Recovery of Hydroxytyrosol from Olive Mill Wastewater Using the Promiscuous Hydrolase/Acyltransferase PestE.

Olive mill wastewater (OMWW) is produced annually during olive oil extraction and contains most of the health-promoting 3-hydroxytyrosol of the olive fruit. To facilitate its recovery, enzymatic transesterification of hydroxytyrosol (HT) was directly performed in an aqueous system in the presence of ethyl acetate, yielding a 3-hydroxytyrosol acetate rich extract. For this, the promiscuous acyltransferase from Pyrobaculum calidifontis VA1 (PestE) was engineered by rational design. The best mutant for the acetylation of hydroxytyrosol (PestE_I208A_L209F_N288A) was immobilized on EziG2 beads, resulting in hydroxytyrosol conversions between 82 and 89 % in one hour, for at least ten reaction cycles in a buffered hydroxytyrosol solution. Due to inhibition by other phenols in OMWW the conversions of hydroxytyrosol from this source were between 51 and 62 %. In a preparative scale reaction, 13.8 mg (57 %) of 3-hydroxytyrosol acetate was extracted from 60 mL OMWW.

Factors contributing to clinically important health utility gains in cochlear implant recipients.

PURPOSE: Cochlear implantation can restore access to sound and speech understanding in subjects with substantial hearing loss. The Health Utilities Index Mark III (HUI3) measures the impact of an intervention on the patient's quality of life and is sensitive to changes in hearing. In the current study we used factor analysis to predict a clinically important gain in HUI3 scores in adult cochlear implant recipients. METHODS: Data were collected in an observational study for 137 adult recipients from a single center who had at least 1-year HUI3 follow-up. Demographic and other baseline parameters were retrospectively analyzed for their association with a clinically important HUI3 scale gain, defined as at least 0.1 points. Data were also collected for the speech spatial qualities (SSQ) scale. RESULTS: Baseline telephone use and HUI3 hearing, speech and emotion attribute levels were significantly associated with clinically important gains in HUI3 scores. However, SSQ scores increased significantly with or without clinically important HUI3 gains. CONCLUSION: Those subjects who were unhappy or experienced difficulties communicating with strangers or in a group were twice as likely to obtain a clinically important gain in health utility compared to those who were happy or had less difficulty communicating. Subjects who were unable to use the telephone prior to cochlear implantation were one and a half times more likely to obtain a clinically important gain. The SSQ scale was more sensitive to hearing improvements due to cochlear implantation. An inability to use the telephone is an easy to assess biomarker for candidacy for cochlear implantation.

Effect of yoga-based lifestyle and dietary modification in overweight individuals with sleep apnea: A randomized controlled trial (ELISA).

BACKGROUND: Obesity is recognised as an important risk factor for obstructive sleep apnea (OSA), with obese individuals at a four times higher risk of being diagnosed with the syndrome. Treating obesity with lifestyle modification is associated with a reduction in the severity of obstructive sleep apnea. Yoga comprises lifestyle modification that includes asana (postures), pranayama (breathing techniques), dhyana (meditation) and guideline principles for healthy living (Yama and Niyama). There is a scarcity of data to evaluate the effect of yoga on OSA. This study was conducted to evaluate the efficacy of Yoga based lifestyle modification on OSA. METHODS: Consenting obese patients (BMI >23) diagnosed with obstructive sleep apnea (OSA) (AHI>5) on Polysomnography (PSG) were enrolled. Eligible patients were randomized into two groups. The control group received counselling for dietary modification (staple Indian) with regular exercise and the active intervention group received Yoga intervention as treatment (OSA module) in addition to similar dietary modification and regular exercise counselling. Polysomnography (PSG) was conducted at baseline and one year follow-up. All patients were evaluated at baseline, six months, and one year for compliance and anthropometric parameters. Additional assessment with Hamilton scales for depression and anxiety, SF-36, and the Pittsburgh sleep quality index was also conducted. RESULTS: A total of 37 eligible patients (19 in the control group and 18 in the yoga group) were recruited for the study. The age [45.73 ± 10.71 vs. 46.22 ± 9.39 years, p = 0.88] and gender [15(78.95%) vs. 12(66.67%), p = 0.48 (males)] distribution was similar in both groups. After adjusting for age and gender, the percentage reduction in weight between the two groups did not reach statistical significance at one year. There was no significant difference in mean AHI between the two groups at one year. However, the number of patients with more than 40% AHI reduction [2/19 (10.52%) vs 8/18 (44.44%), p = 0.02] was significantly higher in the yoga group. Additionally, within the groups, the mean AHI at one year was significantly reduced in the yoga group [51.2 ± 28.0 to 36.8 ± 21.0/hour, p = 0.003], while no significant change was found in the control group [47.2 ± 23 to 38.8 ± 19.9/hour, p = 0.08]. CONCLUSIONS: Lifestyle alteration using Yoga intervention and modification of staple Indian diet may be effective in reducing OSA severity among obese patients. CTRI NUMBER: CTRI/2017/05/008462.

A Survey of Diagnostic Delay in Axial Spondyloarthritis Across Two National Health Service (NHS) Rheumatology Services.

Introduction Diagnostic delay is an ongoing challenge in axial spondyloarthritis (axial SpA). A recent, comprehensive literature review has found a mean average of 8.7 years of delay between symptom onset and formal diagnosis in the United Kingdom (UK). The primary aim of this study was to identify delays to diagnosis experienced by patients with axial SpA under the ongoing care of two urban National Health Service (NHS) rheumatology services. The secondary aims were (a) to count healthcare professional (HCP) interactions after symptom onset but prior to the diagnosis, (b) to compare our data to published delay to diagnosis research and (c) to explore contributing factors locally and the variation between the two UK rheumatology services. Methods A 14-question survey was created to identify the delay to diagnosis and contributing factors across two urban NHS axial SpA services, from the onset of symptoms to diagnosis and commencement of treatment. Participants were recruited from clinic visits between August and November 2021 and completed the survey either on paper or via online survey software, both with HCP support. Results Those completing the survey formed a cohort of 106 participants with an established diagnosis of axial SpA who attended the axial SpA services at either Royal Free NHS Foundation Trust or Salford Care Organisation, Northern Care Alliance NHS Foundation Trust. The mean time from the onset of symptoms to the diagnosis of axial SpA was similar across centres despite the differences in demographics, with Royal Free at 5.72 years and Salford Royal at 5.96 years. When reviewing via median diagnostic delay, there was a notable difference with Royal Free at 6.09 years and Salford Royal at 4.27 years. Across the two sites, between the onset of symptoms and the diagnosis of axial SpA, 90% of the participants saw a general practitioner (GP), of which 63% of the patients saw a GP 1-5 times, 23% saw 5-10 times and 14% saw more than 10 times. Many participants also saw other HCPs, including physiotherapists, other manual therapists and hospital specialists prior to diagnosis. In addition, 32% saw one other HCP, 18% two other HCPs, 9% three, 7% four and 2.7% five other HCPs prior to diagnosis. Close to 80% of the patients stated that they had received adequate axial SpA education at diagnosis, and 76% of the patients were aware of who to contact in the event of a flare. Conclusions These data highlight that the mean average time to diagnosis for both trusts was between five and six years, somewhat lower than the 8.7-year national UK average. However, despite being specialist centres, these data are a long way from the National Axial Spondyloarthritis Society (NASS) "Gold Standard" of one year time to diagnosis. The contributors to this include lack of HCP and community awareness about axial SpA, its recognition and appropriate onwards referral. There is a need for concerted ways of working for the development of patient pathways and public and HCP education to reduce this delay to allow the ambitious NASS Gold Standard of one year time to diagnosis to be achieved.

Recommendations for ethical and responsible use of artificial intelligence in digital agriculture.

Artificial intelligence (AI) applications are an integral and emerging component of digital agriculture. AI can help ensure sustainable production in agriculture by enhancing agricultural operations and decision-making. Recommendations about soil condition and pesticides or automatic devices for milking and apple picking are examples of AI applications in digital agriculture. Although AI offers many benefits in farming, AI systems may raise ethical issues and risks that should be assessed and proactively managed. Poor design and configuration of intelligent systems may impose harm and unintended consequences on digital agriculture. Invasion of farmers' privacy, damaging animal welfare due to robotic technologies, and lack of accountability for issues resulting from the use of AI tools are only some examples of ethical challenges in digital agriculture. This paper examines the ethical challenges of the use of AI in agriculture in six categories including fairness, transparency, accountability, sustainability, privacy, and robustness. This study further provides recommendations for agriculture technology providers (ATPs) and policymakers on how to proactively mitigate ethical issues that may arise from the use of AI in farming. These recommendations cover a wide range of ethical considerations, such as addressing farmers' privacy concerns, ensuring reliable AI performance, enhancing sustainability in AI systems, and reducing AI bias.

Lagging X chromatids specify the orientation of asymmetric organelle partitioning in XX spermatocytes of Auanema rhodensis.

The unequal partitioning of molecules and organelles during cell division results in daughter cells with different fates. An extreme example is female meiosis, in which consecutive asymmetric cell divisions give rise to 1 large oocyte and 2 small polar bodies with DNA and minimal cytoplasm. Here, we test the hypothesis that during an asymmetric cell division during spermatogenesis of the nematode Auanema rhodensis, the late segregating X chromatids orient the asymmetric partitioning of cytoplasmic components. In previous studies, the secondary spermatocytes of wild-type XO males were found to divide asymmetrically to generate functional spermatids that inherit components necessary for sperm viability and DNA-containing residual bodies that inherit components to be discarded. Here we extend that analysis to 2 novel contexts. First, the isolation and analysis of a strain of mutant XX pseudomales revealed that such animals have highly variable patterns of X-chromatid segregation. The pattern of late segregating X chromatids nevertheless predicted the orientation of organelle partitioning. Second, while wild-type XX hermaphrodites were known to produce both 1X and 2X sperm, here, we show that spermatocytes within specific spermatogonial clusters exhibit 2 different patterns of X-chromatid segregation that correlate with distinct patterns of organelle partitioning. Together this analysis suggests that A. rhodensis has coopted lagging X chromosomes during anaphase II as a mechanism for determining the orientation of organelle partitioning.

Design, implementation, and management of an international medical device registry.

BACKGROUND: Registries are powerful clinical investigational tools. Although in hospitals registries may be mandated, industry-sponsored, international registries are voluntary and therefore can require clearer objectives and more planning. The registry also needs sufficient resources and appropriate measurement tools to motivate long-term participation and ensure success. METHODS: We summarize our learnings from 10 years of running a medical device registry that surveys patient-reported benefits of hearing implants. RESULTS: We enlisted 77 participating clinics globally, who actively recruited a total of more than 1500 hearing implant users. We identified the stages in developing a registry specific to hearing loss. Furthermore, we report the challenges and successes in design and implementation and make recommendations for future registries. CONCLUSIONS: Data collection infrastructure needs to be kept up to date throughout the defined registry lifetime, and it is essential to oversee data quality and completeness. Compliance at registry sites is important for data quality and needs to be weighed against the cost of site monitoring. To motivate sites to enter data accurately and expeditiously, we facilitated easy access to their own data which helped to support their clinical routine. TRIAL REGISTRATION: ClinicalTrials.gov NCT02004353. 9th December 2013.

Rational Design for Enhanced Acyltransferase Activity in Water Catalyzed by the Pyrobaculum calidifontis VA1 Esterase.

Biocatalytic transesterification is commonly carried out employing lipases in anhydrous organic solvents since hydrolases usually prefer hydrolysis over acyl transfer in bulk water. However, some promiscuous acyltransferases can catalyze acylation in an aqueous solution. In this study, a rational design was performed to enhance the acyltransferase selectivity and substrate scope of the Pyrobaculum calidifontis VA1 esterase (PestE). PestE wild type and variants were applied for the acylation of monoterpene alcohols. The mutant PestE_I208A is selective for (-)-menthyl acetate (E-Value = 55). Highly active acyltransferases were designed, allowing for complete conversion of (-)-citronellol to citronellyl acetate. Additionally, carvacrol was acetylated but with lower conversions. To the best of our knowledge, this is the first example of the biocatalytic acylation of a phenolic alcohol in bulk water. In addition, a high citronellol conversion of 92% was achieved with the more environmentally friendly and inexpensive acyl donor ethyl acetate using PestE_N288F as a catalyst. PestE_N288F exhibits good acyl transfer activity in an aqueous medium and low hydrolysis activity at the same time. Thus, our study demonstrates an alternative synthetic strategy for acylation of compounds without organic solvents.

Curvature-Induced Anomalous Enhancement in the Work Function of Nanostructures.

An analytical theory to estimate the electronic work function in curved geometries is formulated under Thomas-Fermi approximation. The work function is framed as the work against the electrostatic self-capacitive energy. The contribution of surface curvature is characterized by mean and Gaussian curvature (through multiple scattering expansion). The variation in work function of metal and semimetal nanostructures is shown as the consequence of surface radius of curvature comparable to electronic screening length. For ellipsoidal particles, the maximum value of work function is observed at the equator and poles for oblate and prolate particles, respectively, whereas triaxial ellipsoid shows nonuniform distribution of the work function over the surface. Similarly, theory predicts manifold increase in the work function for a particle with atomic scale roughness. Finally, the theory is validated with experimental data, and it is concluded that the work function of a nanoparticle can be tailored through its shape.