Optimized nanoparticle-mediated base editing of cystic fibrosis-causing variants

Background: Next-generation SARS-CoV-2 vaccines demonstrated that nanoparticle messenger ribonucleic acid (mRNA) delivery is effective and safe for in vivo delivery in humans. Current treatments for cystic fibrosis (CF) primarily focus on modulator drug therapies designed to correct malfunctioning CF transmembrane conductance regulator (CFTR) protein, but these modulators are ineffective for the 10% of people with CF with variants that do not allow protein production. Among these is the splice variant 3120 + 1G >A, the most common CF-causing mutation in native Africans. Gene editing would allow production of CFTR protein and enhancement of function using available CFTR modulators. We have demonstrated that electroporation of a modified CRISPR-Cas9 base editor to primary human bronchial epithelial cells carrying 3120 + 1G >A and F508del mutant alleles achieved 75% genome editing of the splice variant, resulting in approximately 40% wild-type (WT) CFTR function [1]. Here,we evaluate the effectiveness of several new nanoparticle formulations at delivering green fluorescent protein (GFP) mRNA to CF bronchial epithelial (CFBE41o-) cells. Using the optimal formulation,we then tested the efficacy correction of the 3120 + 1G >Avariant in a CFTR expression minigene (EMG) integrated into the genome of isogenic CFBE cells using mRNA and plasmid deoxyribonucleic acid (DNA) encoding adenine base editor (ABE) and guide (g)RNA. Method(s): GFP served as a reporter to evaluate transfection efficiency, cell viability, and mean fluorescence intensity (MFI) for three dosages (150, 75, 32.5 ng of mRNA), four polymer-to-mRNA to weight (w/w) ratios (60, 40, 30, 20), and four polymers (R, Y, G, B). 7-AAD served as a live/dead stain to quantify viability, with flow cytometry results analyzed using FlowJo software. CFBE cells stably expressing the 3120 + 1G >A EMG were transfected with the optimized nanoparticle formulation to deliver ABE and gRNA at two dosages (150, 75 ng) of mRNA and DNA. CFTR function in CFBE cellswas measured by short circuit current, forskolin stimulation, and inh-172 inhibition as a measure of editing efficiency. Result(s): Flow cytometry showed that polymer R achieved more than 85% GFP transfection, compared with a maximum of approximately 35% for the other three polymers at the maximum 150-ng dose, with approximately 80% viability normalized to untreated cells. In addition, polymer R achieved GFP MFI more than one order of magnitude as high as other formulations (~30 000 vs 2700 MFI) for the other three polymers at 150-ng dose and 40 w/w ratio. CFBE cells transfected with polymer R nanoparticles containing ABE and guide RNA at 75 ng and 150 ng showed mean CFTR function increase to 10 muA 6 (standard error of the mean [SEM] 1.1 muA) (~10% of WT) and 6.3 muA (SEM 0.9 muA) (~6% of WT), respectively. Greater toxicity at the higher dose could explain the larger increase in CFTR current at the lower dose. DNA-encoded ABE plasmid and gRNA showed a less robust increase in CFTR function (2.9 muA [SEM 0.4 muA] for 75-ng dose;3.0 muA [SEM 0.4 muA] for 150-ng dose), which was probably a result of the nanoparticle formulation being optimized for RNA instead of DNA cargo or the additional intracellular barriers that must be overcome for successful DNA delivery. Conclusion(s): We demonstrated that an optimized nanoparticle formulation containing ABE and gRNA can correct splicing of isogenic cells bearing the 3120 + 1G >A CFTR variant, resulting in recovery of CFTR function. In ongoing work, we are adapting these nanoparticles for RNA- and DNAencoded ABE and gRNA delivery to primary human bronchial epithelial cells.Copyright © 2022, European Cystic Fibrosis Society. All rights reserved

A dual-functional module cellular electrochemical sensing platform for simultaneous detection guanine and xanthine.

The separation of the superimposed electrochemical signals of intracellular guanine (G) and xanthine (X) is difficult, which is great obstacle to the application of cell electrochemistry. In this paper, independent functional modules, G-functional module (G-FM) and X-functional module (X-FM), were constructed by molecular imprinting technology for sensitive detection of G and X without mutual interference, then integrated in dual-functional module cellular electrochemical sensing platform (DMCEP) as signal sensing units. DMCEP transmitted signals of G and X in cells synchronously to two windows by two signal sensing channels, and achieved the separation of superimposed signals of G and X in cells. DMCEP exhibited satisfactory reproducibility with relative standard deviation (RSD) of 3.10 and 2.22 %, repeatability with RSD of 3.72 and 3.05 % for G and X detection, and detection limit 0.05 µΜ for G and 0.06 µΜ for X. Good linear relationships between cell concentrations and the signals of G and X on DMCEP were shown in range of 0.75-85 × 106 and 3-85 × 106 cells/mL, respectively. The growth of MCF-7 cells was tracked by DMCEP, and showed consistent trend with the cell counting method, while the change of cell viability from lag to logarithmic phase captured by DMCEP was earlier than that of cell counting method. This strategy provided the foundation for the establishment of the cell viability electrochemical detection method, and new insights into the simultaneous recording of other analyses with superimposed peak positions and the simultaneous tracking of multiple biomarkers.

Development of a Novel in-Vitro Hcov-Oc43 Screening Method

Aim: To develop a simple, inexpensive antiviral screening assay, applicable to SARS-CoV-2, using a plate-based bioassay approach to assess the in-vitro activity of compounds against HCoV-OC43. Background(s): Despite the successful deployment of vaccines against SARS-CoV-2 there remains a need for effective antivirals for acute infection treatment. A distinct problem facing the search for new anti-coronavirus compounds is the cost of antiviral screening, compounded by the biosecurity concerns of live SARSCoV- 2 culture. In concert with low pathogenic surrogate virus use, the resazurin reduction assay, which is often employed for compound cytotoxicity assessments can be employed for safe, rapid and inexpensive antiviral screening. Method(s): In-vitro cell based resazurin reduction assays were optimised using remdesivir as a control compound for the assessment of anti-HCoV-OC43 activity. Following optimisation, 246 purified natural compounds from the University of Western Australia's compound collection,were screened using the resazurin bioassay as a primary screen, under pre-treatment and cotreatment conditions. Five compounds, which demonstrated anti- HCoV-OC43 activity, were chosen for secondary screening with dose responses determined using qRT-PCR. Result(s): Primary screens of the 246 compounds using the resazurin bioassay identified five compounds with a relative viral inhibition >60% and a relative cell viability >70% (Table 1). The Z factor of the pre-treatment and co-treatment assays was >0.5 (average +/- SD;0.85 +/- 0.07, 0.91 +/- 0.03 respectively). Further dose response analysis of the top five compounds identified one compound with an IC50 value <10 muM. Conclusion(s): The method developed is an appropriate primary screening tool for the identification of novel compounds with anti-HCoV-OC43 activity.Copyright © 2023 Southern Society for Clinical Investigation.

In the Literature

The article discusses several published infectious diseases studies. It includes a study on the antibody invasion properties of the BQ and XBB subvariants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and two studies that provide information about the mechanisms of HIV silencing, prolonged survival of provirus carrying CD4 cells, and the escape of the cells from immune surveillance.

Raw Honey Of Apiscerana Indica Spp. Induces Transition, In Vitro

On the verge of Covid-19 pandemic accompanied by the vaccination research, proposed research focuses on continuous cell lines which are actually extensively applied as essential as well as economical methods for fundamental scientific analysis, chemical rate of metabolism, analysis about degree of toxicity and formation of biological formula just like vaccines. Mouse spleen cells used are well known to survive only as primary cultures. The availability of non-cancerous long-term cell cultures is scarcely available for in vitro research studies. In order to overcome this limitation, the proposed research attempted to establish a naive, long term-continuous cell culture from Balb/c mouse. This continuous cell culture derived and established from a primary mouse spleen has the potential as a model cell culture for various applications of in vitro testing. This research can be useful for further research in the specified domain.Copyright © 2022 Wolters Kluwer Medknow Publications. All rights reserved.

74th Annual Congress of the Japan Society of Obstetrics and Gynecology

The proceedings contain 429 papers. The topics discussed include: a short peptide encoded by long non-coding RNA small nucleolar RNA host gene 6 promotes cell migration and epithelial-mesenchymal transition by activating transforming growth factor-beta/SMAD signaling pathway in human endometrial cells;a short peptide encoded by long non-coding rna small nucleolar rna host gene 6 promotes cell migration and epithelial-mesenchymal transition by activating transforming growth factor-beta/smad signaling pathway in human endometrial cells;compatible cut-off values for luteinizing hormone and the luteinizing hormone/follicle-stimulating hormone ratio in diagnostic criteria of the Japan society of obstetrics and gynecology for polycystic ovary syndrome;intracytoplasmic sperm injection cycle success in patients under 35 years old with diminished ovarian reserve plus severe male factor;assisted reproductive technology and neonatal intensive care unit: a retrospective observational study from a single center;the value of clinical symptoms, the neutrophil-to-lymphocyte ratio, and ultrasonographic features in predicting adnexal torsion: a case-control study;construction of a diagnostic classifier for cervical intraepithelial neoplasia and cervical cancer based on xgboost feature selection and random forest model;and impact of the COVID-19 pandemic on surgery for benign diseases in gynecology: a nationwide survey by the japan society of obstetrics and gynecology.

Efficacy of Favipiravir and Molnupiravir against Novel Sars-Cov-2 Variants in Vitro and in Vivo

The COVID-19 disease pandemic remains a significant global problem, resulting in hundreds of millions of cases and millions of deaths. The search for specific inhibitors of SARS-CoV-2 for the treatment of this infection remains relevant. Drugs such as Favipiravir and Molnupiravir, which exhibit specific antiviral activity against SARS-CoV-2, are already being used to treat patients. However, there is limited evidence of their effectiveness, especially against novel genetic variants of the COVID-19 pathogen. The aim of this study was to investigate the antiviral effect of these drugs using an in vitro experimental model of SARS-CoV-2 infection in Vero E6 cell culture and an animal model of infection using Syrian hamsters. It has been established that Molnupiravir has an inhibitory effect against variants of the SARS-CoV-2 with IC50 values from 16.51 to 7.88 microM in vitro, and reduces the infectious titer of the virus in the lungs of animals by ~1.5 Log10 in vivo, in while Favipiravir shows lower activity and severe toxicity. Dose selection and frequency of use remain unexplored.Copyright © 2022 Pirogov Russian National Research Medical University. All rights reserved.

Polysaturated fatty acids and their derivatives regulate respiratory infections.

The regulation of inflammation is a complex pathophysiological process that depends on the production of oxygenated lipid derivatives essential polyunsaturated fatty acids, like omega-3 and omega-6, among which are the lipoxins resolvins and protectins, called specialized pro-resolving lipid mediators (SPM). Their activity is associated with the control of respiratory infection processes to modulate the production of proinflammatory cytokines, avoiding damage due to inflammation-associated necrosis, reducing microbial loads, and promoting tissue remodeling. Therefore, we review some of the biochemical, physiological and immunological aspects of SPM in the regulation of inflammation in respiratory infections.Copyright © 2022, Instituto Nacional de Enfermedades Respiratorias. All rights reserved.

Polyoxometalates Impact as Anticancer Agents.

Polyoxometalates (POMs) are oxoanions of transition metal ions, such as V, Mo, W, Nb, and Pd, forming a variety of structures with a wide range of applications. Herein, we analyzed recent studies on the effects of polyoxometalates as anticancer agents, particularly their effects on the cell cycle. To this end, a literature search was carried out between March and June 2022, using the keywords "polyoxometalates" and "cell cycle". The effects of POMs on selected cell lines can be diverse, such as their effects in the cell cycle, protein expression, mitochondrial effects, reactive oxygen species (ROS) production, cell death and cell viability. The present study focused on cell viability and cell cycle arrest. Cell viability was analyzed by dividing the POMs into sections according to the constituent compound, namely polyoxovanadates (POVs), polyoxomolybdates (POMos), polyoxopaladates (POPds) and polyoxotungstates (POTs). When comparing and sorting the IC50 values in ascending order, we obtained first POVs, then POTs, POPds and, finally, POMos. When comparing clinically approved drugs and POMs, better results of POMs in relation to drugs were observed in many cases, since the dose required to have an inhibitory concentration of 50% is 2 to 200 times less, depending on the POMs, highlighting that these compounds could become in the future an alternative to existing drugs in cancer therapy.

Recent advances of nanotechnology in COVID 19: A critical review and future perspective

The global anxiety and economic crisis causes the deadly pandemic coronavirus disease of 2019 (COVID 19) affect millions of people right now. Subsequently, this life threatened viral disease is caused due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, morbidity and mortality of infected patients are due to cytokines storm syndrome associated with lung injury and multiorgan failure caused by COVID 19. Thereafter, several methodological advances have been approved by WHO and US-FDA for the detection, diagnosis and control of this wide spreadable communicable disease but still facing multi-challenges to control. Herein, we majorly emphasize the current trends and future perspectives of nano-medicinal based approaches for the delivery of anti-COVID 19 therapeutic moieties. Interestingly, Nanoparticles (NPs) loaded with drug molecules or vaccines resemble morphological features of SARS-CoV-2 in their size (60–140 nm) and shape (circular or spherical) that particularly mimics the virus facilitating strong interaction between them. Indeed, the delivery of anti-COVID 19 cargos via a nanoparticle such as Lipidic nanoparticles, Polymeric nanoparticles, Metallic nanoparticles, and Multi-functionalized nanoparticles to overcome the drawbacks of conventional approaches, specifying the site-specific targeting with reduced drug loading and toxicities, exhibit their immense potential. Additionally, nano-technological based drug delivery with their peculiar characteristics of having low immunogenicity, tunable drug release, multidrug delivery, higher selectivity and specificity, higher efficacy and tolerability switch on the novel pathway for the prevention and treatment of COVID 19. © 2022 The Author(s)

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

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.

GMP Manufacturing of Cryopreserved peripheral Blood Stem Cell Products

Background: Hematopoietic stem cell preparations are pharmaceuticals that are typically transfused as fresh products. Due to the travel restrictions caused by the Covid-19 pandemic since March 2020, the supply of fresh stem cell preparations to patients became significantly challenging. To maintain global patient supply, a GMP-compliant manufacturing process for cryopreservation of allogeneic stem cell donations was established and validated. Method(s): The donor leukapheresis is transported from the collection center to the manufacturing site at 2-6degreeC using qualified containers. In class D clean rooms, this material is further processed in a completely closed process. If necessary, volume reduction is performed by centrifugation. The suspension is mixed with the cryoprotectant Cryostore CS10 to achieve a final DMSO concentration of 5%. The formulated product is filled into cryobags and cryopreserved under controlled freezing conditions. The frozen products are stored and transported at <= 140 degreeC. The cell counter NC-200 is used to determine the cell count and viability of the product. 97 products in 2020 and 127 products in 2021 were successfully cryopreserved and transported worldwide. Result(s): The mean values of viability of 215 products are 99.3 % (+/- 1.1 % SD) for fresh apheresis products and 93.1 % (+/- 6.2 % SD) for the corresponding cryopreserved final product after thawing. For a good recovery after thawing, the age of the apheresis as well as the DMSO contact time are generally considered to be critical factors. The analysis of the viability of 222 cryopreserved end products after thawing shows a correlation in relation to the age of the apheresis (Fig. 1). As the age of the apheresis increases, the viability decreases. Therefore, care should be taken about the age of the apheresis. A correlation between the DMSO contact time and the viability of the end products after thawing was not observed (Fig. 2). Conclusion(s): GMP-compliant closed system manufacturing of cryopreserved allogeneic stem cell products provides safe and high quality drugs that can be used for transplantation. We have shown that with our manufacturing process, DMSO, which is potentially toxic to cells, has no effect on cell recovery after thawing. Cryopreservation is therefore a suitable and safe method to provide patients with essential therapy worldwide. (Figure Presented).

Imaging tumor-infiltrating CD8+ T-cells in non-small cell lung cancer patients upon neo-adjuvant treatment with durvalumab

Aim/Introduction: Immune checkpoint inhibitors (ICI), like targeting programmed death receptor ligand 1 (PD-L1), have revolutionized anti-cancer treatments, including non-small cell lung cancer (NSCLC) [1, 2]. Assessment of PD-L1 expression on tumor biopsies is current practice, but there is a need for additional biomarkers correlating to the complex mechanism of action of ICI. The presence of tumorinfiltrating CD8+ T-cells (TILs) is a robust biomarker associated with immune therapy success [3]. Tools to track TILs in patients during ICI treatment allow further development of immune-oncology drugs. Material(s) and Method(s): This ongoing single-center prospective study (NCT03853187) includes patients with histologically proven T1b-3N0-1M0 NSCLC eligible for resection. Exclusion criteria are previous anti-cancer therapy and known immune disorders or suppression. Patients receive two courses neo-adjuvant durvalumab (750mg Q2W), after which TIL imaging is performed. Cohort 1 underwent apheresis and magnetic-activated cells sorting to isolate 100 x10e6 autologous CD8+ T-cells for cell labeling with 111In-oxine. Re-injection was followed by 4h post-injection (p.i.) planar imaging, 70h p.i. SPECT imaging, standard-of-care surgery and 78h p.i. uSPECT of the resected lobe. Patients in cohort 2 (ongoing) receive 1.5mg 89Zr-Df-crefmirlimab followed by PET/CT 24h p.i. Result(s): In cohort 1, 8/10 patients underwent apheresis and TIL imaging;one procedure was withdrawn due to COVID-19 restrictions and one due to unsuccessful T-cell isolation. Yield ranged 240-714 x10e6 CD8+ T-cells, purity 84%-97% and cell viability 92%-100%. Labeling efficacy of 100 x10e6 cells for re-injection ranged 42%-64% and injected activity 22,4-36,7 MBq In-111.TIL imaging was completed by 4/5 patients in cohort 2, one subject discontinued neo-adjuvant treatment due to post-obstruction pneumonia.Tumor-to-bloodpool were determined to quantify specific TIL accumulation in the tumor. Our results favor quantification of T-cells on PET over SPECT given its higher sensitivity and spatial resolution. Correlation of imaging findings with density of CD8+ T-cells in the resected tumor is currently ongoing. Conclusion(s): We implemented two methods for tracking CD8+ T-cells in earlystage NSCLC patients after neo-adjuvant durvalumab treatment. Although ex vivo cell labeling perhaps more specifically targets migrating TILs into the tumor, 89Zr-Df-crefmirlimab has the potential to also target residing cells. Quantitative correlation with presence of TILs in the resected tumor will help to determine the role of these imaging tools in the development of immune-oncology drugs.

In vitro antiviral effect of silver nanoparticles against SARS-CoV-2

Introduction: COVID-19 is an acute respiratory tract disease caused by the emerging coronavirus SARS-CoV-2. Although several options for che-moprophylaxis are under development, effective treatment for COVID-19 is not yet available. Objective(s): To investigate the antiviral properties of synthesized silver na-noparticles (AgNPs) against SARS-CoV-2 using in vitro models. Material(s) and Method(s): This work synthesized AgNPs using an electrochemical method and characterized them using physico-chemical techniques (ICP-OES, ultraviolet-visible spectroscopy, and transmission electron microscopy). AgNPs with diameter sizes ranging between 2.6 to 30 nm and an average size of 6.2 nm were obtained by the electrochemical method. The cytotoxic effect and the antiviral activity of prepared AgNPs against SARS-CoV-2 were evaluated in vitro using Vero E6 cells. Cell viability was evaluated by MTT assay in the presence of serial dilutions of AgNPs. The antiviral effect of AgNPs was evaluated before and after the infection of Vero E6 cells by plaque assay. Result(s): Cytotoxic effect was observed at concentrations above 0.07 ppm. AgNPs exhibit a significant reduction of SARS-CoV-2 viral titer after a pre-post treatment strategy with inhibition of 96.5%, 64.13%, and 74.72% at 0.03, 0.017, and 0.008 ppm, respectively. Conclusion(s): Our results suggest that AgNPs could reduce SARS-CoV-2 replication with a low cytotoxic effect. Still, additional in vitro and in vivo studies are required to define its potential therapeutic application in humans. Copyright © 2023 Universidad de Antioquia.

P06-21 MEHP treatment induces a decrease in SIRT1 expression, mitochondrial dysfunction, and transcriptomic alterations in the human trophoblastic cell line HTR-8/svneo

Background: Mono(ethylhexyl) phthalate (MEHP) is the main metabolite of Di(2-ethylhexyl) phthalate (DEHP), a chemical worldwide used as a plastic softener to increase the malleability, flexibility, and durability of several types of plastic, including those employed in bottled water, medical devices, and food wrapping, among others. Importantly, the consumption of these products has dramatically increased during the COVID-19 pandemic. MEHP has been classified as an endocrine disruptor chemical (EDC) and its involuntary intake has been associated with pregnancy complications such as preeclampsia and miscarriages. The placenta is a transitory organ that provides sustainability to the fetus, as well as the transportation of nutrients, hormones, and oxygen. Recent studies have proposed that MEHP may impair placental development and functionality. Nevertheless, little is known about its molecular mechanisms and effects on the placenta. Recent data has suggested that Sirtuin 1 (SIRT1) might be a molecular target. The aim of this study was to analyze the toxic and transcriptomic effects of MEHP in the human trophoblastic cell line HTR-8/svneo, focusing on the SIRT1-related pathways. Methods and Results: The HTR-8/svneo cell line was used as an extravillous trophoblast model to investigate MEHP effects. MEHP concentrations employed in this study were 0.5, 5, 50, 100, and 200 microM. Cell viability was evaluated by two methods: fixable viability staining, using eFluor 780, and MTT assay. Only the MTT assay suggested a significant decrease in cell viability at 48 hours with MEHP treatments of 5, 50, 100, and 200 microM. Mitochondrial biogenesis was analyzed by qPCR amplifying a region of the MT-ND1 mitochondrial gene. GAPDH promoter was used as a reference control. The results suggested a decrease in mitochondrial DNA at 48 hours. The transcriptomic analysis was performed in an Illumina Next-seq 500 with a coverage of 10 million reads. Doses of 5 and 200 microM of MEHP at 48 hours were analyzed. The results show that 41 and 341 genes were differentially expressed, respectively. These genes are involved in trophoblast function and pathophysiology and, according to previous reports, a portion of them are regulated by SIRT1. Finally, the effect of MEHP on SIRT1 was explored at both protein and mRNA levels by western blot and RT-qPCR, respectively. The results for mRNA levels exhibited a significant decline at 24 hours for all treatments, while protein levels were significantly reduced by 200 microM MEHP treatment at 48 hours. Conclusion(s): The present study demonstrates that MEHP treatments promote mitochondrial dysfunction in HTR-8/svneo cells. Moreover, the transcriptome analysis showed that MEHP modifies important signals for placental function and pathophysiology. The decline in SIRT1 levels correlates with the mitochondrial alterations as well as a portion of the transcriptomic changes, suggesting that SIRT1 may have an important role in MEHP effects in trophoblastic cells. Copyright © 2022 Elsevier B.V.

Evaluation of the ChromaCode HDPCRTM RV6 RUO Assay

Background. In December of 2019, an outbreak of pneumonia was reported to theWorld Health Organization. The etiology of this outbreak was identified as novel coronavirus (SARS-CoV-2), which has resulted in a global pandemic. The persistence of circulating SARS-CoV-2 leads to the need for a test that can detect and differentiate SARS-CoV-2 from common causes of respiratory illnesses such as influenza and respiratory syncytial virus (RSV). ChromaCode's HDPCRTMRV6 RUO Assay is a real time PCR test that provides qualitative detection of SARS-CoV-2, Influenza A, Influenza B, and RSV A/B from upper respiratory specimens. An evaluation in an infectious disease molecular laboratory was performed and presented herein. Methods. A combination of residual and contrived samples were used to evaluate the ChromaCode HDPCRTMRV6 RUO assay. Residual upper respiratory samples tested with the standard of care were enrolled in this study to evaluate RV6 RUO. A total of 201 samples originally tested using Diasorin Simplexa tests were enrolled: 17 assisted mid-turbinate, 15 contrived Flu B samples, and 179 nasopharyngeal swabs. The contrived samples were made utilizing dilutions of Zeptometrix Influenza B material (PN 0810253CF LN 325137) in negative NPS matrix to achieve final concentrations of 0.02-0.04 TCID50/mL. Results. The ChromaCode RV6 RUO Assay had comparable results to the standard of care test. The assays had a 99.0% concordance for the Influenza A target, 100% concordance for the Influenza B target, 97.5% concordance for the RSV A/B target, and 99.5% concordance for the SARS-CoV-2 target. Conclusion. There are several tools available for distinguishing between common respiratory illnesses, which now include SARS-CoV-2. This study demonstrated the viability of the HDPCR RV6 RUO Assay to be a solution for distinguishing between common respiratory illnesses. RV6 RUO showed a high degree of concordance with the standard of care test and highlighted the flexibility of the RV6 RUO assay to be used with multiple upper respiratory sample types. The HDPCR RV6 RUO Assay and the accompanying ChromaCode Cloud software is a user-friendly solution with minimal setup time and rapid data analysis.

25 years of commercializing nanomedicine: from tissue engineering to fighting COVID

Nanomedicine has already revolutionized medicine through the development of materials that can prolong circulation in the body, avoid immune system clearance, penetrate cells and bacteria, invade tumors, promote tissue growth, inhibit infection, and so much more. New fields have emerged such as 4D printing which can enhance the performance of nanomaterials by 3D printing them into desirable shapes, implant them, and control their shape through external stimuli (such as near infrared excitation, temperature control, and others). This presentation will provide an overview of 25 years of commercializing University based research into real products helping human health. It will cover the promises and pitfalls of commercializing University based research and even discuss if this is the right model to advance science and research into the medical industry. It will also highlight new areas emerging for commcerialization such as the use of 4D printing in medicine for straightening the spine for scoliosis patients, closing of sphincters that weaken as one ages (for example, to decrease acid reflux from the stomach to the esophagus), promote intervertebral tissue growth by increasing pressure on such tissue during regeneration, deliver stem cells on the same materials in which they are cultured to enhance stem cell viability, and more. It will cover additional new areas like picomedicine, implantable sensors and more. It will cover in vitro and in vivo assessment of such materials and discuss what is needed to experience full application of nanomedicine throughout all of medicine.