A Silicon-Based PDMS-PEG Copolymer Microfluidic Chip for Real-Time Polymerase Chain Reaction Diagnosis.

Polydimethylsiloxane (PDMS) has been widely used to make lab-on-a-chip devices, such as reactors and sensors, for biological research. Real-time nucleic acid testing is one of the main applications of PDMS microfluidic chips due to their high biocompatibility and transparency. However, the inherent hydrophobicity and excessive gas permeability of PDMS hinder its applications in many fields. This study developed a silicon-based polydimethylsiloxane-polyethylene-glycol (PDMS-PEG) copolymer microfluidic chip, the PDMS-PEG copolymer silicon chip (PPc-Si chip), for biomolecular diagnosis. By adjusting the modifier formula for PDMS, the hydrophilic switch occurred within 15 s after contact with water, resulting in only a 0.8% reduction in transmittance after modification. In addition, we evaluated the transmittance at a wide range of wavelengths from 200 nm to 1000 nm to provide a reference for its optical property study and application in optical-related devices. The improved hydrophilicity was achieved by introducing a large number of hydroxyl groups, which also resulted in excellent bonding strength of PPc-Si chips. The bonding condition was easy to achieve and time-saving. Real-time PCR tests were successfully conducted with higher efficiency and lower non-specific absorption. This chip has a high potential for a wide range of applications in point-of-care tests (POCT) and rapid disease diagnosis.

Predictors of Clinical Efficacy of Cytokine Hemoadsorption in COVID-19 (Clinical Trial).

Aim of the study. To evaluate the value of predictors of hemoadsorption clinical efficacy in patients with COVID-19. Materials and methods. This study analyzed the results of treatment of 62 patients with severe COVID-19 in the intensive care unit using selective hemoadsorption of cytokines. All patients with severe COVID-19 were admitted to the intensive care unit within 14 days from the disease onset were subdivided into two groups. Group 1 patients (n=32) received on a top of standard treatment the hemoperfusion (HP) procedure for 4 hours, for 2-3 days in a row, using a cytokine sorption column composed of mesoporous styrene-divinilbenzen copolymer matrix. Group 2 patients were not subjected to extracorporeal blood purification. All patients received IL-6 inhibitors at a baseline in accordance to the temporary guidelines. We evaluated factors of unfavorable outcomes by analyzing changes in biochemical markers of systemic inflammatory response and mortality rates in patients of both groups. Results. Initiation of HP later than 10 days from NCI onset (P < 0.001), length of stay in the ICU, extent of lung damage (P = 0.036) and the SOFA (Sequential Organ Failure Assessment) score (P = 0.009) were the most powerful predictors of unfavorable outcome. Levels of systemic inflammatory response markers (in-terleukin-6, CRP, D-dimer) in both groups did not significantly affect the survival rates and length of hospital stay (P > 0.05). HP group demonstrated better survival (P < 0.05). Mean hospital stay was 31 and 27 days, ICU stay - 11 and 8 days for Groups 1 and 2, respectively (P < 0.05). Conclusion. Treatment of severe COVID-19 patients with HP using novel domestic hemosorbent composed of styrene-divinilbenzen copolymer matrix resulted in decrease in CRP levels on the first day after application and, with early onset, contributed to a significant increase in survival and decreased hospital and ICU stay. Additional studies are warranted to clarify the optimal timing of the initiation of HP in severe COVID-19 patients.Copyright © 2023, V.A. Negovsky Research Institute of General Reanimatology. All rights reserved.

Predictors of Clinical Efficacy of Cytokine Hemoadsorption in COVID-19 (Clinical Trial).

Aim of the study. To evaluate the value of predictors of hemoadsorption clinical efficacy in patients with COVID-19. Materials and methods. This study analyzed the results of treatment of 62 patients with severe COVID-19 in the intensive care unit using selective hemoadsorption of cytokines. All patients with severe COVID-19 were admitted to the intensive care unit within 14 days from the disease onset were subdivided into two groups. Group 1 patients (n=32) received on a top of standard treatment the hemoperfusion (HP) procedure for 4 hours, for 2-3 days in a row, using a cytokine sorption column composed of mesoporous styrene-divinilbenzen copolymer matrix. Group 2 patients were not subjected to extracorporeal blood purification. All patients received IL-6 inhibitors at a baseline in accordance to the temporary guidelines. We evaluated factors of unfavorable outcomes by analyzing changes in biochemical markers of systemic inflammatory response and mortality rates in patients of both groups. Results. Initiation of HP later than 10 days from NCI onset (P < 0.001), length of stay in the ICU, extent of lung damage (P = 0.036) and the SOFA (Sequential Organ Failure Assessment) score (P = 0.009) were the most powerful predictors of unfavorable outcome. Levels of systemic inflammatory response markers (in-terleukin-6, CRP, D-dimer) in both groups did not significantly affect the survival rates and length of hospital stay (P > 0.05). HP group demonstrated better survival (P < 0.05). Mean hospital stay was 31 and 27 days, ICU stay - 11 and 8 days for Groups 1 and 2, respectively (P < 0.05). Conclusion. Treatment of severe COVID-19 patients with HP using novel domestic hemosorbent composed of styrene-divinilbenzen copolymer matrix resulted in decrease in CRP levels on the first day after application and, with early onset, contributed to a significant increase in survival and decreased hospital and ICU stay. Additional studies are warranted to clarify the optimal timing of the initiation of HP in severe COVID-19 patients.Copyright © 2023, V.A. Negovsky Research Institute of General Reanimatology. All rights reserved.

Characterization of Patients with Multisystem Inflammatory Syndrome Temporarily Associated with Covid 19 (Sims-Tac) Experience of the Fundacion Clinica Infantil Club Noel March 2020-December 2021

BACKGROUND AND AIM: descriptive, retrospective METHOD: 39 patients Variables gender, weight, age, mortality, treatment, biomarkers: D-dimer, Troponin, Atrial Natriuretic Peptide, Lactic Dehydrogenase, CRP, ESR, fibrinogen, hemofiltration, hemoadsorption, immunoglobulin pentaglobulin, ivermectin and steroids. RESULT(S): 39 patients were reviewed, 52% female, 48% male, one patient died, the most frequent age range was under 3 years (44%), followed by the age group 3-8 years (20%), weight average was 28.7 kg, 44% received ivermectin, EL 16% immunoglobulin and 56% pentaglobulin, 24% were in therapy. hemoadsorption with OXIRIS filter (oXiris hemofilter (Baxter, Meyzieu, France) to improve the adsorption properties of the AN69ST membrane, which was very impressive in the improvement of the inflammatory picture demonstrated radiologically, 40% of their presentation phenotype was sepsis (HYPERINFLAMATION) 100% received steroids. Regarding the most frequent biomarkers, the average was D-dimer: 1700 ng/dl, troponin I 3.31 ng/ml, Atrial Natriuretic Peptide 1817 pg/ml, procalcitonin 15.57 ng/ ml, C-Reactive Protein of 91 mg/dl and ESR of 65 mm/sec CONCLUSION(S): SIMS-CT is a serious alteration, but its evolution in this series of cases was favorable. The use of haemadsorption is effective in groups where there is severe inflammatory response syndrome and hyperinflammation and this decreases mortality.

Lipid nanoparticle steric stabilization roadmap

Lipid nanoparticles, such as liposomes, cubosomes, hexosomes, solid lipid nanoparticles and nanostructured lipid carriers, have grown in popularity in many biomedical applications including vaccine and nanomedicine development in the last few years. These systems generally require steric stabilizers to maintain colloidal stability in an aqueous medium, as well as to improve in vivo pharmacokinetics and biodistribution profiles. To date, polyethylene glycol (PEG)-lipid conjugates and block copolymers such as Pluronic® F127 are the most commonly employed stabilizers in these systems due to their commercial availability, excellent ability to stabilize the lipid nanoparticles, and the in vivo “stealth” benefit of PEGylated particle surfaces for prolonged blood circulation. However, in recent years, concerns about the immunogenicity and the impeded cell interaction of nanoparticles containing a PEG layer have been rising and driving research interests in designing alternative steric stabilizers, with additional functionalities such as stimuli-responsiveness. This chapter summarizes the classes of steric stabilizers used in lipid nanoparticle formulations, discusses the latest development of custom-designed novel steric stabilizers, identifies current knowledge gaps in steric stabilizer design principles and selection criteria, and presents a development roadmap for next-generation steric stabilizers as a key functional component of lipid nanoparticles.

Fluorescent bioassay for SARS-CoV-2 detection using polypyrene-g-poly(ε-caprolactone) prepared by simultaneous photoinduced step-growth and ring-opening polymerizations.

The construction of a rapid and easy immunofluorescence bioassay for SARS-CoV-2 detection is described. We report for the first time a novel one-pot synthetic approach for simultaneous photoinduced step-growth polymerization of pyrene (Py) and ring-opening polymerization of ε-caprolactone (PCL) to produce a graft fluorescent copolymer PPy-g-PCL that was conjugated to SARS-CoV-2-specific antibodies using EDC/NHS chemistry. The synthesis steps and conjugation products were fully characterized using standard spectral analysis. Next, the PPy-g-PCL was used for the construction of a dot-blot assay which was calibrated for applications to human nasopharyngeal samples. The analytical features of the proposed sensor showed a detection range of 6.03-8.7 LOG viral copy mL-1 (Ct Scores: 8-25), the limit of detection (LOD), and quantification (LOQ) of 1.84 and 6.16 LOG viral copy mL-1, respectively. The repeatability and reproducibility of the platform had a coefficient of variation (CV) ranging between 1.2 and 5.9%. The fluorescence-based dot-blot assay was tested with human samples. Significant differences were observed between the fluorescence intensity of the negative and positive samples, with an overall correct response of 93.33%. The assay demonstrated a high correlation with RT-PCR data. This strategy opens new insights into simplified synthesis procedures of the reporter molecules and their high potential sensing and diagnosis applications.

Teaching a graduate aourse in pharmaceutical excipients: personal experience and perspectives

Excipients are critically important in converting active pharmaceutical ingredients (API) into drug products that have optimal stability, bioavailability, manufacturability, duration of action, and therapeutic benefits. They will play even greater roles in the future to enable drug targeting, delivery of biotech products and vaccines, gene therapy, continuous manufacturing, 3D printing, and so forth. This commentary describes the author’s experience in teaching a graduate course on excipients at St. John’s University to train students on optimal selection and appropriate use of excipients in formulating dosage forms and development of drug delivery systems. The course is offered in 15 two-hour sessions over a semester, and the course materials are divided into 13 modules on chemistry of different classes of polymeric and non-polymeric excipients and their application in dosage form development, including the use as solubilizing agents, lyophilizing agents, cryoprotectants, buffers, biodegradable materials, and carriers for amorphous solid dispersions and 3D printing. The development of coprocessed excipients, the need for new excipients, and the regulatory aspects of excipients are also covered. The course includes presentations by guest speakers from the industry, and the students also watch virtual presentations from experts that are publicly available from the internet. It is a popular course at St. John’s University taken by all graduate students in the pharmaceutics program. It is recommended that such courses are introduced in other pharmacy schools and academic institutions. The course may be adapted to meet specific needs of different academic programs. Professional associations, such as AAPS and CRS, industry groups like IPEC, and the pharmaceutical industry may be able to help in introducing such courses by providing lecture materials and guest lecturers.

TCTAP C-005 Conundrum of a ‘Primary’ Angioplasty: Provisional Stenting of Left Main Stem / Left Anterior Descending During STEMI With Double Bifurcation

Clinical Information Patient Initials or Identifier Number: R Relevant Clinical History and Physical Exam: A 64-year-old lady with underlying dyslipidemia presented to our emergency department with typical chest pain. Immediate electrocardiogram was performed which showed sinus rhythm, ST elevation at lead 1, aVL and V1, hyperacute T wave at V2 till V3 with ST depression at leads II, III and aVF. Hence a diagnosis of acute anterolateral myocardial infarction, Killip 1 was given and urgent referral to cardiologist was made. Subsequently, she was subjected for primary angioplasty. Relevant Test Results Prior to Catheterization: Blood results showed sodium of 134 mmol/L, potassium of 3.5 mmol/L, urea of 3.2 mmol/L and creatinine of 67 mmol/L. Liver enzymes were within normal limits with aspartate transaminase of 38 U/L and alkaline phosphatase of 91 U/L. Creatinine kinase was 330 U/L but increased to 2861 U/L during subsequent day. In addition, COVID-19 RTK antigen was negative. Relevant Catheterization Findings: Coronary angiogram revealed mild disease at proximal right coronary artery and proximal left circumflex. Minimal disease was noted at distal left main stem, but severe disease was observed from proximal left anterior descending till mid left anterior descending. Heterogenous plague suggesting thrombus was seen at ostial first diagonal as well. [Formula presented] [Formula presented] Interventional Management Procedural Step: Right femoral assess was obtained with 7Fr sheath, and SL 3.5 7Fr guiding catheter was engaged to left coronary artery. Intracoronary heparin and tirofiban were given prior to wiring. First diagonal was wired with Sion Blue while left anterior descending was wired with Runthrough Floppy. Post-wiring both vessels, coronary flow remained TIMI 3 and hence we decided to proceed with IVUS. From IVUS, noted fibrous elastic plague with heavy thrombus burden. Intracoronary streptokinase was given and noted improvement of thrombus from IVUS. BMW wired to left circumflex. Lesion predilated with scoring balloon and associated with no reflow events, resolved post vasodilators. Left main stem was stented with Onyx 3.5 x 26 mm and deployed at 16 atm. Both side branches wires were rewired into same branches via Crusade microcatheter. LMS stent was post dilated with NC Euphora 4.5 mm at nominal pressure. Noted impingement of both ostium diagonal and circumflex branches. Balloon kissing inflation was performed for both LAD/Diagonal bifurcation and LMS/LAD/circumflex bifurcation. POT was performed post balloon kissing inflation with NC Euphora 3.5 mm and 4.5 mm for both LAD and LMS respectively. Next, IVUS was repeated for mid LAD stent length and Onyx 3.0 mm X 15 mm was deployed at nominal pressure. IVUS repeated and noted under-expansion of overlapped segments and post dilated with NC Euphora 3.0 mm at high pressure. [Formula presented] [Formula presented] [Formula presented] [Formula presented] Conclusions: Our clinical vignette demonstrated few learning points including utilization of IVUS during primary angioplasty. Understanding of plague characteristic ensures adequate stents expansion especially with fibro elastic plague. In addition, we also demonstrated several precautions in dealing with bifurcation lesions including usage of double lumen microcatheter for wiring the side branches. Even though we opted for provisional stenting, balloon kissing inflation played pivotal role in preserving flow into side branches.

Qatar Biobank in the COVID-19 era and beyond

Background: During the COVID-19 outbreak, Qatar Biobank (QBB) adapted its services to collect, process, and store high-quality data and specimens. This transformation period brought opportunities, risks, and challenges. Methods: QBB holds College of American Pathologists (CAP) accreditation and International Organization for Standardization (ISO) certification for Quality Management (9001:2105) and Information Security Management Systems (27001: 2013)1. These standards helped to effectively convert its operations to facilitate the COVID-19 national project2. The COVID-19 Biorepository is a National Disease-based study aiming to collect adequate health information and biological samples from Qatar residents infected by SARS-CoV-2 virus to enable evidence-based research towards the discovery and development of novel healthcare interventions and to facilitate research projects related to the COVID-19 pandemic. QBB IT Department used open-source Onyx (ObiBa) software to newly develop a COVID-19 electronic system and interface for various clinical devices and Laboratory Information Management System. QBB operational services were adapted to accommodate the COVID-19 project by providing training to the personnel (n=20;i.e. nurses, research assistants etc.) and developing work instructions (n>5) for the participants' recruitment process, data and specimen collection in the different hospitals. A COVID-19 portal was also developed by QBB to facilitate data access for researchers from Qatar and worldwide4. Results: Within 5 weeks QBB transformed to accommodate a disease/virus-based biorepository. At an operational level, multi-adjustments were implemented. QBB IT department developed an electronic system for the data collection and specimen traceability3. Clinically trained staff were transferred to different healthcare facilities to recruit COVID-19 positive patients. QBB laboratory designed special collection kits. QBB Medical Review Office and Scientific and Education departments managed the recruitment process, set up of the study and provided training to the staff. The Communications and Participants Recruitment Department transformed its operations to fit the needs of the COVID-19 initiative. The Research Access Office designed the COVID-19 access portal and supported the project through purchasing of personal protective equipment and other administrative tasks. Conclusion: It is now time to consider lessons learned, as many countries have been affected by this pandemic, and to understand that biobanks are an asset for a country and there is a need to integrate them into a new standard with their sustainability in mind. Qatar Biobank is a good showcase of how a biobank can successfully assist in the collection of important data related to different health crises such as the COVID-19 pandemic.

Antibacterial, antibiofilm, anti-inflammatory, and wound healing effects of nanoscale multifunctional cationic alternating copolymers.

Infectious diseases caused by new or unknown bacteria and viruses, such as anthrax, cholera, tuberculosis and even COVID-19, are a major threat to humanity. Thus, the development of new synthetic compounds with efficient antimicrobial activity is a necessity. Herein, rationally designed novel multifunctional cationic alternating copolymers were directly synthesized through a step-growth polymerization reaction using a bivalent electrophilic cross-linker containing disulfide bonds and a diamine heterocyclic ring. To optimize the activity of these alternating copolymers, several different diamines and cross-linkers were explored to find the highest antibacterial effects. The synthesized nanopolymers not only displayed good to excellent antibacterial activity as judged by minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, and Escherichia coli, but also reduced the number of biofilm cells even at low concentrations, without killing mammalian cells. Furthermore, in vivo experiments using infected burn wounds in mice demonstrated good antibacterial activity and stimulated wound healing, without causing systemic inflammation. These findings suggest that the multifunctional cationic nanopolymers have potential as a novel antibacterial agent for eradication of multidrug resistant bacterial infections.

A highly efficient nanofibrous air filter membrane fabricated using electrospun amphiphilic PVDF-g-POEM double comb copolymer.

Current global emergencies, such as the COVID-19 pandemic and particulate matter (PM) pollution, require urgent protective measures. Nanofibrous air filter membranes that can capture PM0.3 and simultaneously help in preventing the spread of COVID-19 are essential. Therefore, a highly efficient nanofibrous air filter membrane based on amphiphilic poly(vinylidene fluoride)-graft-poly(oxyethylene methacrylate) (PVDF-g-POEM) double comb copolymer was fabricated using atomic transfer radical polymerization (ATRP) and electrospinning. Fourier transform infrared spectroscopy, X-ray diffraction, proton nuclear magnetic resonance, transmission electron microscopy, differential scanning calorimetry, and thermogravimetric analysis were employed to successfully characterize the molecular structure of the fabricated amphiphilic PVDF-g-POEM double comb copolymer. The nanofibrous air filter membrane based on amphiphilic PVDF-g-POEM double comb copolymer achieved a low air resistance of 4.69 mm H2O and a high filtration efficiency of 93.56 % due to enhanced chemical and physical adsorption properties.

Pandemic-Driven Development of a Medical-Grade, Economic and Decentralized Applicable Polyolefin Filament for Additive Fused Filament Fabrication.

A polyolefin with certified biocompatibility according to USP class VI was used by our group as feedstock for filament-based 3D printing to meet the highest medical standards in order to print personal protective equipment for our university hospital during the ongoing pandemic. Besides the chemical resistance and durability, as well as the ability to withstand steam sterilization, this polypropylene (PP) copolymer is characterized by its high purity, as achieved by highly efficient and selective catalytic polymerization. As the PP copolymer is suited to be printed with all common printers in fused filament fabrication (FFF), it offers an eco-friendly cost-benefit ratio, even for large-scale production. In addition, a digital workflow was established focusing on common desktop FFF printers in the medical sector. It comprises the simulation-based optimization of personalized print objects, considering the inherent material properties such as warping tendency, through to validation of the process chain by 3D scanning, sterilization, and biocompatibility analysis of the printed part. This combination of digital data processing and 3D printing with a sustainable and medically certified material showed great promise in establishing decentralized additive manufacturing in everyday hospital life to meet peaks in demand, supply bottlenecks, and enhanced personalized patient treatment.

Disposable DNA Amplification Chips with Integrated Low-Cost Heaters.

Fast point-of-use detection of, for example, early-stage zoonoses, e.g., Q-fever, bovine tuberculosis, or the Covid-19 coronavirus, is beneficial for both humans and animal husbandry as it can save lives and livestock. The latter prevents farmers from going bankrupt after a zoonoses outbreak. This paper describes the development of a fabrication process and the proof-of-principle of a disposable DNA amplification chip with an integrated heater. Based on the analysis of the milling process, metal adhesion studies, and COMSOL MultiPhysics heat transfer simulations, the first batch of chips has been fabricated and successful multiple displacement amplification reactions are performed inside these chips. This research is the first step towards the development of an early-stage zoonoses detection device. Tests with real zoonoses and DNA specific amplification reactions still need to be done.