Paralytic shellfish poisoning (PSP) toxin binders for optical biosensor technology: problems and possibilities for the future: a review

This review examines the developments in optical biosensor technology, which uses the phenomenon of surface plasmon resonance, for the detection of paralytic shellfish poisoning (PSP) toxins. Optical biosensor technology measures the competitive biomolecular interaction of a specific biological recognition element or binder with a target toxin immobilised onto a sensor chip surface against toxin in a sample. Different binders such as receptors and antibodies previously employed in functional and immunological assays have been assessed. Highlighted are the difficulties in detecting this range of low molecular weight toxins, with analogues differing at four chemical substitution sites, using a single binder. The complications that arise with the toxicity factors of each toxin relative to the parent compound, saxitoxin, for the measurement of total toxicity relative to the mouse bioassay are also considered. For antibodies, the cross-reactivity profile does not always correlate to toxic potency, but rather to the toxin structure to which it was produced. Restrictions and availability of the toxins makes alternative chemical strategies for the synthesis of protein conjugate derivatives for antibody production a difficult task. However, when two antibodies with different cross-reactivity profiles are employed, with a toxin chip surface generic to both antibodies, it was demonstrated that the cross-reactivity profile of each could be combined into a single-assay format. Difficulties with receptors for optical biosensor analysis of low molecular weight compounds are discussed, as are the potential of alternative non-antibody-based binders for future assay development in this area

Analysis of spatial variability for the development of reduced lead body surface maps

The spatial frequency of measurement points from standard ECG systems lacks accuracy to diagnose local variability in cardiac activity on the torso. Body Surface Mapping (BSM) improves this accuracy, but lacks the simplicity to be implemented in clinic on a regular basis. Reduced-lead BSM system improves applicability, but currently no standardization of lead reduction has been agreed upon. This research investigates the reduction of BSMs based on Lomb-Scargle Spectral Analysis to determine an appropriate electrode positioning through spatial frequency assessment. Based on the measurement of 13 healthy volunteers, a 128 electrode system could be reduced to a 36 electrode system and an 84 electrode system for ventricular and atrial activity measurements, respectively, with up to 10% loss of the full information provided by the original body surface potential map. Further research will investigate the appropriate positions of these electrodes and the effect of lead reduction for various cardiac abnormalities

A platform to guide catheter ablation of persistent atrial fibrillation using dominant frequency mapping

Dominant frequency (DF) mapping has been widely used to study the pathophysiology of atrial fibrillation (AF). In this study, a DF mapping system was developed to guide catheter ablation on electro-physiology (EP) procedures of persistent AF patients. The proposed platform has an automated graphical user interface (GUI) that processes non-contact unipolar electrograms (EGMs) recorded simultaneously by St. Jude Ensite Velocity System and provides 3D representation of the left atrium with DF behaviours and phase analysis

Spatiotemporal behaviour of high dominant frequency during persistent atrial fibrillation

Atrial electrograms (EGMs) with high dominant frequency (DF) are believed to represent atrial substrates with periodic activation responsible for the maintenance of persistent atrial fibrillation (persAF). This study aimed to assess the DF spatiotemporal behavior using high density noncontact mapping in persAF. For 8 patients undergoing left atrial (LA) persAF ablation, 2048 noncontact virtual unipolar EGMs were simultaneously collected and after the removal of ventricular far-field activity, Fourier based spectral analysis was used to identify DF on each EGM. Atrial areas with the highest DF (HDF, DF ± 0.25 Hz) were delimited in each frame for all EGMs, creating HDF `clouds'. Cumulative HDF clouds found at each frame were counted in the 3-D LA representation. To further assess the temporal stability of the cloud, the number of EGMs not hosting any HDF was determined for each window over time. The results show the number of occurrences of HDF clouds in the LA. The temporal behavior was analyzed by counting the number of positions on the 3-D representation of the LA not visited by HDF along time. Our results show HDF in persAF is not temporally stable and spatial distribution throughout the atria suggests the existence of driver regions with very rapid and regular activity maintaining AF. Therefore mapping the cumulative HDF might be an interesting strategy for ablation

Are atrial fibrillation highest dominant frequency (HDF) areas the source of dominant excitation patterns? A left atrial panoramic view

Atrial fibrillation (AF) catheter ablation success depends on the possibility to accurately determine areas on the atrial endocardium at which AF activation originates. One way to determine if major AF activation pathways originate at identified source is through causality analysis. This work assessed to what extent left atrial highest dominant frequency (HDF) areas can be identified as sources of activation pathways in 10 male subjects suffering from persistent AF. Virtual electrograms were collected from 64 endocardial locations for at least 5 minutes. Frequency and causality were analyzed on 4 s signal segments Causality was assessed using the directed transfer function (DTF) algorithm, and AF activation sources were identified as endocardial locations of which the VEGM signal had high influence on other VEGM signals. Co-localization of high influence and HDF areas was evaluated for different area overlap and spectral organisation (OI) thresholds. Results show that, on average, good overlap only existed in 64.6% (± 8.8%) over all subject using the lowest threshold settings. Good overlap rates reduced with more conservative thresholds. This indicates that HDF areas might not always identify origins of main AF activation pathways

Dynamic behavior of rotors during human persistent atrial fibrillation as observed using non-contact mapping

Rotors have been related to atrial fibrillation (AF) maintenance. We analyzed the behavior of rotors in persistent AF (persAF) utilizing a novel non-contact methodology and compared this to real time dominant frequency (DF) analysis. 2048 noncontact virtual unipolar atrial electrograms (VEGMs) were collected simultaneously (EnSite Array, St. Jude Medical) from 10 persAF patients (duration: 34 ± 25 months) undergoing left atrial (LA) ablation. After QRST-removal, FFT was used to identify the global DF of the LA (range 4-10 Hz; 1 s time-window; 50 % overlap; highest DF (HDF) (DF -0.25 Hz); up to 20 s/patient). The organization index (OI) was measured and phase was found via Hilbert-transform. Phase singularities (PSs) were tracked and were categorized according to their lifespan into short (lifespan 100 ms). A total of 4578 PSs were tracked. 5.05 % (IQR: 2.75 ~ 30.25 %) of the tracked PSs were long-lived and were observed in 11 % (IQR: 2.75 ~ 17.5 %) of the windows. The windows with rotors showed significantly higher HDF (mean ± SD, 8.0 ± 0.43 Hz vs 7.71 ± 0.50 Hz, p<; 0.0001) and lower OI (0.76 ± 0.04 vs 0.79 ± 0.03, p<; 0.0001) when compared with the short-lived PSs windows. During persAF, the LA showed distinct behaviors as characterized by rotors. Often, no rotors were observed during sustained AF and, when present, the rotors continually switched between organized and disorganized behaviors. Long-lived rotors correlated with higher atrial rates. Our results suggest that rotors are not the sole perpetuating mechanism in persAF

Unifying automated fractionated atrial electrogram classification using electroanatomical mapping systems in persistent atrial fibrillation studies

Ablation targeting complex fractionated atrial electrograms (CFAE) for treating persistent atrial fibrillation (persAF) has shown conflicting results. Differences in automated algorithms embedded in NavX (St Jude Medical) and CARTO (Biosense Webster) could influence CF AE target identification for ablation, potentially affecting ablation outcomes. To evaluate this effect, automated CFAE classification performed by NavX and CARTO on the same bipolar electrograms from 18 persAF patients undergoing ablation was compared. Using the default thresholds, NavX classified 69±5% of the electrograms as CFAEs, while CARTO detected 35±5%% (Cohen's kappa κ≈0.3, P<0.0001). Both primary and complementary metrics for each system were optimized to balance CF AE detection for both systems. Using revised thresholds found from receiver operating characteristic curves, NavX classified 45±4%, while CARTO detected 42±5% (κ≈0.5, P<0.0001). Our work takes a first step towards the optimization of CFAE detection between NavX and CARTO by providing revised thresholds to reduce differences in CF AE classification. This would facilitate direct comparisons of persAF CFAE-guided ablation outcome guided by NavX or CARTO

Systematic differences of non-invasive dominant frequency estimation compared to invasive dominant frequency estimation in atrial fibrillation

Non-invasive analysis of atrial fibrillation (AF) using body surface mapping (BSM) has gained significant interest, with attempts at interpreting atrial spectro-temporal parameters from body surface signals. As these body surface signals could be affected by properties of the torso volume conductor, this interpretation is not always straightforward. This paper highlights the volume conductor effects and influences of the algorithm parameters for identifying the dominant frequency (DF) from cardiac signals collected simultaneously on the torso and atrial surface. Bi-atrial virtual electrograms (VEGMs) and BSMs were recorded simultaneously for 5 minutes from 10 patients undergoing ablation for persistent AF. Frequency analysis was performed on 4 s segments. DF was defined as the frequency with highest power between 4-10 Hz with and without applying organization index (OI) thresholds. The volume conductor effect was assessed by analyzing the highest DF (HDF) difference of each VEGM HDF against its BSM counterpart. Significant differences in HDF values between intra-cardiac and torso signals could be observed, independent of OI threshold. This difference increases with increasing endocardial HDF (BSM-VEGM median difference from -0.13 Hz for VEGM HDF at 6.25±0.25 Hz to -4.24 Hz at 9.75±0.25 Hz), thereby confirming the theory of the volume conductor effect in real-life situations. Applying an OI threshold strongly affected the BSM HDF area size and location and atrial HDF area location. These results suggest that volume conductor and measurement algorithm effects must be considered for appropriate clinical interpretation

Investigation on recurrent high dominant frequency spatiotemporal patterns during persistent atrial fibrillation

Atrial regions hosting dominant frequency (DF) may represent potential drivers of persistent atrial fibrillation (persAF). Previous work showed that DF can exhibit cyclic behaviour. This study aims to better understand the spatiotemporal behaviours of persAF over longer time periods. 10 patients undergoing persAF ablation targeted at DF were included. Left atrial (LA) non-contact virtual electrograms (VEGMs, Ensite Array, St Jude Medical) were collected for up to 5 min pre-/post- ablation. DF was identified as the peak from 4-10 Hz, in 4 s windows (50 % overlap). High DF (HDF) map was created and automated pattern recognition algorithm was applied to look for recurring HDF spatial patterns within each patient. Recurring HDF patterns were found in all patients. Patients who changed rhythm to atrial flutter after ablation demonstrated single dominant pattern (DP) among the recorded time period, which might consistent with the higher level of regularity during flutter. Ablation regularized AF as demonstrated by increased DP recurrence after ablation. The time interval (median [IQR]) of DP recurrence for the patients still in atrial fibrillation(AF) after ablation (7 patients) decreased from 21.1 s [11.8~49.7 s] to 15.7s [6.5~18.2 s]. The proposed method quantifies the spatiotemporal regularity of HDF DPs over long time periods and may offer a more comprehensive dynamic overview of persAF behaviour and the impact of ablation

Mycolactone-dependent depletion of endothelial cell thrombomodulin is strongly associated with fibrin deposition in Buruli ulcer lesions

A well-known histopathological feature of diseased skin in Buruli ulcer (BU) is coagulative necrosis caused by the Mycobacterium ulcerans macrolide exotoxin mycolactone. Since the underlying mechanism is not known, we have investigated the effect of mycolactone on endothelial cells, focussing on the expression of surface anticoagulant molecules involved in the protein C anticoagulant pathway. Congenital deficiencies in this natural anticoagulant pathway are known to induce thrombotic complications such as purpura fulimans and spontaneous necrosis. Mycolactone profoundly decreased thrombomodulin (TM) expression on the surface of human dermal microvascular endothelial cells (HDMVEC) at doses as low as 2ng/ml and as early as 8hrs after exposure. TM activates protein C by altering thrombin's substrate specificity, and exposure of HDMVEC to mycolactone for 24 hours resulted in an almost complete loss of the cells' ability to produce activated protein C. Loss of TM was shown to be due to a previously described mechanism involving mycolactone-dependent blockade of Sec61 translocation that results in proteasome-dependent degradation of newly synthesised ER-transiting proteins. Indeed, depletion from cells determined by live-cell imaging of cells stably expressing a recombinant TM-GFP fusion protein occurred at the known turnover rate. In order to determine the relevance of these findings to BU disease, immunohistochemistry of punch biopsies from 40 BU lesions (31 ulcers, nine plaques) was performed. TM abundance was profoundly reduced in the subcutis of 78% of biopsies. Furthermore, it was confirmed that fibrin deposition is a common feature of BU lesions, particularly in the necrotic areas. These findings indicate that there is decreased ability to control thrombin generation in BU skin. Mycolactone's effects on normal endothelial cell function, including its ability to activate the protein C anticoagulant pathway are strongly associated with this. Fibrin-driven tisischemia could contribute to the development of the tissue necrosis seen in BU lesions