Fast one-pot microwave-assisted green synthesis of highly fluorescent plant-inspired S,N-self-doped carbon quantum dots as a sensitive probe for the antiviral drug nitazoxanide and hemoglobin

In this study, we report a one-pot, green, cost-efficient, and fast synthesis of plant-based sulfur and nitrogen self-co-doped carbon quantum dots (S,N-CQDs). By 4-min microwave treatment of onion and cabbage juices as renewable, cheap, and green carbon sources and self-passivation agents, blue emissive S,N-CQDs have been synthesized (λex/λem of 340/418 nm) with a fluorescence quantum yield of 15.2%. A full characterization of the natural biomass-derived quantum dots proved the self-doping with nitrogen and sulfur. The S,N-CQDs showed high efficiency as a fluorescence probe for sensitive determination of nitazoxanide (NTZ), that recently found wide applicability as a repurposed drug for COVID-19, over the concentration range of 0.25–50.0 μM with LOD of 0.07 μM. The nanoprobe has been successfully applied for NTZ determination in pharmaceutical samples with excellent % recovery of 98.14 ± 0.42. Furthermore, the S,N-CQDs proved excellent performance as a sensitive fluorescence nanoprobe for determination of hemoglobin (Hb) over the concentration range of 36.3–907.5 nM with a minimum detectability of 10.30 nM. The probe has been applied for the determination of Hb in blood samples showing excellent agreement with the results documented by a medical laboratory. The greenness of the developed probe has been positively investigated by different greenness metrics and software. The green character of the proposed analytical methods originates from the synthesis of S,N-CQDs from sustainable, widely available, and cheap plants via low energy/low cost microwave-assisted technique. Omission of organic solvents and harsh chemicals beside dependence on mix-and-read analytical approach corroborate the method greenness. The obtained results demonstrated the substantial potential of the synthesized green, safe, cheap, and sustainable S,N-CQDs for pharmaceutical and biological applications. © 2022 Elsevier B.V.

Hybrid Peptide–Agarose Hydrogels for 3D Immunoassays

Recent advances in biosensing analytical platforms have brought relevant outcomes for novel diagnostic and therapy-oriented applications. In this context, 3D droplet microarrays, where hydrogels are used as matrices to stably entrap biomolecules onto analytical surfaces, potentially provide relevant advantages over conventional 2D assays, such as increased loading capacity, lower nonspecific binding, and enhanced signal-to-noise ratio. Here, we describe a hybrid hydrogel composed of a self-assembling peptide and commercial agarose (AG) as a suitable matrix for 3D microarray bioassays. The hybrid hydrogel is printable and self-adhesive and allows analyte diffusion. As a showcase example, we describe its application in a diagnostic immunoassay for the detection of SARS-CoV-2 infection. © 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

CO2-mediated thermal treatment of disposable plastic food containers

In accordance with global economic prosperity, the frequencies of food delivery and takeout orders have been increasing. The pandemic life, specifically arising from COVID-19, rapidly expanded the food delivery service. Thus, the massive generation of disposable plastic food containers has become significant environmental problems. Establishing a sustainable disposal platform for plastic packaging waste (PPW) of food delivery containers has intrigued particular interest. To comprise this grand challenge, a reliable thermal disposable platform has been suggested in this study. From the pyrolysis process, a heterogeneous plastic mixture of PPW was converted into syngas and value-added hydrocarbons (HCs). PPW collected from five different restaurants consisted of polypropylene (36.9 wt%), polyethylene (10.5 wt%), polyethylene terephthalate (18.1 wt%), polystyrene (13.5 wt%), polyvinyl chloride (4.2 wt%), and other composites (16.8 wt%). Due to these compositional complexities, pyrolysis of PPW led to formations of a variety of benzene derivatives and aliphatic HCs. Adapting multi-stage pyrolysis, the different chemicals were converted into industrial chemicals (benzene, toluene, styrene, etc.). To selectively convert HCs into syngas (H2 and CO), catalytic pyrolysis was adapted using supported Ni catalyst (5 wt% Ni/SiO2). Over Ni catalyst, H2 was produced as a main product due to C[sbnd]H bond scission of HCs. When CO2 was used as a co-reactant, HCs were further transformed to H2 and CO through the chemical reactions of CO2 with gas phase HCs. CO2-assisted catalytic pyrolysis also retarded catalyst deactivation inhibiting coke deposition on Ni catalyst. © 2022 Elsevier B.V.

Green Synthesis of Copper Nanoparticles Using Strawberry Leaves and Study of Properties, Anti-cancer Action, and Activity Against Bacteria Isolated from Covid-19 Patients

Severe acute respiratory syndrome type 2 caused by coronavirus 2 is responsible for SARS that led to the emergence of coronavirus disease 2019 (COVID-19). Recent studies have demonstrated a high correlation between secondary bacterial infections and worse outcomes and death in COVID-19 patients. The extensive use of medicines during the last SARS-CoV epidemic led to an increase in the prevalence of multi-drug-resistant germs. Nanoparticles have important characteristics and applications in health, industry, and applied fields, etc. In medical fields, they curb and stop antibiotic-resistant diseases and pathogens. In this study, strawberry leaf extract was used to synthesize copper nanoparticles. The benefits of copper nanoparticles in inhibiting the growth of Pseudomonas aeruginosa and S. aureus bacteria isolated from COVID-19 patients' sputum were tested using the agar well diffusion method. P. aeruginosa and S. aureus bacteria play a significant part in the series of bacterial infections that arise with COVID-19 infection. (1 ml) of strawberry leaf extract was mixed with (50 ml) of copper chloride solution prepared at a concentration of 2 mM at room temperature. The mixture was blended for 7 h to produce copper nanoparticles with a concentration of 2 mM as a stock solution in an environment-friendly manner. The first indication of the production of copper nanoparticles was the increase in the color intensity of the mixture after 7 h. The nanoparticles were detected using UV spectrophotometers, and a scanning electron microscope SEM, XRD, FTIR, and UV-VIS spectral, which appeared at the absorbance of two absorptive peaks, namely: 299 and 804 nm. UV-VIS spectral examination was conducted after a month and was very intense. It also showed two absorbance peaks (300 and 805 nm) with increasing intensity. This is evidence of the insolubility of the nanomaterial and its stability over the month. The scanning electron microscopy results showed that the dimensions of the prepared copper nanoparticles ranged between (46.59 and 58.82 nm). The production of copper nanoparticles in this inexpensive and environmentally friendly biological way has given excellent results in inhibiting the growth of bacteria isolated from COVID-19 patients. The effectiveness of copper nanoparticles was tested against cancerous cells isolated from laryngeal carcinoma, called HeP-2, of a 60-year-old man. The concentration of 50% of the copper nanoparticle solution, which is equivalent to 0.5 mM, gave an inhibition rate of 44.081% in cell cultures. Its effect was compared with the sensitivity of the normal cell line of liver cells (WRL-68);the concentration of 50%, which is equivalent to 0.5 mM, gave an inhibition rate of 5.997% in cell cultures, which showed a good affinity for copper nanoparticles. From this, we conclude that the copper nanoparticles were more effective in inhibiting cancerous cell lines than the normal ones. © 2023 University of Kerbala.

Surface control approach for growth of cerium oxide on flower-like molybdenum disulfide nanosheets enables superior removal of uremic toxins

Due to the high incidence of kidney disease, there is an urgent need to develop wearable artificial kidneys. This need is further exacerbated by the coronavirus disease 2019 pandemic. However, the dialysate regeneration system of the wearable artificial kidney has a low adsorption capacity for urea, which severely limits its application. Therefore, nanomaterials that can effectively remove uremic toxins, especially urea, to regenerate dialysate are required and should be further investigated and developed. Herein, flower-like molybdenum disulphide (MoS2) nanosheets decorated with highly dispersed cerium oxide (CeO2) were prepared (MoS2/CeO2), and their adsorption performances for urea, creatinine, and uric acid were studied in detail. Due to the open interlayer structures and the combination of MoS2 and CeO2, which can provide abundant adsorption active sites, the MoS2/CeO2 nanomaterials present excellent uremic toxin adsorption activities. Further, uremic toxin adsorption capacities were also assessed using a self-made fixed bed device under dynamic conditions, with the aim of developing MoS2/CeO2 for the practical adsorption of uremic toxins. In addition, the biocompatibility of MoS2/CeO2 was systematically analyzed using hemocompatibility and cytotoxicity assays. Our data suggest that MoS2/CeO2 can be safely used for applications requiring close contact with blood. Our findings confirm that novel 2-dimensional nanomaterial adsorbents have significant potential for dialysis fluid regeneration. © 2022

Alternate end-game strategies towards Nirmatrelvir synthesis: Defining a continuous flow process for the preparation of an anti-COVID drug

Scalable alternate end-game strategies for the synthesis of the anti-COVID drug molecule Nirmatrelvir (1, PF-07321332) have been described. The first involves a direct synthesis of 1 via amidation of the carboxylic acid 7 (suitably activated as a mixed anhydride with either pivaloyl chloride or T3P) with the amino-nitrile 10·HCl. T3P was found to be a more practical choice since the reagent promoted efficient and concomitant dehydration of the amide impurity 9 (derived from the amino-amide contaminant 8·HCl invariably present in 10·HCl) to 1. This observation allowed for the development of the second strategy, namely a continuous flow synthesis of 1 from 9 mediated by T3P. Under optimized conditions, this conversion could be achieved within 30 min in flow as opposed to 12–16 h in a traditional batch process. The final API had quality attributes comparable to those obtained in conventional flask processes.

Scheelite-type rare earth vanadates TVO4 (T = Ho, Y, Dy) electrocatalysts: Investigation and comparison of T site variations towards bifunctional electrochemical sensing application

In the wake of the recent COVID-19 pandemic, antibiotics are now being used in unprecedented quantities across the globe, raising major concerns regarding pharmaceutical pollution and antimicrobial resistance (AMR). In view of the incoming tide of alarming apprehensions regarding their aftermath, it is critical to investigate control strategies that can halt their spread. Rare earth vanadates notable for their fundamental and technological significance are increasingly being used as electrochemical probes for the precise quantification of various pharmaceutical compounds. However, a comprehensive study of the role of the cationic site in tailoring the response mechanism is relatively unexplored. Hence, in this work we present a facile hydrothermal synthesis route of rare earth vanadates TVO4 (T = Ho, Y, Dy) as efficient electrocatalyst for the simultaneous detection of nitrofurazone (NF) and roxarsone (RX). There appears to be a significant correlation between T site substitution, morphological and the electrochemical properties of rare earth metal based vanadates. Following a comparative study of the electrochemical activity, the three rare-earth vanadates were found to respond differently depending on their composition of T sites. The results demonstrate that Dy-based vanadate displays increased electrical conductivity and rapid charge transfer characteristics. Thus, under optimal reaction conditions DyVO4- based electrodes imparts outstanding selectivity towards the detection of NF and RX with an extensive detection window of NF = 0.01–264 µM & RX = 0.01–21 µM and 36–264 µM and low detection limit (0.002, 0.0009 µM for NF and RX, respectively). In real-time samples, the proposed sensor reveals itself to be a reliable electrode material capable of detecting residues such as NF and RX. © 2022 Elsevier B.V.

AMICOUMACIN-BASED PRODRUG DEVELOPMENT APPROACH

Coronavirus disease COVID-19, caused by the SARS-CoV-2 virus, is highly contagious and has a severe morbidity. Providing care to patients with COVID-19 requires the development of new types of antiviral drugs. The aim of this work is to develop a prodrug for the treatment of coronavirus disease using the antibiotic Amicoumacin A (Ami), the mechanism of action of which is based on translation inhibition. Enzymatic hydrolysis of an inactivated prodrug by the SARS-CoV-2 main protease can lead to the release of the active Ami molecule and, as a consequence, the suppression of protein biosynthesis in infected cells. To test the proposed hypothesis, a five-stage synthesis of an inactivated analogue of Amicoumacin A was carried out. Its in vitro testing with the SARS-CoV-2 recombinant protease MPro showed a low percentage of hydrolysis. Further optimization of the peptide fragment of the inactivated analog recognized by the SARS-CoV-2 MPro protease may lead to an increase in proteolysis and the release of Amicoumacin A.

Synthesis and Crystal Structure of Adamantylated 4,5,6,7-Tetrahalogeno-1H-benzimidazoles Novel Multi-Target Ligands (Potential CK2, M2 and SARS-CoV-2 Inhibitors); X-ray/DFT/QTAIM/Hirshfeld Surfaces/Molecular Docking Study.

A series of new congeners, 1-[2-(1-adamantyl)ethyl]-1H-benzimidazole (AB) and 1-[2-(1-adamantyl)ethyl]-4,5,6,7-tetrahalogeno-1H-benzimidazole (Hal=Cl, Br, I; tClAB, tBrAB, tIAB), have been synthesized and studied. These novel multi-target ligands combine a benzimidazole ring known to show antitumor activity and an adamantyl moiety showing anti-influenza activity. Their crystal structures were determined by X-ray, while intermolecular interactions were studied using topological Bader's Quantum Theory of Atoms in Molecules, Hirshfeld Surfaces, CLP and PIXEL approaches. The newly synthesized compounds crystallize within two different space groups, P-1 (AB and tIAB) and P21/c (tClAB and tBrAB). A number of intramolecular hydrogen bonds, C-H⋯Hal (Hal=Cl, Br, I), were found in all halogen-containing congeners studied, but the intermolecular C-H⋯N hydrogen bond was detected only in AB and tIAB, while C-Hal⋯π only in tClAB and tBrAB. The interplay between C-H⋯N and C-H⋯Hal hydrogen bonds and a shift from the strong (C-H⋯Cl) to the very weak (C-H⋯I) attractive interactions upon Hal exchange, supplemented with Hal⋯Hal overlapping, determines the differences in the symmetry of crystalline packing and is crucial from the biological point of view. The hypothesis about the potential dual inhibitor role of the newly synthesized congeners was verified using molecular docking and the congeners were found to be pharmaceutically attractive as Human Casein Kinase 2, CK2, inhibitors, Membrane Matrix 2 Protein, M2, blockers and Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2, inhibitors. The addition of adamantyl moiety seems to broaden and modify the therapeutic indices of the 4,5,6,7-tetrahalogeno-1H-benzimidazoles.

Weekly updating of guideline recommendations was feasible: the Australian National COVID-19 Clinical Evidence Taskforce.

OBJECTIVE: To investigate how quickly evidence was incorporated into the Australian living guidelines for COVID-19 during the first 12 months of the pandemic. STUDY DESIGN AND SETTING: For each study concerning drug therapies included in the guideline from April 3, 2020 to April 1, 2021 we extracted the publication date of the study and the guideline version the study was included in. We analysed two subgroups of studies: those published in high impact factor journals and those with 100 or more participants. RESULTS: In the first year we published 37 major versions of the guidelines, incorporating 129 studies that investigated 48 drug therapies informing 115 recommendations. The median time from first publication of a study to incorporation in the guideline was 27 days (IQR 16 to 44), ranging from 9 to 234 days. For the 53 studies in the highest impact factor journals, the median was 20 days (IQR 15 to 30) and for the 71 studies with 100 or more participants the median was 22 days (IQR 15 to 36). CONCLUSION: Developing and sustaining living guidelines where evidence is rapidly incorporated is a resource- and time-intensive undertaking, however this study demonstrates that it is feasible, even over a long period.

metaCOVID: A web-application for living meta-analyses of COVID-19 trials.

Outputs from living evidence syntheses projects have been used widely during the pandemic by guideline developers to form evidence-based recommendations. However, the needs of different stakeholders cannot be accommodated by solely providing pre-defined non amendable numerical summaries. Stakeholders also need to understand the data and perform their own exploratory analyses. This requires resources, time, statistical expertise, software knowledge as well as relevant clinical expertise to avoid spurious conclusions. To assist them, we created the metaCOVID application which, based on automation processes, facilitates the fast exploration of the data and the conduct of sub-analyses tailored to end-users needs. metaCOVID has been created in R and is freely available as an R-Shiny application. Based on the COVID-NMA platform (https://covid-nma.com/) the application conducts living meta-analyses of randomized controlled trials related to COVID-19 treatments and vaccines for several outcomes. Several options are available for subgroup and sensitivity analyses. The results are presented in downloadable forest plots. We illustrate metaCOVID through three examples involving well-known treatments and vaccines for COVID-19. The application is freely available from https://covid-nma.com/metacovid/.

LC-MS/MS-PRM Quantification of IgG Glycoforms Using Stable Isotope Labeled IgG1 Fc Glycopeptide Standard.

Targeted quantification of proteins is a standard methodology with broad utility, but targeted quantification of glycoproteins has not reached its full potential. The lack of optimized workflows and isotopically labeled standards limits the acceptance of glycoproteomics quantification. In this work, we introduce an efficient and streamlined chemoenzymatic synthesis of a library of isotopically labeled glycopeptides of IgG1 which we use for quantification in an energy optimized LC-MS/MS-PRM workflow. Incorporation of the stable isotope labeled N-acetylglucosamine enables an efficient monitoring of all major fragment ions of the glycopeptides generated under the soft higher-energy C-trap dissociation (HCD) conditions, which reduces the coefficients of variability (CVs) of the quantification to 0.7-2.8%. Our results document, for the first time, that the workflow using a combination of stable isotope labeled standards with intrascan normalization enables quantification of the glycopeptides by an electron transfer dissociation (ETD) workflow, as well as the HCD workflow, with the highest sensitivity compared to traditional workflows. This was exemplified by a rapid quantification (13 min) of IgG1 Fc glycoforms from COVID-19 patients.

A rapid priority setting exercise combining existing, emergent evidence with stakeholder knowledge identified broad topic uncertainties.

OBJECTIVE: The project aimed to rapidly identify priority topic uncertainties as a first step to identify future systematic review questions of pertinence to key international faecal incontinence (FI) stakeholders (patients, carers, healthcare professionals, policy makers and voluntary, community or social enterprise representatives). The paper aim is to share our methods, experience and learning with other groups planning to deliver a rapid priority setting exercise. STUDY DESIGN: An evidence gap map incorporated three evidence streams: emerging evidence identified through horizon scanning; existing evidence identified through systematic searches of bibliographic databases; and FI stakeholder insights collected through an international survey. The evidence gap map was presented during an online workshop with stakeholders, where they shared their expertise to expand, refine and rank topic uncertainties using ideation techniques, focus group discussions, consensus techniques and online polling. RESULTS: The multi-step methods used to deliver this priority-setting exercise resulted in identification of broad priority topic uncertainties. The methods appear to have high acceptability and engagement with participants but await full evaluation. CONCLUSIONS: This project successfully followed robust methodology, building upon frameworks from published priority setting and evidence gap mapping projects whilst incorporating strong patient and public involvement components.

Clinical experiences of final-year nursing students during the COVID-19 pandemic: A systematic review and meta-synthesis.

OBJECTIVES: This systematic review aimed to qualitatively synthesise existing literature to examine the clinical nursing experiences of final-year nursing students during the COVID-19 pandemic and provide recommendations for the effective management of clinical placement of nursing students. DESIGN: A qualitative systematic review was conducted and reported following Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. DATA SOURCES: Five electronic databases were searched and qualitative studies were included for analysis if they focussed on the clinical nursing experiences of final-year nursing students during the COVID-19 pandemic. REVIEW METHODS: Data synthesis was conducted by extracting all findings, developing categories, and producing synthesised findings. RESULTS: Four synthesised findings were concluded: 1) facing the unknown and willingness to help, 2) challenging the clinical environment, 3) transition improving professional identity, and 4) finding ways out of the pandemic. CONCLUSIONS: The transition of nursing students to clinical nursing practice during the pandemic is a personally and professionally challenging process, while nursing students try to adapt to the changing clinical environment and enhance their professional identity. Nursing managers and health policymakers should acknowledge the challenges encountered by nursing students during the pandemic and support the professional growth of future nursing teams by providing high-quality supervision.

Green Synthesis of Copper Nanoparticles by Using Plant Extracts and their Biomedical Applications – An Extensive Review

In recent years, the green synthesis of different metal nanoparticles has become a substan-tial technique for the synthesis of different essential nanoparticles and their potential applications in technological, industrial along with biomedical fields. Among the several essential nanoparticles, copper nanoparticles (CuNPs) have attracted enormous attention for their wide range of applications like the production of gas sensors, solar cells, high-temperature superconductors as well as drug delivery materials and catalysis owing to its distinctive optical, electrical, dielectric, imaging and catalytic, etc. properties. Herein, in this review, our aim is to find out the recent progress of synthesis, as well as different optical and structural characterizations of green, synthesized CuNPs along with their broad-spectrum biomedical applications, mainly antibacterial, antifungal, antiviral and anticancer as well as the future perspective of research trends in the green synthesis of CuNPs. CuNPs have been synthesized by different researchers using three methods, namely, physical, chemical, and biological. In this review, the eco-friendly, efficient and low cost different established biological/green synthesis methods of CuNPs using different plant extracts like leaves, flowers, fruits, seeds, latex, etc., as capping and reducing agents have been briefly discussed, along with reaction conditions together with their optical as well as structural analysis. Effects of different parameters on the green synthesis of CuNPs like the presence of phytochemicals and confirmation of phytochemicals, temperature, pH, etc., are eluci-dated. Studies of the antibacterial activity of biomolecules capped CuNPs by different researchers against both Gram-positive and Gram-negative bacterial strains along with minimum inhibitory concentration (MIC) values have been summarized. Furthermore, antifungal and antiviral effects of green synthesized CuNPs studied by different researchers are mentioned with minimum inhibitory concentration (MIC) values. The anticancer activity of green synthesized CuNPs against different cancer cells studied by different researchers is summarized with correlation sizes of CuNPs on anticancer activity. The review also focuses on in vivo applications of green synthesized CuNPs along with clinical trails. Furthermore, an emphasis is given to the effectiveness of CuNPs in combating COVID-19. © 2023 Bentham Science Publishers.

Feasibility of targeted antiviral therapy for acute respiratory viral infections in the COVID-19 pandemic.

The aim of this study was to analyze the efficacy and safety of using etiotropic therapy with favipiravir and molnupiravir that can selectively bind and inhibit not only SARS-CoV-2 proteins but also other RNA-containing pathogens of acute respiratory diseases. High transmission of pathogens, the risk of becoming chronic, frequent complications, cases of co-infection with several pathogens, which can lead to a more severe course of the disease, insufficient vaccination effectiveness, all this requires additional strategies for both prevention and treatment of acute respiratory viral infections. RNA-dependent RNA polymerase (RdRp), which has no equivalent in human cells, is involved in RNA synthesis and is an excellent therapeutic target for diseases caused by RNA viruses, including SARS-CoV-2. The long process of drug development and the "reuse" of drugs approved for other indications or successfully tested in terms of safety and tolerability pose the challenge of rapid establishment of an effective drug, including for the treatment of severe cases of COVID-19. Copyright © 2022, Dynasty Publishing House.

Direct One-Pot Synthesis of Propofol from Paracetamol Tablets

The COVID-19 pandemic has affected millions of people in the entire world and caused a shortage of several drugs, including propofol. Therefore, several protocols for propofol synthesis have been published in recent years. Herein, we present a process starting from paracetamol, a very common and abundant active pharmaceutical ingredient. Since the first three steps (Friedel-Crafts double alkylation, acetyl deprotection, and diazotization) are done in acidic media, a one-pot approach was developed. Furthermore, we observed that the extraction of the final product can be simplified by steam-distillation, leading to propofol in 47% isolated yield with high purity. This presented process could be an example of active pharmaceutical ingredient reuse since similar results were observed with commercial paracetamol tablets (with excipients) regardless of expiration date. © 2023 American Chemical Society.

Screening of Asian Natural Materials to Promote beta-Endorphin Synthesis

Since the recent coronavirus disease 2019 pandemic and the lifestyle changes it necessitated, the demand for mental health treatment has skyrocketed, with long wait lists for both psychological and psychiatric care. Over-the-counter supplements and home remedies are increasingly sought. In this study, we screened natural materials and blended supplements from Asia that may improve the mood and mental health of humans by testing cell viability and expression of the proopiomelanocortin gene as a marker of beta-endorphin production in rat hypothalamus neuron cells. Among 23 tested samples, 3 samples produced significantly higher cell viability in R-HTH-507 cells than the control treatment. In a real-time-polymerase chain reaction (RT-PCR) experiment, 7 samples showed significant beta-endorphin synthesis activity. This is the first report that the Asian natural materials Areca catechu, Moringa oleifera, Lignosis rhinocerus, and Aegle marmelos promote beta-endorphin synthesis;further investigation will identify the active ingredients in the blended samples. These results suggested that these Asian natural materials have great potential to expand the range of treatments for mental health. Copyright © The Author(s) 2023.

Acute flaccid myelitis diagnosed after IVIG: a diagnostic dilemma

Case Report: A 4-year-old African American male presented to an outside emergency department (ED) following sudden inability to move left upper extremity. Past medical history was unremarkable and routine vaccinations were up to date. Radiograph of affected extremity ruled out fractures and patient was discharged to follow up with primary care physician. Two days later mother brought him to our ED due to persistent left upper extremity paralysis, poor appetite, and subjective fever. On exam his left arm was warm and tender to dull and sharp touch;he had definite loss of active movement, hypotonia and absence of deep tendon reflexes. The patient had winging of left scapula and could not shrug left shoulder. MRI of cervical and thoracic spine showed enlargement of spinal cord from C2-C6 level with gray matter hyperintensity, slightly asymmetric to the left. Laboratory studies showed leukocytosis (14 000/mcL) and CSF studies showed pleocytosis of 89 WBC/mcL (93.3% mononuclear cells and 6.7% polymorphonuclear cells), 0 RBCs, normal glucose and protein, and a negative CSF meningoencephalitis multiplex PCR panel. Due to high suspicion of demyelinating or autoimmune condition he was treated with high dose steroids and IVIG. Subsequently neuromyelitis optica was ruled out as aquaporin-4 receptor antibodies (AB) and myelin oligodendrocyte glycoprotein AB were normal. CSF myelin basic protein and oligoclonal bands were absent ruling out demyelinating disorders. CSF arboviruses IgM and West Nile IgM were negative. He showed minimal improvement in left upper extremity movement but repeat spinal cord MRI one week later showed improved cord thickness with less hyperintensity. Respiratory multiplex PCR was negative including enteroviruses. Repeat CSF studies after IVIG showed increased IgG index and IgG synthesis suggestive of recent spinal cord infection, consistent with acute flaccid myelitis (AFM). Pre-IVIG blood PCR was invalid for enteroviruses due to PCR inhibitors found in the sample. Blood post-IVIG was negative for mycoplasma IgM, West Nile IgM, and arboviruses IgM. Enterovirus panel titers (post-IVIG) were positive for coxsackie A (1:32), coxsackie B type 4 (1:80) and 5 (1:320), echovirus type 11 (1:160) and 30 (1:80) as well as positive for poliovirus type 1 and 3. These titers could not distinguish acute infection from patient's immunity or false-positives as a result of IVIG. He was discharged with outpatient follow-up visits with neurology, infectious disease, occupational and physical therapy, showing only mild improvement after discharge. Discussion(s):With the anticipated resurgence of AFM after the peak of COVID-19 pandemic, our case illustrates the need to consider this diagnostic possibility in patients with flaccid paralysis. It is important to remember CSF IgG synthesis is not affected by IVIG. In addition when treatment plans include IVIG, appropriate samples should be collected before IVIG to facilitate accurate work-up for infectious diseases. Copyright © 2023 Southern Society for Clinical Investigation.

Process Intensification and Increased Safety for the On-Demand Continuous Flow Synthesis of Dithiothreitol, a Crucial Component in Polymerase Chain Reaction Testing Kits

The importance of rapid access to diagnostics tools in the identification of pathogens-including their crucial component, bioreagents-was recently underscored in the COVID-19 pandemic. The currently adopted synthesis of dithiothreitol (DTT) involves four steps in batch with long reaction times and which generates a highly carcinogenic and mutagenic bis-epoxide intermediate. In this work, we have developed an intensified telescoped three-step continuous flow synthesis of DTT involving a base-mediated ring closure epoxidation, a nucleophilic epoxide opening with thioacetic acid, and an acid-mediated deacetylation. One of the key features is that the first two steps are conducted in a telescoped continuous flow fashion, allowing generation and consumption of the hazardous intermediate in situ, suppressing the need for its isolation, and improving the overall safety of the synthesis. The process is completed by an acid-catalyzed deacetylation and a subsequent recrystallization to afford the desired DTT. Flow chemistry allows here to intensify the process by using high temperatures and high pressures while minimizing the number of unit operations and improving the overall safety of the process. Our protocol permits the on-demand production of DTT in case of future outbreaks.