A Newly Developed Web-Based Resource on Genetic Eye Disorders for Users With Visual Impairment (Gene.Vision): Usability Study

Background: Despite the introduction of the Web Content Accessibility Guidelines and legislations, many websites remain poorly accessible to users with disability, especially those with visual impairment, as the internet has become a more visually complex environment. With increasing reliance on the internet and almost 2 million people in the United Kingdom being affected by vision loss, it is important that they are not overlooked when developing web-based materials. A significant proportion of those affected have irreversible vision loss due to rare genetic eye disorders, and many of them use the internet as a primary source of information for their conditions. However, access to high-quality web-based health information with an inclusive design remains a challenge for many. We have developed a new web-based resource for genetic eye disorders called Gene.Vision that aims to provide a holistic guide for patients, relatives, and health care professionals. Objective: Through a usability testing session of our website prototype, this study aims to identify key web-based accessibility features for internet users with vision impairment and to explore whether the contents provided in Gene.Vision are relevant and comprehensible. Methods: A face-to-face testing session with 8 participants (5 patients, 2 family members, and 1 member of the public) and 8 facilitators was conducted on a prototype website. Remote testing was performed with another patient due to COVID-19 restrictions. Home page design, navigation, content layout and quality, language, and readability were explored through direct observation and task completion using the think-aloud method. A patient focus group was organized to elicit further feedback. Qualitative data were gathered and analyzed to identify core themes through open and axial coding. Results: All participants had good computer literacy; 6 patients with visual impairment used visual aid software including iOS VoiceOver and Speak Screen, iOS Classic Invert, ZoomText 2020, Job Access With Speech, and Nonvisual Desktop Access. The features identified by the participants that will enhance accessibility and usability for users with visual impairment were a consistent website layout, a structured information hierarchy with a clear description of links, good chromatic and luminance contrast, a simple home page with predictable and easy navigation, adaptability to various assistive software, and readable and relevant content. They reported that dynamic content (such as carousels) and large empty spaces reduced accessibility. Information on research, support available, practical advice, and links to charities were incentives for repeated website visits. Conclusions: We demonstrated the importance of developing a website with a user-based approach. Through end user testing, we identified several key web-based accessibility features for people with visual impairment. Target end users should always be involved early and throughout the design process to ensure their needs are met. Many of these steps can be implemented easily and will aid in search engine optimization

Structural Adaptability Facilitates Histidine Heme Ligation in a Cytochrome P450

Almost all known members of the cytochrome P450 (CYP) superfamily conserve a key cysteine residue that coordinates the heme iron. Although mutation of this residue abolishes monooxygenase activity, recent work has shown that mutation to either serine or histidine unlocks non-natural carbene- and nitrene-transfer activities. Here we present the first crystal structure of a histidine-ligated P450. The T213A/C317H variant of the thermostable CYP119 from Sulfolobus acidocaldarius maintains heme iron coordination through the introduced ligand, an interaction that is accompanied by large changes in the overall protein structure. We also find that the axial cysteine C317 may be substituted with any other amino acid without abrogating folding and heme cofactor incorporation. Several of the axial mutants display unusual spectral features, suggesting that they have active sites with unique steric and electronic properties. These novel, highly stable enzyme active sites will be fruitful starting points for investigations of non-natural P450 catalysis and mechanisms

Influence of Soldiers' Cardiorespiratory Fitness on Physiological Responses and Dropouts During a Loaded Long-distance March.

Introduction: In military service, marching is an important, common, and physically demanding task. Minimizing dropouts, maintaining operational readiness during the march, and achieving a fast recovery are desirable because the soldiers have to be ready for duty, sometimes shortly after an exhausting task. The present field study investigated the influence of the soldiers' cardiorespiratory fitness on physiological responses during a long-lasting and challenging 34 km march. Materials and methods: Heart rate (HR), body core temperature (BCT), total energy expenditure (TEE), energy intake, motivation, and pain sensation were investigated in 44 soldiers (20.3 ± 1.3 years, 178.5 ± 7.0 cm, 74.8 ± 9.8 kg, body mass index: 23.4 ± 2.7 kg × m-2, peak oxygen uptake ($\dot{\rm{V}}$O2peak): 54.2 ± 7.9 mL × kg-1 × min-1) during almost 8 hours of marching. All soldiers were equipped with a portable electrocardiogram to record HR and an accelerometer on the hip, all swallowed a telemetry pill to record BCT, and all filled out a pre- and post-march questionnaire. The influence of aerobic capacity on the physiological responses during the march was examined by dividing the soldiers into three fitness groups according to their $\dot{\rm{V}}$O2peak. Results: The group with the lowest aerobic capacity ($\dot{\rm{V}}$O2peak: 44.9 ± 4.8 mL × kg-1 × min-1) compared to the group with the highest aerobic capacity ($\dot{\rm{V}}$O2peak: 61.7 ± 2.2 mL × kg-1 × min-1) showed a significantly higher (P < .05) mean HR (133 ± 9 bpm and 125 ± 8 bpm, respectively) as well as peak BCT (38.6 ± 0.3 and 38.4 ± 0.2 °C, respectively) during the march. In terms of recovery ability during the break, no significant differences could be identified between the three groups in either HR or BCT. The energy deficit during the march was remarkably high, as the soldiers could only replace 22%, 26%, and 36% of the total energy expenditure in the lower, middle, and higher fitness group, respectively. The cardiorespiratory fittest soldiers showed a significantly higher motivation to perform when compared to the least cardiorespiratory fit soldiers (P = .002; scale from 1 [not at all] to 10 [extremely]; scale difference of 2.3). A total of nine soldiers (16%) had to end marching early: four soldiers (21%) in the group with the lowest aerobic capacity, five (28%) in the middle group, and none in the highest group. Conclusion: Soldiers with a high $\dot{\rm{V}}$O2peak showed a lower mean HR and peak BCT throughout the long-distance march, as well as higher performance motivation, no dropouts, and lower energy deficit. All soldiers showed an enormous energy deficit; therefore, corresponding nutritional strategies are recommended

Textile additive manufacturing: An overview

Additive manufacturing (3D-printing) is a rapidly emerging technology grouped under the heading of Industry 4.0 and revolutionises the way products are created. One emerging area is textiles (apparel) where additive manufacturing allows the rapid fabrication of products that are not easily produced using traditional manufacturing approaches. In this review, we provide an overview of recent developments in textile additive manufacturing highlighting this exciting and rapidly growing research area and offer insights into the future perspectives of this promising and innovative technology

Detection and diversity of a putative novel heterogeneous polymorphic proline-glycine repeat (Pgr) protein in the footrot pathogen Dichelobacter nodosus

Dichelobacter nodosus, a Gram-negative anaerobic bacterium, is the essential causative agent of footrot in sheep. Currently, depending on the clinical presentation in the field, footrot is described as benign or virulent; D. nodosus strains have also been classified as benign or virulent, but this designation is not always consistent with clinical disease. The aim of this study was to determine the diversity of the pgr gene, which encodes a putative proline-glycine repeat protein (Pgr). The pgr gene was present in all 100 isolates of D. nodosus that were examined and, based on sequence analysis had two variants, pgrA and pgrB. In pgrA, there were two coding tandem repeat regions, R1 and R2: different strains had variable numbers of repeats within these regions. The R1 and R2 were absent from pgrB. Both variants were present in strains from Australia, Sweden and the UK, however, only pgrB was detected in isolates from Western Australia. The pgrA gene was detected in D. nodosus from tissue samples from two flocks in the UK with virulent footrot and only pgrB from a flock with no virulent or benign footrot for >10 years. Bioinformatic analysis of the putative PgrA protein indicated that it contained a collagen-like cell surface anchor motif. These results suggest that the pgr gene may be a useful molecular marker for epidemiological studies

Evaluation of pulse rate measurement with a wrist worn device during different tasks and physical activity

The purpose of this study was to evaluate the wrist-worn device Mio FUSE, which estimates heart rate (HR) based on photo-plethysmography, 1) in a large study group during a standardised activity, 2) in a small group during a variety of activities and 3) to investigate factors affecting HR accuracy in a real-world setting. First, 53 male participants (20 ±1 years; 1.79 ±0.07 m; 76.1 ±10.5 kg) completed a 35-km march wearing the Equivital EQ-02 as a criterion measure. Second, 5 participants (whereof 3 female; 29 ±5 years; 1.74 ±0.07 m; 67.8 ±11.1 kg) independently performed 25 activities, categorised as sitting passive, sitting active, standing, cyclic and anti-cyclic activities with the Polar H7 as a criterion device. Equivalence testing and Bland-and-Altman analyses were undertaken to assess the accuracy to the criterion devices. Third, confounders affecting HR accuracy were investigated using multiple backwards regression analyses. The Mio FUSE was equivalent to the respective criterion measures with only small systematic biases of -3.5 bpm (-2.6%) and -1.7 bpm (-1.3%) with limits of agreements of ±10.1 bpm and ±10.8 bpm during the 35-km march and during different activities, respectively. Confounding factors negatively affecting the accuracy of the Mio FUSE were found to include larger wrist size and intensified arm and/or wrist movement. The wrist-worn Mio FUSE can be recommended to estimate overall HR accurately for different types of activities in healthy adults. However, during sporting activities involving intensified arm and/or wrist movement or for detailed continuous analysis, a chest strap is preferred to the Mio FUSE to optimise HR estimation accuracy

Validation of ambulatory monitoring devices to measure energy expenditure and heart rate in a military setting

Objectives.; To investigate the validity of different devices and algorithms used in military organizations worldwide to assess physical activity energy expenditure (PAEE) and heart rate (HR) among soldiers.; Design.; Device validation study.; Methods; . Twenty-three male participants serving their mandatory military service accomplished, firstly, nine different military specific activities indoors, and secondly, a normal military routine outdoors. Participants wore simultaneously an ActiHeart, Everion, MetaMax 3B, Garmin Fenix 3, Hidalgo EQ02, and PADIS 2.0 system. The PAEE and HR data of each system were compared to the criterion measures MetaMax 3B and Hidalgo EQ02, respectively.; Results; . Overall, the recorded systematic errors in PAEE estimation ranged from 0.1 (±1.8) kcal.min; -1; to -1.7 (±1.8) kcal.min; -1; for the systems PADIS 2.0 and Hidalgo EQ02 running the Royal Dutch Army algorithm, respectively, and in the HR assessment ranged from -0.1 (±2.1) b.min; -1; to 0.8 (±3.0) b.min; -1; for the PADIS 2.0 and ActiHeart systems, respectively. The mean absolute percentage error (MAPE) in PAEE estimation ranged from 29.9% to 75.1%, with only the Everion system showing an overall MAP

The catalytic role of glutathione transferases in heterologous anthocyanin biosynthesis

Anthocyanins are ubiquitous plant pigments used in a variety of technological applications. Yet, after over a century of research, the penultimate biosynthetic step to anthocyanidins attributed to the action of leucoanthocyanidin dioxygenase has never been efficiently reconstituted outside plants, preventing the construction of heterologous cell factories. Through biochemical and structural analysis, here we show that anthocyanin-related glutathione transferases, currently implicated only in anthocyanin transport, catalyse an essential dehydration of the leucoanthocyanidin dioxygenase product, flavan-3,3,4-triol, to generate cyanidin. Building on this knowledge, introduction of anthocyanin-related glutathione transferases into a heterologous biosynthetic pathway in baker's yeast results in >35-fold increased anthocyanin production. In addition to unravelling the long-elusive anthocyanin biosynthesis, our findings pave the way for the colourants' heterologous microbial production and could impact the breeding of industrial and ornamental plants

Multiple locus VNTR analysis highlights that geographical clustering and distribution of Dichelobacter nodosus, the causal agent of footrot in sheep, correlates with inter-country movements

Dichelobacter nodosus is a Gram-negative, anaerobic bacterium and the causal agent of footrot in sheep. Multiple locus variable number tandem repeat (VNTR) analysis (MLVA) is a portable technique that involves the identification and enumeration of polymorphic tandem repeats across the genome. The aims of this study were to develop an MLVA scheme for D. nodosus suitable for use as a molecular typing tool, and to apply it to a global collection of isolates. Seventy-seven isolates selected from regions with a long history of footrot (GB, Australia) and regions where footrot has recently been reported (India, Scandinavia), were characterised. From an initial 61 potential VNTR regions, four loci were identified as usable and in combination had the attributes required of a typing method for use in bacterial epidemiology: high discriminatory power (D > 0.95), typeability and reproducibility. Results from the analysis indicate that D. nodosus appears to have evolved via recombinational exchanges and clonal diversification. This has resulted in some clonal complexes that contain isolates from multiple countries and continents; and others that contain isolates from a single geographic location (country or region). The distribution of alleles between countries matches historical accounts of sheep movements, suggesting that the MLVA technique is sufficiently specific and sensitive for an epidemiological investigation of the global distribution of D. nodosus

Cofactor specificity motifs and the induced fit mechanism in class I ketol-acid reductoisomerases

Although most sequenced members of the industrially important ketol-acid reductoisomerase (KARI) family are class I enzymes, structural studies to date have focused primarily on the class II KARIs, which arose through domain duplication. In the present study, we present five new crystal structures of class I KARIs. These include the first structure of a KARI with a six-residue β2αB (cofactor specificity determining) loop and an NADPH phosphate-binding geometry distinct from that of the seven- and 12-residue loops. We also present the first structures of naturally occurring KARIs that utilize NADH as cofactor. These results show insertions in the specificity loops that confounded previous attempts to classify them according to loop length. Lastly, we explore the conformational changes that occur in class I KARIs upon binding of cofactor and metal ions. The class I KARI structures indicate that the active sites close upon binding NAD(P)H, similar to what is observed in the class II KARIs of rice and spinach and different from the opening of the active site observed in the class II KARI of Escherichia coli. This conformational change involves a decrease in the bending of the helix that runs between the domains and a rearrangement of the nicotinamide-binding site