric and duodenal ulcer disease. DESIGN Follow-up of up to 2 years in patients with healed ulcers who had participated in randomized, controlled trials. SETTING A Veterans Affairs hospital. PARTICIPANTS A total of 109 patients infected with H. pylori who had a recently healed duodenal (83 patients) or gastric ulcer (26 patients) as confirmed by endoscopy. INTERVENTION Patients received ranitidine, 300 mg, or ranitidine plus triple therapy. Triple therapy consisted of tetracycline, 2 g; metronidazole, 750 mg; and bismuth subsalicylate, 5 or 8 tablets (151 mg bismuth per tablet) and was administered for the first 2 weeks of treatment; ranitidine therapy was continued until the ulcer had healed or 16 weeks had elapsed. After ulcer healing, no maintenance antiulcer therapy was given. MEASUREMENTS Endoscopy to assess ulcer recurrence was done at 3-month intervals or when a patient developed symptoms, for a maximum of 2 years. RESULTS The probability of recurrence for patients who received triple therapy plus ranitidine was significantly lower than that for patients who received ranitidine alone: for patients with duodenal ulcer, 12% (95% CI, 1% to 24%) compared with 95% (CI, 84% to 100%); for patients with gastric ulcer, 13% (CI, 4% to 31%) compared with 74% (44% to 100%). Fifty percent of patients who received ranitidine alone for healing of duodenal or gastric ulcer had a relapse within 12 weeks of healing. Ulcer recurrence in the triple therapy group was related to the failure to eradicate H. pylori and to the use of nonsteroidal anti-inflammatory drugs. CONCLUSIONS Eradication of H. pylori infection markedly changes the natural history of peptic ulcer in patients with duodenal or gastric ulcer. Most peptic ulcers associated with H. pylori infection are curable.

Helicobacter pylori infection has been associated with gastritis, duodenal ulcer, gastric ulcer, and the epidemic form of gastric carcinoma. Eradication of H. pylori infection has proven to be difficult. Recently, combinations of antimicrobial drugs have been shown to eradicate greater than 50% of infections; however, the results have proven variable, and the factors influencing effectiveness of therapy are unclear. In the present study, the effectiveness of a triple therapy for eradication of H. pylori infection was evaluated. Triple therapy consisted of 2 g tetracycline, 750 mg metronidazole, and five or eight tablets of bismuth subsalicylate daily in 93 patients (70 with duodenal ulcer, 17 with gastric ulcer, and 6 with simple H. pylori gastritis). Combinations of a sensitive urea breath test, serology, culture, and histology were used to confirm the presence of infection, eradication, or relapse. Eradication was defined as inability to show H. pylori greater than or equal to 1 month after ending therapy. The overall eradication rate was 87%. The factors evaluated for their effect on predicting eradication included age, gender, type of disease, duration of therapy, amount of bismuth subsalicylate [five or eight Pepto-Bismol tablets daily (Procter & Gamble, Cincinnati, OH)], and compliance with the prescribed medications. Stepwise regression showed that compliance was the most important factor predicting success; the success rate was 96% for patients who took greater than 60% of the prescribed medications and 69% for patients who took less. For those taking greater than 60% of the prescribed therapy, the eradication rates were similar (a) for patients receiving therapy for 14 days or when tetracycline and bismuth subsalicylate were taken for an additional 14 days; (b) for patients with duodenal ulcer, gastric ulcer, and simple H. pylori gastritis; and (c) whether five or eight bismuth subsalicylate tablets were taken. It is concluded that triple therapy is effective for eradication of H. pylori and that future studies need to take compliance into account for comparisons between regimens.

Glaucoma is a progressive optic nerve disease characterized by a loss of retinal ganglion cell (RGC) axons and thinning of the retinal nerve fiber layer (RNFL). Axon loss can be as much as 50% before the disease is clinically detectable.1 Imaging devices have been developed to quantitatively measure RNFL thinning and birefringence loss. For example, optical coherence tomography (OCT) produces cross-sectional images of the retina allowing measurement of RNFL thickness.2 Polarization-sensitive OCT (PS-OCT)3 and scanning laser polarimetry (GDx)4 measure phase retardation in the RNFL which arises from anisotropic structures in the RNFL.5 Changes in RNFL birefringence may precede RGC death. Birefringence (Δn) is a dimensionless measure of the anisotropy of the refractive index (n) of a material and is given by the difference of the extraordinary index (ne) and ordinary index (no) where no and ne are the refractive indices for polarization perpendicular and parallel to the axis of anisotropy respectively (Δn = ne - no). Phase retardation is proportional to birefringence according to the expression Δn = λ0PR/360°ΔZ where λ0 is the free-space wavelength, PR is the phase retardation, and ΔZ is the thickness of the material.6 In general, Δn of the RNFL is weakly wavelength dependent across visible and near infrared wavelengths.7 The RNFL demonstrates form birefringence that originates from anisotropic cylindrically shaped cellular structures in the RGC axons.8,9 The major cylindrical structures of the RGC axonal cytoskeleton are neurotubules (NT), neurofilaments, and neurotubule-associated proteins.10 Huang et al.5 identify neurotubules as the source of birefringence from the RNFL. Theoretical analysis has attributed RNFL reflectance to the cylindrically shaped mitochondria and axonal membranes.9 It is postulated that either changes in neurotubule density of RNFL bundles or neurotubule packing density within axons themselves are responsible for the differences in birefringence surrounding the ONH.11 Neurotubule density (ρRNFL) is defined here as a scalar estimate of the number of neurotubules per unit area of RNFL tissue (NT/μm2). There is strong evidence that the regional characteristic ρRNFL surrounding the optic nerve head (ONH) is a source of birefringence,5,12,13 but it has not been clear if ρRNFL is the only source of birefringence.9 RNFL birefringence has been investigated as a possible diagnostic to detect early sub-cellular changes in glaucoma before there is any measurable change in RNFL thickness.5,12,13 Fortune et al.13 injected colchicine into the vitreous of non-human primate eyes and observed neurotubule disruption with a reduction of RNFL birefringence without any accompanying change in RNFL thickness. These results show that decreases in neurotubule number occur prior to thinning of the retinal nerve fiber layer and in situ measurement of ρRNFL can be used as a diagnostic for cytoskeletal changes in RGC axons.13 This study compares the measured birefringence signal in the peripapillary retina with the corresponding measured anatomical features to explore the origin of the RNFL birefringence.

OBJECTIVE To determine whether antimicrobial therapy for Helicobacter pylori infection accelerates the healing of duodenal ulcers. DESIGN Single-blind, randomized, controlled trial. SETTING Veterans Affairs hospital. PARTICIPANTS One hundred and five patients with endoscopically verified duodenal ulcers. INTERVENTION Patients received either ranitidine, 300 mg/d, or ranitidine, 300 mg/d, plus "triple therapy" (2 g/d of tetracycline, 750 mg/d of metronidazole, and 5 or 8 bismuth subsalicylate tablets per day). Triple therapy was administered for only the first 2 weeks of ulcer treatment. MEASUREMENTS Videoendoscopic assessment of ulcer status was done until ulcer healing was complete. Evaluations were done after 2, 4, 8, 12, and 16 weeks of therapy. MAIN RESULTS Ulcer healing was more rapid in patients receiving ranitidine plus triple therapy than in patients receiving ranitidine alone (P less than 0.01). The cumulative percentages of patients with healed ulcers in the group receiving ranitidine plus triple therapy and in the group receiving ranitidine alone were as follows: 37% and 18% after week 2; 74% and 53% after week 4; 84% and 68% after week 8; 96% and 80% after week 12; and 98% and 84% after week 16. CONCLUSION Combined therapy with anti-H. pylori agents and ranitidine was superior to ranitidine alone for duodenal ulcer healing. Our results indicate that H. pylori plays a role in duodenal ulcer disease.

Summary Invariant natural killer T (iNKT) cells are innate T lymphocytes that promote host defense against a variety of microbial pathogens. Whether microbial ligands are required for their protective effects remains unclear. Here, we show that iNKT cells stimulate human-monocyte-derived dendritic cells (DCs) to produce inflammatory mediators in a manner that does not require the presence of microbial compounds. Interleukin 2 (IL-2)-exposed iNKT cells selectively induced repeated cytoplasmic Ca 2+ fluxes in DCs that were dependent on signaling by the P2X 7 purinergic receptor and mediated by ATP released during iNKT-DC interactions. Exposure to iNKT cells led to DC cyclooxygenase 2 ( PTGS2 ) gene transcription, and release of PGE 2 that was associated with vascular permeabilization in vivo. Additionally, soluble factors were released that induced neutrophil recruitment and activation and enhanced control of Candida albicans . These results suggest that sterile interactions between iNKT cells and monocyte-derived DCs lead to the production of non-redundant inflammatory mediators that promote neutrophil responses.

Review of all hepatitis B surface antigen-positive patients reported to the county health department over the past 3 years showed that 60% of patients had been diagnosed at our hospital, which serves an inner city, predominantly black, indigent population. Sera from 524 adult patients admitted to the hospital were prospectively collected and tested for hepatitis B surface antigen, eight (1.5%) of which were antigen-positive. Census tract data allowed us to perform selective screening of sera from 95 other patients who were subsequently hospitalized and deemed “high risk” for hepatitis B infection. Nine of 95 (9.5%) selectively screened samples were positive for hepatitis B surface antigen, which represented a significantly higher rate of antigen carriage (P

Magnetic- and fluorescent-activated cell sorting (MACS and FACS) are used for isolation of distinct cell populations for subsequent studies including transcriptomics. The latter allows for the analysis of infection-induced alterations in gene expression profiles. MACS and FACS both use antibodies against cell surface molecules to isolate populations of interest. Standardized methods for both approaches exist for use in mouse models. These protocols, however, do not account for the fact that infection-associated immunopathology can significantly modulate the cell surface expression of targeted molecules. This is the case for Trypanosoma brucei brucei infection, where downregulation of CD23 surface expression on B cells has been reported. This hallmark of progressing infection interferes with the commercially available MACS technique for B cell purification, as CD23 expression is the target for the separation between Marginal Zone (MZ) and Follicular (Fo) B cells. Here, we provide a robust alternative method for isolation of infection-derived MZ B cells using CD1d and B220 surface molecules in a two-step MACS-FACS approach. The method yields 99% pure viable infection-derived MZ B cells, allowing extraction of a high quality total RNA suitable for subsequent RNA sequencing.