Machine Learning Color Feature Analysis of a High Throughput Nanoparticle Conjugate Sensing Assay.

Plasmonic nanoparticles are finding applications within the single molecule sensing field in a "dimer" format, where interaction of the target with hairpin DNA causes a decrease in the interparticle distance, leading to a localized surface plasmon resonance shift. While this shift may be detected using spectroscopy, achieving statistical relevance requires the measurement of thousands of nanoparticle dimers and the timescales required for spectroscopic analysis are incompatible with point-of-care devices. However, using dark-field imaging of the dimer structures, simultaneous digital analysis of the plasmonic resonance shift after target interaction of thousands of dimer structures may be achieved in minutes. The main challenge of this digital analysis on the single-molecule scale was the occurrence of false signals caused by non-specifically bound clusters of nanoparticles. This effect may be reduced by digitally separating dimers from other nanoconjugate types. Variation in image intensity was observed to have a discernible impact on the color analysis of the nanoconjugate constructs and thus the accuracy of the digital separation. Color spaces wherein intensity may be uncoupled from the color information (hue, saturation, and value (HSV) and luminance, a* vector, and b* vector (LAB) were contrasted to a color space which cannot uncouple intensity (RGB) to train a classifier algorithm. Each classifier algorithm was validated to determine which color space produced the most accurate digital separation of the nanoconjugate types. The LAB-based learning classifier demonstrated the highest accuracy for digitally separating nanoparticles. Using this classifier, nanoparticle conjugates were monitored for their plasmonic color shift after interaction with a synthetic RNA target, resulting in a platform with a highly accurate yes/no response with a true positive rate of 88% and a true negative rate of 100%. The sensor response of tested single stranded RNA (ssRNA) samples was well above control responses for target concentrations in the range of 10 aM-1 pM.

Effects of an evidence-based model for cochlear implant aftercare delivery on clinical efficiency and patient outcomes.

OBJECTIVES: To assess an evidence-based model (EBM) approach to cochlear implant (CI) aftercare that includes a modified, reduced treatment schedule for newly-implanted adult CI recipients consisting of four appointments (initial activation, 1-, 3- and 6- months postactivation) in the first year post-surgery. METHOD: This prospective multicenter proof-of-concept study was conducted across three clinics in the United States by five experienced CI clinicians. Seventeen newly-implanted adult patients with postlingual hearing loss enrolled in the study. Hearing outcomes were measured using objective speech testing and subjective self-report measures. RESULTS: Most recipients (14/17; 82%) were able to follow the four-appointment EBM schedule. The reduced number of visits translated into an average time savings of 3 hours per patient. Significant improvements in speech perception were observed at both 3- and 6-months postactivation, as measured by CNC words in quiet and AzBio sentences at +10 dB SNR, consistent with published results achieved by traditional practices. Recipients were significantly satisfied with telephone, music, small group conversation, and television listening at 6 months postactivation. Recipient satisfaction with overall service was rated as "excellent" by 14/14 (100%) respondents. CONCLUSION: The four-appointment EBM approach delivered efficient and effective audiological aftercare to CI recipients in the first year following CI implantation.

Dizziness, Falls, and Hearing Loss in Adults Living With Sickle Cell Disease.

OBJECTIVE: The aim of this study was to assess the prevalence of reported dizziness/imbalance, frequency of falls, and hearing loss in adults with sickle cell disease (SCD) and determine the relationship of these self-reported problems both during and in the absence of an SCD crisis. Also, the impact of educational level and health insurance on seeking treatment services, as well as the relationship of falls to stroke, legal blindness, and other orthopedic problems, was assessed. Interrupted blood flow to shared vestibular and auditory arteries supports the notion of increased likelihood of balance deficits and increased falls in this population. DESIGN: A cross-sectional survey study design was used. Adults living with SCD responded to a questionnaire that was distributed online and through traditional mail. STUDY SAMPLE: Adults living with SCD (N = 135) participated in the study. RESULTS: Responses revealed 70% of participants with SCD experienced dizziness/imbalance and 23% reported hearing loss. Furthermore, 33% of participants reported falling one or more times in the last year. The prevalence of dizziness/imbalance, falling, and hearing loss in the respondents with SCD was much higher than that of the general population of the United States. Additionally, for dizziness and falling, the prevalence was higher not only than the national average but also for persons over 65 years of age. A significant association was demonstrated between dizziness/imbalance and hearing loss as well as dizziness/imbalance and falls for adults living with SCD. In fact, participants with self-reported hearing loss were 5.2 times more likely to also report dizziness/imbalance. They were 4.9 times more likely to fall if they also reported dizziness/imbalance. Numbness of the feet was revealed to significantly impact the likelihood of falling in this disease population and should be further studied. Regarding SCD crisis status, dizziness/imbalance and falls were more likely to occur outside of SCD crisis than during a crisis. Furthermore, pain levels were significantly associated with dizziness/imbalance only when respondents were in crisis and not in the absence of a crisis. In crisis, higher pain levels were reported from respondents who also reported dizziness/imbalance than from those who did not report dizziness. No significant relationship was revealed between hearing loss and falls. DISCUSSION: These results provide justification for patient and health care provider education regarding appropriate referrals for vestibular/balance assessments and provision of fall prevention strategies. Future studies on balance and SCD are encouraged.

Single particle detection of protein molecules using dark-field microscopy to avoid signals from nonspecific adsorption.

A massively parallel single particle sensing method based on core-satellite formation of Au nanoparticles was introduced for the detection of interleukin 6 (IL-6). This method exploits the fact that the localized plasmon resonance (LSPR) of the plasmonic nanoparticles will change as a result of core-satellite formation, resulting in a change in the observed color. In this method, the hue (color) value of thousands of 67 nm Au nanoparticles immobilized on a glass coverslip surface is analyzed by a Matlab code before and after the addition of reporter nanoparticles containing IL-6 as target protein. The average hue shift as the result of core-satellite formation is used as the basis to detect small amount of proteins. This method enjoys two major advantages. First it is able to analyze the hue values of thousands of nanoparticles in parallel in less than a minute. Secondly the method is able to circumvent the effect of non-specific adsorption, a major issue in the field of biosensing.

Optical tweezers-based characterisation of gold core-satellite plasmonic nano-assemblies incorporating thermo-responsive polymers.

We report on the characterisation of the optical properties and dynamic behaviour of optically trapped single stimuli-responsive plasmonic nanoscale assemblies. Nano-assemblies consist of a core-satellite arrangement where the constituent nanoparticles are connected by the thermoresponsive polymer, poly(DEGA-co-OEGA). The optical tweezers allow the particles to be held isolated in solution and interrogated using dark-field spectroscopy. Additionally, controlling the optical trapping power provides localised heating for probing the thermal response of the nanostructures. Our results identify a number of distinct core-satellite configurations that can be stably trapped, which are verified using finite element modelling. Laser heating of the nanostructures through the trapping laser yields irreversible modification of the arrangement, as observed through the scattering spectrum. We consider which factors may be responsible for the observed behaviour in the context of the core-satellite geometry, polymer-solvent interaction, and the bonding of the nanoparticles.

How Nanoparticles Transform Single Molecule Measurements into Quantitative Sensors.

Single molecule measurements are revolutionizing the understanding of the stochastics of behavior of single molecules. There is a common theme referred to as a near-field approach, in how many single molecule measurements are being performed in assays. The term near field is used because the measurement volume is typically very small such that a single molecule, or a single molecule binding pair, within that volume is of an appreciable concentration. The next development in detection will be performing many single molecule measurements at one time such that single molecule measurements can be used as the basis for quantitative analysis. There have already been some notable developments in this direction. Again, all have a common theme in that nanoparticles are used to create many near-field volumes that can be measured simultaneously. Herein, the coupled developments in nanoparticles and measurement strategies that allow nanoparticles to be the backbone of the next generation of sensing technologies are discussed.

A rapid readout for many single plasmonic nanoparticles using dark-field microscopy and digital color analysis.

The integration of plasmonic nanoparticles into biosensors has the potential to increase the sensitivity and dynamic range of detection, through the use of single nanoparticle assays. The analysis of the localized surface plasmon resonance (LSPR) of plasmonic nanoparticles has allowed the limit of detection of biosensors to move towards single molecules. However, due to complex equipment or slow analysis times, these technologies have not been implemented for point-of-care detection. Herein, we demonstrate an advancement in LSPR analysis by presenting a technique, which utilizes an inexpensive CMOS-equipped digital camera and a dark-field microscope, that can analyse the λmax of over several thousand gold nanospheres in less than a second, without the use of a spectrometer. This improves the throughput of single particle spectral analysis by enabling more nanoparticles to be probed and in a much shorter time. This technique has been demonstrated through the detection of interleukin-6 through a core-satellite binding assay. We anticipate that this technique will aid in the development of high-throughput, multiplexed and point-of-care single nanoparticle biosensors.

The Recognizability and Localizability of Auditory Alarms: Setting Global Medical Device Standards.

Objective Four sets of eight audible alarms matching the functions specified in IEC 60601-1-8 were designed using known principles from auditory cognition with the intention that they would be more recognizable and localizable than those currently specified in the standard. Background The audible alarms associated with IEC 60601-1-8, a global medical device standard, are known to be difficult to learn and retain, and there have been many calls to update them. There are known principles of design and cognition that might form the basis of more readily recognizable alarms. There is also scope for improvement in the localizability of the existing alarms. Method Four alternative sets of alarms matched to the functions specified in IEC 60601-1-8 were tested for recognizability and localizability and compared with the alarms currently specified in the standard. Results With a single exception, all prototype sets of alarms outperformed the current IEC set on both recognizability and localizability. Within the prototype sets, auditory icons were the most easily recognized, but the other sets, using word rhythms and simple acoustic metaphors, were also more easily recognized than the current alarms. With the exception of one set, all prototype sets were also easier to localize. Conclusion Known auditory cognition and perception principles were successfully applied to an existing audible alarm problem. Application This work constitutes the first (benchmarking) phase of replacing the alarms currently specified in the standard. The design principles used for each set demonstrate the relative ease with which different alarm types can be recognized and localized.

Fetal Bovine Collagen Matrix in the Treatment of a Full Thickness Burn Wound: A Case Report With Long-Term Follow-Up.

The treatment of full thickness skin wounds commonly associated with large burns continues to represent a challenging clinical entity. The current treatment for large TBSA burns is split thickness autologous skin grafting; however, this treatment often results in poor textural durability, hypertrophic scarring, and fibrotic contractures. In this case report, we describe our experience and long-term follow-up results after the application of fetal bovine collagen (FBC) matrix (PriMatrix, TEI Biosciences, Boston, MA) to burn wounds clinically assessed as full thickness that healed without the need for subsequent skin grafting. The patient presented with 25% TBSA burns and was debrided and covered with FBC on postburn day 7. By postoperative day 12, the patient had large areas of reepithelialization distributed throughout the wound bed. By postoperative day 26, the patient had significantly more areas of wound closure and was discharged. Reepithelialization and repigmentation continued, and long-term follow-up after 26 months demonstrated complete reepithelialization and nearly complete repigmentation, without the appearance of contractures or hypertrophic scarring. This case report highlights the use of FBC as a scaffold capable of dermal regeneration and spontaneous reepithelialization with an excellent long-term functional and cosmetic outcome.

Bison meat has a lower atherogenic risk than beef in healthy men.

The rearing method of bison and the nutrient content of the meat may make bison a healthier alternative to beef. We hypothesized that the acute and chronic effects of bison consumption, in comparison to beef, will result in a less perturbed blood lipid panel and a reduced inflammatory and oxidative stress response which will minimize the detrimental effect on vascular function. A double-blind, cross-over randomized trial was employed to examine the consequence of a single 12 oz serving (n = 14) and 7 weeks of chronic consumption (n = 10) (12 oz/d, 6 d/wk) of each meat. Measurements included blood lipids, interleukin-6, plasminogen activator inhibitor 1, C-reactive protein, oxidized low-density lipoprotein, protein carbonyl, hydroperoxides, flow-mediated dilation (FMD) and FMD/shear rate. Following a single beef meal, triglycerides and oxidized low-density lipoprotein were elevated (67% ± 45% and 18% ± 17% respectively); there was a tendency for hydroperoxides to be elevated (24% ± 37%); and FMD/shear rate was reduced significantly (30% ± 38%). Following a single meal of bison: there was a smaller increase in triglycerides (30% ± 27%), and markers of inflammation and oxidative stress and FMD/shear rate were unchanged. Chronic consumption of either meat did not influence body weight, % body fat, or blood lipids. Protein carbonyl (24% ± 45%), plasminogen activator inhibitor 1 (78% ± 126%), interleukin-6 (59% ± 76%) and C-reactive protein (72% ± 57%) were significantly elevated and FMD/shear rate was significantly reduced (19% ± 28%) following 7 weeks of beef consumption, but not bison consumption. Based on our findings, the data suggest that bison consumption results in a reduced atherogenic risk compared to beef.

Training the Trainers of Tomorrow Today - driving excellence in medical education.

Training the Trainers of Tomorrow Today (T4) is a new way to deliver "Training for Trainers". Responding to local dissatisfaction with existing arrangements, T4 builds on 3 essential requirements for a future shape of training: 1. Clinical Leadership and a Collaborative Approach 2. Cross-Specialty Design and Participation 3. Local Delivery and Governance Networks Design principles also included: 3 levels of training to reflect differing needs of clinical supervisors, educational supervisors and medical education leader, mapping to GMC requirements and the London Deanery's Professional Development Framework; alignment of service, educational theory and research; recognition of challenges in delivering and ensuring attendance in busy acute and mental health settings, and the development of a faculty network. The delivery plan took into account census of professional development uptake and GMC Trainee Surveys. Strong engagement and uptake from the 11 Trusts in NW London has been achieved, with powerful penetration into all specialties. Attendance has exceeded expectations. Against an initial 12 month target of 350 attendances, 693 were achieved in the first 8 months. Evaluation of content demonstrates modules are pitched appropriately to attendees needs, with positive feedback from trainers new to the role. Delivery style has attracted high ratings of satisfaction: 87% attendees rating delivery as "good\excellent". External evaluation of impact demonstrated improved training experiences through changes in supervision, the learning environment and understanding of learning styles. We have addressed sustainability of the programme by advertising and recruiting Local Faculty Development Trainers. Volunteer consultants and higher trainees are trained to deliver the programme on a cascade model, supported by the Specialty Tutors, individual coaching and educational bursaries. The Trainers are local champions for excellence in training, provide a communication between the programme and local providers, are a repository of expertise in their service, and trouble shoot local barriers to engagement.