Subtyping clinical specimens of influenza A virus by use of a simple method to amplify RNA targets.

This work presents the clinical application of a robust and unique approach for RNA amplification, called a simple method for amplifying RNA targets (SMART), for the detection and identification of subtypes of H1N1 pandemic, H1N1 seasonal, and H3N2 seasonal influenza virus. While all the existing amplification techniques rely on the diffusion of two molecules to complex RNA structures, the SMART achieves fast and efficient amplification via single-molecule diffusion. The SMART utilizes amplifiable single-stranded DNA (ssDNA) probes, which serve as reporter molecules for capturing specific viral RNA (vRNA) sequences and are subsequently separated on a microfluidic chip under zero-flow conditions. The probe amplification and detection are performed using an isothermal (41°C) amplification scheme via a modified version of nucleic acid sequence-based amplification (NASBA). In our study, 116 consecutive, deidentified, clinical nasopharyngeal swab samples were analyzed independently in a blinded fashion using the SMART, reverse transcription-PCR (RT-PCR), antigen (Ag) testing, and viral culture. The SMART was shown to have a limit of detection (LOD) of approximately 10(5) vRNA copies/ml, corresponding with a time-to-positivity (TTP) value of 70 min for real-time detection. The SMART correctly detected influenza virus in 98.3% of the samples with a subtyping accuracy of 95.7%. This work demonstrates that the SMART represents a highly accurate diagnostic platform for the detection and subtyping of influenza virus in clinical specimens and offers significant advantages over the current commercially available diagnostic tools.

Sepsis and scientific revolutions.

HYPOTHESIS: The therapeutic approach to sepsis is following an evolutionary process of scientific discovery as articulated in the landmark work by Kuhn, The Structure of Scientific Revolutions, first published 50 years ago. BACKGROUND: Incremental advances, beginning with the introduction of antimicrobials and most recently highlighted by revised, evidence-based guidelines for the management of sepsis, have been accompanied by episodic paradigm shifts. Although some of these have shown success, there are numerous, noteworthy failures, largely involving immune- and coagulation-based therapeutic strategies. CONCLUSION: A sustained paradigm shift in the approach to treating sepsis has yet to emerge, but recent data suggest that an open-minded posture informed by novel pathobiologic findings may eventually bear fruit.

Novel approaches to the treatment of systemic anthrax.

Anthrax continues to generate concern as an agent of bioterrorism and as a natural cause of sporadic disease outbreaks. Despite the use of appropriate antimicrobial agents and advanced supportive care, the mortality associated with the systemic disease remains high. This is primarily due to the pathogenic exotoxins produced by Bacillus anthracis as well as other virulence factors of the organism. For this reason, new therapeutic strategies that target events in the pathogenesis of anthrax and may potentially augment antimicrobials are being investigated. These include anti-toxin approaches, such as passive immune-based therapies; non-antimicrobial drugs with activity against anthrax toxin components; and agents that inhibit binding, processing, or assembly of toxins. Adjunct therapies that target spore germination or downstream events in anthrax intoxication are also under investigation. In combination, these modalities may enhance the management of systemic anthrax.

A half-century of meningococcal vaccines.

The first safe and effective vaccine for the prevention of invasive meningococcal disease was created fifty years ago. The vaccine employed a novel platform, polysaccharide capsular antigen, based on the discovery that anticapsular antibody conferred protective immunity in humans. As with most new paradigms in vaccinology, it derived from important basic research from other scientific disciplines over the preceding years. The success of the first monovalent polysaccharide vaccine in nearly eliminating invasive meningococcal disease in military settings led to accelerated advances in polysaccharide vaccine development against other serogroups of meningococcus and other encapsulated pathogens. As gaps in vaccine efficacy arose over the past half-century, new vaccine technologies and approaches were developed to address the challenges. Several of these, including conjugate vaccines and "reverse vaccinology" led to other novel, successful vaccines that have had a significant, favorable global impact on invasive meningococcal disease. The history of meningococcal vaccine discovery may provide insights into the future of vaccine efforts against other infectious threats.

An Integrated Mobile Acute Care Service Enhances Value.

Reducing the cost of care while enhancing its quality and experience are essential components to success in value-based care. Because emergency department (ED) and hospital settings represent high-cost environments, the authors sought to reduce their unnecessary use by deploying a novel care delivery service that offers mobile, on-demand care for high-acuity conditions in patient homes. This study is a retrospective quality improvement evaluation of the initial year of the mobile acute care model in a health system with a substantial penetration of value-based care. Although all patients were eligible for mobile services as clinically indicated, those in accountable care organizations were prioritized by the care management teams. A variety of operational, clinical, and financial metrics were assessed to determine the program's performance and value. There were 3436 patient encounters during the study period, a utilization rate of 71% that trended upward throughout the year. Of these visits, 44% involved patients in value-based payment models; 80% of these represented patients in Medicare risk agreements. Throughout the year, progressively improving operational and clinical performance were observed, as were consistently high patient satisfaction scores. An estimated 63.8% of total mobile visits resulted in ED avoidance; 21.6% were emergency medical transport avoidant; 14.1% led to avoided hospital observation or inpatient stays. Patients were highly satisfied with the service. In-home mobile care for high-acuity illness can prevent unnecessary ED and hospital use for some patients and is associated with high patient satisfaction. Acute mobile care is a useful component of a value-based care strategy.

Direct sequence detection of structured h5 influenza viral RNA.

We describe the development of sequence-specific molecular beacons (dual-labeled DNA probes) for identification of the H5 influenza subtype, cleavage motif, and receptor specificity when hybridized directly with in vitro transcribed viral RNA (vRNA). The cloned hemagglutinin segment from a highly pathogenic H5N1 strain, A/Hanoi/30408/2005(H5N1), isolated from humans was used as template for in vitro transcription of sense-strand vRNA. The hybridization behavior of vRNA and a conserved subtype probe was characterized experimentally by varying conditions of time, temperature, and Mg2+ to optimize detection. Comparison of the hybridization rates of probe to DNA and RNA targets indicates that conformational switching of influenza RNA structure is a rate-limiting step and that the secondary structure of vRNA dominates the binding kinetics. The sensitivity and specificity of probe recognition of other H5 strains was calculated from sequence matches to the National Center for Biotechnology Information influenza database. The hybridization specificity of the subtype probes was experimentally verified with point mutations within the probe loop at five locations corresponding to the other human H5 strains. The abundance frequencies of the hemagglutinin cleavage motif and sialic acid recognition sequences were experimentally tested for H5 in all host viral species. Although the detection assay must be coupled with isothermal amplification on the chip, the new probes form the basis of a portable point-of-care diagnostic device for influenza subtyping.

A simulation-based biodefense and disaster preparedness curriculum for internal medicine residents.

AIMS: Disaster and bioterrorism preparedness is poorly integrated into the curricula of internal medicine residency programs. Given that victims may present to a variety of healthcare venues, including primary care practices, inpatient hospital wards, and intensive care units, we developed a curriculum to address this need. METHODS: The curriculum consisted of four didactic sessions with supplemental readings covering biologic, chemical, and radiologic agents, as well as public health infrastructure. All 30 internal medicine resident participants also underwent a four hour training seminar at a high fidelity human simulation center. Instruction included the use of personal protective equipment (PPE)and participation in simulated scenarios utilizing technologically sophisticated mannequins with monitoring and interactive capability. Sessions were videotaped, reviewed with participants, and followed by self-evaluation and constructive feedback. RESULTS: Compared to a control group of residents who did not undergo training, the participants' level of knowledge was significantly better, with mean objective test scores of 66.8%+/-11.8% SD vs. 50%+/-13.1% SD, p < 0.0001. Although there was a trend toward increasing knowledge with increasing level of training in the control group, this difference was not significant. Subjective preparedness was also significantly better in the intervention group (p < 0.0001). Objective improvements were not maintained after one year. CONCLUSIONS: In this pilot study, a disaster-preparedness curriculum including simulation-based training had a positive effect on residents' knowledge base and ability to respond to disaster. However, this effect had diminished after one year, indicating the need for reinforcement at regular intervals.

A Predictive Model Facilitates Early Recognition of Spinal Epidural Abscess in Adults.

INTRODUCTION: Spinal epidural abscess (SEA), a highly morbid and potentially lethal deep tissue infection of the central nervous system has more than tripled in incidence over the past decade. Early recognition at the point of initial clinical presentation may prevent irreversible neurologic injury or other serious, adverse outcomes. To facilitate early recognition of SEA, we developed a predictive scoring model. METHODS: Using data from a 10-year, retrospective, case-control study of adults presenting for care at a tertiary-care, regional, academic medical center, we used the Integrated Discrimination Improvement Index (IDI) to identify candidate discriminators and created a multivariable logistic regression model, refined based on p-value significance. We selected a cutpoint that optimized sensitivity and specificity. RESULTS: The final multivariable logistic regression model based on five characteristics -patient age, fever and/or rigor, antimicrobial use within 30 days, back/neck pain, and injection drug use - shows excellent discrimination (AUC 0.88 [95% confidence interval {0.84, 0.92}]). We used the model's ß coefficients to develop a scoring system in which a cutpoint of six correctly identifies cases 89% of the time. Bootstrapped validation measures suggest this model will perform well across samples drawn from this population. CONCLUSION: Our predictive scoring model appears to reliably discriminate patients who require emergent spinal imaging upon clinical presentation to rule out SEA and should be used in conjunction with clinical judgment.

Decreasing Emergency Department Walkout Rate and Boarding Hours by Improving Inpatient Length of Stay.

INTRODUCTION: Patient progress, the movement of patients through a hospital system from admission to discharge, is a foundational component of operational effectiveness in healthcare institutions. Optimal patient progress is a key to delivering safe, high-quality and high-value clinical care. The Baystate Patient Progress Initiative (BPPI), a cross-disciplinary, multifaceted quality and process improvement project, was launched on March 1, 2014, with the primary goal of optimizing patient progress for adult patients. METHODS: The BPPI was implemented at our system's tertiary care, academic medical center, a high-volume, high-acuity hospital that serves as a regional referral center for western Massachusetts. The BPPI was structured as a 24-month initiative with an oversight group that ensured collaborative goal alignment and communication of operational teams. It was organized to address critical aspects of a patient's progress through his hospital stay and to create additional inpatient capacity. The specific goal of the BPPI was to decrease length of stay (LOS) on the inpatient adult Hospital Medicine service by optimizing an interdisciplinary plan of care and promoting earlier departure of discharged patients. Concurrently, we measured the effects on emergency department (ED) boarding hours per patient and walkout rates. RESULTS: The BPPI engaged over 300 employed clinicians and non-clinicians in the work. We created increased inpatient capacity by implementing daily interdisciplinary bedside rounds to proactively address patient progress; during the 24 months, this resulted in a sustained rate of discharge orders written before noon of more than 50% and a decrease in inpatient LOS of 0.30 days (coefficient: -0.014, 95% CI [-0.023, -0.005] P< 0.005). Despite the increase in ED patient volumes and severity of illness over the same time period, ED boarding hours per patient decreased by approximately 2.1 hours (coefficient: -0.09; 95% CI [-0.15, -0.02] P = 0.007). Concurrently, ED walkout rates decreased by nearly 32% to a monthly mean of 0.4 patients (coefficient: 0.4; 95% CI [-0.7, -0.1] P= 0.01). CONCLUSION: The BPPI realized significant gains in patient progress for adult patients by promoting earlier discharges before noon and decreasing overall inpatient LOS. Concurrently, ED boarding hours per patient and walkout rates decreased.

Spinal Epidural Abscess in Adults: A 10-Year Clinical Experience at a Tertiary Care Academic Medical Center.

Background. Delayed recognition of spinal epidural abscess (SEA) contributes to poor outcomes from this highly morbid and potentially lethal infection. We performed a case-control study in a regional, high-volume, tertiary care, academic medical center over the years 2005-2015 to assess the potential changing epidemiology, clinical and laboratory manifestations, and course of this disorder and to identify factors that might lead to early identification of SEA. Methods. Diagnostic billing codes consistent with SEA were used to identify inpatient admissions for abstraction. Subjects were categorized as cases or controls based on the results of spinal imaging studies. Characteristics were compared using Fisher's exact or Kruskal-Wallis tests. All P values were 2-sided with a critical threshold of <.05. Results. We identified 162 cases and 88 controls during the study period. The incidence of SEA increased from 2.5 to 8.0 per 10 000 admissions, a 3.3-fold change from 2005 to 2015 (P < .001 for the linear trend). Compared with controls, cases were significantly more likely to have experienced at least 1 previous healthcare visit or received antimicrobials within 30 days of admission; to have comorbidities of injection drug use, alcohol abuse, or obesity; and to manifest fever or rigors. Cases were also more likely to harbor coinfection at a noncontiguous site. When available, inflammatory markers were noted to be markedly elevated in cases. Focal neurologic deficits were seen with similar frequencies in both groups. Conclusions. Based on our analysis, it appears that selected factors noted at the time of clinical presentation may facilitate early recognition of SEA.

History of U.S. military contributions to the study of vaccines against infectious diseases.

The U.S. military has a long and illustrious history of involvement with vaccines against infectious diseases. For more than 200 years, the military has been actively engaged in vaccine research and has made many important contributions to the development of these products for use in disease prevention and control. Through the efforts of military researchers, numerous serious threats to the health of American troops and their families have been mitigated.

Promoting high value inpatient care via a coaching model of structured, interdisciplinary team rounds.

The professional development of early career hospital physicians may be improved by embedding an experienced physician in a coaching role during structured, interdisciplinary team rounds. This article gives a descriptive report of such a model and discusses how it may promote delivery of high-value care to adult inpatients.

Human immunodeficiency virus (HIV)-specific antibody in cervicovaginal lavage specimens obtained from women infected with HIV type 1.

To evaluate correlates of anti-human immunodeficiency virus (HIV) type 1 (HIV-1) immunoglobulin (Ig) in the genital tract, anti-HIV-gp120 IgA and IgG titers in cervicovaginal lavage specimens obtained from 104 HIV-1-infected women were measured by enzyme-linked immunosorbent assay. Overall, 24% and 94% of women had detectable anti-gp120 IgA and IgG, respectively. CD4 cell count correlated negatively with total IgA concentration (r=-0.301; P=.0027) and positively with specific IgA activity (anti-gp120 IgA titer/total IgA concentration, r=0.306; P=.0023). Women with bacterial vaginosis had 5-fold lower anti-gp120 IgG titer (P=.0042), 5-fold lower total IgG concentration (P< or =.0001), and 4-fold higher specific IgG activity (P=.0474) compared with women who did not have bacterial vaginosis. Enhanced understanding of correlates of mucosal immunity to HIV-1 may assist in the design of vaccine strategies or in the prevention of vertical transmission of HIV-1.

The "forgotten zone": acquired disorders of the trachea in adults.

The upper airway is generally defined as the air passage segment that extends between the naso- or oropharynx and the carina. The longest segment of the upper airway-the trachea-begins at the inferior portion of the larynx and extends to the branch point of the main carina. The trachea has the potential to be a "forgotten zone" in differential diagnoses, as pathological processes involving this portion may not receive prominent clinical consideration in disorders presenting with respiratory symptoms and signs. Unlike the oropharynx, this anatomical area is beyond visualization on routine inspection; unlike the mediastinum and lung fields, it is a potential "blind spot" on initial, plain radiographic examination of the chest. Nonetheless, the adult trachea is affected by a number of primary disorders and is also a target organ of a variety of systemic diseases. This review will focus on both primary and systemic diseases involving the adult trachea with specific attention to their clinical manifestations and diagnostic hallmarks.

Microdroplet sandwich real-time rt-PCR for detection of pandemic and seasonal influenza subtypes.

As demonstrated by the recent 2012/2013 flu epidemic, the continual emergence of new viral strains highlights the need for accurate medical diagnostics in multiple community settings. If rapid, robust, and sensitive diagnostics for influenza subtyping were available, it would help identify epidemics, facilitate appropriate antiviral usage, decrease inappropriate antibiotic usage, and eliminate the extra cost of unnecessary laboratory testing and treatment. Here, we describe a droplet sandwich platform that can detect influenza subtypes using real-time reverse-transcription polymerase chain reaction (rtRT-PCR). Using clinical samples collected during the 2010/11 season, we effectively differentiate between H1N1p (swine pandemic), H1N1s (seasonal), and H3N2 with an overall assay sensitivity was 96%, with 100% specificity for each subtype. Additionally, we demonstrate the ability to detect viral loads as low as 10(4) copies/mL, which is two orders of magnitude lower than viral loads in typical infected patients. This platform performs diagnostics in a miniaturized format without sacrificing any sensitivity, and can thus be easily developed into devices which are ideal for small clinics and pharmacies.