Development and Evaluation of Brief Web-Based Education for Primary Care Providers to Address Inequities in Lung Cancer Screening and Smoking Cessation Treatment.

Annual lung cancer screening (LCS) is recommended for individuals at high risk for lung cancer. However, primary care provider-initiated discussions about LCS and referrals for screening are low overall, particularly among Black or African Americans and other minoritized racial and ethnic groups. Disparities also exist in receiving provider advice to quit smoking. Effective methods are needed to improve provider knowledge about LCS and tobacco-related disparities, and to provide resources to achieve equity in LCS rates. We report the feasibility and impact of pairing a self-directed Lung Cancer Health Disparities (HD) Web-based course with the National Training Network Lung Cancer Screening (LuCa) course on primary care providers' knowledge about LCS and the health disparities associated with LCS. In a quasi-experimental study, primary care providers (N = 91) recruited from the MedStar Health System were assigned to complete the LuCa course only vs. the LuCa + HD courses. We measured pre-post-LCS-related knowledge and opinions about the courses. The majority (60.4%) of providers were resident physicians. There was no significant difference between groups on post-test knowledge (p > 0.05). However, within groups, there was an improvement in knowledge from pre- to post-test (LuCa only (p = 0.03); LuCa + HD (p < 0.001)). The majority of providers (81%) indicated they planned to improve their screening and preventive practices after having reviewed the educational modules. These findings provide preliminary evidence that this e-learning course can be used to educate providers on LCS, smoking cessation, and related disparities impacting patients.

Iron availability enhances the cellular energetics of aerobic Escherichia coli cultures while upregulating anaerobic respiratory chains.

Aerobic Escherichia coli growth at restricted iron concentrations (≤ 1.75 ± 0.04 µM) is characterized by lower biomass yield, higher acetate accumulation and higher activation of the siderophore iron-acquisition systems. Although iron homeostasis in E. coli has been studied intensively, previous studies focused only on understanding the regulation of the iron import systems and the iron-requiring enzymes. Here, the effect of iron availability on the energy metabolism of E. coli has been investigated. It was established that aerobic cultures growing under limiting iron conditions showed lower ATP yield per glucose, lower growth rate and lower TCA cycle activity and respiration, at the same time as increased glucose consumption, acetate and pyruvate accumulation, practically mimicking microaerobic growth. However, at excess iron, independent of oxygen availability, the cultures showed high cellular energetics (5.8 ATP/mol of glucose) by using pathways requiring iron-rich complex proteins found in the TCA cycle and respiratory chain. In conditions of iron excess, some iron-requiring terminal reductases of the respiratory chain, that were thought to function only under anaerobiosis, were used by the E. coli, when in aerobic conditions, to maintain high respiratory activity. This allowed it to produce more biomass and more reactive oxygen species that were controlled by the higher activity of the antioxidant defenses (SOD, peroxidase and catalase) and the iron-sulfur cluster repair systems.

A Randomized Trial of Telephone-Based Smoking Cessation Treatment in the Lung Cancer Screening Setting.

BACKGROUND: Lung cancer mortality is reduced via low-dose computed tomography screening and treatment of early-stage disease. Evidence-based smoking cessation treatment in the lung screening setting can further reduce mortality. We report the results of a cessation trial from the National Cancer Institute's Smoking Cessation at Lung Examination collaboration. METHODS: Eligible patients (n = 818) aged 50-80 years were randomly assigned (May 2017-January 2021) to the intensive vs minimal arms (8 vs 3 phone sessions plus 8 vs 2 weeks of nicotine patches, respectively). Bio-verified (primary) and self-reported 7-day abstinence rates were assessed at 3, 6, and 12 months post random assignment. Logistic regression analyses evaluated the effects of study arm. All statistical tests were 2-sided. RESULTS: Participants reported 48.0 (SD = 17.2) pack-years, and 51.6% were not ready to quit in less than 30 days. Self-reported 3-month quit rates were statistically significantly higher in the intensive vs minimal arm (14.3% vs 7.9%; odds ratio [OR] = 2.00, 95% confidence interval [CI] = 1.26 to 3.18). Bio-verified abstinence was lower but with similar relative differences between arms (9.1% vs 3.9%; OR = 2.70, 95% CI = 1.44 to 5.08). Compared with the minimal arm, the intensive arm was more effective among those with greater nicotine dependence (OR = 3.47, 95% CI = 1.55 to 7.76), normal screening results (OR = 2.58, 95% CI = 1.32 to 5.03), high engagement in counseling (OR = 3.03, 95% CI = 1.50 to 6.14), and patch use (OR = 2.81, 95% CI = 1.39 to 5.68). Abstinence rates did not differ statistically significantly between arms at 6 months (OR = 1.2, 95% CI = 0.68 to 2.11) or 12 months (OR = 1.4, 95% CI = 0.82 to 2.42). CONCLUSIONS: Delivering intensive telephone counseling and nicotine replacement with lung screening is an effective strategy to increase short-term smoking cessation. Methods to maintain short-term effects are needed. Even with modest quit rates, integrating cessation treatment into lung screening programs may have a large impact on tobacco-related mortality.

Comparison of the GenCut Core Biopsy System to Transbronchial Biopsy Forceps for Flexible Bronchoscopic Lung Biopsy.

BACKGROUND: Pulmonary nodules are increasingly detected on screening and routine chest imaging, leading to an increase in diagnostic procedures. Bronchoscopy with transbronchial biopsy (TBBx) is the most common diagnostic modality, with diagnostic yield between 43% and 86%, largely dependent on the use of navigational modalities. In 2015 a new biopsy tool by Medtronic, the GenCut core biopsy system [GenCut transbronchial needle aspiration (TBNA)], was developed with the intention of improving yield in lung nodule biopsies. Our goal was to determine the efficacy of this new device when used in addition to TBBx. PATIENTS AND METHODS: This is a prospective observational study of 324 consecutive bronchoscopic lung biopsies in which both TBBx and GenCut TBNA were performed on the same lesion. We recorded patient and nodule characteristics, along with the bronchoscopic modalities used. The primary outcome was the diagnostic yield with the addition of the GenCut TBNA, and the key secondary outcome was the complication rate. RESULTS: Of the 324 nodule biopsies analyzed, 164 (50.6%) were diagnostic via TBBx or GenCut TBNA.In all, 97 (59%) were positive in both TBBx and GenCut TBNA, 43 (26.2%) were positive only in TBBx, and 24 (14.6%) were positive only in GenCut TBNA. Overall, the addition of the GenCut TBNA increased the diagnostic yield by 7.4% (P<0.01). There were 7 complications: 5 pneumothoraxes and 2 episodes of bleeding. CONCLUSION: The diagnostic yield is improved by using the GenCut core biopsy system in addition to traditional TBBx forceps when performing bronchoscopy for pulmonary nodules, without an increase in complications. These biopsy methods should be used in tandem for the greatest yield.

Change in amount smoked and readiness to quit among patients undergoing lung cancer screening.

BACKGROUND: There is mixed evidence regarding whether undergoing computed tomography lung cancer screening (LCS) can serve as a "teachable moment" that impacts smoking behavior and attitudes. The study aim was to assess whether the standard procedures of undergoing LCS and receiving free and low-cost evidence-based cessation resources impacted short-term smoking-related outcomes. METHODS: Participants were smokers (N=87) who were registered to undergo lung screening and were enrolled in a cessation intervention trial. We conducted two phone interviews, both preceding trial randomization: the first interview was conducted prior to lung screening, and the second interview followed lung screening (median =12.5 days post-screening) and participants' receipt of their screening results. The interviews assessed demographic characteristics, interest in evidence-based cessation intervention methods, and tobacco-related characteristics, including cigarettes per day and readiness to quit. Participants received minimal evidence-based cessation resources following the pre-lung screening interview. RESULTS: Participants were 60.3 years old, 56.3% female, and reported a median of 40 pack-years. Participants were interested in using several evidence-based strategies, including counseling from a healthcare provider (76.7%) and receiving nicotine replacement therapy (69.8%). Pre-lung screening, 25.3% smoked ≤10 cigarettes per day, and 29.9% were ready to quit in the next 30 days. We conducted two McNemar binomial distribution tests to assess change from pre- to post-screening. At the post-lung screening assessment, approximately three-quarters reported no change on these variables. However, 23.3% reported smoking fewer cigarettes per day, whereas 4.7% reported smoking more cigarettes per day (McNemar P=0.002), and 17.2% reported increased readiness to quit, whereas 6.9% reported decreased readiness to quit (McNemar P=0.078). CONCLUSIONS: Following receipt of cessation resources and completion of lung screening, most participants reported no change in smoking outcomes. However, there was a significant reduction in cigarettes per day, and there was a trend for increased readiness to quit. This setting may provide a potential "teachable moment" and an opportunity to assist smokers with quitting. However, more proactive and intensive interventions will be necessary to capitalize on these changes and to support abstinence in the long-term.

Predictors of attrition in a smoking cessation trial conducted in the lung cancer screening setting.

SIGNIFICANCE: Although it is a requirement that tobacco treatment is offered to cigarette smokers undergoing low-dose computed tomographic lung cancer screening (LCS), not all smokers engage in treatment. To understand the barriers to tobacco treatment in this setting, we evaluated predictors of attrition in a smoking cessation trial among individuals undergoing LCS. METHODS: Prior to LCS, 926 participants, 50-80 years old, completed the baseline (T0) phone assessment, including demographic, clinical, tobacco, and psychological characteristics. Following LCS and receipt of the results, participants completed the pre-randomization (T1) assessment. RESULTS: At the T1 assessment, 735 (79%) participants were retained and 191 (21%) dropped out. In multivariable analyses, attrition was higher among those who: smoked >1 pack per day (OR = 1.44, CI 1.01, 2.06) or had undergone their first (vs. annual) LCS scan (OR = 1.70, CI 1.20, 2.42). Attrition was lower among those with: more education (associates (OR = 0.67, CI = 0.46, 0.98) or bachelor's degree (OR = 0.56, CI 0.35, 0.91) vs. high school/GED), some (vs. none/a little) worry about lung cancer (OR = 0.60, CI 0.39, 0.92), or a screening result that was benign (OR = 0.57, CI 0.39, 0.82) or probably benign (OR = 0.38, CI 0.16, 0.90) vs. negative. CONCLUSIONS: This study illuminated several LCS-related factors that contributed to trial attrition. Increasing tobacco treatment in this setting will require targeted strategies for those who report little lung cancer worry, are undergoing their first LCS exam, and/or who have a negative LCS result. Addressing attrition and reducing barriers to tobacco treatment will increase the likelihood of cessation, thereby reducing the risk of developing lung cancer.

Bayesian latent class model estimates of diagnostic accuracy for three test methods designed to detect spring viremia of carp virus.

Spring viremia of carp virus (SVCV) causes a systemic hemorrhagic disease that poses a significant risk to wild and cultured fish and is listed as notifiable by the World Organization for Animal Health. Validated molecular diagnostic tools for SVCV are required to accurately describe and analyze the ecology of the virus. Here, the diagnostic specificity (DSp) and sensitivity (DSe) (i.e. accuracy) of three SVCV diagnostic tests - 2 reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays Q1G and Q2N and virus isolation by cell culture (VI) - were evaluated using 2-class latent class models run in maximum likelihood (ML) and Bayesian frameworks. Virus-free or experimentally-infected koi were sorted into three populations with low, moderate or high prevalence levels of SVCV (n = 269 fish in total). Koi kidney tissues were tested using Q2N and Q1G and for the VI assay, pools of kidney, spleen and gill tissues were used. All samples were blinded and analyzed in one laboratory. The ML and Bayesian approaches successfully estimated the diagnostic accuracy of the 3 tests with the exception of 1 ML model. The estimates were consistent across the two frameworks. The DSe estimates were higher for Q1G (>98 %) and Q2N (>96 %) compared to VI (>60 %). The DSp of all three tests varied by 12-15 % (79-91 % for Q1G, 79-94 % for Q2N and 81-97 % for VI) across same-fish samples revealing the potential range in test performance for one sample. The 3 fish populations had distinct SVCV prevalence levels estimated at 0-3 % (low), 70-73 % (moderate) and 95-96 % (high). The Bayesian covariance models revealed minor DSe dependence between Q1G and Q2N. The results suggested that SVCV diagnostic tests Q2N and Q1G are suitable for use as diagnostic assays and are fit for presumptive diagnosis, surveillance, and certification of populations or individuals as SVCV free.

Measures of diagnostic precision (repeatability and reproducibility) for three test methods designed to detect spring viremia of carp virus.

Spring viremia of carp virus (SVCV) is a rhabdovirus of the Sprivivirus genus and the etiological agent of an internationally regulated aquatic animal disease in several fish species, including koi carp Cyprinus carpio L. The virus has a complex lifecycle with both acute and persistent stages of infection and can cause high mortality in affected populations. In this study, the diagnostic repeatability (within laboratory agreement) and reproducibility (between laboratory agreement) of 3 tests were investigated to assess their fitness as SVCV diagnostic tools. The tests, reverse transcription quantitative polymerase chain reaction (RT-qPCR) assays targeting either the SVCV glycoprotein (Q1G) or nucleoprotein (Q2N) genes and virus isolation by cell culture (VI), were performed in a blinded study with four Canadian laboratories. Test panels consisted of duplicate sets of 100 tissue samples collected from 3 SVCV prevalence populations of koi: a low-prevalence negative reference population (n = 20 fish) as well as moderate- (n = 50 fish) and high-prevalence (n = 30 fish) populations of koi experimentally infected with SVCV. The Q1G and Q2N tests were performed with kidney tissue in 3 laboratories and with brain tissue in 1 laboratory whereas pools of kidney, spleen and gill tissues were tested with the VI assay in 2 laboratories. Agreement of binary results was evaluated using the observed proportion of agreement, Cohen's kappa and Gwet's agreement coefficient (AC1) whereas the concordance correlation coefficient (ccc) and Bland Altman's limit of agreement were used to evaluate agreement of the RT-qPCR continuous data. Gwet's AC1 provided a more stable estimate of agreement than Cohen's kappa. Overall, high repeatability (AC1, 0.78-0.90) and reproducibility (AC1, 0.74-0.89) were observed for the Q1G and Q2N tests when kidney tissue was used. Lower agreement estimates of repeatability (AC1, 0.54-0.77) and reproducibility (AC1, 0.50-0.80) were obtained for the VI test. RT-qPCR reproducibility was low with kidney-brain tissue pairs (AC1, 0.09-0.46) and high with inter-test pairs of brain (AC1, 0.76-0.86) or kidney tissue (0.75-0.86). Tissue-specific differences in virus load affected test precision and informed final tissue selection. Repeatability (ccc, 0.94-0.97) and reproducibility (ccc, 0.91-0.97) estimates of agreement for paired continuous data from the RT-qPCR assays were similarly high with kidney tissue and lower with paired brain (ccc, 0.15-0.83) and kidney-brain tissues (ccc, 0.01-0.55). The high precision of Q1G and Q2N with kidney tissue suggests that the tests are performing similarly and are suitable candidates for assessment of their diagnostic accuracy.

Analytical validation of two RT-qPCR tests and detection of spring viremia of carp virus (SVCV) in persistently infected koi Cyprinus carpio.

Spring viremia of carp virus (SVCV) ia a carp sprivivirus and a member of the genus Sprivivirus within the family Rhabdoviridae. The virus is the etiological agent of spring viremia of carp, a disease of cyprinid species including koi Cyprinus carpio L. and notifiable to the World Organisation for Animal Health. The goal of this study was to explore hypotheses regarding inter-genogroup (Ia to Id) SVCV infection dynamics in juvenile koi and contemporaneously create new reverse-transcription quantitative PCR (RT-qPCR) assays and validate their analytical sensitivity, specificity (ASp) and repeatability for diagnostic detection of SVCV. RT-qPCR diagnostic tests targeting the SVCV nucleoprotein (Q2N) or glycoprotein (Q1G) nucleotides were pan-specific for isolates typed to SVCV genogroups Ia to Id. The Q2N test had broader ASp than Q1G because Q1G did not detect SVCV isolate 20120450 and Q2N displayed occasional detection of pike fry sprivivirus isolate V76. Neither test cross-reacted with other rhabdoviruses, infectious pancreatic necrosis virus or co-localizing cyprinid herpesvirus 3. Both tests were sensitive with observed 50% limits of detection of 3 plasmid copies and high repeatability. Test analysis of koi immersed in SVCV showed that the virus could be detected for at least 167 d following exposure and that titer, prevalence, replicative rate and persistence in koi were correlated significantly with virus virulence. In this context, high virulence SVCV isolates were more prevalent, reached higher titers quicker and persisted in koi for longer periods of time relative to moderate and low virulence isolates.

Molecular phylogeny of sturgeon mimiviruses and Bayesian hierarchical modeling of their effect on wild Lake Sturgeon (Acipenser fulvescens) in Central Canada.

Sturgeon mimiviruses can cause a lethal disease of the integumentary systems of sturgeon (Acipenseridae). Here we provide phylogeographic evidence that sturgeon mimivirus is endemic in endangered populations of wild Lake Sturgeon within Canada's Hudson Bay drainage basin. Namao virus (NV) variants were diagnosed in 24% of Lake Sturgeon samples (n = 1329) collected between 2010-2015. Lake Sturgeon populations with the highest virus prevalence were from the Nelson River (58%) in 2015, Saskatchewan River (41%) in 2010 and South Saskatchewan River (36%) in 2011. Bayesian phylogenetic reconstructions suggested that four NV variants, designated HBDB I-IV, co-circulate temporally and spatially within and between the genetically and biogeographically distinct Lake Sturgeon populations. Evidence from recapture studies suggested that Lake Sturgeon across the basin are persistently infected with NV at prevalence and titer (103.6 equivalent plasmid copies per µg DNA) levels consistent with the hypothesis that wild Lake Sturgeon populations serve as a maintenance population and reservoir for sturgeon mimiviruses. Bayesian hierarchical modeling of NV in the Landing River population of Lake Sturgeon suggested that host weight and age were the best predictors of sturgeon mimivirus presence and titer, respectively, whereas water flow rate, level and temperature, and number of previous captures did not significantly improve model fit. A negative relationship was estimated between sturgeon mimivirus presence and Lake Sturgeon weight and between virus titer and Lake Sturgeon age.

Effect of restricted dissolved oxygen on expression of Clostridium difficile toxin A subunit from E. coli.

The repeating unit of the C. difficile Toxin A (rARU, also known as CROPS [combined repetitive oligopeptides]) C-terminal region, was shown to elicit protective immunity against C. difficile and is under consideration as a possible vaccine against this pathogen. However, expression of recombinant rARU in E. coli using the standard vaccine production process was very low. Transcriptome and proteome analyses showed that at restricted dissolved oxygen (DO) the numbers of differentially expressed genes (DEGs) was 2.5-times lower than those expressed at unrestricted oxygen. Additionally, a 7.4-times smaller number of ribosome formation genes (needed for translation) were down-regulated as compared with unrestricted DO. Higher rARU expression at restricted DO was associated with up-regulation of 24 heat shock chaperones involved in protein folding and with the up-regulation of the global regulator RNA chaperone hfq. Cellular stress response leading to down-regulation of transcription, translation, and energy generating pathways at unrestricted DO were associated with lower rARU expression. Investigation of the C. difficile DNA sequence revealed the presence of cell wall binding profiles, which based on structural similarity prediction by BLASTp, can possibly interact with cellular proteins of E. coli such as the transcriptional repressor ulaR, and the ankyrins repeat proteins. At restricted DO, rARU mRNA was 5-fold higher and the protein expression 27-fold higher compared with unrestricted DO. The report shows a strategy for improved production of C. difficile vaccine candidate in E. coli by using restricted DO growth. This strategy could improve the expression of recombinant proteins from anaerobic origin or those with cell wall binding profiles.

Effect of amino acids on transcription and translation of key genes in E. coli K and B grown at a steady state in minimal medium.

Growing E. coli to high densities is a common strategy for biologicals production. The process is implemented by using complex or minimal media with different feeding strategies. To understand the effect of amino acids, E. coli B and K were grown at a steady state of 0.35 h-1 in glucose minimal medium with and without amino acids, while their metabolism, protein abundance and gene expression were compared. The results showed that amino acids promoted higher acetate excretion, higher fatty acid biosynthesis (K strain), repressed glucose uptake rate, and decreased expression of proteins associated with the TCA cycle, glyoxylate shunt and amino acid biosynthesis. In presence of amino acids, E. coli K upregulated fatty acid biosynthesis and repressed more genes and proteins involved in amino acid biosynthesis than E. coli B. These findings are correlated with higher yield on glucose (Yx/s) and high specific biomass production rate (qx) in K strain in the presence of amino acids. In contrast, pre-formed precursor molecules such as amino acids did not affect fatty acid biosynthesis in E. coli B or Yx/s and qx, which were higher than those of E. coli K, suggesting that constitutive synthesis of energetically demanding precursors and higher fatty acid ß-oxidation activity is key for high biomass-performer E. coli B. Both strains turned off unnecessary pathways and directed their metabolism to proteome efficient overflow metabolism likely to generate energy and provide protein to functions supporting higher growth rate.

Cognitive Variability Predicts Incident Alzheimer's Disease and Mild Cognitive Impairment Comparable to a Cerebrospinal Fluid Biomarker.

BACKGROUND: Alzheimer's disease (AD) biomarkers are emerging as critically important for disease detection and monitoring. Most biomarkers are obtained through invasive, resource-intense procedures. A cognitive marker, intra-individual cognitive variability (IICV) may provide an alternative or adjunct marker of disease risk for individuals unable or disinclined to undergo lumbar puncture. OBJECTIVE: To contrast risk of incident AD and mild cognitive impairment (MCI) associated with IICV to risk associated with well-established biomarkers: cerebrospinal fluid (CSF) phosphorylated tau protein (p-tau181) and amyloid-ß 42 (Aß42) peptide. METHODS: Dispersion in cognitive performance, IICV, was estimated with a published algorithm, and included Trail Making Test A and B, Rey Auditory Verbal Learning Test (RAVLT), and the American National Adult Reading Test (ANART). CSF biomarkers were expressed as a ratio: p-tau181/Aß42, wherein high values signified pathognomonic profiles. Logistic regression models included longitudinal data from 349 Alzheimer's Disease Neuroimaging Initiative (ADNI) participants who completed lumbar puncture. All subjects were cognitively healthy (n = 105) or diagnosed with MCI (n = 244) at baseline. We examined odds of conversion associated with baseline elevations in IICV and/or ratio of CSF p-tau181/Aß42. RESULTS: When included in models alone or in combination with CSF p-tau181/Aß42, one standard IICV unit higher was associated with an estimated odds ratio for incident AD or MCI of 2.81 (95% CI: 1.83-4.33) in the most inclusive sample, and an odds ratio of 3.41 (95% CI: 2.03-5.73) when restricted to participants with MCI. Iterative analyses suggested that IICV independently improved model fit even when individual index components were included in comparative models. CONCLUSIONS: These analyses provide preliminary support for IICV as a marker of incident AD and MCI. This easily-disseminated, non-invasive marker compared favorably to well-established CSF biomarkers.

Diagnostic validation of three test methods for detection of cyprinid herpesvirus 3 (CyHV-3).

Cyprinid herpesvirus 3 (CyHV-3) is the aetiological agent of koi herpesvirus disease in koi and common carp. The disease is notifiable to the World Organisation for Animal Health. Three tests-quantitative polymerase chain reaction (qPCR), conventional PCR (cPCR) and virus isolation by cell culture (VI)-were validated to assess their fitness as diagnostic tools for detection of CyHV-3. Test performance metrics of diagnostic accuracy were sensitivity (DSe) and specificity (DSp). Repeatability and reproducibility were measured to assess diagnostic precision. Estimates of test accuracy, in the absence of a gold standard reference test, were generated using latent class models. Test samples originated from wild common carp naturally exposed to CyHV-3 or domesticated koi either virus free or experimentally infected with the virus. Three laboratories in Canada participated in the precision study. Moderate to high repeatability (81 to 99%) and reproducibility (72 to 97%) were observed for the qPCR and cPCR tests. The lack of agreement observed between some of the PCR test pair results was attributed to cross-contamination of samples with CyHV-3 nucleic acid. Accuracy estimates for the PCR tests were 99% for DSe and 93% for DSp. Poor precision was observed for the VI test (4 to 95%). Accuracy estimates for VI/qPCR were 90% for DSe and 88% for DSp. Collectively, the results show that the CyHV-3 qPCR test is a suitable tool for surveillance, presumptive diagnosis and certification of individuals or populations as CyHV-3 free.

Long-term outcomes of stereotactic body radiation therapy (SBRT) with fiducial tracking for inoperable stage I non-small cell lung cancer (NSCLC).

BACKGROUND: Stereotactic body radiation therapy (SBRT) for stage I non-small cell lung cancer (NSCLC) is considered standard of care in the medically inoperable patient population. Multiple methods of SBRT delivery exist including fiducial-based tumor tracking, which allows for smaller treatment margins and avoidance of patient immobilization devices. We explore the long-term clinical outcomes of this novel fiducial-based SBRT method. METHODS: In this single institutional retrospective review, we detail the outcomes of medically inoperable pathologically confirmed stage I NSCLC. Patients were treated with the Cyberknife SBRT system using a planning target volume (PTV) defined as a 5-mm expansion from gross tumor volume (GTV) without creation of an internal target volume (ITV). Dose was delivered in three or five equal fractions of 10 to 20 Gy. Pretreatment and posttreatment pulmonary function test (PFT) changes and evidence of late radiological rib fractures were analyzed for the majority of patients. Actuarial local control, locoregional control, distant control, and overall survival were calculated using the Kaplan-Meier method. RESULTS: Sixty-one patients with a median age of 75 years were available for analysis. The majority (80 %) of patients were deemed to be medically inoperable due to underlying pulmonary dysfunction. Eleven patients (18 %) developed symptomatic pneumothoraces secondary to fiducial placement under CT guidance, which precipitously dropped to 0 % following transition to bronchoscopic fiducial placement. The 2-year rib fracture risk was 21.4 % with a median time to rib fracture of 2.9 years. PFTs averaged over all patients and parameters demonstrated small absolute declines, 5.7 % averaged PFT decline, at approximately 1 year of follow-up, but only the diffusing capacity of lung for carbon monoxide (DLCO) demonstrated a statistically significant decline (10.29 vs. 9.01 mL/min/mmHg, p = 0.01). Five-year local control, locoregional control, and overall survival were 87.6, 71.8, and 39.3 %, respectively. CONCLUSIONS: Despite reduced treatment margins and lack of patient immobilization, SBRT with fiducial-based tumor tracking achieves clinically comparable long-term outcomes to other linac-based SBRT approaches.

Cognitive variability-A marker for incident MCI and AD: An analysis for the Alzheimer's Disease Neuroimaging Initiative.

INTRODUCTION: The potential of intra-individual cognitive variability (IICV) to predict incident mild cognitive impairment (MCI) or Alzheimer's disease (AD) was examined and compared to well-established neuroimaging and genetic predictors. METHODS: IICV was estimated using four neuropsychological measures for n = 1324 Alzheimer's Disease Neuroimaging Initiative (ADNI) participants who were cognitively healthy or diagnosed with MCI at baseline. IICV was used to predict time to incident MCI or AD, and compared to hippocampal volume loss and APOE ε4 status via survival analysis. RESULTS: In survival analyses, controlling for age, education, baseline diagonosis, and APOE ε4 status, likelihood ratio tests indicate that IICV is associated with time to cognitive status change in the full sample (P < .0001), and when the sample was restricted to individuals with MCI at baseline (P < .0001). DISCUSSION: These findings suggest IICV may be a low-cost, noninvasive alternative to traditional AD biomarkers.

Stereotactic body radiotherapy (SBRT) for high-risk central pulmonary metastases.

BACKGROUND AND PURPOSE: Radiotherapy of central lung tumors carries a higher risk of treatment-related toxicity and local failure. In the era of aggressive oligometastic management the exploration of the proper dose-fractionation for metastatic central lung tumors is essential. MATERIALS AND METHODS: Patients diagnosed with high-risk metastatic lesions of the central pulmonary tree comprised this single-institutional retrospective analysis. "High-risk" central pulmonary lesions were defined as those with abutment and/or invasion of the mainstem bronchus. All patients were treated using the CyberKnife SBRT system in 5 fractions to a total dose of 35 or 40 Gy. RESULTS: Twenty patients were treated from 2008 to 2011 at Georgetown University Hospital. At a median follow up of 19 months, 1-year Kaplan-Meier local control and overall survival was 70 and 75 %, respectively. Late grade 2 or higher atelectasis was the most common treatment-related toxicity and was significantly associated with maximum dose to the mainstem bronchus. Gross endobronchial involvement was associated with significantly lower overall survival. CONCLUSIONS: Five-fraction SBRT to a total dose of 35 or 40 Gy appears to be a safe and effective management strategy for high-risk central pulmonary metastatic lesions, though care should be taken to limit the maximum point dose to the mainstem bronchus.

Sturgeon nucleo-cytoplasmic large DNA virus phylogeny and PCR tests.

Sturgeon epitheliotropic nucleo-cytoplasmic large DNA viruses (NCLDVs) can cause a lethal disease of the integumentary system. These viruses have not been assigned to any currently recognized family or genus. In this study, phylogenetic analyses using the major capsid protein (MCP) showed that the sturgeon NCLDVs formed a cohesive taxonomic group, could be identified to the species or possibly sub-species level and formed a distinct evolutionary lineage within the Megavirales. The genetic relatedness of the sturgeon virus MCP allowed design of 3 PCR diagnostic tests with analytical specificity (ASp) inclusive of this group of viruses. The conventional PCR test, C1, had broader ASp than the 2 quantitative PCR tests, Q1 and Q2, and was inclusive of the sturgeon viruses as well as some viruses belonging to the families Mimi-, Phycodna-, or Iridoviridae. Q2 had broader specificity than Q1 but both tests recognized the sturgeon NCLDVs and did not cross-react with co-localizing sturgeon herpesviruses. Analytical test performance characteristics evaluated for Q1 and Q2 revealed sensitive assays with observed 50% limits of detection between 3 and 6.25 plasmid copies and high intra- and inter-assay repeatability. Q1 was used to test for sturgeon viruses in endangered populations of lake sturgeon Acipenser fulvescens within the Winnipeg River or Nelson River drainage systems of Manitoba, Canada. Test results indicated that namao virus is endemic in the Nelson River water basin. These tests meet the analytical requirements for diagnostic testing in Canada and are useful tools for disease management in sturgeon conservation stocking programs in North America.

Incorporating a Smoking Cessation Intervention into Lung Cancer Screening Programs: Preliminary Studies.

Two preliminary studies assessed whether telephone counseling (TC) is a feasible smoking cessation intervention following lung cancer screening. Seven older smokers undergoing lung cancer screening (pack years = 61.5) completed three TC sessions, which incorporated the screening result as motivation to quit. Participation (87.5%) and retention (85.7%) rates were good, and four smokers quit smoking (three of whom received abnormal results). We conducted four focus groups with 16 current and former older smokers (pack years = 55). Most believed that an abnormal scan would motivate them to quit and expressed interest in TC. TC may be feasible and potentially efficacious within lung screening programs.

A new species of nucleo-cytoplasmic large DNA virus (NCLDV) associated with mortalities in Manitoba lake sturgeon Acipenser fulvescens.

A newly discovered virus, Namao virus, associated with morbidity and mortality, was detected among juvenile lake sturgeon Acipenser fulvescens being propagated by a conservation stocking program for this endangered species in Manitoba, Canada. The outbreaks resulted in cumulative mortalities of 62 to 99.6% among progeny of wild Winnipeg River or Nelson River lake sturgeon and occurred at 2 geographically separate facilities. Namao virus was detected in almost 94% of the moribund or dead lake sturgeon according to a conventional polymerase chain reaction (cPCR) test that is based upon amplification of a 219 bp fragment of the virus major capsid protein (MCP). The virus itself was large (242 to 282 nm) and icosahedral-shaped with 2 capsids and a condensed bar-shaped core. It was found in virus factories within the host cell cytoplasm and displayed a tropism for the integument. Namao virus caused cellular changes characterized by enlarged eosinophilic epithelial cells in the gills and skin. Samples suspected of containing Namao virus did not have cytopathic effects on primary lake sturgeon or established white sturgeon cell lines. However, viral nucleic acid was detected in the former after prolonged incubation periods. Using primers designed from conserved regions of the MCP from NCLDVs, an estimated 95 to 96% of the Namao virus MCP open reading frame was captured. Phylogenetic analysis using the MCP of Namao virus and 27 other NCLDVs suggested that Namao virus and white sturgeon iridovirus share a common evolutionary past and might be members of the family Mimiviridae or a new, as yet unrecognized, virus family.