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The COVID-19 Pandemic has revolutionized the field of mRNA vaccine technology, highlighting its utility and rapidly expanding potential, propelling it to the forefront of future vaccines & therapeutics for cryptic diseases. T7 RNA Polymerase (T7 RNAP) is a high-fidelity enzyme used for the synthesis of single-stranded mRNA from DNA templates using In vitro transcription (IVT). T7 RNAP is now the mainstay for the development of mRNA vaccines and therapeutics, accounting for 34 % of the overall raw material cost for mRNA manufacturing. Although small-scale purification processes for T7 RNAP exist, scalable process platforms and comprehensive analytical characterization are still lacking. Moreover, to address global raw material supply chain issues and to facilitate the efficient and large-scale production of mRNA vaccines & therapeutics, here we report a three-step purification process platform, yielding ∼ 85–100 mg/L of T7 RNAP from E. coli fermentation harvest. Also, to demonstrate its high-level purity and structural integrity, we have characterized it using orthogonal analytical techniques. Furthermore, when evaluated in an IVT system, this highly pure T7 RNAP showed activity that was comparable to the commercial T7 RNAP in producing integral RNA (10 kb) from a linearized plasmid DNA template. Collectively, these findings establish a purification process platform for the commercial production of T7 RNAP enzyme to support the future development of mRNA-based vaccines and therapeutics. [Display omitted] • 85–100 mg of T7 RNAP purified from 1 L of E. Coli fermentation harvest. • Purified T7 RNAP showed 99 % and 94 % purity in SE and RP-HPLC respectively. • Purified T7 RNAP demonstrated an intact secondary structure in CD. • Purified T7 RNAP produced 10 kb of integral RNA in IVT. • T7 RNAP production platform for mRNA vaccines and therapeutics. [ABSTRACT FROM AUTHOR]

Background: Frailty, defined as multidimensional prognostic index (MPI), has been recently identified as strong predictor of disability and mortality in the elderly with acute ischemic stroke (AIS). The stress hyperglycemia ratio (SHR) is a recently introduced biomarker significantly associated with poor outcome in AIS. Objectives: This study aimed to investigate in what extent frailty, measured by MPI, and SHR affects the 3-months outcome of patients > 65 years-old with AIS. Methods: Consecutive patients with AIS >65 years-old who underwent intravenous thrombolysis (IVT) from 2015 to 2019 were enrolled in a German and an Italian Stroke Unit. The SHR was calculated by dividing the fasting plasma glucose at admission with glycated hemoglobin. Demographics and clinical premorbid data, stroke-related variables, including baseline and post-treatment NIHSS score were included in a logistic regression model. The 3-months functional outcome was evaluated by using modified Rankin scale (mRS); good outcome was defined as mRS 0–2, poor as mRS ≥ 3. Results: One hundred and fifty-five AIS patients were enrolled in the study. Median MPI was 0.19 [0.13–0.31]; 118 (76.1%) patients were classified as "robust" and 37 (23.9%) as "frail." In regression analysis, age, NIHSS, and MPI demonstrated as the most significant predictor of 3-months good outcome in the whole cohort. In robust patients, SHR values were significantly associated with the outcome. Conclusions: MPI is associated with the 3-months outcome in our cohort, in particular with good outcome. Conversely, SHR seems to be associated with a 3-months poor outcome in "robust" patients but not in frail patients. [ABSTRACT FROM AUTHOR]

This study analyzes the dynamics of short-term treatment response to the first intravitreal faricimab injection in eyes with diabetic macular edema (DME). This retrospective, single-center, clinical trial was conducted at the Department of Ophthalmology, University Hospital Zurich. Patients with treatment-naïve and pretreated DME were included. Patient chart data and imaging were analyzed. Safety and efficacy (corrected visual acuity (CVA), central subfield thickness (CST), and signs of intraocular inflammation (IOI)) of the first faricimab intravitreal therapy (IVT) were evaluated weekly until 4 weeks after injection. Forty-three eyes (81% pretreated) of 31 patients were included. Four weeks after the first faricimab IVT, CVA remained stable and median CST (µm) decreased significantly (p < 0.001) from 325.0 (293.5–399.0) at baseline to 304.0 (286.5–358.0). CVA at week 4 was only associated with baseline CVA (p < 0.001). CST was the only predictive variable (p = 0.002) between baseline and week 4 CST. Weekly safety assessments did not show any sign of clinically significant IOI. This study suggests faricimab is an effective treatment for (pretreated) DME, showing structural benefit 1 month following the first injection without short-term safety signals. [ABSTRACT FROM AUTHOR]

Intravenous thrombolysis (IVT) and dual antiplatelet therapy (DAPT) have been widely used in minor ischemic stroke (MIS) treatment. However, the clinical outcomes and safety of these two treatments have not been compared within the early thrombolytic time window. Here, we conducted a multicenter, ambispective cohort study involving patients with MIS presenting within 4.5 h of symptom onset at 3 affiliated hospitals of Jinan University from 2018–2022. The patients were divided into the IVT group and DAPT group. The primary outcome was a 90-day excellent outcome (mRS ≤ 1). A total of 1,026 patients were enrolled, of whom 492 were assigned to the IVT group and 534 were assigned to the DAPT group. The IVT group had better 90-day excellent outcomes (mRS ≤ 1) than the DAPT group (OR 1.69, 95% CI 1.14–2.52, P = 0.010). Among the 623 patients with nondisabling stroke, the proportion of mRS ≤ 1 in the IVT group was higher than the DAPT group (P = 0.009). In the subtypes of MIS with large vessel occlusion/stenosis and with isolated symptoms, the 90-day outcomes of the IVT group and DAPT group were not different (P > 0.05). In conclusion, compared with DAPT, IVT was associated with better 90-day clinical outcomes in patients with MIS (in particular, for those with mRS > 1), including earlier clinical improvement.IVT also benefited the early neurological improvement of patients with severe stenosis/occlusion of intracranial large vessels, nondisabling mild stroke, nondisabling mild stroke with isolated symptoms. [ABSTRACT FROM AUTHOR]

mRNA vaccines are leading a medical revolution. mRNA technologies utilize the host's own cells as bio-factories to produce proteins that serve as antigens. This revolutionary approach circumvents the complicated processes involved in traditional vaccine production and empowers vaccines with the ability to respond to emerging or mutated infectious diseases rapidly. Additionally, the robust cellular immune response elicited by mRNA vaccines has shown significant promise in cancer treatment. However, the inherent instability of mRNA and the complexity of tumor immunity have limited its broader application. Although the emergence of pseudouridine and ionizable cationic lipid nanoparticles (LNPs) made the clinical application of mRNA possible, there remains substantial potential for further improvement of the immunogenicity of delivered antigens and preventive or therapeutic effects of mRNA technology. Here, we review the latest advancements in mRNA vaccines, including but not limited to target selection and delivery systems. This review offers a multifaceted perspective on this rapidly evolving field. [ABSTRACT FROM AUTHOR]

BackgroundFrailty, defined as multidimensional prognostic index (MPI), has been recently identified as strong predictor of disability and mortality in the elderly with acute ischemic stroke (AIS). The stress hyperglycemia ratio (SHR) is a recently introduced biomarker significantly associated with poor outcome in AIS.ObjectivesThis study aimed to investigate in what extent frailty, measured by MPI, and SHR affects the 3-months outcome of patients > 65 years-old with AIS.MethodsConsecutive patients with AIS >65 years-old who underwent intravenous thrombolysis (IVT) from 2015 to 2019 were enrolled in a German and an Italian Stroke Unit. The SHR was calculated by dividing the fasting plasma glucose at admission with glycated hemoglobin. Demographics and clinical premorbid data, stroke-related variables, including baseline and post-treatment NIHSS score were included in a logistic regression model. The 3-months functional outcome was evaluated by using modified Rankin scale (mRS); good outcome was defined as mRS 0–2, poor as mRS ≥ 3.ResultsOne hundred and fifty-five AIS patients were enrolled in the study. Median MPI was 0.19 [0.13–0.31]; 118 (76.1%) patients were classified as “robust” and 37 (23.9%) as “frail.” In regression analysis, age, NIHSS, and MPI demonstrated as the most significant predictor of 3-months good outcome in the whole cohort. In robust patients, SHR values were significantly associated with the outcome.ConclusionsMPI is associated with the 3-months outcome in our cohort, in particular with good outcome. Conversely, SHR seems to be associated with a 3-months poor outcome in “robust” patients but not in frail patients.

In the absence of clinical trials, the benefit of endovascular therapy (EVT) on the treatment of acute ischemic stroke (AIS) with primary distal and medium vessel occlusions (DMVO) is still not well defined. The aim of the study is to evaluate EVT with or without intravenous thrombolysis (EVT ± IVT) in primary DMVO stroke in comparison with a control cohort treated with IVT alone.

Objectives: This study entailed a weekly analysis of real-world data (RWD) on the safety and efficacy of intravitreal (IVT) faricimab in neovascular age-related macular degeneration (nAMD). Methods: A retrospective, single-centre clinical trial was conducted at the Department of Ophthalmology, University Hospital Zurich, University of Zurich, Switzerland, approved by the Cantonal Ethics Committee of Zurich, Switzerland. Patients with nAMD were included. Data from patient charts and imaging were analysed. The safety and efficacy of the first faricimab injection were evaluated weekly until 4 weeks after injection. Results: Sixty-three eyes with a complete 4-week follow-up were enrolled. Six eyes were treatment-naïve; fifty-seven eyes were switched to faricimab from another treatment. Neither group showed signs of retinal vasculitis during the 4 weeks after injection. Central subfield thickness (CST) and volume (CSV) showed a statistically significant decrease compared to the baseline in the switched group (CST: p = 0.00383; CSV: p = 0.00702) after 4 weeks. The corrected visual acuity returned to the baseline level in both groups. The macular neovascularization area decreased in both groups, but this was not statistically significant. A complete resolution of sub- and intraretinal fluid after 4 weeks was found in 40% (switched) and 75% (naïve) of the treated patients. Conclusions: The weekly follow-ups reflect the structure–function relationship beginning with a fast functional improvement within two weeks after injection followed by a return to near-baseline levels after week 3. The first faricimab injection in our cohort showed a high safety profile and a statistically significant reduction in macular oedema in switched nAMD patients. [ABSTRACT FROM AUTHOR]

Background: There are three main methods of mechanical thrombectomy (MT): using a stent retriever (SR) only (SO), aspiration catheter (AC) only (AO) and the combined method (CM) using both the SR and AC. This paper describes a real-life, single-center experience using SO, AO and CM during 276 consecutive MTs. Methods: The primary endpoint was the frequency of first-pass complete (FPE TICI3). The secondary endpoints were final mTICI 2b-3, procedure duration, clinical outcome and the total number of device passes. The third aim of this study was to test the association between the clinical outcomes in patients treated with each method and various factors. Results: There was a significant difference (p = 0.016) between the groups' FPE TICI3 rates with 46% mTICI 3 in the AO group, 41% in the CM group and 21% in the SO group. AO resulted in procedure time shortening to a mean duration of 43 min, and the scores were 56 min for CM and 63 min for SO (p < 0.0001). There were no significant differences in clinical outcomes or in-hospital mortality. The analysis showed a correlation between good clinical outcomes and the administration of IVT: OR 1.71 (1.03–2.84) p = 0.039. Patients ≥66 years old had higher odds of a bad outcome compared to younger patients in general (OR, 1.99 95% CI, 1.17–3.38; p = 0.011). FPE TICI3 was associated with good functional outcomes in the whole treated cohort (OR, 1.98; 95% CI, 1.21–3.25; p = 0.006). Conclusions: In our series, AO proved to be the best starting point in most cases. It demonstrates good technical efficacy regarding FPE, it is fast and clinical outcomes seem to be the least age- and FPE TICI3-dependent. It can be easily converted into the combined method, which had the second-best outcomes in our cohort. [ABSTRACT FROM AUTHOR]

mRNA vaccines are leading a medical revolution. mRNA technologies utilize the host’s own cells as bio-factories to produce proteins that serve as antigens. This revolutionary approach circumvents the complicated processes involved in traditional vaccine production and empowers vaccines with the ability to respond to emerging or mutated infectious diseases rapidly. Additionally, the robust cellular immune response elicited by mRNA vaccines has shown significant promise in cancer treatment. However, the inherent instability of mRNA and the complexity of tumor immunity have limited its broader application. Although the emergence of pseudouridine and ionizable cationic lipid nanoparticles (LNPs) made the clinical application of mRNA possible, there remains substantial potential for further improvement of the immunogenicity of delivered antigens and preventive or therapeutic effects of mRNA technology. Here, we review the latest advancements in mRNA vaccines, including but not limited to target selection and delivery systems. This review offers a multifaceted perspective on this rapidly evolving field.

Synoptic fingerprinting of flood driving rainfall events over the Hyderabad city, India were assessed through an unsupervised machine learning technique—self‐organizing maps (SOM) for the period 1979–2020. Flood dates were identified using nonscientific data sources, and then large‐scale climate variables, that is, integrated vapour transport (IVT) and geopotential height at 500 hPa (Z500) along with rainfall at daily scale were considered. SOM was applied on IVT and results indicated large spatial scale‐ and season‐specific mechanisms in addition to local circulation systems—this highlights SOM's ability to cluster circulation mechanisms as per the relevance. The rainy seasons, that is, monsoon and postmonsoon showed well‐known moisture pathways. While synoptic systems can be easily linked with monsoon and postmonsoon mechanisms, it is interesting to note that the local systems exist even during the monsoon and postmonsoon season in addition to the summer season. Spatial patterns of Z500 corresponding to SOM nodes of IVT showed the presence of low‐pressure systems (LPSs), tropical cyclones and trough systems. Further analysis was performed, and a total of 35 atmospheric rivers (ARs) were identified. Plots of back trajectories for the selected AR events indicated that Arabian Sea is the main source of moisture in the monsoon season followed by Bay of Bengal for postmonsoon and tropical cyclones in summer season events. Additionally, moisture from land is also observed as another source of moisture. Back‐trajectory analysis further indicated AR‐LPS interaction during the monsoon and ARs feeding moisture to LPSs. Overall, well‐known and season‐specific synoptic systems that brought significant rainfall to the study region were identified by the SOM. The proposed framework is adaptable for different locations, and as floods in cities across the globe are on the rise, these findings will have a prominent role in urban flood forecasting and management. [ABSTRACT FROM AUTHOR]

Background: Despite the high incidence of acute ischemic stroke (AIS) in cancer patients, there is still no consensus about the safety of recanalization therapies in this cohort. Objectives: In this observational study, our aim was to investigate the bleeding risk after acute recanalization therapy in AIS patients with active malignancy. Methods and Study Design: We retrospectively analyzed observational data of 1016 AIS patients who received intravenous thrombolysis with rtPA (IVT) and/or endovascular therapy (EVT) between January 2017 and December 2020 with a focus on patients with active malignancy. The primary safety endpoint was the occurrence of stroke treatment-related major bleeding events, that is, symptomatic intracranial hemorrhage (SICH) and/or relevant systemic bleeding. The primary efficacy endpoint was neurological improvement during hospital stay (NI). Results: None of the 79 AIS patients with active malignancy suffered from stroke treatment-related systemic bleeding. The increased rate (7.6% versus 4.7%) of SICH after therapy compared to the control group was explained by confounding factors. A total of nine patients with cerebral tumor manifestation received acute stroke therapy, two of them suffered from stroke treatment-related intracranial hemorrhage remote from the tumor, both asymptomatic. The group of patients with active malignancy and the control group showed comparable rates of NI. Conclusion: Recanalization therapy in AIS patients with active malignancy was not associated with a higher risk for stroke treatment-related systemic or intracranial bleeding. IVT and/or EVT can be regarded as a safe therapy option for AIS patients with active malignancy. [ABSTRACT FROM AUTHOR]

Objectives: This study entailed a weekly analysis of real-world data (RWD) on the safety and efficacy of intravitreal (IVT) faricimab in neovascular age-related macular degeneration (nAMD). Methods: A retrospective, single-centre clinical trial was conducted at the Department of Ophthalmology, University Hospital Zurich, University of Zurich, Switzerland, approved by the Cantonal Ethics Committee of Zurich, Switzerland. Patients with nAMD were included. Data from patient charts and imaging were analysed. The safety and efficacy of the first faricimab injection were evaluated weekly until 4 weeks after injection. Results: Sixty-three eyes with a complete 4-week follow-up were enrolled. Six eyes were treatment-naïve; fifty-seven eyes were switched to faricimab from another treatment. Neither group showed signs of retinal vasculitis during the 4 weeks after injection. Central subfield thickness (CST) and volume (CSV) showed a statistically significant decrease compared to the baseline in the switched group (CST: p = 0.00383; CSV: p = 0.00702) after 4 weeks. The corrected visual acuity returned to the baseline level in both groups. The macular neovascularization area decreased in both groups, but this was not statistically significant. A complete resolution of sub- and intraretinal fluid after 4 weeks was found in 40% (switched) and 75% (naïve) of the treated patients. Conclusions: The weekly follow-ups reflect the structure–function relationship beginning with a fast functional improvement within two weeks after injection followed by a return to near-baseline levels after week 3. The first faricimab injection in our cohort showed a high safety profile and a statistically significant reduction in macular oedema in switched nAMD patients.

Purpose: The increased use of neuroimaging and innovations in ischemic stroke (IS) treatment have improved outcomes, but the impact on median hospital costs is not well understood.Methods: A retrospective study was conducted using Medicare 5% claims data for 75,525 consecutive index IS hospitalizations for patients aged ≥65 years from 2012 to 2019 (values in 2019 dollars). IS episode cost was calculated in each year for trend analysis and stratified by cost components, including neuroimaging (CT angiography [CTA], CT perfusion [CTP], MRI, and MR angiography [MRA]), treatment (endovascular thrombectomy [EVT] and/or intravenous thrombolysis), and patient sociodemographic factors. Logistic regression was performed to analyze the drivers of high-cost episodes and median regression to assess drivers of median costs.Results: The median IS episode cost increased by 4.9% from $9,509 in 2012 to $9,973 in 2019 (P = .0021). Treatment with EVT resulted in the greatest odds of having a high-cost (>$20,000) hospitalization (odds ratio [OR], 71.86; 95% confidence interval [CI], 54.62-94.55), as did intravenous thrombolysis treatment (OR, 3.19; 95% CI, 2.90-3.52). Controlling for other factors, neuroimaging with CTA (OR, 1.72; 95% CI, 1.58-1.87), CTP (OR, 1.32; 95% CI, 1.14-1.52), and/or MRA (OR, 1.26; 95% CI, 1.15-1.38) had greater odds of having high-cost episodes than those without CTA, CTP, and MRA. Length of stay > 4 days (OR, 4.34; 95% CI, 3.99-4.72) and in-hospital mortality (OR, 1.85; 95% CI, 1.63-2.10) were also associated with high-cost episodes.Conclusions: From 2012 to 2019, the median IS episode cost increased by 4.9%, with EVT as the main cost driver. However, the increasing treatment cost trends have been partially offset by decreases in median length of stay and in-hospital mortality. [ABSTRACT FROM AUTHOR]

The cellular automaton (CA) and an integral value transformation (IVT) evolving in discrete time steps are two mathematical models that are well established. Theoretically, it can be suggested that a CA possesses the capacity to produce varieties of evolutionary patterns. However, computing a CA in higher dimensions or computing a nonlinear CA may be complex. In such cases, an IVT can be conveniently used. This paper presents the relation between the transition functions of a one-dimensional CA and an IVT. It also highlights the algebraic structures on the basis of binary operations for a set of transition functions of a one-dimensional CA and for a set of IVTs. The suitability of using an IVT over a CA is discussed. Also, we present the evolutionary models of two deoxyribonucleic acid (DNA) sequences through IVTs and their spacetime diagrams. This can eventually bring out some characteristic features of the evolutionary sequences. [ABSTRACT FROM AUTHOR]

After the COVID-19 pandemic, messenger RNA (mRNA) has revolutionized traditional vaccine manufacturing. With the increasing number of RNA-based therapeutics, valuable new scientific insights into these molecules have emerged. One fascinating area of study is the formation of double-stranded RNA (dsRNA) during in vitro transcription (IVT) which is considered a significant impurity, as it has been identified as a major trigger in the cellular immune response pathway. Therefore, there is a growing importance placed to develop and optimize purification processes for the removal of this by-product. Traditionally, efforts have primarily focused on mRNA purification after IVT through chromatographic separations, with anion exchange and reverse phase chromatography emerging as effective tools for this purpose. However, to the best of our knowledge, the influence and significance of the quality of the linearized plasmid have not been thoroughly investigated. Plasmids production involves the growth of bacterial cultures, bacterial harvesting and lysis, and multiple filtration steps for plasmid DNA purification. The inherent complexity of these molecules, along with the multitude of purification steps involved in their processing, including the subsequent linearization and the less-developed purification techniques for linearized plasmids, often result in inconsistent batches with limited control over by-products such as dsRNA. This study aims to demonstrate how the purification process employed for linearized plasmids can impact the formation of dsRNA. Several techniques for the purification of linearized plasmids based on both, resin filtration and chromatographic separations, have been studied. As a result of that, we have optimized a chromatographic method for purifying linearized plasmids using monolithic columns with C4 chemistry (butyl chains located in the surface of the particles), which has proven successful for mRNAs of various sizes. This chromatographic separation facilitates the generation of homogeneous linearized plasmids, leading to mRNA batches with lower levels of dsRNA during subsequent IVT processes. This finding reveals that dsRNA formation is influenced not only by RNA polymerase and IVT conditions but also by the quality of the linearized template. The results suggest that plasmid impurities may contribute to the production of dsRNA by providing additional templates that can be transcribed into sequences that anneal with the mRNA molecules. This highlights the importance of considering the quality of plasmid purification in relation to dsRNA generation during transcription. Further investigation is needed to fully understand the mechanisms and implications of plasmid-derived dsRNA. This discovery could shift the focus in mRNA vaccine production, placing more emphasis on the purification of linearized plasmids and potentially saving, in some instances, a purification step for mRNA following IVT.

RNA-based therapeutics and vaccines are opening up new avenues for modern medicine. To produce these useful RNA-based reagents, in vitro transcription (IVT) is an important reaction that primarily determines the yield and quality of the product. Therefore, IVT condition should be well optimized to achieve high yield and purity of transcribed RNAs. To this end, real-time monitoring of RNA production during IVT, which allows for fine tuning of the condition, would be required. Currently, light-up RNA aptamer and fluorescent dye pairs are considered as useful strategies to monitor IVT in real time. Fluorophore-labeled antisense probe-based methods can also be used for real-time IVT monitoring. In addition, a high-performance liquid chromatography (HPLC)-based method that can monitor IVT reagent consumption has been developed as a powerful tool to monitor IVT reaction in near real-time. This mini-review briefly introduces some strategies and examples for real-time IVT monitoring and discusses pros and cons of IVT monitoring methods.

Background: Despite the high incidence of acute ischemic stroke (AIS) in cancer patients, there is still no consensus about the safety of recanalization therapies in this cohort. Objectives: In this observational study, our aim was to investigate the bleeding risk after acute recanalization therapy in AIS patients with active malignancy. Methods and Study Design: We retrospectively analyzed observational data of 1016 AIS patients who received intravenous thrombolysis with rtPA (IVT) and/or endovascular therapy (EVT) between January 2017 and December 2020 with a focus on patients with active malignancy. The primary safety endpoint was the occurrence of stroke treatment-related major bleeding events, that is, symptomatic intracranial hemorrhage (SICH) and/or relevant systemic bleeding. The primary efficacy endpoint was neurological improvement during hospital stay (NI). Results: None of the 79 AIS patients with active malignancy suffered from stroke treatmentrelated systemic bleeding. The increased rate (7.6% versus 4.7%) of SICH after therapy compared to the control group was explained by confounding factors. A total of nine patients with cerebral tumor manifestation received acute stroke therapy, two of them suffered from stroke treatment-related intracranial hemorrhage remote from the tumor, both asymptomatic. The group of patients with active malignancy and the control group showed comparable rates of NI. Conclusion: Recanalization therapy in AIS patients with active malignancy was not associated with a higher risk for stroke treatment-related systemic or intracranial bleeding. IVT and/or EVT can be regarded as a safe therapy option for AIS patients with active malignancy. [ABSTRACT FROM AUTHOR]

Summary: Recently, several healthcare organizations use decision‐making systems based on electronic health record (EHR) data in order to guarantee patient's safety and improve the quality of healthcare. In essence, the evolutions of Internet of Things (IoT) technologies have been of great help for implementing an integrated and interoperable decision‐making system based on EHR and medical devices (MDs). Those IoT‐based systems allow Clinicians collecting real‐time health data and provide accurate patient's monitoring. Nevertheless, several studies have shown that it is hard to improve the quality of healthcare using the current EHR IoT‐based systems since they do not allow to easily express clinician needs. Interactive visualization tools have been proposed to improve the efficacy and utility of these EHR based systems. However, there is no framework that provides a visual summary of patient data to clinician for planning specific clinical tasks, subsequently evaluating clinician responses, visually exploring EHR data and MDs data, gaining insights, supporting dynamic coordination processes care, and forming and validating hypotheses and risks. This article addresses this problem and introduces SIMCard, an aggregation‐based connected EHR visualization framework for patient monitoring, interpreting and predicting with MDs. The proposed framework aims to synthesize patient's clinical data into a single aggregating model for both EHR and MD conforming to health standard and terminologies. It also allows to link the aggregating model to the relevant medical knowledge in order to provide a connected and dynamic care and preventive plan. Last but not least, it provides an aggregated visualization model capable of displaying graphically a patient's personal data from databases, healthcare devices and sensors to reduce cognitive barriers related to the complexity of medical information and interpretation of health data. To demonstrate the refinement and design of our system and to observe user's actual practice of visualizing and analyzing real‐world dataset, we evaluated our system and compare to existing ones. [ABSTRACT FROM AUTHOR]

The superiority of mechanical thrombectomy and intravenous thrombolysis versus intravenous thrombolysis alone for acute ischemic stroke caused by large vessel occlusions has been established. This treatment can be organized into 2 models: drip and ship (DS) versus mothership (MS). We analyzed the National Inpatient Sample (NIS) data to compare the outcomes between these models in real-world settings. NIS data were queried for 2017–2018 and propensity matching was used to match the differences. Outcomes for each group (disability at discharge and procedural complications) were compared. A total of 1226 patients were included in analysis (DS, n = 540; MS, n = 686) and groups were matched with respect to age, gender, and comorbidities. A total of 930 patients were included in the final analysis after propensity matching (DS, n = 465, MS, n = 465). The mean age in the DS group was 68.9 years (standard deviation [SD], 14.7) and 69.4 years (SD, 14) in the MS group (P = 0.752). The mean National Institutes of Health Stroke Scale score was 16.75 (SD, 6.07) in the DS group and 16.54 (SD, 5.99) in the MS group (P = 0.478). At discharge, minimal disability was noted in 22.4% in the DS group versus 26.2% in the MS group (P = 0.293). In-hospital mortality was lower in the MS group (8.8% vs. 7.1%; P = 0.32). The intracerebral hemorrhage (ICH) and intraventricular hemorrhage (IVH) rates were higher in the DS group (ICH, 24.3% vs. 18.7%; IVH, 2.4% vs. 0.9%) (ICH, P = 0.038; IVH, P = 0.068). Analyzing the efficacy and safety profile of DS versus MS models with the NIS database showed a trend toward better discharge outcomes and lower mortality for the MS group, although it did not reach statistical significance. [ABSTRACT FROM AUTHOR]

Background: The dramatic increase in intravitreal injections (IVTs) has been accompanied by a greater need for safer procedures. The ongoing debate about topical antibiotic prophylaxis after IVTs emphasizes the importance of large-scale studies. We aimed to study the role of topical antibiotic prophylaxis in reducing the risk of acute endophthalmitis after IVTs. Methods: Population-based cohort study, in France, from 2009 to 2018, including all French patients receiving IVTs of corticosteroids or anti-VEGF agents. Results: A total of 5,291,420 IVTs were performed on 605,434 patients. The rate of topical antibiotic prophylaxis after IVTs progressively decreased during the study period, with a sharp drop in 2014 (from 84.6% in 2009 to 27.4% in 2018). Acute endophthalmitis occurred in 1274 cases (incidence rate = 0.0241%). Although antibiotic prophylaxis did not alter the risk of endophthalmitis (p = 0.06), univariate analysis showed an increased risk after fluoroquinolone and aminoglycoside prophylaxis. This increased risk was not found in multivariate analysis. However, we observed an increased risk related to the use of fixed combinations of fluoroquinolones and aminoglycosides with corticosteroids (IRR = 1.89; 95% CI = 1.57–2.27%, antibiotics combined with corticosteroids). Conclusion: These results are consistent with the literature. Endophthalmitis rates after IVTs did not decrease with topical antibiotic prophylaxis. The use of a combination of antibiotics and corticosteroids doubles the risk of endophthalmitis and should be avoided. Avoiding antibiotic prophylaxis would reduce the costs and the potential risks of antibiotic resistance. [ABSTRACT FROM AUTHOR]

Background. Flow reactor synthesis of RNA using immobilized T7 RNA polymerase is a promising technological approach for large-scale therapeutic RNA production. Immobilization of T7 RNAP for flow reactor systems requires protocol for site-specific immobilization that allows controling the immobilization position to maximize activity and half-life of the enzyme. Aim. To develop protocol for site-specific introduction of bioorthogonal reaction groups into T7 RNAP while preserving activity of the enzyme. Methods. N-terminal 8xHisTag, FGE motif and G3S linker were introduced into T7RNAP CDS by long-overhang PCR and modified CDS was cloned into pGEX-2T backbone under lac-promoter. TAG codon was introduced into 8xHisTag-FGEmotif-GS3-T7RNAP in place of Cys codon of FGE motif by SDM. T5pLacOdriven cassette for over-expression of C-terminal fragment of L11 ribosome protein was cloned downstream of T7RNAP cassette by Golden Gate assembly. CDSs for mtbFGE, pAzFRS.2.t1 and CTA-tRNA were synthesized from oligo pools (Yuria-Pharm, Ukraine) by OE-PCR. Helper plasmids carrying expression cassette for mtbFGE and for pAzFRS.t1/CTA-tRNA were assembled by Golden Gate cloning according to reference constructs (AddGene ID102615, AddGene ID73546). In both systems, the expression and helper vectors were co-transformed into E.coli BL21 strain, ON cultures were inoculated into fresh media (in presence of arabinose and 1mM of pAzF for pAzF system) and induced at OD 0.6 by addition of IPTG/arabinose (and 100uM of CuSO4 for FGly system). The cultures were incubated for 16h at RT, harvested and purified by Ni-NTA spin columns. The label incorporations were quantified by RP-LC-MS of an intact protein or modification-containing peptide, cleaved by TEV protease. Results. FGly system. Investigation of initial unsuccessful attempts to obtain modified protein identified in cellulo aggregation of mtbFGE as a root cause. Induction and cultivation of producer cells at reduced temperatures (18 °C) allowed to obtain protein with >94% modification rate at final titers of 6 mg/L. Quantitative in vitro modification of FGE motif by purified mtbFGE has also been demonstrated for T7RNAP and can be used as well. pAzF system. Initial attempts of co-translational pAzF installation failed to produce detectable level of T7 RNAP. Investigation confirmed issues with helper plasmid toxicity, possibly due to sequestration of ribosomes by uncharged CTA-tRNA. Addition of pAzF and arabinose into culture media during cultivation (at 18 °C) allowed to rescue production of fully modified T7RNAP achieving final titers of 3mg/L. Mass-spectrometry of purified products showed presence of ~35% of reduced azide form, which was lowered by removal of DTT from purification buffers. Conclusion. For both systems, the optimized protocols were developed, which allow successful incorporation of the bioorthogonal groups with near-quantitative modification efficacy and practical yields. Due to a greater flexibility of pAzF system for tether group positioning it will be used for immobilization studies. [ABSTRACT FROM AUTHOR]

The functionality of messenger RNA, such as stability and translation, is determined by several elements. In Eukaryotes, the 5' end of the mRNA is modified to contain a 5' cap structure, the presence of which protects the mRNA from degradation by 5' to 3' exoribonucleases and promotes mRNA translation. The in vitro synthesis of RNA has recently attracted ample attention for its application as a source of therapeutic agents or research tools. Although in vitro mRNA synthesis (IVT) methodology is well established and is still being improved, not all synthesized RNA molecules have the expected properties. For example, the co-transcriptional addition of a 5' cap is frequently incomplete. Yet, the uncapped mRNA molecules are undesirable and must be removed before further processing. Here, we present a protocol for the enzymatic removal of uncapped RNA molecules. This approach offers an excellent opportunity to enrich IVT products with fully competent, 5' cap-containing mRNA molecules., (© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Self-amplifying RNA (saRNA) vaccines are able to induce a higher antigen-specific immune response with a more cost-effective and rapid production process compared to plasmid DNA vaccines. saRNAs are synthesized through in vitro transcription (IVT); however, this process has mainly been optimized for relatively short mRNAs. Methods: Here, we optimized the IVT process for long saRNAs, approximately 9.4 kb, through a design of experiment (DoE) approach to produce a maximal RNA yield and validated the optimal IVT method on various sizes of RNA. Results: We found that magnesium has the highest impact on RNA yield with acetate ions enabling a higher yield than chloride ions. In addition, the interaction between magnesium and nucleoside triphosphates (NTPs) is highly essential for IVT. Further addition of sodium acetate (NaOAc) during IVT provided no added benefit in RNA yield. Moreover, pyrophosphatase was not essential for productive IVT. The optimal IVT method can be used to synthesize different lengths of RNA. Conclusions: These findings emphasize the ability to synthesize high quality and quantity of saRNA through IVT and that the optimal amount of each component is essential for their interactions to produce a high RNA yield.

Objective: This study investigates the mismatch between the National Institutes of Health Stroke Scale (NIHSS) score and the computed tomography (CT) findings measured by the Alberta Stroke Program Early CT Score (ASPECTS) for predicting the functional outcome and safety of intravenous thrombolysis (IVT) treatment in patients with acute ischemic stroke (AIS). Methods: This prospective observational study includes patients with AIS who underwent CT imaging within 4.5 h of the onset of symptoms. Patients were divided into the NIHSS–ASPECTS mismatch (NAM)-positive and NAM-negative groups (group P and N, respectively). The clinical outcome was assessed using the Modified Rankin Scale (mRS). Safety outcomes included progression, symptomatic intracerebral hemorrhage (sICH), intracerebral hemorrhage (ICH), adverse events, clinical adverse events, and mortality. Results: A total of 208 patients were enrolled in the study. In group P, IVT treatment was associated with a good functional outcome at 3 months (p = 0.005) and 1 year (p = 0.001). A higher percentage of patients with favorable mRS (0–2) (p = 0.01) and excellent mRS (0–1) (p = 0.011) functional outcomes was obtained at 1 year in group P with IVT treatment. Group N did not benefit from the same treatment (p = 0.352 and p = 0.480 at 3 months and 1 year, respectively). There were no statistically significant differences in sICH, ICH, mortality rates, or other risks between the IVT and conventional treatment groups. Conclusion: IVT treatment is associated with a good functional outcome in patients with NAM, without increasing the risks of sICH, ICH, mortality, or other negative outcomes. NAM promises to be an easily obtained indicator for guiding the treatment decisions of AIS. [ABSTRACT FROM AUTHOR]

In the present study, the performances of the NWP models on two heavy rainfalls on 20 July and 22 August 2021 over Henan Province were investigated. The impacts of the water vapor transport to the extreme rainfall were further discussed. The results showed that the regional model system in the Guangzhou Meteorological Service generally showed high scores on the extreme rainfall over Henan. The maximum 24h accumulative rainfall by the 24h forecasts by the CMA-GD reached 556 mm over Henan Province. The 24-h and 48-h Threat Score (TS) of heavy rainfall reached 0.56 and 0.64. The comparisons of the Fraction Skill Score (FSS) verifications of the heavy rainfall by CMA-GD and CMA-TRAMS at the radium of 40km reached 0.96 and 0.87. The water vapor transport to the extreme rainfall showed that the vertically integrated water vapor transport (IVT) of the whole layer before the occurrence of the heavy rainfall exhibited a double-eyes distribution in case 7 · 20. The north eye over Henan reached the same magnitude of IVT as the typhoon eye (Cempaka) over south China. The IVT over the lower troposphere (<500 hPa) showed an overwhelming magnitude than the upper level, especially in the planetary boundary layer (<700 hPa). More practical technical needs to be developed to improve its performances on the forecasting of extreme rainfall, as well as more experiments need to be conducted to examine the effects of the specific terrain and physical schemes on the extreme rainfall. [ABSTRACT FROM AUTHOR]

Background: The dramatic increase in intravitreal injections (IVTs) has been accompanied by a greater need for safer procedures. The ongoing debate about topical antibiotic prophylaxis after IVTs emphasizes the importance of large-scale studies. We aimed to study the role of topical antibiotic prophylaxis in reducing the risk of acute endophthalmitis after IVTs. Methods: Population-based cohort study, in France, from 2009 to 2018, including all French patients receiving IVTs of corticosteroids or anti-VEGF agents. Results: A total of 5,291,420 IVTs were performed on 605,434 patients. The rate of topical antibiotic prophylaxis after IVTs progressively decreased during the study period, with a sharp drop in 2014 (from 84.6% in 2009 to 27.4% in 2018). Acute endophthalmitis occurred in 1274 cases (incidence rate = 0.0241%). Although antibiotic prophylaxis did not alter the risk of endophthalmitis (p = 0.06), univariate analysis showed an increased risk after fluoroquinolone and aminoglycoside prophylaxis. This increased risk was not found in multivariate analysis. However, we observed an increased risk related to the use of fixed combinations of fluoroquinolones and aminoglycosides with corticosteroids (IRR = 1.89; 95% CI = 1.57–2.27%, antibiotics combined with corticosteroids). Conclusion: These results are consistent with the literature. Endophthalmitis rates after IVTs did not decrease with topical antibiotic prophylaxis. The use of a combination of antibiotics and corticosteroids doubles the risk of endophthalmitis and should be avoided. Avoiding antibiotic prophylaxis would reduce the costs and the potential risks of antibiotic resistance.

The IVT for power frequency is an essential facility in the power system. Due to the requirement for high-voltage insulation, the sensing part of the conventional IVTs is bulky and costly. The purpose of this article is to propose a new voltage measurement method based on capacitive voltage divider for GIS. The equivalent circuit of the measurement method is built. The numerical simulation model for the measurement system is built based on the actual size of GIS with rated voltage of 220 kV. And FEM is employed to analyze the transfer relation and the accuracy of the proposed measurement method. The experiments are conducted in the laboratory and at an actual GIS platform, respectively. The transfer relation between the applied voltage and the induced voltage is obtained. Compared with conventional IVTs, the dimension of the measurement system is very small. Due to the low voltage of the sensing part, the insulation problem of the sensing part is avoided. And due to simple structure and linear characteristics, the transfer relation of the proposed measurement system can be obtained with high accuracy. [ABSTRACT FROM AUTHOR]

Days of extreme precipitation over California are evaluated in Coupled Model Intercomparison Project Phase 6 (CMIP6) models and the ERA‐Interim reanalysis. In the current climate, the model spread in composited precipitation on extreme precipitation days is closely related to the magnitude of composited integrated vapor transport (IVT) across models, a proxy for the intensity of atmospheric rivers. Most models underestimate the magnitude of IVT associated with extreme precipitation, according to ERA‐Interim. This is due mostly to the contribution of moisture, which almost all models overestimate, while the contribution of lower‐tropospheric wind speed is generally closer to the reanalyses. Moreover, most models underestimate the variance in the latitude of maxima of numerous variables among days of extreme California precipitation. That is, in the general circulation models there is a lack of diversity in the latitude of the disturbances bringing winter precipitation to California. In the future climate, most models project a decrease in the frequency of southward‐displaced disturbances among California extreme precipitation days. Hence, the greatest increases in extreme precipitation are over northern California. However, the historical underestimate of the latitudinal variance of disturbances calls into question the reliability of these projections. This bias should be especially considered for dynamical downscaling efforts over the region. Key Points: Coupled Model Intercomparison Project Phase 6 models overestimate moisture and hence moisture flux associated with extreme precipitation over California, according to reanalysesGeneral circulation models (GCMs) underestimate the variance in latitude of disturbances on California extreme precipitation daysGCMs project future California extremes to be increasingly associated with disturbances centered farther north [ABSTRACT FROM AUTHOR]

Purpose: Leptomeningeal collaterals can slow down infarction growth; however, despite good collaterals in the DAWN and DEFUSE 3 trials, outcomes were devastating if reperfusion was not attempted. The aim of this study was to compare the influence of collaterals on morphological and functional outcome in patients with acute middle cerebral artery (MCA) stroke undergoing intravenous thrombolysis (IVT) vs. supportive care (non-IVT). Methods: Out of 1639 consecutive patients examined with multiparametric computed tomography (CT) for suspected ischemic stroke, all patients with confirmed MCA stroke who did not undergo endovascular thrombectomy were selected. Propensity score matching (PSM) was used to match IVT and non-IVT treated patients for potential confounders including age, sex, National Institutes of Health Stroke Scale (NIHSS) score on admission, Alberta Stroke Program Early CT Score (ASPECTS), and occlusion site. Regression analysis after PSM was performed to identify independent associations. Results: After PSM, 90 IVT patients were matched with 90 non-IVT patients. In multivariable regression analysis, a high regional leptomeningeal collateral (rLMC) score was independently associated with lower final infarction volume (FIV) in the IVT group (b = −0.472, p < 0.001) but not in the non-IVT group (b = −0.116, p = 0.327). The trichotomized rLMC scores predicted functional outcome in IVT treated patients (adjusted odds ratio, aOR = 4.57, 95% confidence interval, CI, 1.03–20.32, p = 0.046) but showed no independent association with outcome in the non-IVT group (aOR = 0.69, 95% CI 0.07–6.80, p = 0.753). Conclusion: Good collaterals favored smaller FIV and good functional outcome in IVT treated patients but not in non-IVT treated patients. Good collateral flow may have limited prognostic value if IVT is not administered to attempt reperfusion [ABSTRACT FROM AUTHOR]

In vitro transcribed (IVT) mRNA therapies is a promising approach for the effective and safe treatment of various diseases. However, IVT‐mRNA triggers immune responses due to recognition by human RNA sensors, but incorporation of chemically modified nucleosides have been shown to reduce such responses. Nonetheless, an assay reflecting the complexity of the human immune system is still needed. Here, we present a simple and fast ex vivo method called "RNA Immunogenic Assay" for measuring the immunogenicity of IVT‐mRNA in human whole blood. Chemically modified and unmodified mRNA were complexed with a transfection reagent (TransIT), and co‐incubated in human whole blood and specific cytokines were quantified (TNF‐α, INF‐α, IL‐6, and IL‐12p70) using ELISAs. The qPCR analysis was conducted to unveil the activation of specific immune pathway. Our findings demonstrated that the complete replacement of uridine with pseudouridine reduced TNF‐α, IL‐6, and IL‐12p70 levels and did not elevate the expression of genes involved in immune response against RNA including IRF7/3, EIF2A, & RNASEL. In addition, we observed that the transcript length was not found to be a confounding factor in RNA immunogenicity generation and our assay provide the feasibility to dissect donor specific immune response against mRNA therapeutics. [ABSTRACT FROM AUTHOR]

The COVID-19 pandemic has escalated the occurrence of hypoxia including thrombotic stroke worldwide, for which nitric oxide (NO) therapy seems very promising and translatable. Therefore, various modes/routes of NO-delivery are now being tested in different clinical trials for safer, faster, and more effective interventions against ischemic insults. Intravenous (IV) infusion of S-Nitrosoglutathione (GSNO), the major endogenous molecular pool of NO, has been reported to protect against mechanical cerebral ischemia-reperfusion (IR); however, it has been never tested in any kind of “clinically” relevant thromboembolic stroke models with or without comorbidities and in combination with the thrombolytic reperfusion therapy. Moreover, “IV-effects” of higher dose of GSNO following IR-injury have been contradicted to augment stroke injury. Herein, we tested the hypothesis that nebulization of low-dose GSNO will not alter blood pressure (BP) and will mitigate stroke injury in diabetic mice via enhanced cerebral blood flow (CBF) and brain tissue oxygenation (PbtO2). GSNO-nebulization (200 μg/kgbwt) did not alter BP, but augmented the restoration of CBF, improved behavioral outcomes and reduced stroke injury. Moreover, GSNO-nebulization increased early reoxygenation of brain tissue/PbtO2 as measured at 6.5 h post-stroke following thrombolytic reperfusion, and enervated unwanted effects of late thrombolysis in diabetic stroke. We conclude that the GSNO-nebulization is safe and effective for enhancing collateral microvascular perfusion in the early hours following stroke. Hence, nebulized-GSNO therapy has the potential to be developed and translated into an affordable field therapy against ischemic events including strokes, particularly in developing countries with limited healthcare infrastructure.

Introduction: Intravitreal dexamethasone (DEX) implant is indicated for the treatment of macular oedema due to diabetic retinopathy, retinal vein occlusion and uveitis. The most common complications are cataract and elevated intraocular pressure (IOP). Accidental injection of DEX implant into the lens is a rare complication and only few papers presented it., Case Presentation: A 40-year-old man was treated with DEX implant for diabetic macular oedema in both eyes. At 1 week follow-up visit, slit lamp examination showed the DEX implant was located in the crystalline lens of the right eye (RE) without any sign of inflammation, cataract or elevated IOP, so we decided to plan a normal follow-up schedule. Macular oedema relapsed 5 months after the injection in the left eye (LE), whereas the RE did not show any sing of intraretinal or subretinal fluid. Six months after DEX implantation an uneventful phacoemulsification and intraocular lens placement were performed in the RE because of IOP elevation., Conclusions: The therapeutic effect of DEX implant can be maintained for a longer period of time than intravitreal implant, determining complete reabsorption of macular oedema. Intralenticular implant can be maintained inside the lens until either IOP increases, cataract progresses, or other complications occur., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Background: Retinal artery occlusion (RAO) may lead to irreversible blindness. For acute RAO, intravenous thrombolysis (IVT) can be considered as treatment. However, due to the rarity of RAO, data about IVT safety and effectiveness is limited., Methods: From the multicenter database ThRombolysis for Ischemic Stroke Patients (TRISP), we retrospectively analyzed visual acuity (VA) at baseline and within 3 months in IVT and non-IVT treated RAO patients. Primary outcome was difference of VA between baseline and follow up (∆VA). Secondary outcomes were rates of visual recovery (defined as improvement of VA ⩾ 0.3 logMAR), and safety (symptomatic intracranial hemorrhage (sICH) according to ECASS II criteria, asymptomatic intracranial hemorrhage (ICH) and major extracranial bleeding). Statistical analysis was performed using parametric tests and a linear regression model adjusted for age, sex and baseline VA., Results: We screened 200 patients with acute RAO and included 47 IVT and 34 non-IVT patients with complete information about recovery of vision. Visual Acuity at follow up significantly improved compared to baseline in IVT patients (∆VA 0.5 ± 0.8, p  < 0.001) and non-IVT patients (∆VA 0.40 ± 1.1, p  < 0.05). No significant differences in ∆VA and visual recovery rate were found between groups at follow up. Two asymptomatic ICH (4%) and one (2%) major extracranial bleeding (intraocular bleeding) occurred in the IVT group, while no bleeding events were reported in the non-IVT group., Conclusion: Our study provides real-life data from the largest cohort of IVT treated RAO patients published so far. While there is no evidence for superiority of IVT compared to conservative treatment, bleeding rates were low. A randomized controlled trial and standardized outcome assessments in RAO patients are justified to assess the net benefit of IVT in RAO., Competing Interests: Declaration of conflicting interestsThe author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: PB: Research funds by the Gottfried and Julia Bangerter-Rhyner Foundation and Swiss Academy of Medical Sciences and funding for travel and conference fees from BMS/Pfizer. SW: Research funds by the Swiss National Science Foundation, the UZH Clinical research priority program (CRPP) stroke, the Swiss Heart foundation, the Zurich Neuroscience Center (ZNZ), speaker honoraria from Springer, Teva Pharma and consultancy fees from Bayer and Novartis. STE has received funding for travel or speaker honoraria from Bayer, Boehringer Ingelheim and Daiichi-Sankyo. He has served on scientific advisory boards for Bayer, Boehringer Ingelheim, BMS/Pfizer, and MindMaze and on the editorial board of Stroke. His institutions have received an educational grant from Pfizer, compensation from Stago for educational efforts and research support from Daiichi-Sankyo, the Science Funds [Wissenschaftsfonds] of the University Hospital Basel, the University Basel, from the “Wissenschaftsfonds Rehabilitation” of the University Department for Geriatric Medicine Felix Platter, the “Freiwillige Akademische Gesellschaft Basel,” the Swiss Heart Foundation, and the Swiss National Science Foundation. PR has received honoraria for advisory board participation from Boehringer Ingelheim and Pfizer and lecture fees from Boehringer Ingelheim, Bayer, Pfizer and Daiichi Sankyo. His institution has received a grant from Boehringer Ingelheim for the ECASS-4 Study. RRL received funding for speaker honoraria from Pfizer, Boehringer Ingelheim, Biogen and Iscehma View. He has served on scientific advisory boards for Novo-Nordisk and Bayer.

Intravitreal injection (IVI) of a drug allows for immediate intraocular concentrations of active ingredients higher than those obtained by intravenous injection while reducing the risk of systemic side effects. Today, IVI's play a central role in the treatment of many vitreoretinal diseases. With the aging of the population and the advent of vascular endothelial growth factor (VEGF) antagonists, their indications have increased exponentially, creating structural, organizational and economic difficulties. IVI is now one of the most widely performed medical procedures in industrialized countries, and its indications are expected to expand further in the near future with the development of new molecules. Although the overall safety of this practice is proven, an IVI exposes the patient to a 0.05 % risk of endophthalmitis, the consequences of which are often dramatic. This article details the current recommendations, in particular regarding asepsis and antisepsis, and proposes a typical sequence for performing an IVI., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

In this work, we present a novel architecture of the flywheel-infinitely variable transmission (F-IVT) actuator with increased energy saving performance. The F-IVT is an actuator for artificial knee joints with energy recovery capabilities. It uses a flywheel and an IVT with the purpose of storing and releasing energy and stabilizing the motor working point. The architecture proposed herein, named “linear F-IVT”, is compared with the rotating F-IVT and with the clutchable-series elastic actuator, through a model-based approach. By simulating the operation of these actuators in walking at different speed and stairs climbing of different inclination, the linear F-IVT results to be the most effective in reducing the peak of electric power and motor torque, also achieving a reduction of the electric energy demanded. These results suggest the possibility to achieve a significant motor downsizing, possibly leading to the development of a lightweight actuator to improve the portability and the operating range of wearable robots for lower limbs. Abbreviations: BS: ball screw; C-SEA: clutchable - series elastic actuator; CVT: continuously variable transmission; F-IVT: flywheel-infinitely variable transmission; HD: harmonic drive; IVT: infinitely variable transmission [ABSTRACT FROM AUTHOR]

After the COVID-19 pandemic, messenger RNA (mRNA) has revolutionized traditional vaccine manufacturing. With the increasing number of RNA-based therapeutics, valuable new scientific insights into these molecules have emerged. One fascinating area of study is the formation of double-stranded RNA (dsRNA) during in vitro transcription (IVT) which is considered a significant impurity, as it has been identified as a major trigger in the cellular immune response pathway. Therefore, there is a growing importance placed to develop and optimize purification processes for the removal of this by-product. Traditionally, efforts have primarily focused on mRNA purification after IVT through chromatographic separations, with anion exchange and reverse phase chromatography emerging as effective tools for this purpose. However, to the best of our knowledge, the influence and significance of the quality of the linearized plasmid have not been thoroughly investigated. Plasmids production involves the growth of bacterial cultures, bacterial harvesting and lysis, and multiple filtration steps for plasmid DNA purification. The inherent complexity of these molecules, along with the multitude of purification steps involved in their processing, including the subsequent linearization and the less-developed purification techniques for linearized plasmids, often result in inconsistent batches with limited control over by-products such as dsRNA. This study aims to demonstrate how the purification process employed for linearized plasmids can impact the formation of dsRNA. Several techniques for the purification of linearized plasmids based on both, resin filtration and chromatographic separations, have been studied. As a result of that, we have optimized a chromatographic method for purifying linearized plasmids using monolithic columns with C4 chemistry (butyl chains located in the surface of the particles), which has proven successful for mRNAs of various sizes. This chromatographic separation facilitates the generation of homogeneous linearized plasmids, leading to mRNA batches with lower levels of dsRNA during subsequent IVT processes. This finding reveals that dsRNA formation is influenced not only by RNA polymerase and IVT conditions but also by the quality of the linearized template. The results suggest that plasmid impurities may contribute to the production of dsRNA by providing additional templates that can be transcribed into sequences that anneal with the mRNA molecules. This highlights the importance of considering the quality of plasmid purification in relation to dsRNA generation during transcription. Further investigation is needed to fully understand the mechanisms and implications of plasmid-derived dsRNA. This discovery could shift the focus in mRNA vaccine production, placing more emphasis on the purification of linearized plasmids and potentially saving, in some instances, a purification step for mRNA following IVT., Competing Interests: JM, VL, AL, HM, and DC were employed by the Company Certest Pharma, Certest Biotec., (Copyright © 2023 Martínez, Lampaya, Larraga, Magallón and Casabona.)

RNA-based therapeutics and vaccines are opening up new avenues for modern medicine. To produce these useful RNA-based reagents, in vitro transcription (IVT) is an important reaction that primarily determines the yield and quality of the product. Therefore, IVT condition should be well optimized to achieve high yield and purity of transcribed RNAs. To this end, real-time monitoring of RNA production during IVT, which allows for fine tuning of the condition, would be required. Currently, light-up RNA aptamer and fluorescent dye pairs are considered as useful strategies to monitor IVT in real time. Fluorophore-labeled antisense probe-based methods can also be used for real-time IVT monitoring. In addition, a high-performance liquid chromatography (HPLC)-based method that can monitor IVT reagent consumption has been developed as a powerful tool to monitor IVT reaction in near real-time. This mini-review briefly introduces some strategies and examples for real-time IVT monitoring and discusses pros and cons of IVT monitoring methods., Competing Interests: KHL, JS, SK, and SRH are employee of Rznomics Inc. S-WL is CEO of Rznomics Inc., (Copyright © 2023 Lee, Song, Kim, Han and Lee.)

Background: This study evaluates the effect of a mass media intervention (MMI) on the number of admissions to the emergency room (ER) with a suspected acute-onset stroke and on the number of patients treated with intravenous thrombolysis (IVT). The MMI was run for 1 month, with a precampaign month, priming the treatment chain and raising in-hospital awareness.Methods: This study conducted a prospective analysis of all patients admitted to the ER before, during, and after the MMI to assess changes in admission rate and IVT treatment rate. The primary end point was to assess any change in the IVT treatment rate, and the secondary end point was to determine the number of acute stroke admissions to the ER within 4.5 hours of symptom onset.Results: The MMI resulted in an increased IVT treatment rate from 7.3 to 11.3 patients per month, an increment of 54% (P = .002). This also resulted in IVT treatment percentages as high as 42%. The number of suspected acute stroke admissions to the ER increased from 37.3 to 72.8 patients per month, a 97% increase (P < .0001). The Epinion interviews showed that stroke symptom recognition increased from 66% to 75%.Conclusion: The MMI produced a significant increase in IVT treatment rate. This increased treatment rate is largely due to the increase in the number of acute stroke admissions to the ER. Targeting the treatment chain before the MMI led to unprecedented IVT treatment percentages. The effect of our MMI lasted for approximately 6 months before tapering off; repeated campaigns are a prerequisite for sustained IVT treatment rates. [ABSTRACT FROM AUTHOR]

Introduction: Intravitreal injections (IVT) of a drug aim to rapidly obtain effective concentrations greater than those that would be obtained by a periocular or intravenous injection. The objective of the present study is to demonstrate the contribution of IVT in the treatment of pathologies of the posterior segment of the eye., Methodology: We conducted a 21-month ambidirectional observational study from January 1, 2020, to September 30, 2021. We included by nonprobability sampling all consenting patients admitted for IVT at IOTA Teaching Hospital., Results: During our study, 201 patients were collected out of 30 739 patients seen in consultation. The hospital frequency of IVT was 0.65%. There were 111 women and 90 men. The M/F ratio was 0.82. The number of patients who received IVT antibiotics was 135. Anti-VEGF was injected in 64 patients. Two patients received IVT corticosteroids. Complications frequently encountered were pain at upon injection (94.03%), IOP spike (11.94%) and cataract (7.46%). In our sample, we observed an improvement in visual acuity in 56.21% of cases and a reduction in macular edema in 45.16% of cases., Discussion: The contribution of IVT in the treatment of vitreoretinal diseases is extraordinary. Rigorous observation of aseptic technique and good practices protects against infectious complications., Conclusion: IVT has allowed us to effectively treat various diseases of the vitreous and retina. IVT is a simple procedure, but it must be performed with the same aseptic technique as surgical procedures., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

We present the effects of an In-Se surface treatment of Cu-rich ([Cu]/[In] > 1) CuInSe2 absorbers in order to utilize the superior transport properties of Cu-rich absorbers for high-efficiency solar cells. A Cu-poor surface was successfully formed on a Cu-rich absorber by co-evaporation of indium and selenium under appropriate conditions. The aim is to suppress the interface recombination, which is generally observed at the interface between CdS and Cu-rich CuInSe2. A surface-treated device achieved an efficiency of 13.1%, improved from 9.5% obtained with an untreated Cu-rich device. The open-circuit voltage of In-Se treated devices reach the same level as devices made from Cu-poor absorbers grown by a three-stage process. By comparing the results of the experiments and simulations, it is shown that the role of the Cu-poor surface on Cu-rich absorbers is to prevent interface recombination, mainly due to a lower doping level of the Cu-poor surface layer than in the Cu-rich CuInSe2. Thus, it becomes possible to uncouple the interface from the bulk properties in Cu-rich solar cells. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Water quality monitoring requires a reliable and practical on-site detection method for heavy metal ions. Combining an in vitro transcription (IVT) technology with allosteric transcription factors (aTFs), we developed a cell-free paper-based biosensor for on-site detection of Hg2+ and Pb2+ in water. Suitable aTFs screened using surface plasmon resonance (SPR) were employed for building biosensors. ATFs could disassociate from DNA due to their specific affinity to metal ions, and fluorescent RNA was transcribed as a signal. The developed biosensor could quantitatively detect Hg2+ in a linear dynamic range of 0.5–500 nM and Pb2+ in a 1–250 nM range in a 1 h period. The LOD of the biosensor was 0.5 nM for Hg2+ and 0.1 nM for Pb2+. The recoveries ranged from 91.09% to 123.24% for actual water samples detection. Furthermore, freeze-drying was used to create a paper-based biosensor that could detect Hg2+ and Pb2+ simultaneously on-site. This research presents a useful technique for various heavy metal ion detections. [Display omitted] • Novel screening strategy of aTFs with BIAcore assay. • The rapid and visual signal output by the 3WJdB aptamer. • High sensitive and selective detection of Hg2+ and Pb2+ by aTFs-based biosensors. • Fast and reliable water quality monitoring and heavy metal risk management. [ABSTRACT FROM AUTHOR]

Abstract: In this paper, the power re-circulation and the torque ratio of Infinitely Variable Transmissions (IVT) are investigated. The focus is on the gear neutral condition, which is very useful in many applications but actually critical in terms of efficiency, reliability and control. An experimental investigation of the output to input torque ratio as a function of the speed ratio close to the neutral gear is shown, the results of which are compared with a theoretical model of the IVT transmission in steady-state. It is shown that the power split transmission can be utilized as a torque multiplier for vanishing output angular velocity, as requested by application to terrestrial vehicles. It is also shown that the mathematical model predicts the overall trend of the torque ratio and its value with vanishing speed ratio, a result that can be of some help for optimal control of the transmission. Furthermore, the power re-circulation in the neutral gear is discussed in detail for vanishing output torque. [Copyright &y& Elsevier]