Circular Economy Electrochemistry: Recycling Old Mixed Material Additively Manufactured Sensors into New Electroanalytical Sensing Platforms.

Recycling used mixed material additively manufactured electroanalytical sensors into new 3D-printing filaments (both conductive and non-conductive) for the production of new sensors is reported herein. Additively manufactured (3D-printed) sensing platforms were transformed into a non-conductive filament for fused filament fabrication through four different methodologies (granulation, ball-milling, solvent mixing, and thermal mixing) with thermal mixing producing the best quality filament, as evidenced by the improved dispersion of fillers throughout the composite. Utilizing this thermal mixing methodology, and without supplementation with the virgin polymer, the filament was able to be cycled twice before failure. This was then used to process old sensors into an electrically conductive filament through the addition of carbon black into the thermal mixing process. Both recycled filaments (conductive and non-conductive) were utilized to produce a new electroanalytical sensing platform, which was tested for the cell's original application of acetaminophen determination. The fully recycled cell matched the electrochemical and electroanalytical performance of the original sensing platform, achieving a sensitivity of 22.4 ± 0.2 µA µM-1, a limit of detection of 3.2 ± 0.8 µM, and a recovery value of 95 ± 5% when tested using a real pharmaceutical sample. This study represents a paradigm shift in how sustainability and recycling can be utilized within additively manufactured electrochemistry toward promoting circular economy electrochemistry.

Immunological and Structural Characterization of Titin Main Immunogenic Region; I110 Domain Is the Target of Titin Antibodies in Myasthenia Gravis.

Myasthenia gravis (MG) is an autoimmune disease caused by antibodies targeting the neuromuscular junction (NJ) of skeletal muscles. The major MG autoantigen is nicotinic acetylcholine receptor. Other autoantigens at the NJ include MuSK, LRP4 and agrin. Autoantibodies to the intra-sarcomeric striated muscle-specific gigantic protein titin, although not directed to the NJ, are invaluable biomarkers for thymoma and MG disease severity. Thymus and thymoma are critical in MG mechanisms and management. Titin autoantibodies bind to a 30 KDa titin segment, the main immunogenic region (MIR), consisting of an Ig-FnIII-FnIII 3-domain tandem, termed I109-I111. In this work, we further resolved the localization of titin epitope(s) to facilitate the development of more specific anti-titin diagnostics. For this, we expressed protein samples corresponding to 8 MIR and non-MIR titin fragments and tested 77 anti-titin sera for antibody binding using ELISA, competition experiments and Western blots. All anti-MIR antibodies were bound exclusively to the central MIR domain, I110, and to its containing titin segments. Most antibodies were bound also to SDS-denatured I110 on Western blots, suggesting that their epitope(s) are non-conformational. No significant difference was observed between thymoma and non-thymoma patients or between early- and late-onset MG. In addition, atomic 3D-structures of the MIR and its subcomponents were elucidated using X-ray crystallography. These immunological and structural data will allow further studies into the atomic determinants underlying titin-based autoimmunity, improved diagnostics and how to eventually treat titin autoimmunity associated co-morbidities.

Circular Economy Electrochemistry: Creating Additive Manufacturing Feedstocks for Caffeine Detection from Post-Industrial Coffee Pod Waste.

The recycling of post-industrial waste poly(lactic acid) (PI-PLA) from coffee machine pods into electroanalytical sensors for the detection of caffeine in real tea and coffee samples is reported herein. The PI-PLA is transformed into both nonconductive and conductive filaments to produce full electroanalytical cells, including additively manufactured electrodes (AMEs). The electroanalytical cell was designed utilizing separate prints for the cell body and electrodes to increase the recyclability of the system. The cell body made from nonconductive filament was able to be recycled three times before the feedstock-induced print failure. Three bespoke formulations of conductive filament were produced, with the PI-PLA (61.62 wt %), carbon black (CB, 29.60 wt %), and poly(ethylene succinate) (PES, 8.78 wt %) chosen as the most suitable for use due to its equivalent electrochemical performance, lower material cost, and improved thermal stability compared to the filaments with higher PES loading and ability to be printable. It was shown that this system could detect caffeine with a sensitivity of 0.055 ± 0.001 µA µM-1, a limit of detection of 0.23 µM, a limit of quantification of 0.76 µM, and a relative standard deviation of 3.14% after activation. Interestingly, the nonactivated 8.78% PES electrodes produced significantly better results in this regard than the activated commercial filament toward the detection of caffeine. The activated 8.78% PES electrode was shown to be able to detect the caffeine content in real and spiked Earl Grey tea and Arabica coffee samples with excellent recoveries (96.7-102%). This work reports a paradigm shift in the way AM, electrochemical research, and sustainability can synergize and feed into part of a circular economy, akin to a circular economy electrochemistry.

All-in-One Single-Print Additively Manufactured Electroanalytical Sensing Platforms.

This manuscript provides the first report of a fully additively manufactured (AM) electrochemical cell printed all-in-one, where all the electrodes and cell are printed as one, requiring no post-assembly or external electrodes. The three-electrode cell is printed using a standard non-conductive poly(lactic acid) (PLA)-based filament for the body and commercially available conductive carbon black/PLA (CB/PLA, ProtoPasta) for the three electrodes (working, counter, and reference; WE, CE, and RE, respectively). The electrochemical performance of the cell is evaluated first against the well-known near-ideal outer-sphere redox probe hexaamineruthenium(III) chloride (RuHex), showing that the cell performs well using an AM electrode as the pseudo-RE. Electrochemical activation of the WE via chronoamperometry and NaOH provides enhanced electrochemical performances toward outer-sphere probes and for electroanalytical performance. It is shown that this activation can be completed using either an external commercial Ag|AgCl RE or through simply using the internal AM CB/PLA pseudo-RE and CE. This all-in-one electrochemical cell (AIOEC) was applied toward the well-known detection of ascorbic acid (AA) and acetaminophen (ACOP), achieving linear trends with limits of detection (LODs) of 13.6 ± 1.9 and 4.5 ± 0.9 µM, respectively. The determination of AA and ACOP in real samples from over-the-counter effervescent tablets was explored, and when analyzed individually, recoveries of 102.9 and 100.6% were achieved against UV-vis standards, respectively. Simultaneous detection of both targets was also achieved through detection in the same sample exhibiting 149.75 and 81.35% recoveries for AA and ACOP, respectively. These values differing from the originals are likely due to electrode fouling due to the AA oxidation being a surface-controlled process. The cell design produced herein is easily tunable toward different sample volumes or container shapes for various applications among aqueous electroanalytical sensing; however, it is a simple example of the capabilities of this manufacturing method. This work illustrates the next step in research synergising AM and electrochemistry, producing operational electrochemical sensing platforms in a single print, with no assembly and no requirements for exterior or commercial electrodes. Due to the flexibility, low-waste, and rapid prototyping of AM, there is scope for this work to be able to span and impact a plethora of research areas.

Conformational changes in twitchin kinase in vivo revealed by FRET imaging of freely moving C. elegans.

The force-induced unfolding and refolding of proteins is speculated to be a key mechanism in the sensing and transduction of mechanical signals in the living cell. Yet, little evidence has been gathered for its existence in vivo. Prominently, stretch-induced unfolding is postulated to be the activation mechanism of the twitchin/titin family of autoinhibited sarcomeric kinases linked to the mechanical stress response of muscle. To test the occurrence of mechanical kinase activation in living working muscle, we generated transgenic Caenorhabditis elegans expressing twitchin containing FRET moieties flanking the kinase domain and developed a quantitative technique for extracting FRET signals in freely moving C. elegans, using tracking and simultaneous imaging of animals in three channels (donor fluorescence, acceptor fluorescence, and transmitted light). Computer vision algorithms were used to extract fluorescence signals and muscle contraction states in each frame, in order to obtain fluorescence and body curvature measurements with spatial and temporal precision in vivo. The data revealed statistically significant periodic changes in FRET signals during muscle activity, consistent with a periodic change in the conformation of twitchin kinase. We conclude that stretch-unfolding of twitchin kinase occurs in the active muscle, whereby mechanical activity titrates the signaling pathway of this cytoskeletal kinase. We anticipate that the methods we have developed here could be applied to obtaining in vivo evidence for force-induced conformational changes or elastic behavior of other proteins not only in C. elegans but in other animals in which there is optical transparency (e.g., zebrafish).

Use of sodium zirconium cyclosilicate for up-titration of renin-angiotensin-aldosterone system inhibitor therapy in patients with heart failure: a case series.

BACKGROUND: Patients often receive suboptimal dosing of renin-angiotensin-aldosterone system inhibitor (RAASi) therapy over concerns of hyperkalaemia. However, studies have shown associations between suboptimal dosing or interruptions to therapy and adverse clinical events. Therefore, effective treatments for hyperkalaemia that can enable optimal RAASi therapy are needed. This case series examines eight patients whose commencement on the novel potassium binder sodium zirconium cyclosilicate (SZC) allowed for the initiation and/or up-titration of RAASi therapy. CASE SUMMARY: Eight patients aged 64-87 years with heart failure (HF) with reduced ejection fraction all developed hyperkalaemia (serum potassium (sK+) >5.0 mmol/L) while receiving RAASi therapy. Following initiation of SZC, all patients experienced eventual stabilization of sK+ levels. All patients were able to initiate, restart, or up-titrate RAASi therapy with five patients achieving optimal medical therapy. Left ventricular ejection fraction improved in four patients, two patients are now re-classified as New York Heart Association Class I, and an additional patient had improved exercise tolerance. Follow-up for Patient 8 is still ongoing. DISCUSSION: These real-world cases demonstrate that use of SZC to manage hyperkalaemia in patients with HF is feasible and allows optimization of RAASi therapy.

Health-Related Quality of Life Among Patients With HR+/HER2- Early Breast Cancer.

PURPOSE: The goal of this study was to characterize health-related quality of life (HRQOL) among patients diagnosed with early-stage, hormone receptor-positive (HR+)/human epidermal growth factor receptor 2 negative (HER2-) breast cancer. METHODS: A multinational (United States, Japan, France, Germany, Italy, Spain, and United Kingdom) study of patients diagnosed with stage I to III HR+/HER2- breast cancer, either receiving adjuvant treatment or under postadjuvant surveillance, was conducted between June and October 2019. Patients were identified by their consulting physician and invited to complete the Functional Assessment of Cancer Therapy-Breast (FACT-B) and the EQ-5D-5L pen and paper questionnaires. EQ-5D-5L index scores were derived by using available country-specific health state value sets, where available, and numerically compared with general population scores derived from published normative and population data. Descriptive summary statistics were reported for FACT-B, Functional Assessment of Cancer Therapy-General (FACT-G) (total and specific subscales), the EQ-5D index scores, and the EQ-VAS scores for each country. Results were stratified according to disease-free treatment status (active adjuvant treatment or postadjuvant surveillance), age (25-44, 45-54, 55-64, or ≥65 years), stage (I, II, or III), and menopausal status at the time of questionnaire completion (pre-/peri-menopausal or postmenopausal). FINDINGS: Overall, 1110 patients completed the HRQOL questionnaires (mean age, 59 years; 79% active adjuvant treatment, and 21% under surveillance postadjuvant treatment at time of questionnaire administration; 31% stage I, 48% stage II, and 20% stage III at diagnosis). Of these, 1102 completed the FACT-B and 1083 completed the EQ-5D-5L questionnaires. The mean (SD) FACT-B total score was 99.0 (21.9). The mean FACT-G total score was 72.5 (17.8), which was comparable to the published normative score. The mean EQ-5D index and EQ-VAS scores for each country were similar to corresponding population means; EQ-5D index scores ranged from 0.842 (0.098) in Japan to 0.916 (0.109) in France, and EQ-VAS scores from 68.0 (18.4) in Germany to 78.6 (16.4) in the United States. In addition, mean scores were comparable between the active adjuvant treatment and postadjuvant surveillance groups for the FACT-B total (99.4 [22.5] and 97.7 [19.7], respectively), FACT-G total (72.8 [18.3] and 71.3 [16.0]), EQ-5D index score (0.868 [0.135] and 0.869 [0.142]), and EQ-VAS (74.9 [17.2] and 74.4 [16.1]). IMPLICATIONS: Patient-reported HRQOL among patients with HR+/HER2- early breast cancer who were disease-free was high, with reported scores comparable to normative scores. These results improve our understanding of HRQOL among patients with early disease and may facilitate future studies examining the potential impact of adjuvant treatment and disease recurrence, including metastasis.

Roles of ATM and ATR in DNA double strand breaks and replication stress.

The maintenance of genome integrity is critical for the faithful replication of the genome during cell division and for protecting cells from accumulation of DNA damage, which if left unrepaired leads to a loss of genetic information, a breakdown in cell function and ultimately cell death and cancer. ATM and ATR are master kinases that are integral to homologous recombination-mediated repair of double strand breaks and preventing accumulation of dangerous DNA structures and genome instability during replication stress. While the roles of ATM and ATR are heavily intertwined in response to double strand breaks, their roles diverge in the response to replication stress. This review summarises our understanding of the players and their mode of actions in recruitment, activation and activity of ATM and ATR in response to DNA damage and replication stress and discusses how controlling localisation of these kinases and their activators allows them to orchestrate a stress-specific response.

Roles of ATM and ATR in DNA double strand breaks and replication stress.

The maintenance of genome integrity is critical for the faithful replication of the genome during cell division and for protecting cells from accumulation of DNA damage, which if left unrepaired leads to a loss of genetic information, a breakdown in cell function and ultimately cell death and cancer. ATM and ATR are master kinases that are integral to homologous recombination-mediated repair of double strand breaks and preventing accumulation of dangerous DNA structures and genome instability during replication stress. While the roles of ATM and ATR are heavily intertwined in response to double strand breaks, their roles diverge in the response to replication stress. This review summarises our understanding of the players and their mode of actions in recruitment, activation and activity of ATM and ATR in response to DNA damage and replication stress and discusses how controlling localisation of these kinases and their activators allows them to orchestrate a stress-specific response.

Systemic capillary leak syndrome: a rare but potentially life-threatening cause of protein loss and oedema in B cell prolymphocytic leukaemia.

A 64-year-old man had a several year history of B prolymphocytic leukaemia (PLL) which behaved indolently and had not required any treatment. Five years after diagnosis, he developed hypoalbuminaemia associated with severe lower-limb oedema, consistent with systemic capillary leak syndrome (SCLS). He recovered spontaneously but went on to have three further increasingly severe and protracted episodes over the subsequent 18 months. There was no identifiable precipitating factor for these episodes, but his peripheral lymphocyte count continued to increase slowly. The start of treatment for his PLL with chemoimmunotherapy was followed by a rapid resolution of residual oedema and normalisation of serum albumin. He has had no further attacks of SCLS in the 14 months since he started therapy for PLL. SCLS is a rare consequence of haematological malignancy which may show an excellent response to treatment of the haematological disease.

Multidisciplinary, patient-centred approach to improving compliance with venous thromboembolism (VTE) prophylaxis in a district general hospital.

Hospital-acquired venous thromboembolism (VTE) accounts for an estimated 25 000 preventable deaths per annum in the UK and is associated with significant healthcare costs. The National Institute for Health and Care Excellence guidelines on the prevention of VTE in hospitalised patients highlight the clinical and cost-effectiveness of VTE prevention strategies. A multidisciplinary quality improvement team (MD QIT) based in a district general hospital sought to improve compliance with VTE prophylaxis prescription to greater than 85% of patients within a 3-month time frame. Quality improvement methodology was adopted over three cycles of the project. Interventions included the introduction of a 'VTE sticker' to prompt risk assessment; educational material for medical staff and allied healthcare professionals; and patient information raising the awareness of the importance of VTE prophylaxis. Implementation of these measures resulted in significant and sustained improvements in rates of risk assessment within 24 hours of admission to hospital from 51% compliance to 94% compliance after cycle 2 of the project. Improvements were also observed in medication dose adjustment for the patient weight from 69% to 100% compliance. Dose adjustments for renal function showed similar trends with compliance with guidelines improving from 80% to 100%. These results were then replicated in a different clinical environment. In conclusion, this project exemplifies the benefits of MD QITs in terms of producing sustainable and replicable improvements in clinical practice and in relation to meeting approved standards of care for VTE risk assessment and prescription. It has been demonstrated that the use of educational material in combination with a standardised risk assessment tool, the 'VTE sticker', significantly improved clinical practice in the context of a general medical environment.

Evidence for Self-healing Benign Grain Boundaries and a Highly Defective Sb2Se3-CdS Interfacial Layer in Sb2Se3 Thin-Film Photovoltaics.

The crystal structure of Sb2Se3 gives rise to unique properties that cannot otherwise be achieved with conventional thin-film photovoltaic materials, such as CdTe or Cu(In,Ga)Se2. It has previously been proposed that grain boundaries can be made benign provided only the weak van der Waals forces between the (Sb4Se6)n ribbons are disrupted. Here, it is shown that non-radiative recombination is suppressed even for grain boundaries cutting across the (Sb4Se6)n ribbons. This is due to a remarkable self-healing process, whereby atoms at the grain boundary can relax to remove any electronic defect states within the band gap. Grain boundaries can, however, impede charge transport due to the fact that carriers have a higher mobility along the (Sb4Se6)n ribbons. Because of the ribbon misorientation, certain grain boundaries can effectively block charge collection. Furthermore, it is shown that CdS is not a suitable emitter to partner Sb2Se3 due to Sb and Se interdiffusion. As a result, a highly defective Sb2Se3 interfacial layer is formed that potentially reduces device efficiency through interface recombination.

Diabetes and global ageing among 65-99-year-old adults: Findings from the International Diabetes Federation Diabetes Atlas, 9th edition.

AIMS: The main aims are: (a) to draw attention to the nature and pattern of recent global and regional prevalence estimates and projections of diabetes in older adults (65-99 years), and (b) to describe the societal health implications of these changes on a global scale. METHODS: Diabetes prevalences and projections were estimated using a logistic regression method to generate smoothed age- and sex-specific prevalence estimates for 5-year age groups. RESULTS: In 2019, it is estimated that 19.3% of people aged 65-99 years (135.6 million, 95% CI: 107.6-170.6 million) live with diabetes. It is projected that the number of people older than 65 years (65-99 years) with diabetes will reach 195.2 million by 2030 and 276.2 million by 2045. For the regional distribution, the highest prevalence in 2019 being the North America and Caribbean Region at 27.0%. Countries with the highest number of people older than 65 years with diabetes are China, the United States of America and India. CONCLUSIONS: There is a need for more data from national and regional sources on those aged 65 years and over, but the prevailing evidence points to diabetes being a considerable global chronic illness burden in ageing societies.

Mortality attributable to diabetes in 20-79 years old adults, 2019 estimates: Results from the International Diabetes Federation Diabetes Atlas, 9th edition.

AIMS: To estimate the number of deaths attributable to diabetes in 20-79-year-old adults in 2019. METHODS: The following were used to estimate the number of deaths attributable to diabetes: all-cause mortality estimates from the World Health Organization life table, country level age- and sex-specific estimates of diabetes prevalence in 2019 and relative risks of death in people with diabetes compared to people without diabetes. RESULTS: An estimated 4.2 million deaths among 20-79-year-old adults are attributable to diabetes. Diabetes is estimated to contribute to 11.3% of deaths globally, ranging from 6.8% (lowest) in the Africa Region to 16.2% (highest) in the Middle East and North Africa. About half (46.2%) of the deaths attributable to diabetes occur in people under the age of 60 years. The Africa Region has the highest (73.1%) proportion of deaths attributable to diabetes in people under the age of 60 years, while the Europe Region has the lowest (31.4%). CONCLUSIONS: Diabetes is estimated to contribute to one in nine deaths among adults aged 20-79 years. Prevention of diabetes and its complications is essential, particularly in middle-income countries, where the current impact is estimated to be the largest. Contemporary data from diverse populations are needed to validate these estimates.

Global and regional estimates and projections of diabetes-related health expenditure: Results from the International Diabetes Federation Diabetes Atlas, 9th edition.

AIMS: Diabetes and its complications have a significant economic impact on individuals and their families, health systems and national economies. METHODS: The direct health expenditure of diabetes was calculated relying on the following inputs: diagnosed and undiagnosed diabetes prevalence estimates, United Nations population estimates, World Health Organization health expenditure per capita and ratios of health expenditure for people with diabetes compared to people without diabetes. RESULTS: The estimated global direct health expenditure on diabetes in 2019 is USD 760 billion and is expected to grow to a projected USD 825 billion by 2030 and USD 845 billion by 2045. There is a wide variation in annual health expenditures on diabetes. The United States of America has the highest estimated expenditure with USD 294.6 billion, followed by China and Brazil, with USD 109.0 billion and USD 52.3 billion, respectively. The age group with the largest annual diabetes-related health expenditure is 60-69 years with USD 177.7 billion, followed by 50-59 years, and 70-79 years with USD 173.0 billion and USD 171.5 billion, respectively. Slightly higher diabetes-related health expenditure is seen in women than in men (USD 382.6 billion vs. USD 377.6 billion, respectively). The same difference is expected to be present in 2030 and 2045. CONCLUSIONS: There were large disparities between high-, middle- and low-income countries with total health expenditures in high-income countries being over 300 times those in low-income countries. The ratio for annual direct health expenditure per person between these groups of countries is more than 38-fold.

Structures and regulations of ATM and ATR, master kinases in genome integrity.

Homologous recombination (HR) is a faithful repair mechanism for double stranded DNA breaks. Two highly homologous master kinases, the tumour suppressors ATM and ATR (Tel1 and Mec1 in yeast), coordinate cell cycle progression with repair during HR. Despite their importance, our molecular understanding of these apical coordinators has been limited, in part due to their large sizes. With the recent development in cryo-electron microscopy, significant advances have been made in structural characterisation of these proteins in the last two years. These structures, combined with new biochemical studies, now provide a more detailed understanding of how a low basal activity is maintained and how activation may occur. In this review, we summarize recent advances in the structural and molecular understanding of these key components in HR, compare the common and distinct features of these kinases and suggest aspects of structural components that are likely to be involved in regulating its activity.

Cryo-EM Structure of Nucleotide-Bound Tel1ATM Unravels the Molecular Basis of Inhibition and Structural Rationale for Disease-Associated Mutations.

Yeast Tel1 and its highly conserved human ortholog ataxia-telangiectasia mutated (ATM) are large protein kinases central to the maintenance of genome integrity. Mutations in ATM are found in ataxia-telangiectasia (A-T) patients and ATM is one of the most frequently mutated genes in many cancers. Using cryoelectron microscopy, we present the structure of Tel1 in a nucleotide-bound state. Our structure reveals molecular details of key residues surrounding the nucleotide binding site and provides a structural and molecular basis for its intrinsically low basal activity. We show that the catalytic residues are in a productive conformation for catalysis, but the phosphatidylinositol 3-kinase-related kinase (PIKK) regulatory domain insert restricts peptide substrate access and the N-lobe is in an open conformation, thus explaining the requirement for Tel1 activation. Structural comparisons with other PIKKs suggest a conserved and common allosteric activation mechanism. Our work also provides a structural rationale for many mutations found in A-T and cancer.