Evaluation of Alinity m CMV assay performance for detecting CMV in plasma, cerebrospinal fluid, and bronchoalveolar lavage specimens.

Accurate detection and quantification of cytomegalovirus (CMV) is crucial to preventing adverse outcomes in immunocompromised individuals. Current assays were developed for use with plasma specimens, but CMV may be present in bronchoalveolar lavage (BAL) fluid and cerebrospinal fluid (CSF). We evaluated the performance of the Abbott Alinity m CMV assay compared to the Abbott RealTime CMV assay for quantification of CMV in plasma, BAL, and CSF specimens. To evaluate clinical performance, 190 plasma, 78 BAL, and 20 CSF specimens were tested with the Alinity m assay and compared to the RealTime assay. The Alinity m CMV assay showed high precision (SD <0.01 to 0.13) for all 3 specimen types. Clincal plasma and BAL specimens with quantifiable CMV DNA demonstrated strong correlation to RealTime CMV assay results (r2 = 0.9779 for plasma, r2 = 0.9373 for BAL). The Alinity m CMV assay may be useful for quantification of CMV in plasma, BAL, and CSF specimens.

Engagement to collaboration ensures success.

Disruption to our daily business and functional lives is becoming more frequent, complex and costly. As leaders, what do we do with what we know, the support and tools we have, and our knowledge regarding the resources we need to acquire to navigate this disrupted world? One thing is clear: no one can do it alone. This is not a new concept - the ancient Greeks understood the power of the group. This paper argues that collaboration is the key to amplified knowledge, ability, energy, foresight and innovation, as there is obvious synergy when individuals, groups or organisations join together in a shared vision and with a dedicated purpose. This paper describes a process model developed by the Mid-Atlantic Center for Emergency Management & Public Safety to transform operational functions and spark quality engagement, the synergy of ideas and outcomes, and enhanced sustainability of purpose. This model uses a blend of new knowledge and experiences to build on collaboration models of the past, and has proven to be a success.

Postanesthesia complications in pediatric patients with previous SARS-CoV-2 infection: A cohort study.

BACKGROUND: Children with SARS-CoV-2 infection are at increased risk for postanesthesia complications. There is minimal data regarding how long that elevated complication risk persists beyond initial SARS-CoV-2 diagnosis. AIMS: We investigated postanesthesia complications in children with SARS-CoV-2 infection within 90 days of diagnosis. METHODS: We completed a single-center, retrospective, case-control study of pediatric patients with confirmed SARS-CoV-2 infection within 90 days undergoing anesthesia between January 3-October 7, 2020. Each SARS-CoV-2 positive patient was matched 1:2 by age and type of procedure with a non-SARS-CoV-2 cohort. The primary outcome was the rate of all postanesthesia complications within 30 days of the procedure, defined as unplanned escalations of care within 48 h, cardiac, respiratory, thrombotic, and hemorrhagic events within 30 days. Secondary outcomes were 30-day mortality and hospital length of stay. RESULTS: Of the 341 patients included, 114 patients were SARS-CoV-2 positive and 227 were SARS-CoV-2 negative. Patients with a positive test 0-7 days prior to anesthesia had an increased risk difference in all postanesthesia complications within 30 days (19.9, 95% CI [4.7, 35.1], p = .001) and increased risk difference in length of hospital stay (7.8, 95% CI [1.2, 14.4], p < .001). Patients who underwent anesthesia greater than 42 days from SARS-CoV-2 diagnosis had an increased risk difference in cardiac complications within 30 days (4.3, 95% CI [0.9, 10.0], p = .029). There was no increased hospital length of stay among SARS-CoV-2 positive patients diagnosed greater than 8 days before anesthetic. There were no deaths within 30 days of anesthetic. CONCLUSIONS: Postanesthesia complications are higher in children who undergo anesthesia within 7 days of SARS-CoV-2 diagnosis. Additional cardiac risk may persist beyond the immediate period of initial diagnosis. Larger samples are needed to further evaluate the risk of delayed postanesthesia complications and guide optimal timing of surgery.

Cohort profile for the Loma Linda University Health BREATHE programme: a model to study continuously incentivised employee smoking cessation.

PURPOSE: The purpose of the Loma Linda University Health (LLUH) BREATHE cohort is to test the efficacy of a novel method of continuously incentivising participation in workplace smoking cessation on participation, long-term abstinence, health outcomes, healthcare costs and healthcare utilisation. PARTICIPANTS: In 2014, LLUH-a US academic medical centre and university-incentivised participation in a workplace smoking cessation programme (LLUH BREATHE) by lowering health plan costs. Specifically, LLUH introduced a Wholeness Health Plan (WHP) option that, for the smokers, continuously incentivises participation in nicotine screening and the LLUH BREATHE smoking cessation programme by offering an 'opt-in wellness discount' that consisted of 50%-53% lower out of pocket health plan costs (ie, monthly employee premiums, copayments). This novel 'continuously incentivised' model lowers annual health plan costs for smokers who, on an annual basis, attempt or maintain cessation from tobacco use. The annual WHP cost savings for smokers far exceed the value of short-term incentives that have been tested in workplace cessation trials to date. This ongoing health plan option offered to over 16 000 employees has created an open, dynamic LLUH BREATHE cohort of current and former smokers (n=1092). FINDINGS TO DATE: Our profile of the LLUH BREATHE cohort indicates that after 5 years of follow-up in a prospective cohort study (2014-2019), continuously incentivised smoking cessation produced a 74% participation (95% CI (71% to 77%)) in employer-sponsored smoking cessation attempts that were occurring less than a year after the incentive was offered. The cohort can be purposed to examine the effect of continuously incentivised cessation on cessation outcomes, health plan utilisation/costs, use of electronic nicotine delivery systems, and COVID-19 outcomes.

Medical planning for disaster response: Identifying risk factors for developing adult respiratory distress syndrome among trauma patients.

INTRODUCTION: Adult respiratory distress syndrome (ARDS) is a well-described complication of critical illness. We hy-pothesized that rates of comorbid diseases in a population may influence the risk for developing ARDS in trauma pa-tients. This can help plan medical responses. METHODS: Patients from the 2017 National Trauma Databank were analyzed. Inclusion criteria were an injury sever-ity score (ISS) of ≥ 2 and 1 or more documented days of mechanical ventilation. Data were analyzed using χ2, Student's t test, Mann-Whitney U test, or logistic regression as indicated. RESULTS: Diabetes (odds ratio [OR] 1.33, 95 percent confidence interval [CI] 1.17-1.52), smoking (OR 1.26, 95 per-cent CI 1.13-1.40), transfusion (OR 1.20, 95 percent CI 1.09-1.32), ISS (OR 1.02, 95 percent CI 1.02-1.03), male gen-der (OR 1.22, 95 percent CI 1.10-1.35), decreasing Glasgow coma score (OR 1.04, 95 percent CI 1.03-1.05), and in-creasing abbreviated injury score of the thorax (OR 1.12, 95 percent CI 1.09-1.16) were associated with an increase in risk for developing ARDS. CONCLUSION: Diabetes and smoking are risk factors for developing ARDS after trauma. Medical response planning in countries with high rates of diabetes mellitus or smoking should take into account a greater need for intensive care and longer patient admissions to field hospitals.

Participation in a workplace smoking cessation program incentivized by lowering the cost of health care coverage: Findings from the LLUH BREATHE cohort.

INTRODUCTION: Systematic analyses of workplace smoking cessation programs indicate that efficacy can be enhanced by using incentives. There is variation in the type of incentives used and their effect on participation and efficacy. The aim of our study was to examine whether lowering employee health plan costs (employee contributions, co-pays) encourage employee smokers to participate in workplace smoking cessation. METHODS: We conducted a 2014-2015 prospective cohort study of 415 employee smokers of Loma Linda University Health (LLUH). The employees were offered participation in a workplace smoking cessation program (LLUH BREATHE Initiative) with the incentive of enrollment in an employer-provided health plan that had a 50% lower employee monthly contribution and co-payment relative to the employer-provided health plan for non-participants. Participation rates and variables associated with participation were analyzed. RESULTS: In the LLUH BREATHE cohort, we found a very high rate of participation (72.7%; 95% CI: 69-77%) in workplace smoking cessation that was encouraged by a lower out-of-pocket health plan cost for the participating employee and/or spouse. Participation did, however, vary by gender and spouse, whereby female employee households with a qualifying smoker were more than two times more likely (employee: OR=2.89, 95% CI: 1.59-5.24; or spouse: OR=2.71, 95% CI: 1.47-5.00) to participate in smoking cessation than male employee households. The point prevalence, at four months, of abstinence from smoking among the participants was 48% (95% CI: 42-54%). CONCLUSIONS: Our findings indicate that a workplace smoking cessation program that uses a novel reward-based incentive of lower out-of-pocket health plan costs results in a participation rate that is much higher than US norms.

Association Between Asthma Hospital Visits and Ozone Concentration in Maricopa County, Arizona (2007-2012).

The objective of this study was to evaluate the association of asthma hospital visits with ozone concentrations in Maricopa County, Arizona. We used time plots and distributed lag nonlinear models to achieve these objectives while accounting for some potential confounders including temperature and day of the week. A total of 90,381 asthma hospitalizations were retrieved from the dataset (daily median = 39, range: 8-122). Asthma hospitalizations were highest in 2008 (16,949), during the months of November through December, and lowest in 2011 (13,213), during the months of June through July. By contrast, the average daily ozone concentration ranged from 27.05 parts per billion (ppb) in 2012 to 30.15 ppb in 2008 and from 13.96 ppb in December to 40.58 ppb in May. The association between asthma hospitalizations (relative risk [RR/per 10 ppb increase of ozone]) start at -1.046 (95% confidence interval [1.029, 1.064] at lag 0) and gradually decrease over several days. Our findings suggest exposure to ozone is associated with increased RR of asthma hospital visits in Maricopa County lasting several days. This study used recently developed methods that are freely available and could be used to evaluate other health events that are measured over time.

Esophagogastric Fistula Caused by an Angelchik Antireflux Prosthesis.

The Angelchik prosthesis is an antireflux device that was popular in the 1980s for treatment of refractory gastroesophageal reflux disease (GERD). We present a patient who developed a gastroesophageal fistula 17 years after Angelchik prosthesis placement. The incidence of late complications continues to grow, and clinicians should consider device malfunction in patients with history of Angelchik placement presenting with abdominal symptoms.

Burkholderia pseudomallei Capsular Polysaccharide Recognition by a Monoclonal Antibody Reveals Key Details toward a Biodefense Vaccine and Diagnostics against Melioidosis.

Burkholderia pseudomallei is the bacterium responsible for melioidosis, an infectious disease with high mortality rates. Since melioidosis is a significant public health concern in endemic regions and the organism is currently classified as a potential biothreat agent, the development of effective vaccines and rapid diagnostics is a priority. The capsular polysaccharide (CPS) expressed by B. pseudomallei is a highly conserved virulence factor and a protective antigen. Because of this, CPS is considered an attractive antigen for use in the development of both vaccines and diagnostics. In the present study, we describe the interactions of CPS with the murine monoclonal antibody (mAb) 4C4 using a multidisciplinary approach including organic synthesis, molecular biology techniques, surface plasmon resonance, and nuclear magnetic spectroscopy. Using these methods, we determined the mode of binding between mAb 4C4 and native CPS or ad hoc synthesized capsular polysaccharide fragments. Interestingly, we demonstrated that the O-acetyl moiety of CPS is essential for the interaction of the CPS epitope with mAb 4C4. Collectively, our results provide important insights into the structural features of B. pseudomallei CPS that enable antibody recognition that may help the rational design of CPS-based vaccine candidates. In addition, our findings confirm that the mAb 4C4 is suitable for use in an antibody-based detection assay for diagnosis of B. pseudomallei infections.

Immunoassay for Capsular Antigen of Bacillus anthracis Enables Rapid Diagnosis in a Rabbit Model of Inhalational Anthrax.

Inhalational anthrax is a serious biothreat. Effective antibiotic treatment of inhalational anthrax requires early diagnosis; the further the disease has progressed, the less the likelihood for cure. Current means for diagnosis such as blood culture require several days to a result and require advanced laboratory infrastructure. An alternative approach to diagnosis is detection of a Bacillus anthracis antigen that is shed into blood and can be detected by rapid immunoassay. The goal of the study was to evaluate detection of poly-γ-D-glutamic acid (PGA), the capsular antigen of B. anthracis, as a biomarker surrogate for blood culture in a rabbit model of inhalational anthrax. The mean time to a positive blood culture was 26 ± 5.7 h (mean ± standard deviation), whereas the mean time to a positive ELISA was 22 ± 4.2 h; P = 0.005 in comparison with blood culture. A lateral flow immunoassay was constructed for detection of PGA in plasma at concentrations of less than 1 ng PGA/ml. Use of the lateral flow immunoassay for detection of PGA in the rabbit model found that antigen was detected somewhat earlier than the earliest time point at which the blood culture became positive. The low cost, ease of use, and rapid time to result of the lateral flow immunoassay format make an immunoassay for PGA a viable surrogate for blood culture for detection of infection in individuals who have a likelihood of exposure to B. anthracis.

Development of a prototype lateral flow immunoassay (LFI) for the rapid diagnosis of melioidosis.

Burkholderia pseudomallei is a soil-dwelling bacterium and the causative agent of melioidosis. Isolation of B. pseudomallei from clinical samples is the "gold standard" for the diagnosis of melioidosis; results can take 3-7 days to produce. Alternatively, antibody-based tests have low specificity due to a high percentage of seropositive individuals in endemic areas. There is a clear need to develop a rapid point-of-care antigen detection assay for the diagnosis of melioidosis. Previously, we employed In vivo Microbial Antigen Discovery (InMAD) to identify potential B. pseudomallei diagnostic biomarkers. The B. pseudomallei capsular polysaccharide (CPS) and numerous protein antigens were identified as potential candidates. Here, we describe the development of a diagnostic immunoassay based on the detection of CPS. Following production of a CPS-specific monoclonal antibody (mAb), an antigen-capture immunoassay was developed to determine the concentration of CPS within a panel of melioidosis patient serum and urine samples. The same mAb was used to produce a prototype Active Melioidosis Detect Lateral Flow Immunoassay (AMD LFI); the limit of detection of the LFI for CPS is comparable to the antigen-capture immunoassay (∼0.2 ng/ml). The analytical reactivity (inclusivity) of the AMD LFI was 98.7% (76/77) when tested against a large panel of B. pseudomallei isolates. Analytical specificity (cross-reactivity) testing determined that 97.2% of B. pseudomallei near neighbor species (35/36) were not reactive. The non-reactive B. pseudomallei strain and the reactive near neighbor strain can be explained through genetic sequence analysis. Importantly, we show the AMD LFI is capable of detecting CPS in a variety of patient samples. The LFI is currently being evaluated in Thailand and Australia; the focus is to optimize and validate testing procedures on melioidosis patient samples prior to initiation of a large, multisite pre-clinical evaluation.

Constant domains influence binding of mouse-human chimeric antibodies to the capsular polypeptide of Bacillus anthracis.

Our laboratory previously described the binding characteristics of the murine IgG3 monoclonal antibody (MuAb) F26G3. This antibody binds the poly-glutamic acid capsule (PGA) of Bacillus anthracis, an essential virulence factor in the progression of anthrax. F26G3 IgG3 MuAb binds PGA with a relatively high functional affinity (10 nM), produces a distinct "rim" quellung reaction, and is protective in a murine model of pulmonary anthrax. This study engineered an IgG subclass family of F26G3 mouse-human chimeric antibodies (ChAb). The F26G3 ChAbs displayed 9- to 20-fold decreases in functional affinity, as compared with the parent IgG3 MuAb. Additionally, the quellung reactions that were produced by the ChAbs all differed from the parent IgG3 MuAb in that they appeared "puffy" in nature. This study demonstrates that human constant domains may influence multiple facets of antibody binding to microbial capsular antigens despite their spatial separation from the traditional antigen-binding site.

IgG subclass and heavy chain domains contribute to binding and protection by mAbs to the poly γ-D-glutamic acid capsular antigen of Bacillus anthracis.

Bacterial capsules are common targets for antibody-mediated immunity. The capsule of Bacillus anthracis is unusual among capsules because it is composed of a polymer of poly-γ-d-glutamic acid (γdPGA). We previously generated murine IgG3 monoclonal antibodies (mAbs) to γdPGA that were protective in a murine model of pulmonary anthrax. IgG3 antibodies are characteristic of the murine response to polysaccharide antigens. The goal of the present study was to produce subclass switch variants of the γdPGA mAbs (IgG3 → IgG1 → IgG2b → IgG2a) and assess the contribution of subclass to antibody affinity and protection. Subclass switch antibodies had identical variable regions but differed in their heavy chains. The results showed that a switch from the protective IgG3 to IgG1, IgG2b or IgG2a was accompanied by i) a loss of protective activity ii) a change in mAb binding to the capsular matrix, and iii) a loss of affinity. These results identify a role for the heavy chain constant region in mAb binding. Hybrid mAbs were constructed in which the CH1, CH2 or CH3 heavy chain constant domains from a non-protective, low binding IgG2b mAb were swapped into the protective IgG3 mAb. The IgG3 mAb that contained the CH1 domain from IgG2b showed no loss of affinity or protection. In contrast, swapping the CH2 or CH3 domains from IgG2b into IgG3 produced a reduction in affinity and a loss of protection. These studies identify a role for the constant region of IgG heavy chains in affinity and protection against an encapsulated bacterial pathogen.

Stereo-selective binding of monoclonal antibodies to the poly-γ-D-glutamic acid capsular antigen of Bacillus anthracis.

Bacillus anthracis is surrounded by an anti-phagocytic capsule that is entirely composed of γ-linked D-glutamic acid (γDPGA). γDPGA is required for virulence and is produced in large quantities following spore germination. We have previously described the isolation of several γDPGA-reactive mAbs. The reagents are effective in both immunoprotection and diagnostic applications. The current work was done to further investigate the specificity of γDPGA-reactive mAbs. The specificity of each mAb was characterized using surface plasmon resonance. Our results indicate that each mAb is stereoselective for binding to D-glutamic acid oligomers, but to varying degrees. In particular, mAb F26G3 is highly selective for γDPGA; alterations in stereochemistry disrupted recognition. These differences in mAb reactivity suggest that binding of γDPGA by mAb F26G3 is more specific than non-directional ionic interactions between a negatively charged antigen and a positively charged antibody.

Data File Standard for Flow Cytometry, version FCS 3.1.

The flow cytometry data file standard provides the specifications needed to completely describe flow cytometry data sets within the confines of the file containing the experimental data. In 1984, the first Flow Cytometry Standard format for data files was adopted as FCS 1.0. This standard was modified in 1990 as FCS 2.0 and again in 1997 as FCS 3.0. We report here on the next generation flow cytometry standard data file format. FCS 3.1 is a minor revision based on suggested improvements from the community. The unchanged goal of the standard is to provide a uniform file format that allows files created by one type of acquisition hardware and software to be analyzed by any other type.The FCS 3.1 standard retains the basic FCS file structure and most features of previous versions of the standard. Changes included in FCS 3.1 address potential ambiguities in the previous versions and provide a more robust standard. The major changes include simplified support for international characters and improved support for storing compensation. The major additions are support for preferred display scale, a standardized way of capturing the sample volume, information about originality of the data file, and support for plate and well identification in high throughput, plate based experiments. Please see the normative version of the FCS 3.1 specification in Supporting Information for this manuscript (or at http://www.isac-net.org/ in the Current standards section) for a complete list of changes.

The water-polo intermittent shuttle test: a match-fitness test for water-polo players.

PURPOSE: To develop and validate an intermittent match-fitness test for water-polo players. METHODS: Eight male junior players performed the Water Polo Intermittent Shuttle Test (WIST) twice to assess test reliability. To assess test sensitivity and validity, 104 male and female players from different competition standards and playing positions were tested. Eighteen players performed the WIST 5 times throughout a season to track fitness changes. Twelve players performed the WIST 48 hours before 4 consecutive National League games, and coaches awarded individual match-fitness scores based on game performances to assess the relationship between match fitness and test results. Heart rate (HR) and blood lactate (La(blood)) were measured during and after each test, respectively. RESULTS: Test-retest performance values were 216 +/- 90 vs 229 +/- 96 m (r = .98, P = .0001, coefficient of variation [CV] = 5.4%), peak HR 190 +/- 8 vs 192 +/- 10 bpm (r = .96, P = .0002, CV = 1.2%), and La(blood) 7.0 +/- 1.8 vs 6.4 +/- 1.6 mmol/L (r = .84, P = .0092, CV = 8.8%). Significant differences were observed among different standards of play (range junior regional females 102 +/- 10 m, senior international males 401 +/- 30 m) and playing positions (field players 305 +/- 154 m, center forwards 255 +/- 118, goal keepers 203 +/- 135 m). Test performance was lower in the early season (344 +/- 118 m) than the remainder of the season (range 459 +/- 138 to 550 +/- 176 m). WIST performance and match-fitness scores correlated for all field players (r = .57, P = .054) but more highly for field players other than center forwards (r = .83, P = .0027). CONCLUSIONS: The WIST is a reliable, sensitive, and valid match-fitness test for water-polo players. It could become a useful tool to assess the effects of different interventions on match fitness.

The wages of healing: ethical issues in the compensation of physicians.

Though not often addressed as a moral issue, the incomes of physicians and other health professionals are fraught with ethical significance. While there is no perfect means of determining appropriate income level, possible approaches include market worth, comparable worth, societal worth, and fairness. In fact, however, income is not the only reward that makes a medical career worthwhile, and a preoccupation with wealth can impede the pursuit of other equally worthy objectives. In an era when financial incentives and disincentives tend to receive the lionshare of attention, it is vital that physicians continue to bear these other objectives in mind.

Quantitative vibration threshold testing in carpal tunnel syndrome: analysis strategies for optimizing reliability.

Tuning forks and electronic vibrometers have been used to quantify vibration sensation thresholds, which are thought to be affected early in carpal tunnel syndrome (CTS). The purpose of this study was to identify a reliable testing procedure for a newly designed, computer-controlled vibrometer (PCV50; Ztech, Salt Lake City, UT). Fifty-two patients (mean age 48+/-8 years) with electromyographically confirmed CTS were tested on one occasion. The computer-controlled vibrometer, with a fixed frequency of 50 Hz, used stepwise changes in amplitude to determine vibration sensation threshold. Each patient had three vibrometer measures (trials) taken on the pulp of the third digit of their right and left hands during the first test session and were retested by a single repetition 40 to 60 minutes later (retest). Intraclass correlation coefficients (ICCs) were used to examine several data analysis strategies. The strategy that generated the highest ICCs for both the right and left hands assumed that the first trial was a learning or practice attempt, and compared the average of the second and third trials with the score from the second session (ICC=0.86 and 0.89, respectively). The computer-controlled vibrometer offered an easily administered, quantitative, and comfortable means to assess median nerve function. Using this reliable testing procedure will allow for additional investigations to determine its usefulness in the early detection and accurate quantification of CTS-related impairment.