Antigen-based multiplex strategies to discriminate SARS-CoV-2 natural and vaccine induced immunity from seasonal human coronavirus humoral responses.

Sensitive and specific SARS-CoV-2 antibody assays remain critical for community and hospital-based SARS-CoV-2 sero-surveillance. With the rollout of SARS-CoV-2 vaccines, such assays must be able to distinguish vaccine from natural immunity to SARS-CoV-2 and related human coronaviruses. Here, we developed and implemented multiplex microsphere-based immunoassay strategies for COVD-19 antibody studies that incorporates spike protein trimers of SARS-CoV-2 and the endemic seasonal human coronaviruses (HCoV), enabling high throughout measurement of pre-existing cross-reactive antibodies. We varied SARS-CoV-2 antigen compositions within the multiplex assay, allowing direct comparisons of the effects of spike protein, receptor-binding domain protein (RBD) and nucleocapsid protein (NP) based SARS-CoV-2 antibody detection. Multiplex immunoassay performance characteristics are antigen-dependent, and sensitivities and specificities range 92-99% and 94-100%, respectively, for human subject samples collected as early as 7-10 days from symptom onset. SARS-CoV-2 spike and RBD had a strong correlative relationship for the detection of IgG. Correlation between detectable IgG reactive with spike and NP also had strong relationship, however, several PCR-positive and spike IgG-positive serum samples were NP IgG-negative. This spike and NP multiplex immunoassay has the potential to be useful for differentiation between vaccination and natural infection induced antibody responses. We also assessed the induction of de novo SARS-CoV-2 IgG cross reactions with SARS-CoV and MERS-CoV spike proteins. Furthermore, multiplex immunoassays that incorporate spike proteins of SARS-CoV-2 and HCoVs will permit investigations into the influence of HCoV antibodies on COVID-19 clinical outcomes and SARS-CoV-2 antibody durability.

A betacoronavirus multiplex microsphere immunoassay detects early SARS-CoV-2 seroconversion and antibody cross reactions.

Sensitive and specific SARS-CoV-2 antibody assays remain critical for community and hospital-based SARS-CoV-2 surveillance. Here, we developed and applied a multiplex microsphere-based immunoassay (MMIA) for COVD-19 antibody studies that incorporates spike protein trimers of SARS-CoV-2, SARS-CoV-1, MERS-CoV, and the seasonal human betacoronaviruses, HCoV-HKU1 and HCoV-OC43, that enables measurement of off-target pre-existing cross-reactive antibodies. The MMIA performances characteristics are: 98% sensitive and 100% specific for human subject samples collected as early as 10 days from symptom onset. The MMIA permitted the simultaneous identification of SARS-CoV-2 seroconversion and the induction of SARS-CoV-2 IgG antibody cross reactions to SARS-CoV-1 and MERS-CoV. Further, synchronous increases of HCoV-OC43 IgG antibody levels was detected with SARS-CoV-2 seroconversion in a subset of subjects for whom early infection sera were available prior to their SARS-CoV-2 seroconversion, suggestive of an HCoV-OC43 memory response triggered by SARS-CoV-2 infection.

Orbiting and In-Situ Lidars for Earth and Planetary Applications.

At NASA Goddard Space Flight Center, we have been developing spaceborne lidar instruments for space sciences. We have successfully flown several missions in the past based on mature diode pumped solid-state laser transmitters. In recent years, we have been developing advanced laser technologies for applications such as laser spectroscopy, laser communications, and interferometry. In this article, we will discuss recent experimental progress on these systems and instrument prototypes for ongoing development.

Accuracy of Dynamic Navigation for Dental Implant Placement-Model-Based Evaluation.

The purpose of this model-based study was to determine the accuracy of placing dental implants using a new dynamic navigation system. This investigation focuses on measurements of overall accuracy for implant placement relative to the virtual plan in both dentate and edentulous models, and provides a comparison with a meta-analysis of values reported in the literature for comparable static guidance, dynamic guidance, and freehand placement studies. This study involves 1 surgeon experienced with dynamic navigation placing implants in models under clinical simulation using a dynamic navigation system (X-Guide, X-Nav Technologies, LLC, Lansdale, Pa) based on optical triangulation tracking. Virtual implants were placed into planned sites using the navigation system computer. Post-implant placement cone-beam scans were taken. These scans were mesh overlaid with the virtual plan and used to determine deviations from the virtual plan. The primary outcome variables were platform and angular deviations comparing the actual placement to the virtual plan. The angular accuracy of implants delivered using the tested device was 0.89° ± 0.35° for dentate case types and 1.26° ± 0.66° for edentulous case types, measured relative to the preoperative implant plan. Three-dimensional positional accuracy was 0.38 ± 0.21 mm for dentate and 0.56 ± 0.17 mm for edentulous, measured from the implant apex.

Toxoplasma gondii seroprevalence: 30-year trend in an HIV-infected US military cohort.

To determine if Toxoplasma gondii IgG antibody prevalence is declining in HIV-infected persons, we analyzed data (1984-2013) from the US Military HIV Natural History Study. We found that T. gondii seroprevalence at enrollment was associated with age and decreased significantly after 1995 (P=0.004), similar to the general US population.

Interactive CT-video registration for the continuous guidance of bronchoscopy.

Bronchoscopy is a major step in lung cancer staging. To perform bronchoscopy, the physician uses a procedure plan, derived from a patient's 3D computed-tomography (CT) chest scan, to navigate the bronchoscope through the lung airways. Unfortunately, physicians vary greatly in their ability to perform bronchoscopy. As a result, image-guided bronchoscopy systems, drawing upon the concept of CT-based virtual bronchoscopy (VB), have been proposed. These systems attempt to register the bronchoscope's live position within the chest to a CT-based virtual chest space. Recent methods, which register the bronchoscopic video to CT-based endoluminal airway renderings, show promise but do not enable continuous real-time guidance. We present a CT-video registration method inspired by computer-vision innovations in the fields of image alignment and image-based rendering. In particular, motivated by the Lucas-Kanade algorithm, we propose an inverse-compositional framework built around a gradient-based optimization procedure. We next propose an implementation of the framework suitable for image-guided bronchoscopy. Laboratory tests, involving both single frames and continuous video sequences, demonstrate the robustness and accuracy of the method. Benchmark timing tests indicate that the method can run continuously at 300 frames/s, well beyond the real-time bronchoscopic video rate of 30 frames/s. This compares extremely favorably to the ≥ 1 s/frame speeds of other methods and indicates the method's potential for real-time continuous registration. A human phantom study confirms the method's efficacy for real-time guidance in a controlled setting, and, hence, points the way toward the first interactive CT-video registration approach for image-guided bronchoscopy. Along this line, we demonstrate the method's efficacy in a complete guidance system by presenting a clinical study involving lung cancer patients.

Image-guided bronchoscopy for peripheral lung lesions: a phantom study.

BACKGROUND: Ultrathin bronchoscopy guided by virtual bronchoscopy (VB) techniques show promise for the diagnosis of peripheral lung lesions. In a phantom study, we evaluated a new real-time, VB-based, image-guided system for guiding the bronchoscopic biopsy of peripheral lung lesions and compared its performance to that of standard bronchoscopy practice. METHODS: Twelve bronchoscopists of varying experience levels participated in the study. The task was to use an ultrathin bronchoscope and a biopsy forceps to localize 10 synthetically created lesions situated at varying airway depths. For route planning and guidance, the bronchoscopists employed either standard bronchoscopy practice or the real-time image-guided system. Outcome measures were biopsy site position error, which was defined as the distance from the forceps contact point to the ground-truth lesion boundary, and localization success, which was defined as a site identification having a biopsy site position error of < or = 5 mm. RESULTS: Mean (+/- SD) localization success more than doubled from 43 +/- 16% using standard practice to 94 +/- 7.9% using image guidance (p < 10(-15) [McNemar paired test]). The mean biopsy site position error dropped from 9.7 +/- 9.1 mm for standard practice to 2.2 +/- 2.3 mm for image guidance. For standard practice, localization success decreased from 56% for generation 3 to 4 lesions to 31% for generation 6 to 8 lesions and also decreased from 51% for lesions on a carina vs 23% for lesions situated away from a carina. These factors were far less pronounced when using image guidance, as follows: success for generation 3 to 4 lesions, 97%; success for generation 6 to 8 lesions, 91%; success for lesions on a carina, 98%; success for lesions away from a carina, 86%. Bronchoscopist experience did not significantly affect performance using the image-guided system. CONCLUSIONS: Real-time, VB-based image guidance can potentially far exceed standard bronchoscopy practice for enabling the bronchoscopic biopsy of peripheral lung lesions.

Interbronchoscopist variability in endobronchial path selection: a simulation study.

BACKGROUND: Endobronchial path selection is important for the bronchoscopic diagnosis of focal lung lesions. Path selection typically involves mentally reconstructing a three-dimensional path by interpreting a stack of two-dimensional (2D) axial plane CT scan sections. The hypotheses of our study about path selection were as follows: (1) bronchoscopists are inaccurate and overly confident when making endobronchial path selections based on 2D CT scan analysis; and (2) path selection accuracy and confidence improve and become better aligned when bronchoscopists employ path-planning methods based on virtual bronchoscopy (VB). METHODS: Studies of endobronchial path selection comparing three path-planning methods (ie, the standard 2D CT scan analysis and two new VB-based techniques) were performed. The task was to navigate to discrete lesions located between the third-order and fifth-order bronchi of the right upper and middle lobes. Outcome measures were the cumulative accuracy of making four sequential path selection decisions and self-reported confidence (1, least confident; 5, most confident). Both experienced and inexperienced bronchoscopists participated in the studies. RESULTS: In the first study involving a static paper-based tool, the mean (+/- SD) cumulative accuracy was 14 +/- 3% using 2D CT scan analysis (confidence, 3.4 +/- 1.3) and 49 +/- 15% using a VB-based technique (confidence, 4.2 +/- 1.1; p = 0.0001 across all comparisons). For a second study using an interactive computer-based tool, the mean accuracy was 40 +/- 28% using 2D CT scan analysis (confidence, 3.0 +/- 0.3) and 96 +/- 3% using a dynamic VB-based technique (confidence, 4.6 +/- 0.2). Regardless of the experience level of the bronchoscopist, use of the standard 2D CT scan analysis resulted in poor path selection accuracy and misaligned confidence. Use of the VB-based techniques resulted in considerably higher accuracy and better aligned decision confidence. CONCLUSIONS: Endobronchial path selection is a source of error in the bronchoscopy workflow. The use of VB-based path-planning techniques significantly improves path selection accuracy over use of the standard 2D CT scan section analysis in this simulation format.

3D CT-video fusion for image-guided bronchoscopy.

Bronchoscopic biopsy of the central-chest lymph nodes is an important step for lung-cancer staging. Before bronchoscopy, the physician first visually assesses a patient's three-dimensional (3D) computed tomography (CT) chest scan to identify suspect lymph-node sites. Next, during bronchoscopy, the physician guides the bronchoscope to each desired lymph-node site. Unfortunately, the physician has no link between the 3D CT image data and the live video stream provided during bronchoscopy. Thus, the physician must essentially perform biopsy blindly, and the skill levels between different physicians differ greatly. We describe an approach that enables synergistic fusion between the 3D CT data and the bronchoscopic video. Both the integrated planning and guidance system and the internal CT-video registration and fusion methods are described. Phantom, animal, and human studies illustrate the efficacy of the methods.