In Vivo mRNA CAR T Cell Engineering via Targeted Ionizable Lipid Nanoparticles with Extrahepatic Tropism.

With six therapies approved by the Food and Drug Association, chimeric antigen receptor (CAR) T cells have reshaped cancer immunotherapy. However, these therapies rely on ex vivo viral transduction to induce permanent CAR expression in T cells, which contributes to high production costs and long-term side effects. Thus, this work aims to develop an in vivo CAR T cell engineering platform to streamline production while using mRNA to induce transient, tunable CAR expression. Specifically, an ionizable lipid nanoparticle (LNP) is utilized as these platforms have demonstrated clinical success in nucleic acid delivery. Though LNPs often accumulate in the liver, the LNP platform used here achieves extrahepatic transfection with enhanced delivery to the spleen, and it is further modified via antibody conjugation (Ab-LNPs) to target pan-T cell markers. The in vivo evaluation of these Ab-LNPs confirms that targeting is necessary for potent T cell transfection. When using these Ab-LNPs for the delivery of CAR mRNA, antibody and dose-dependent CAR expression and cytokine release are observed along with B cell depletion of up to 90%. In all, this work conjugates antibodies to LNPs with extrahepatic tropism, evaluates pan-T cell markers, and develops Ab-LNPs capable of generating functional CAR T cells in vivo.

Women are Underrepresented in Non-small Cell Lung Cancer Clinical Trials: A Systematic Review.

PURPOSE: To perform a systematic review of clinical trials examining non-small cell lung cancer (NSCLC) to better understand the equity afforded to women in the study of lung cancer. METHODS: An electronic search was conducted for all NSCLC clinical trials published between 2010 and 2020 with included words "carcinoma, non-small cell, lung" and "non-small cell lung cancer." Studies from PubMed, Cochrane, and SCOPUS were included and were uploaded into Covidence to assist with systematic review. All articles were screened by two separate individuals and reviewed for location, study type, cancer stage, field of study of the research team, and percentage of females included. Student's t-test was used to compare the means of males and females. RESULTS: Across the 269 studies that met inclusion criteria, fewer females than males were enrolled (38.7% vs. 61.1%; p < 0.0001). Compared with studies from 2010 to 2015, those from 2016 to 2020 had greater representation of females (36.7% vs. 41.4%, p = 0.0091, respectively). Both nonsurgical and surgical studies enrolled fewer female than male patients (38.1% vs. 61.7%, p < 0.0001; 43.1% vs. 57.2%, p = 0.0002, respectively). Clinical trials from the USA had the least difference between sexes with an average of 46.7% females enrolled. Less females compared with males were enrolled in early-stage NSCLC (37.6% female vs. 62.6% male, p < 0.0001) and late-stage NSCLC trials (37.6% female vs. 62.0% male, p < 0.0001). CONCLUSIONS: Despite recent improvement, there continues to be significant underrepresentation of females compared with males in NSCLC clinical trials.

Conjugated Coordination Nanosheets with Molecular Rotors for Pseudocapacitors: Nanoarchitectonics and Enhanced Performance.

High-level pseudocapacitive materials require incorporations of significant redox regions into conductive and penetrable skeletons to enable the creation of devices capable of delivering high power for extended periods. Coordination nanosheets (CNs) are appealing materials for their high natural electrical conductivities, huge explicit surface regions, and semi-one-layered adjusted pore clusters. Thus, rational design of ligands and topological networks with desired electronic structure is required for the advancement in this field. Herein, we report three novel conjugated CNs (RV-10-M, M = Zn, Ni, and Co), by utilizing the full conjugation of the terpyridine-attached flexible tetraphenylethylene (TPE) units as the molecular rotors at the center. We prepare binder-free transparent nanosheets supported on Ni-foam with outstanding pseudocapacitive properties via a hydrothermal route followed by facile exfoliation. Among three CNs, the high surface area of RV-10-Co facilitates fast transport of ions and electrons and could achieve a high specific capacity of 670.8 C/g (1677 F/g) at 1 A/g current density. Besides, the corresponding flexible RV-10-Co possesses a maximum energy density of 37.26 Wh kg-1 at a power density of 171 W kg-1 and 70% capacitance retention even after 1000 cycles.

Orthogonal Design of Experiments for Optimization of Lipid Nanoparticles for mRNA Engineering of CAR T Cells.

Viral engineered chimeric antigen receptor (CAR) T cell therapies are potent, targeted cancer immunotherapies, but their permanent CAR expression can lead to severe adverse effects. Nonviral messenger RNA (mRNA) CAR T cells are being explored to overcome these drawbacks, but electroporation, the most common T cell transfection method, is limited by cytotoxicity. As a potentially safer nonviral delivery strategy, here, sequential libraries of ionizable lipid nanoparticle (LNP) formulations with varied excipient compositions were screened in comparison to a standard formulation for improved mRNA delivery to T cells with low cytotoxicity, revealing B10 as the top formulation with a 3-fold increase in mRNA delivery. When compared to electroporation in primary human T cells, B10 LNPs induced comparable CAR expression with reduced cytotoxicity while demonstrating potent cancer cell killing. These results demonstrate the impact of excipient optimization on LNP performance and support B10 LNPs as a potent mRNA delivery platform for T cell engineering.

Pericardioesophageal fistulae after left atrial ablation: a case series.

Pericardioesophageal fistula is an uncommon, yet serious complication that can occur after left atrial ablation for cardiac arrhythmias. Timing of this complication is variable; however, it has been reported to occur from a week to over a month post-ablation. The incidence of this complication after ablation is <0.05%; however with increasing rates of left atrial ablations, early recognition is imperative. Nonspecific symptoms, including chest pain, dysphagia, and fever, can indicate the presence of a fistula within the first month after ablation. Early drainage with subsequent definitive treatment is key to limiting morbidity. Here we report four cases of pericardioesophageal fistula all occurring ~1 month post-ablation, with two patients surviving after prompt diagnosis and surgical treatment. Successful treatment in these two cases was achieved with fistula takedown and intercostal muscle flap interposition and esophageal stenting.

Minimally Invasive Pneumonectomy vs Open Pneumonectomy: Outcomes and Predictors of Conversion.

BACKGROUND: In the modern era, it is unknown if minimally invasive pneumonectomy for non-small cell lung cancer (NSCLC) provides a survival advantage over open pneumonectomy. METHODS: Patients who underwent pneumonectomy for NSCLC between 2015 and 2020 were queried from the National Cancer Database. Surgical approach was categorized as robot-assisted (RATS), video-assisted thoracoscopic (VATS), or open pneumonectomy on an intention-to-treat basis. Propensity score matching was performed to balance patient cohorts. Univariate and multivariate regression analyses were used to examine the association between surgical approach and 30 and 90-day mortality, and a Cox proportional hazards model was used to assess overall survival. RESULTS: Overall, 3784 patients were identified, including 73% open (n=2776), 19% VATS (n=725), and 8% RATS (n=283). The overall conversion rate from minimally invasive to open was 29.5% (n=298). After propensity matching 212 patients per cohort, there were no differences between open, VATS, and RATS 30-day (9.4% vs 8.5% vs 7.5%, p=0.807, respectively) or 90-day mortality (14.2% vs 12.3% vs 10.4%, p=0.516, respectively). Median overall survival was similar among open (48 months, 95% CI 35.6-64.1), VATS (51.0 months, 95% CI 34.9-72.3), and RATS approaches (50 months, 95% CI 42.6-NA, p=0.560). On multivariate analysis of the matched cohort, there was no association between approach and overall survival. RATS (OR 0.67, 95% CI 0.47-0.94, p=0.020) and neoadjuvant chemotherapy (OR 0.52, 95% CI 0.27-0.98, p=0.045) were found to be protective against conversion to open. CONCLUSIONS: Minimally-invasive pneumonectomy can be performed with short-term and long-term survival that are equivalent to open pneumonectomy.

Self-Assembled Conjugated Coordination Polymer Nanorings: Role of Morphology and Redox Sites for the Alkaline Electrocatalytic Oxygen Evolution Reaction.

Electrocatalytic water splitting provides a sustainable method for storing intermittent energies, such as solar energy and wind, in the form of hydrogen fuel. However, the oxygen evolution reaction (OER), constituting the other half-cell reaction, is often considered the bottleneck in overall water splitting due to its slow kinetics. Therefore, it is crucial to develop efficient, cost-effective, and robust OER catalysts to enhance the water-splitting process. Transition-metal-based coordination polymers (CPs) serve as promising electrocatalysts due to their diverse chemical architectures paired with redox-active metal centers. Despite their potential, the rational use of CPs has faced obstacles including a lack of insights into their catalytic mechanisms, low conductivity, and morphology issues. Consequently, achieving success in this field requires the rational design of ligands and topological networks with the desired electronic structure. This study delves into the design and synthesis of three novel conjugated coordination polymers (CCPs) by leveraging the full conjugation of terpyridine-attached flexible tetraphenylethylene units as electron-rich linkers with various redox-active metal centers [Co(II), Ni(II), and Zn(II)]. The self-assembly process is tuned for each CCP, resulting in two distinct morphologies: nanosheets and nanorings. The electrocatalytic OER performance efficiency is then correlated with factors such as the nanostructure morphology and redox-active metal centers in alkaline electrolytes. Notably, among the three morphologies studied, nanorings for each CCP exhibit a superior OER activity. Co(II)-integrated CCPs demonstrate a higher activity between the redox-active metal centers. Specifically, the Co(II) nanoring morphology displays exceptional catalytic activity for OER, with a lower overpotential of 347 mV at a current density of 10 mA cm-2 and small Tafel slopes of 115 mV dec-1. The long-term durability is demonstrated for at least 24 h at 1.57 V vs RHE during water splitting. This is presumably the first proof that links the importance of nanostructure morphologies to redox-active metal centers in improving the OER activity, and it may have implications for other transdisciplinary energy-related applications.

Bile acid-containing lipid nanoparticles enhance extrahepatic mRNA delivery.

Lipid nanoparticles (LNPs) have emerged as a viable, clinically-validated platform for the delivery of mRNA therapeutics. LNPs have been utilized as mRNA delivery systems for applications including vaccines, gene therapy, and cancer immunotherapy. However, LNPs, which are typically composed of ionizable lipids, cholesterol, helper lipids, and lipid-anchored polyethylene glycol, often traffic to the liver which limits the therapeutic potential of the platform. Several approaches have been proposed to resolve this tropism such as post-synthesis surface modification or the addition of synthetic cationic lipids. Methods: Here, we present a strategy for achieving extrahepatic delivery of mRNA involving the incorporation of bile acids, a naturally-occurring class of cholesterol analogs, during LNP synthesis. We synthesized a series of bile acid-containing C14-4 LNPs by replacing cholesterol with bile acids (cholic acid, chenodeoxycholic acid, deoxycholic acid, or lithocholic acid) at various ratios. Results: Bile acid-containing LNPs (BA-LNPs) were able to reduce delivery to liver cells in vitro and improve delivery in a variety of other cell types, including T cells, B cells, and epithelial cells. Our subsequent in vivo screening of selected LNP candidates injected intraperitoneally or intravenously identified a highly spleen tropic BA-LNP: CA-100, a four-component LNP containing cholic acid and no cholesterol. These screens also identified BA-LNP candidates demonstrating promise for other mRNA therapeutic applications such as for gastrointestinal or immune cell delivery. We further found that the substitution of cholic acid for cholesterol in an LNP formulation utilizing a different ionizable lipid, C12-200, also shifted mRNA delivery from the liver to the spleen, suggesting that this cholic acid replacement strategy may be generalizable. Conclusion: These results demonstrate the potential of a four-component BA-LNP formulation, CA-100, for extrahepatic mRNA delivery that could potentially be utilized for a range of therapeutic and vaccine applications.

Multivariate modeling and process optimization of Hg(II) remediation using solvothermal synthesized 2D MX/Fe3O4 by response surface methodology: characteristics and mechanism study.

Two-dimensional MXene with layered structure has recently emerged as a nanomaterial with fascinating characteristics and applicability. Herein, we prepared the newly modified magnetic MXene (MX/Fe3O4) nanocomposite using solvothermal approach and investigated its adsorption behavior to study the removal efficiency of Hg(II) ions from aqueous solution. The effect of adsorption parameters such as adsorbent dose, time, concentration, and pH were optimized using response surface methodology (RSM). The experimental data fitted well with quadratic model to predict the optimum conditions for maximum Hg(II) ion removal efficiency which were found to be at adsorbent dose 0.871 g/L, time 103.6 min, concentration 40.17 mg/L, and 6.5 pH respectively. To determine the adequacy of the developed model, a statistical analysis of variance (ANOVA) was used, which demonstrated high agreement between the experimental data and the suggested model. According to isotherm result, the experimental data were following the best agreement with the Redlich-Peterson isotherm model. The results of the experiments revealed that the maximum Langmuir adsorption capacity of 699.3 mg/g was obtained at optimum conditions, which was closed to the experimental adsorption capacity of 703.57 mg/g. The adsorption phenomena was well represented by the pseudo-second-order model (R2 = 0.9983). On the whole, it was clear that MX/Fe3O4 has lot of potential as a Hg(II) ion impurity removal agent in aqueous solutions.

Green-synthesized, pH-stable and biocompatible carbon nanosensor for Fe3+: An experimental and computational study.

Brightly fluorescent Carbon Dots (CDs) were synthesized by green hydrothermal method using commonly available biomass (Aloe vera) as carbon precursor. Their physiochemical and optical characterization was done by standard microscopic and spectroscopic techniques. Photophysical features of their aqueous dispersion were investigated in detail. The influence of wide pH range (2-12), high ionic load (2M) and temperature on their photoluminescence behavior was investigated. Their in-vitro cytotoxicity examination was conducted on Human Cervical Cancer Cells (HeLa) using MTT assay. Testing of their ion-recognition property for common metal ions was done in aqueous medium. These CDs exhibited preferential interaction with Fe3+ over other tested metal ions, without any functionalization. Interaction between CDs and Fe3+ was analyzed in the light of Density Functional Theory (DFT). The work demonstrates that these CDs are acting as nanoprobe for Fe3+ and sensing it at ultra-trace level (5 nM).

Hydroxycholesterol substitution in ionizable lipid nanoparticles for mRNA delivery to T cells.

Delivery of nucleic acids, such as mRNA, to immune cells has become a major focus in the past decade with ionizable lipid nanoparticles (LNPs) emerging as a clinically-validated delivery platform. LNPs-typically composed of ionizable lipids, cholesterol, phospholipids, and polyethylene glycol lipids -have been designed and optimized for a variety of applications including cancer therapies, vaccines, and gene editing. However, LNPs have only recently been investigated for delivery to T cells, which has various therapeutic applications including the engineering of T cell immunotherapies. While several LNP formulations have been evaluated for mRNA delivery, recent work has demonstrated that the utilization of cholesterol analogs may enhance mRNA delivery. Other studies have shown that cholesterols modified with hydroxyl groups can alter endocytic recycling mechanisms. Here, we engineered a library of LNPs incorporating hydroxycholesterols to evaluate their impact on mRNA delivery to T cells by leveraging endosomal trafficking mechanisms. Substitution of 25% and 50% 7α-hydroxycholesterol for cholesterol in LNPs enhanced mRNA delivery to primary human T cells ex vivo by 1.8-fold and 2.0-fold, respectively. Investigation of endosomal trafficking revealed that these modifications also increase late endosome production and reduce the presence of recycling endosomes. These results suggest that hydroxyl modification of cholesterol molecules incorporated into LNP formulations provides a mechanism for improving delivery of nucleic acid cargo to T cells for a range of immunotherapy applications.

Amniotic fluid stabilized lipid nanoparticles for in utero intra-amniotic mRNA delivery.

Congenital disorders resulting in pathological protein deficiencies are most often treated postnatally with protein or enzyme replacement therapies. However, treatment of these disorders in utero before irreversible disease onset could significantly minimize disease burden, morbidity, and mortality. One possible strategy for the prenatal treatment of congenital disorders is in utero delivery of messenger RNA (mRNA). mRNA is a nucleic acid therapeutic that has previously been investigated as a platform for protein replacement therapies and gene editing technologies. While viral vectors have been explored to induce intracellular expression of mRNA, they are limited in their clinical application due to risks associated with immunogenicity and genomic integration. As an alternative to viral vectors, safe and efficient in utero mRNA delivery can be achieved using ionizable lipid nanoparticles (LNPs). While LNPs have demonstrated potent in vivo mRNA delivery to the liver following intravenous administration, intra-amniotic delivery has the potential to deliver mRNA to cells and tissues beyond those in the liver, such as in the skin, lung, and digestive tract. However, LNP stability in fetal amniotic fluid and how this stability affects mRNA delivery has not been previously investigated. Here, we engineered a library of LNPs using orthogonal design of experiments (DOE) to evaluate how LNP structure affects their stability in amniotic fluid ex utero and whether a lead candidate identified from these stability measurements enables intra-amniotic mRNA delivery in utero. We used a combination of techniques including dynamic light scattering (DLS), transmission electron microscopy (TEM), and chromatography followed by protein content quantification to screen LNP stability in amniotic fluids. These results identified multiple lead LNP formulations that are highly stable in amniotic fluids ranging from small animals to humans, including mouse, sheep, pig, and human amniotic fluid samples. We then demonstrate that stable LNPs from the ex utero screen in mouse amniotic fluid enabled potent mRNA delivery in primary fetal lung fibroblasts and in utero following intra-amniotic injection in a murine model. This exploration of ex utero stability in amniotic fluids demonstrates a means by which to identify novel LNP formulations for prenatal treatment of congenital disorders via in utero mRNA delivery.

Postoperative Air Leaks After Lung Surgery: Predictors, Intraoperative Techniques, and Postoperative Management.

Postoperative air leak is one of the most common complications after pulmonary resection and contributes to postoperative pain, complications, and increased hospital length of stay. Several risk factors, including both patient and surgical characteristics, increase the frequency of air leaks. Appropriate intraoperative tissue handling is the most important surgical technique to reduce air leaks. Digital drainage systems have improved the management of postoperative air leak via objective data, portability, and ease of use in the outpatient setting. Several treatment strategies have been used to address prolonged air leak, including pleurodesis, blood patch, placement of endobronchial valves, and reoperative surgery.

Synthesis of highly fluorescent and water soluble graphene quantum dots for detection of heavy metal ions in aqueous media.

Fluorescent graphene quantum dots (GQDs) are nanomaterials which possess unique properties that show great potential in different applications. In this work, GQDs were synthesized using graphene oxide (GO) as precursor via thermal treatment at high temperature. The obtained GQDs were highly fluorescent and were suitable for the determination of heavy metal ions. X-ray diffraction, FTIR spectroscopy, and UV visible spectroscopy confirm the formation of GQDs. TEM images show that formed GQDs have size ranging from 2 to 10 nm. Emission profile of aqueous GQDs was taken by exciting GQDs at different wavelength. The intensity of GQDs remains the same for 4-5 months. Furthermore, as prepared, GQDs were used for selective recognition of Fe3+, Pb+2, and Cr3+ from the bunch of different metal ions in aqueous media. Lower limit of detection obtained for Fe3+, Cr3+ and Pb2+ using GQDs were 50, 100 and 100 nM, respectively, which indicates that the GQDs can be utilized as a promising material for sensing of the heavy metal ions. Graphical abstract.

The coexistence of anal incontinence in women with urinary incontinence.

PURPOSE: To investigate the coexistentence of anal incontinence (AI) in patients with urinary incontinence (UI). MATERIALS AND METHODS: UDI 6 questionnaire was applied to consecutive 800 patients for the determination of symptomatic UI. Cases were also questioned for AI. The patients with the complaint of UI were evaluated for the coexistence of AI. Each patient with AI, had completed the Cleveland Clinic Florida Fecal Incontinence questionnaire for the determination of the severity of AI. Chi-square and Mann-Whitney U test were used for the evaluation of qualitative data. Multivariable analysis using logistic regression was done to test the overall significance of all variables that were significantly associated with UI using univariate analysis. RESULTS: There were 233 (29.1%) cases of UI and 105 (13.1%) cases of AI. 60 (7.5%) of these patients were suffering from both UI and AI. 60 (57.1%) of 105 patients with AI also had UI. In patients with UI, the coexistence of AI was found in 25.7%. There was a positive correlation between UI and AI (R = 0.240, Chi-square P < 0.001). In logistic regression model, AI [(OR) 3.5, 95% confidence interval 2.26, 5.49], parity and episiotomy were found to be associated with UI. Parity and vaginal delivery by episiotomy were risk factors for the occurrence of AI. CONCLUSION: AI is positively correlated with UI patients. Symptoms of AI must be questioned in all gynecology patients. Unnecessary episiotomy should be avoided.

Serum adiponectin levels in high school girls with polycystic ovary syndrome and hyperandrogenism.

OBJECTIVE: To compare the serum adiponectin levels together with metabolic and hormonal parameters among teenage girls at the early onset of polycystic ovary syndrome (PCOS) and hyperandrogenism with controls. DESIGN: Prospective study. SETTINGS: Education and research hospital, outpatient gynecological endocrinology clinic. PARTICIPANTS: Four hundred seventy-nine teenage girls from a school of nursing were interviewed for the signs and symptoms of PCOS. Among them, 42 cases who had a definitive diagnosis of PCOS with hyperandrogenism based on Rotterdam diagnostic criteria were recruited for the study and other causes of hyperandrogenemia had been excluded. The controls were recruited from regularly cycling healthy teenage girls from the same high school of nursing; none of those who agreed to join the study met any of the diagnostic criteria for PCOS (n = 44). INTERVENTIONS: Cases were selected as group I: PCOS with body mass index (BMI) < 25 kg/m² (n = 20), group II: PCOS with BMI > 25 kg/m² (n = 22), group III: Controls with BMI < 25 kg/m² (n = 21) and group IV: Controls with BMI > 25 kg/m² (n = 23). Serum adiponectin, metabolic and hormonal parameters were compared in PCOS patients with BMI matched controls. MAIN OUTCOME MEASURES: Difference of serum adiponectin levels, metabolic and hormonal parameters between teenage girls with PCOS and controls. RESULTS: Serum adiponectin levels were not significantly different in group I and group II. Serum adiponectin levels were significantly decreased in group I and group II compared with both control groups (III and IV). CONCLUSION: Serum adiponectin levels were lower in teenage girls with PCOS and this reduction was independent from BMI.

Can increased nuchal translucency in the first trimester of pregnancy predict gestational diabetes mellitus.

The current study was designed to evaluate whether increased nuchal translucency can predict gestational diabetes mellitus. This was a prospective observational study. Among the pregnant women at 11-14 weeks of pregnancy who came to our prenatal unit for a first trimester screening test, 389 pregnant women whose nuchal translucency above 95th centile were selected as the study group and 386 age-matched pregnant women whose nuchal translucency were within the normal range were enrolled as a control group. First, subjects underwent a 50 g glucose screening test; if it was positive then a 100 g oral glucose tolerance test was performed. The main outcome measures were the prevalence of gestational diabetes mellitus and impaired glucose tolerance and the number of macrosomic infants. Impaired glucose tolerance was more common in pregnant women whose nuchal translucency was above the 95th centile (p = 0.048). In addition, macrosomic infants were also more common in pregnant women with a fetal nuchal translucency above the 95th centile (p = 0.045). Macrosomia was more common in the study group with gestational diabetes mellitus (p = 0.046). In conclusion, increased nuchal translucency seems to be predictive for impaired glucose tolerance and macrosomia, which are associated with gestational diabetes mellitus.