Optimization and Characterization of Quercetin Vitamin C Nano-Phytosome Formulation

Objective: To design an optimal formulation for quercetin and vitamin C nano-phytosome. Method(s): Nano-phytosomes are prepared by the thin layer hydration technique using a 2-level-5-factor design experimental. A total of 32 experimental formulas were used for data analysis. The ratio of quercetin: soy lecithin (X1), the ratio of quercetin: cholesterol (X2), stirring speed (X3), stirring temperature (X4), and stirring time (X5) were used as independent factors, while globule size as a dependent factor. Data analysis was carried out by Design Expert12 application. Characterization of the optimal formula included physicochemical evaluation, globule size analysis, zeta potential, polydispersity index, entrapment efficiency, Transition Electron Microscopy (TEM) analysis, and FTIR analysis. Result(s): The optimal formula consisted of quercetin: vitamin C: lecithin: cholesterol ratio of 1: 1: 1.046: 0.105 mol;stirring speed 763.986 rpm;stirring time of 59 min, at temperature 51.73 degreeC which produced 59.26 nm average globule size, PDI value 0.66;zeta potential value-35.93+/-0.95 mV and average SPAN value 0.61. This formulation showed entrapment efficiency of quercetin 91.69+/-0.18 % and vitamin C 90.82+/-0.13 %. The TEM and FITR analysis showed the morphological of the globules and interactions between the drugs, soy lecithin, and cholesterol to form nano-phytosomes. Conclusion(s): The conditions to obtain the optimal formula for quercetin vitamin C nano-phytosome consisted of quercetin: vitamin C: lecithin: cholesterol ratio of 1: 1: 1.046: 0.105 mol;stirring speed 763.986 rpm;stirring time of 59 min, and at temperature 51.73 degreeC.Copyright © 2023 The Authors.

Ventilator-associated events: From surveillance to optimizing management

Mechanical ventilation (MV) is a life-support therapy that may predispose to morbid and lethal complications, with ventilator-associated pneumonia (VAP) being the most prevalent. In 2013, the Center for Disease Control (CDC) defined criteria for ventilator-associated events (VAE). Ten years later, a growing number of studies assessing or validating its clinical applicability and the potential benefits of its inclusion have been published. Surveillance with VAE criteria is retrospective and the focus is often on a subset of patients with higher than lower severity. To date, it is estimated that around 30% of ventilated patients in the intensive care unit (ICU) develop VAE. While surveillance enhances the detection of infectious and non-infectious MV-related complications that are severe enough to impact the patient's outcomes, there are still many gaps in its classification and management. In this review, we provide an update by discussing VAE etiologies, epidemiology, and classification. Preventive strategies on optimizing ventilation, sedative and neuromuscular blockade therapy, and restrictive fluid management are warranted. An ideal VAE bundle is likely to minimize the period of intubation. We believe that it is time to progress from just surveillance to clinical care. Therefore, with this review, we have aimed to provide a roadmap for future research on the subject.Copyright © 2022 The Author(s)

Liquid Chromatographic Approach for Analysis of Favipiravir–A Repurpose Drug for COVID-19

Favipiravir is a potential repurpose moiety to treat COVID-19 by depletion of virus load in infectious patients. To analyze and separate Favipiravir with remarkable efficiency, X-Bridge C8 column (150 x 4.6 mm, 5 µ) and a solvent phase of 0.1% TEA and acetonitrile (40:60 v/v) with 1-mL/min flow rate were used. The eluted favipiravir and possible degradants were detected at 225 nm. Further, the process was validated by using ICH (Q2R1) guidelines to ensure the method's suitability in the pharmaceutical sector. The RT of Favipiravir was observed at 3.7 min with good linearity of 2 to 30 µg/mL. %RSD of both system and method precision was assessed in the series of 0.32 to 0.98. The mean percentage recovery of Favipiravir was in the range of 99.0–100.4%. The limit of detection (LoD) and limit of quantification (LoQ) were assessed to be 0.024 and 0.084 μg/mL for favipiravir. The outcomes confirmed that the projected approach was economical, insightful, simple and precise with better sensitivity. Investigation of Favipiravir in the incidence of a variety of stressed or forced degradation environments ensures stability indicating quality of the developed approach. © 2023, Dr. Yashwant Research Labs Pvt. Ltd.. All rights reserved.

Optimization of SARS-CoV-2 spike protein receptor binding domain expression in Pichia pastoris and evaluation of its immunogenicity.

Objective To effectively express the receptor binding domain (RBD) of SARS-CoV-2 spike protein in Pichia pastoris and to evaluate its immunogenicity. Methods The gene encoding the RBD protein was synthesized and cloned into the pPICZalphaA plasmid. After linearization, the plasmid was transferred and integrated into the genome of Pichia pastoris. The expressed RBD protein in culture supernatant was analyzed by Western blot and Biolayer interferometry. After screening, a single clone expressing the RBD protein was selected. The high-level expression of RBD protein was achieved by optimizing the fermentation process, including the salt concentration adjusting of the medium and induction condition optimization (pH, temperature and duration) . The immunogenicity of the expressed RBD protein was evaluated in a mouse model. Results A single clone with a high expression level of RBD protein was obtained and named RBD-X33. The expression level of RBD protein in the fermentation supernatant reached up to 240 mg / L after optimization of the induction condition (HBSM medium, pH = 6. 5 +/- 0. 3, 22 and 120 h) . In the mouse experiment, the recombinant RBD protein was formulated with Alum + CpG dual adjuvant and injected into mice. The binding IgG antibody levels were up to 2. 7 x 106 tested by ELISA and the neutralizing antibody levels were up to 726. 8 tested by live virus neutralizing antibody assay (prototype) . Conclusions The RBD protein could be efficiently expressed in Pichia pastoris and induce stronger immune response in animals. This study suggested that the recombinant SARS-CoV-2 RBD protein expressed in Pichia pastoris could serve as a candidate antigen in the development of SARS-CoV-2 vaccine.Copyright © 2022 Society of Microbiology and Immunology. All rights reserved.

Strategies to Improve Time to Activation within an Ambulatory Remote Patient Monitoring Program

Remote patient monitoring (RPM) programs have been shown to effectively decrease rates of healthcare utilization among patients with chronic conditions. Immediately enrolling a patient and activating them in the RPM program either upon or soon after discharge is an important step in achieving these benefits. We tested interventions across three Plan-Do-Study-Act quality improvement cycles to understand the extent to which operational improvements would lead to timely activation. Each improvement cycle resulted in decreased time to activation, with the cumulative effect (as applied to patients on the COVID-19 RPM program) resulting in a reduction that was overall greater than the sum of the individual improvements. As additional healthcare systems develop and deploy RPM programs, the learnings from this project can help to provide insight into the operational and logistical challenges encountered in providing these services as well as potential interventions that can be used to achieve timely activation.

A Case Study on Waiting Line Management for the Land Transportation Office of the Philippines

The Philippines' Land Transportation Office (LTO) has the task of evaluating and processing citizen's applications for driver's license for professional and non-professional purposes. The office sees many customers each day and the need for an efficient waiting sitting was needed. Several process optimization and productivity improvements will be discussed in this research. Improvements along the lines of robotic process optimization, social media bots, existing efficient and effective processes, and the like, will be considered in developing suggestions for optimizing waiting line management in the LTO. In addition, existing congestion surveillance methods are explored to consider more solutions. The LTO currently implements a multi-channel, multi-phase line management system for all branches in the country. The research aims to evaluate its effectiveness in processing new driver's license applications in the National Capital Region within a set period of days. The system has proven inefficient based on recent data taken between 2019-2020 [1], even considering the decline in applications because of the COVID-19 pandemic. There is an expected spike in citizens applying for driver's licenses once the quarantine restrictions are lifted entirely, and an effective waiting line management system will be needed. A solution can be created by taking inspiration from efficient process systems from the United States and United Kingdom. © 2021 IEEE.

Simulation and Optimization of Isopropanol Production by Acetone Hydrogenation Process

Isopropanol is an extremely widely used chemical product. With the outbreak of COVID-19 worldwide, the export demand of isopropanol keeps increasing, and the supply is in short supply. In order to meet the demand of isopropyl alcohol in the international market, the existing production process faces new challenges and needs to be improved and upgraded. In this design, Aspen Plus was used to simulate the existing hydrogenation process of acetone to isopropanol, and it was improved and optimized to establish an annual output of 5 kT isopropanol and a variety of by-products of different specifications. The process adopts double-effect distillation and heat pump distillation to reduce energy consumption, and a set of waste liquid recovery device is designed to effectively recover the by-products and achieve the purpose of improving economic benefits. Simulation results of Aspen Plus show that energy consumption, carbon emission, water consumption and solid waste utilization rate can be reduced by 36.73%, 40%, 45% and 80% respectively, meeting the requirements of “Made in China 2025”. © 2022 SPIE. All rights reserved.

Monitoring and Optimisation of Ag Nanoparticle Spray-Coating on Textiles.

An automatic lab-scaled spray-coating machine was used to deposit Ag nanoparticles (AgNPs) on textile to create antibacterial fabric. The spray process was monitored for the dual purpose of (1) optimizing the process by maximizing silver deposition and minimizing fluid waste, thereby reducing suspension consumption and (2) assessing AgNPs release. Monitoring measurements were carried out at two locations: inside and outside the spray chamber (far field). We calculated the deposition efficiency (E), finding it to be enhanced by increasing the spray pressure from 1 to 1.5 bar, but to be lowered when the number of operating sprays was increased, demonstrating the multiple spray system to be less efficient than a single spray. Far-field AgNPs emission showed a particle concentration increase of less than 10% as compared to the background level. This finding suggests that under our experimental conditions, our spray-coating process is not a critical source of worker exposure.

Patient Empowerment During the COVID-19 Pandemic by Ensuring Safe and Fast Communication of Test Results: Implementation and Performance of a Tracking System.

BACKGROUND: Overcoming the COVID-19 crisis requires new ideas and strategies for online communication of personal medical information and patient empowerment. Rapid testing of a large number of subjects is essential for monitoring and delaying the spread of SARS-CoV-2 in order to mitigate the pandemic's consequences. People who do not know that they are infected may not stay in quarantine and, thus, risk infecting others. Unfortunately, the massive number of COVID-19 tests performed is challenging for both laboratories and the units that conduct throat swabs and communicate the results. OBJECTIVE: The goal of this study was to reduce the communication burden for health care professionals. We developed a secure and easy-to-use tracking system to report COVID-19 test results online that is simple to understand for the tested subjects as soon as these results become available. Instead of personal calls, the system updates the status and the results of the tests automatically. This aims to reduce the delay when informing testees about their results and, consequently, to slow down the virus spread. METHODS: The application in this study draws on an existing tracking tool. With this open-source and browser-based online tracking system, we aim to minimize the time required to inform the tested person and the testing units (eg, hospitals or the public health care system). The system can be integrated into the clinical workflow with very modest effort and avoids excessive load to telephone hotlines. RESULTS: The test statuses and results are published on a secured webpage, enabling regular status checks by patients; status checks are performed without the use of smartphones, which has some importance, as smartphone usage diminishes with age. Stress tests and statistics show the performance of our software. CTest is currently running at two university hospitals in Germany-University Hospital Ulm and University Hospital Tübingen-with thousands of tests being performed each week. Results show a mean number of 10 (SD 2.8) views per testee. CONCLUSIONS: CTest runs independently of existing infrastructures, aims at straightforward integration, and aims for the safe transmission of information. The system is easy to use for testees. QR (Quick Response) code links allow for quick access to the test results. The mean number of views per entry indicates a reduced amount of time for both health care professionals and testees. The system is quite generic and can be extended and adapted to other communication tasks.

High cell density perfusion process for high yield of influenza A virus production using MDCK suspension cells.

Similar to the recent COVID-19 pandemic, influenza A virus poses a constant threat to the global community. For the treatment of flu disease, both antivirals and vaccines are available with vaccines the most effective and safest approach. In order to overcome limitations in egg-based vaccine manufacturing, cell culture-based processes have been established. While this production method avoids egg-associated risks in face of pandemics, process intensification using animal suspension cells in high cell density perfusion cultures should allow to further increase manufacturing capacities worldwide. In this work, we demonstrate the development of a perfusion process using Madin-Darby canine kidney (MDCK) suspension cells for influenza A (H1N1) virus production from scale-down shake flask cultivations to laboratory scale stirred tank bioreactors. Shake flask cultivations using semi-perfusion mode enabled high-yield virus harvests (4.25 log10(HAU/100 µL)) from MDCK cells grown up to 41 × 106 cells/mL. Scale-up to bioreactors with an alternating tangential flow (ATF) perfusion system required optimization of pH control and implementation of a temperature shift during the infection phase. Use of a capacitance probe for on-line perfusion control allowed to minimize medium consumption. This contributed to a better process control and a more economical performance while maintaining a maximum virus titer of 4.37 log10(HAU/100 µL) and an infectious virus titer of 1.83 × 1010 virions/mL. Overall, this study clearly demonstrates recent advances in cell culture-based perfusion processes for next-generation high-yield influenza vaccine manufacturing for pandemic preparedness. KEY POINTS: • First MDCK suspension cell-based perfusion process for IAV produciton was established. • "Cell density effect" was overcome and process was intensified by reduction of medium use and automated process control. • The process achieved cell density over 40 × 106 cells/mL and virus yield over 4.37 log10(HAU/100 µL).