The BTLA-HVEM axis restricts CAR T cell efficacy in cancer.

The efficacy of T cell-based immunotherapies is limited by immunosuppressive pressures in the tumor microenvironment. Here we show a predominant role for the interaction between BTLA on effector T cells and HVEM (TNFRSF14) on immunosuppressive tumor microenvironment cells, namely regulatory T cells. High BTLA expression in chimeric antigen receptor (CAR) T cells correlated with poor clinical response to treatment. Therefore, we deleted BTLA in CAR T cells and show improved tumor control and persistence in models of lymphoma and solid malignancies. Mechanistically, BTLA inhibits CAR T cells via recruitment of tyrosine phosphatases SHP-1 and SHP-2, upon trans engagement with HVEM. BTLA knockout thus promotes CAR signaling and subsequently enhances effector function. Overall, these data indicate that the BTLA-HVEM axis is a crucial immune checkpoint in CAR T cell immunotherapy and warrants the use of strategies to overcome this barrier.

Streamlined measurement of chimeric antigen receptor T-cell concentration, size, viability and two-color phenotyping during manufacturing.

BACKGROUND AIMS: The successful development of CD19-targeted chimeric antigen receptor (CAR) T-cell therapies has led to an exponential increase in the number of patients recieving treatment and the advancement of novel CAR T products. Therefore, there is a strong need to develop streamlined platforms that allow rapid, cost-effective, and accurate measurement of the key characteristics of CAR T cells during manufacturing (i.e., cell number, cell size, viability, and basic phenotype). METHODS: In this study, we compared the novel benchtop cell analyzer Moxi GO II (ORFLO Technologies), which enables simultaneous evaluation of all the aforementioned parameters, with current gold standards in the field: the Multisizer Coulter Counter (cell counter) and the BD LSRFortessa (flow cytometer). RESULTS: Our results demonstrated that the Moxi GO II can accurately measure cell number and cell size (i.e., cell volume) while simultaneously assessing simple two-color flow cytometry parameters, such as CAR T-cell viability and CD4 or CAR expression. CONCLUSIONS: These measurements are comparable with those of gold standard instruments, demonstrating that the Moxi GO II is a promising platform for quickly monitoring CAR T-cell growth and phenotype in research-grade and clinical samples.

Safety and efficacy of a novel anti-CD19 chimeric antigen receptor T cell product targeting a membrane-proximal domain of CD19 with fast on- and off-rates against non-Hodgkin lymphoma: a first-in-human study.

BACKGROUND: Commercial anti-CD19 chimeric antigen receptor T-cell therapies (CART19) are efficacious against advanced B-cell non-Hodgkin lymphoma (NHL); however, most patients ultimately relapse. Several mechanisms contribute to this failure, including CD19-negative escape and CAR T dysfunction. All four commercial CART19 products utilize the FMC63 single-chain variable fragment (scFv) specific to a CD19 membrane-distal epitope and characterized by slow association (on) and dissociation (off) rates. We hypothesized that a novel anti-CD19 scFv that engages an alternative CD19 membrane-proximal epitope independent of FMC63 and that is characterized by faster on- and off-rates could mitigate CART19 failure and improve clinical efficacy. METHODS: We developed an autologous CART19 product with 4-1BB co-stimulation using a novel humanized chicken antibody (h1218). This antibody is specific to a membrane-proximal CD19 epitope and harbors faster on/off rates compared to FMC63. We tested h1218-CART19 in vitro and in vivo using FMC63-CART19-resistant models. We conducted a first-in-human multi-center phase I clinical trial to test AT101 (clinical-grade h1218-CART19) in patients with relapsed or refractory (r/r) NHL. RESULTS: Preclinically, h1218- but not FMC63-CART19 were able to effectively eradicate lymphomas expressing CD19 point mutations (L174V and R163L) or co-expressing FMC63-CAR19 as found in patients relapsing after FMC63-CART19. Furthermore, h1218-CART19 exhibited enhanced killing of B-cell malignancies in vitro and in vivo compared with FMC63-CART19. Mechanistically, we found that h1218-CART19 had reduced activation-induced cell death (AICD) and enhanced expansion compared to FMC63-CART19 owing to faster on- and off-rates. Based on these preclinical results, we performed a phase I dose-escalation trial, testing three dose levels (DL) of AT101 (the GMP version of h1218) using a 3 + 3 design. In 12 treated patients (7 DLBCL, 3 FL, 1 MCL, and 1 MZL), AT101 showed a promising safety profile with 8.3% grade 3 CRS (n = 1) and 8.3% grade 4 ICANS (n = 1). In the whole cohort, the overall response rate was 91.7%, with a complete response rate of 75.0%, which improved to 100% in DL-2 and -3. AT101 expansion correlates with CR and B-cell aplasia. CONCLUSIONS: We developed a novel, safe, and potent CART19 product that recognizes a membrane-proximal domain of CD19 with fast on- and off-rates and showed significant efficacy and promising safety in patients with relapsed B-cell NHL. TRIAL REGISTRATION: NCT05338931; Date: 2022-04-01.

A novel approach to measure carbonate alkalinity in aqueous solutions using a total organic carbon analyzer.

Carbonate alkalinity is crucial in regulating the pH and buffering capacity of natural water systems. Thus, its accurate measurement is essential to understand various water environments that affect water quality and ecosystem health. However, conventional potentiometric titration has some limitations. It results in inaccurate measurements of carbonate alkalinity when the alkalinity levels are low or when high dissolved organic matter or inorganic ion levels exist. Herein, we propose a novel approach to accurately measure carbonate alkalinity using a total organic carbon (TOC) analyzer. An extensive study comparing the accuracy and reliability of the conventional potentiometric titration method with those of the newly developed TOC method was conducted to develop and verify highly accurate measurements of carbonate alkalinity. The TOC method has several advantages over the conventional potentiometric titration methods, such as its ability to accurately measure carbonate alkalinity in the presence of high dissolved organic matter or inorganic ion levels and its ability to provide rapid and automated measurements with high reproducibility. Because, the limit of detection, limit of quantification, and the variation coefficient of the measurements was 0.016 mM (0.2 mgC/L), 0.050 mM (0.6 mgC/L), and 3.68 % respectively. Thus, the development of a novel TOC method has significant environmental implications as it provides a reliable and accurate means to measure carbonate alkalinity in solutions containing various organic matter types.

Mitigation of benzoic acid-driven autotoxicity in waste nutrient solution using O3 and O3/H2O2 treatments: Seed germination and root growth of Lactuca sativa L.

Benzoic acid (BA), a secondary metabolite released through root exudates, is considered to be the most common inhibitor that leads to plant autotoxicity, even at low concentrations in closed hydroponic systems. In this study, to mitigate BA-driven autotoxicity, the effects of O3 and O3/H2O2 oxidation treatment (O3 concentration: 1, 2, 4, 8 mg L-1, H2O2 concentration: 4, 8 mg L-1) on waste nutrient solution (WNS) were investigated in terms of BA degradation, the rate of germination inhibition (GI), and the rate of root growth inhibition (RI). In the case of O3 treatment, the BA degradation rate improved up to 14.1% as the O3 concentration increased, while alleviation of GI was insignificant (94.6-100%), confirming that a single O3 treatment was unsuitable for mitigating autotoxicity. On the other hand, O3/H2O2 treatment increased BA degradation by up to 24.8%, thereby significantly reducing GI (up to 7.69%) and RI (up to 0.88%). Both the highest BA mineralization rate and phytotoxicity mitigation was observed at BA125 (4-4) (BA mineralization: 16.7%, GI: 12.82%, RI: 11.69%) and BA125 (1-8) (BA mineralization: 17.7%, GI: 7.69%, RI: 0.88%) at each H2O2 concentration. In addition, the operating costs were evaluated by a chemical and electricity cost analysis at the different treatments. As a result, the operating costs of BA125 (4-4) and BA125 (1-8) were calculated to be 0.40 and 0.42 $ L-1 mg-1 of mineralized BA, respectively. After consideration of the mineralization rate, autotoxicity mitigation, and operating cost, BA125 (1-8) was suggested for the optimal treatment condition and our findings would contribute to the alleviation of BA-driven autotoxicity.

Enhanced selectivity and recovery of phosphate and nitrate ions onto coffee ground waste biochars via co-precipitation of Mg/Al layered double hydroxides: A potential slow-release fertilizer.

In this study, the feasibility of Mg/Al layered double hydroxides (LDH) functionalized coffee ground waste biochars (LDHMgAl@CWGB) as a potential adsorbent to selectively recover phosphate (PO43-) and nitrate (NO3-) ions in aqueous phases and their consecutive uses as a slow-release fertilizer for stimulating the plant growth were identified. The higher adsorption capacity of PO43- and NO3- ions by LDHMgAl@CWGB (PO43- = 6.98 mgP/g, NO3- = 2.82 mgN/g) compared with pristine coffee ground waste biochars (CWGB; PO43- = 0.19 mgP/g, NO3- = 0.32 mgN/g) was mainly due to the incorporation of Mg/Al mixed oxides and Cl contents. Chemisorption and intra-particle mainly controlled the adsorptive recovery of PO43- and NO3- ions by CWGB and LDHMgAl@CWGB in aqueous phases and their adsorption toward CWGB and LDHMgAl@CWGB proceeded endothermically and spontaneously. The changes in the major adsorption mechanisms of PO43- and NO3- ions from ligand exchange (CWGB) to electrostatic surface complexation and anion-exchange (LDHMgAl@CWGB) supported the conclusion that the alternation of the surface features through Mg/Al LDH functionalization might improve selectivity and adsorption capacity of PO43- and NO3- ions onto CWGB under the co-existence of Cl-, SO42-, and HCO3- ions. Since PO43-- and NO3--loaded LDHMgAl@CWGB exhibited much higher seed germination, root and shoot growth rates of garden cress seeds (Lepidium sativum L) than other liquid and solid matrices, including 5 mgP/L PO43- and 5 mgN/L NO3-, 10 mgP/L PO43- and 10 mgN/L NO3-, and LDHMgAl@CWGB, it can be postulated that PO43-- and NO3--loaded LDHMgAl@CWGB could be practically applicable to the agricultural field as a slow-release fertilizer to facilitate the seed germination, root and shoot growth of the plants.

Apoptosis: a Janus bifrons in T-cell immunotherapy.

Immunotherapy has revolutionized the treatment of cancer. In particular, immune checkpoint blockade, bispecific antibodies, and adoptive T-cell transfer have yielded unprecedented clinical results in hematological malignancies and solid cancers. While T cell-based immunotherapies have multiple mechanisms of action, their ultimate goal is achieving apoptosis of cancer cells. Unsurprisingly, apoptosis evasion is a key feature of cancer biology. Therefore, enhancing cancer cells' sensitivity to apoptosis represents a key strategy to improve clinical outcomes in cancer immunotherapy. Indeed, cancer cells are characterized by several intrinsic mechanisms to resist apoptosis, in addition to features to promote apoptosis in T cells and evade therapy. However, apoptosis is double-faced: when it occurs in T cells, it represents a critical mechanism of failure for immunotherapies. This review will summarize the recent efforts to enhance T cell-based immunotherapies by increasing apoptosis susceptibility in cancer cells and discuss the role of apoptosis in modulating the survival of cytotoxic T lymphocytes in the tumor microenvironment and potential strategies to overcome this issue.

Functionalization of pine sawdust biochars with Mg/Al layered double hydroxides to enhance adsorption capacity of synthetic azo dyes: Adsorption mechanisms and reusability.

This study determined that the adsorption of azo dyes, Methyl Orange (MO) and Sunset Yellow FCF (SYF), using the pristine pine sawdust biochar (PSB) and post-modified PSB with Mg/Al layered double hydroxides (PSB-LDHMgAl) was examined to offer valuable information into the differences in their adsorption mechanisms. Although a lower specific surface area of PSB-LDHMgAl (147.2 m2 g-1) than PSB (495.7 m2 g-1), LDHMgAl were successfully functionalized on the PSB surface through co-precipitation, which was highly related to the improvements of adsorption capacity of PSB-LDHMgAl toward MO and SYF. The MO and SYF adsorption kinetics by PSB and PSB-LDHMgAl were confirmed to the pseudo-second-order and considered chemisorption. The adsorption capacity of MO and SYF adsorbed onto PSB-LDHMgAl (MO = 21.8 mg g-1, SYF = 23.6 mg g-1) were significantly higher than that of PSB (MO = 2.2 mg g-1, SYF = 1.6 mg g-1). The adsorption isotherms of MO and SYF by PSB were well fitted by Freundlich isotherm, whereas the MO and SYF via PSB-LDHMgAl were by Langmuir isotherm. Even after 3 adsorption-desorption cycles using desorbents, the PSB-LDHMgAl remained excellent reusability (reuse efficiency: >81.2%). These findings suggest that post-modification with LDHMgAl might accelerate the adsorption performance (i.e., electrostatic interaction) of azo dyes to PSB in water.

Modulation of the gut microbiota engages antigen cross-presentation to enhance antitumor effects of CAR T cell immunotherapy.

Several studies have shown the influence of commensal microbes on T cell function, specifically in the setting of checkpoint immunotherapy for cancer. In this study, we investigated how vancomycin-induced gut microbiota dysbiosis affects chimeric antigen receptor (CAR) T immunotherapy using multiple preclinical models as well as clinical correlates. In two murine tumor models, hematopoietic CD19+-A20 lymphoma and CD19+-B16 melanoma, mice receiving vancomycin in combination with CD19-directed CAR T cell (CART-19) therapy displayed increased tumor control and tumor-associated antigens (TAAs) cross-presentation compared with CART-19 alone. Fecal microbiota transplant from human healthy donors to pre-conditioned mice recapitulated the results obtained in naive gut microbiota mice. Last, B cell acute lymphoblastic leukemia patients treated with CART-19 and exposed to oral vancomycin showed higher CART-19 peak expansion compared with unexposed patients. These results substantiate the role of the gut microbiota on CAR T cell therapy and suggest that modulation of the gut microbiota using vancomycin may improve outcomes after CAR T cell therapy across tumor types.

An autopsy study of hollow fiber and multibore ultrafiltration membranes from a pilot-scale ultra high-recovery filtration system for surface water treatment.

The organic fouling characteristics of hollow fiber ultrafiltration (HFUF) and multibore ultrafiltration (MBUF) membranes from long-term ultrafiltration (UF) membrane systems were systemically investigated in this study. The objective was to obtain insights into the fouling behavior of dissolved organic matter (DOM) in a pilot-scale ultra-high-recovery membrane filtration system (p-UHMS) used for surface water treatment. The pilot system consisted of a series of two different UF membranes (1st stage: polyvinylidene fluoride (PVDF) HFUF and 2nd stage: polyethersulfone (PES) MBUF). It was designed to feed the HFUF concentrate to the MBUF membranes to achieve ≥99.5 % total water recovery for surface water treatment, as these advances might enhance the production efficiencies of drinking water. The experimental results confirmed that hydrophobic DOM controlled the formation of HFUF membrane organic fouling, whereas hydrophilic DOM, including polysaccharide-like and protein-like matter, promoted MBUF membrane fouling. These opposing trends were attributed to the hydrophilic characteristics of the MBUF membrane surfaces (contact angle: PVDF = 90-130° and PES ≤ 80°), which reduced the hydrophobic interactions between the UF membrane surfaces and foulants. The performance declines of the MBUF membrane due to fouling layer formation was considerably severer than those of the HFUF membrane, decreasing total permeate water in the p-UHMS. Moreover, the quantity of the desorbed MBUF membrane foulants via 0.1 N NaOH was roughly 7.2 times larger than that of the desorbed HFUF membrane foulants through 0.1 N NaOH, indicating that alkaline-based cleaning agent could much more efficiently recover the performance of the fouled MBUF membranes. Hence, adequate cleaning strategies using alkaline-based agent for the MBUF membrane appeared to be essential for preventing the performance deterioration of the p-UHMS.

Synthesis and applications of bismuth-impregnated biochars originated from spent coffee grounds for efficient adsorption of radioactive iodine: A mechanism study.

The adsorption of radioactive iodine, which is capable of presenting high mobility in aquatic ecosystems and generating undesirable health effects in humans (e.g., thyroid gland dysfunction), was comprehensively examined using pristine spent coffee ground biochar (SCGB) and bismuth-impregnated spent coffee ground biochar (Bi@SCGB) to provide valuable insights into the variations in the adsorption capacity and mechanisms after pretreatment with Bi(NO3)3. The greater adsorption of radioactive iodine toward Bi@SCGB (adsorption capacity (Qe) = 253.71 µg/g) compared to that for SCGB (Qe = 23.32 µg/g) and its reduced adsorption capability at higher pH values provide evidence that the adsorption of radioactive iodine with SCGB and Bi@SCGB is strongly influenced by the presence of bismuth materials and the electrostatic repulsion between their negatively charged surfaces and negatively charged radioactive iodine (IO3-). The calculated R2 values for the adsorption kinetics and isotherms support that chemisorption plays a crucial role in the adsorption of radioactive iodine by SCGB and Bi@SCGB in aqueous phases. The adsorption of radioactive iodine onto SCGB was linearly correlated with the contact time (h1/2), and the diffusion of intra-particle predominantly determined the adsorption rate of radioactive iodine onto Bi@SCGB (Cstage II (129.20) > Cstage I (42.33)). Thermodynamic studies revealed that the adsorption of radioactive iodine toward SCGB (ΔG° = -8.47 to -7.83 kJ/mol; ΔH° = -13.93 kJ/mol) occurred exothermically and that for Bi@SCGB (ΔG° = -15.90 to -13.89 kJ/mol; ΔH° = 5.88 kJ/mol) proceeded endothermically and spontaneously. The X-ray photoelectron spectroscopy (XPS) analysis of SCGB and Bi@SCGB before and after the adsorption of radioactive iodine suggest the conclusion that the change in the primary adsorption mechanism from electrostatic attraction to surface precipitation upon the impregnation of bismuth materials on the surfaces of spent coffee ground biochars is beneficial for the adsorption of radioactive iodine in aqueous phases.

Modulation of BCL-2 in Both T Cells and Tumor Cells to Enhance Chimeric Antigen Receptor T-cell Immunotherapy against Cancer.

Chimeric antigen receptor T-cell (CART) immunotherapy led to unprecedented responses in patients with refractory/relapsed B-cell non-Hodgkin lymphoma (NHL); nevertheless, two thirds of patients experience treatment failure. Resistance to apoptosis is a key feature of cancer cells, and it is associated with treatment failure. In 87 patients with NHL treated with anti-CD19 CART, we found that chromosomal alteration of B-cell lymphoma 2 (BCL-2), a critical antiapoptotic regulator, in lymphoma cells was associated with reduced survival. Therefore, we combined CART19 with the FDA-approved BCL-2 inhibitor venetoclax and demonstrated in vivo synergy in venetoclax-sensitive NHL. However, higher venetoclax doses needed for venetoclax-resistant lymphomas resulted in CART toxicity. To overcome this limitation, we developed venetoclax-resistant CART by overexpressing mutated BCL-2(F104L), which is not recognized by venetoclax. Notably, BCL-2(F104L)-CART19 synergized with venetoclax in multiple lymphoma xenograft models. Furthermore, we uncovered that BCL-2 overexpression in T cells intrinsically enhanced CART antitumor activity in preclinical models and in patients by prolonging CART persistence. SIGNIFICANCE: This study highlights the role of BCL-2 in resistance to CART immunotherapy for cancer and introduces a novel concept for combination therapies-the engineering of CART cells to make them resistant to proapoptotic small molecules, thereby enhancing the therapeutic index of these combination therapies. This article is highlighted in the In This Issue feature, p. 2221.

Effects of two-step cleaning sequences on foulant extraction from multibore ultrafiltration membranes in a pilot-scale membrane filtration system for surface water treatment.

The cleaning efficiencies of fouled multibore ultrafiltration membrane (UFMB) operated from a pilot-scale UF process for surface water treatment were systemically investigated according to the sequences of two different cleaning solutions. The experimental results decisively confirmed that HPI DOM and HPO DOM/multivalent ions complexation significantly resulted in the fouling formations on UFMB due to their neutral charge characteristic. The basic cleaning agent effectively extracted the organic foulants attached on UFMB, indicating that the type of cleaning agent was a critical factor influencing on the cleaning efficiency of fouled UFMB. However, the cleaning sequence 1 (CS-1: 0.1 M NaOH >0.1 M HCl; the total DOC = 725.77 mgC∙m-2; the total TN = 146.35 mgN∙m-2, total inorganic contents = 132.62 mg m-2) much more effectively extracted the foulants on the UFMB surfaces than the cleaning sequence 2 (CS-2: 0.1 M HCl >0.1 M NaOH; the total DOC = 604.49 mgC∙m-2; the total of TN = 121.79 mgN∙m-2, total inorganic contents = 73.43 mg m-2). The morphological results also clearly showed that the cleaned UFMB surface using CS-1 were effectively recovered, as compared with those using CP-2. Overall, this study implied that the hydroxide ions from the basic cleaning agent promoted the infiltration of the acidic cleaning agent into the densely formed fouling layers on the UFMB surfaces and demonstrated that the cleaning sequences strategy could significantly govern the restoration of UFMB performance during the pilot-scale surface water treatment system operation.

Repolarization of Tumor-Infiltrating Myeloid Cells for Augmentation of CAR T Cell Therapies.

Although CAR T cell therapies have proven to be effective in treating hematopoietic cancers, their abilities to regress solid tumors have been less encouraging. Mechanisms to explain these disparities have focused primarily on differences in cancer cell heterogeneity, barriers to CAR T cell penetration of solid tumors, and immunosuppressive microenvironments. To evaluate the contributions of immunosuppressive tumor-associated macrophages (TAMs) and myeloid-derived suppressor cells (MDSCs) on CAR T cell efficacies, we have exploited the ability of a folate-targeted Toll-like receptor 7 agonist (FA-TLR7-1A) to specifically reactivate TAMs and MDSCs from an immunosuppressive to pro-inflammatory phenotype without altering the properties of other immune cells. We report here that FA-TLR7-1A significantly augments standard CAR T cell therapies of 4T1 solid tumors in immune competent mice. We further show that co-administration of the FA-TLR7-1A with the CAR T cell therapy not only repolarizes TAMs and MDSCs from an M2-like anti-inflammatory to M1-like pro-inflammatory phenotype, but also enhances both CAR T cell and endogenous T cell accumulation in solid tumors while concurrently increasing their states of activation. Because analogous myeloid cells in healthy tissues ar not altered by administration of FA-TLR7-1A, no systemic activation of the immune system nor accompanying weight loss is observed. These data argue that immunosuppressive myeloid cells contribute prominently to the failure of CAR T cells to eradicate solid tumors and suggest that methods to reprogram tumor associated myeloid cells to a more inflammatory phenotype could significantly augment the potencies of CAR T cell therapies.

Intravenous to Oral Transition of Amiodarone (IOTA): Effect of Various Durations of Overlap on Atrial Fibrillation Recurrence After Cardiothoracic Surgery.

ABSTRACT: The use of amiodarone for postoperative atrial fibrillation (AF) is widespread; however, there is a paucity of data on the optimal duration of overlap when transitioning from intravenous (IV) to oral amiodarone. The objective of this study was to evaluate the safety and efficacy of varying durations of overlap when amiodarone IV infusion is transitioned to oral administration in cardiothoracic surgery patients. This retrospective, observational, single-center study included cardiothoracic surgery patients who were initiated on IV amiodarone for supraventricular arrhythmia and subsequently transitioned to oral amiodarone. The primary outcome was AF recurrence within 24 hours after IV amiodarone discontinuation. Safety outcomes include occurrence of bradycardia or hypotension while on amiodarone. A total of 184 patients were included for analysis. AF recurrence occurred in 24.5% of patients (n = 45). No significant association was found between various overlap durations and AF recurrence (odds ratio (OR) 1.00, 95% CI 1.00-1.01, P = 0.9). In addition, no significant association was found between duration of overlap and rates of bradycardia (OR 1.00, 95% confidence interval (CI) 0.99-1.00, P = 0.08) or hypotension (OR 1.00, 95% CI 0.99-1.00, P = 0.21), which occurred in 35.9% and 47.3% of patients, respectively. Our study suggests following conversion to normal sinus rhythm; cardiothoracic surgery patients can effectively and safely be transitioned from IV to oral amiodarone without the need for specific overlap duration or transition strategy.

Improving the performance of machine learning models for early warning of harmful algal blooms using an adaptive synthetic sampling method.

Many countries have attempted to monitor and predict harmful algal blooms to mitigate related problems and establish management practices. The current alert system-based sampling of cell density is used to intimate the bloom status and to inform rapid and adequate response from water-associated organizations. The objective of this study was to develop an early warning system for cyanobacterial blooms to allow for efficient decision making prior to the occurrence of algal blooms and to guide preemptive actions regarding management practices. In this study, two machine learning models: artificial neural network (ANN) and support vector machine (SVM), were constructed for the timely prediction of alert levels of algal bloom using eight years' worth of meteorological, hydrodynamic, and water quality data in a reservoir where harmful cyanobacterial blooms frequently occur during summer. However, the proportion imbalance on all alert level data as the output variable leads to biased training of the data-driven model and degradation of model prediction performance. Therefore, the synthetic data generated by an adaptive synthetic (ADASYN) sampling method were used to resolve the imbalance of minority class data in the original data and to improve the prediction performance of the models. The results showed that the overall prediction performance yielded by the caution level (L1) and warning level (L2) in the models constructed using a combination of original and synthetic data was higher than the models constructed using original data only. In particular, the optimal ANN and SVM constructed using a combination of original and synthetic data during both training (including validation) and test generated distinctively improved recall and precision values of L1, which is a very critical alert level as it indicates a transition status from normalcy to bloom formation. In addition, both optimal models constructed using synthetic-added data exhibited improvement in recall and precision by more than 33.7% while predicting L-1 and L-2 during the test. Therefore, the application of synthetic data can improve detection performance of machine learning models by solving the imbalance of observed data. Reliable prediction by the improved models can be used to aid the design of management practices to mitigate algal blooms within a reservoir.

Antiviral Nanomaterials for Designing Mixed Matrix Membranes.

Membranes are helpful tools to prevent airborne and waterborne pathogenic microorganisms, including viruses and bacteria. A membrane filter can physically separate pathogens from air or water. Moreover, incorporating antiviral and antibacterial nanoparticles into the matrix of membrane filters can render composite structures capable of killing pathogenic viruses and bacteria. Such membranes incorporated with antiviral and antibacterial nanoparticles have a great potential for being applied in various application scenarios. Therefore, in this perspective article, we attempt to explore the fundamental mechanisms and recent progress of designing antiviral membrane filters, challenges to be addressed, and outlook.

Design of Neuraminidase-Targeted Imaging and Therapeutic Agents for the Diagnosis and Treatment of Influenza Virus Infections.

The last step in influenza virus replication involves the assembly of viral components on the infected cell's plasma membrane followed by budding of intact virus from the host cell surface. Because viral neuraminidase and hemagglutinin are both inserted into the host cell's membrane during this process, influenza virus-infected cells are distinguished from uninfected cells by the presence of viral neuraminidase and hemagglutinin on their cell surfaces. In an effort to exploit this difference in cell surface markers for development of diagnostic and therapeutic agents, we have modified an influenza neuraminidase inhibitor, zanamivir, for targeting of attached imaging and therapeutic agents selectively to influenza viruses and virus-infected cells. We have designed here a zanamivir-conjugated rhodamine dye that allows visual monitoring of binding, internalization, and intracellular trafficking of the fluorescence-labeled neuraminidase in virus-infected cells. We also synthesize a zanamivir-99mTc radioimaging conjugate that permits whole body imaging of the virus's biodistribution and abundance in infected mice. Finally, we create both a zanamivir-targeted cytotoxic drug (i.e., zanamivir-tubulysin B) and a viral neuraminidase-targeted CAR T cell and demonstrate that they are both able to kill viral neuraminidase-expressing cells without damaging healthy cells. Taken together, these data suggest that the influenza virus neuraminidase inhibitor, zanamivir, can be exploited to improve the diagnosis, imaging, and treatment of influenza virus infections.

Changes in adsorption mechanisms of radioactive barium, cobalt, and strontium ions using spent coffee waste biochars via alkaline chemical activation: Enrichment effects of O-containing functional groups.

The single adsorption of radioactive barium (Ba(II)), cobalt (Co(II)), and strontium (Sr(II)) ions using pristine (SCWB-P) and chemically activated spent coffee waste biochars with NaOH (SCWB-A) were thoroughly explored in order to provide deeper insights into the changes in their adsorption mechanisms through alkaline chemical activation. The greater removal efficiencies of SCWB-A (76.6-97.3%) than SCWB-P (45.6-75.2%) and the consistency between the adsorptive removal patterns (Ba(II) > Sr(II) > Co(II)) and oxygen bond dissociation enthalpies (BaO (562 kJ/mol) > SrO (426 kJ/mol) > CoO (397 kJ/mol)) of radioactive species supported the assumption that the adsorption removal of radioactive species with spent coffee waste biochars highly depended on the abundances of O-containing functional groups. The calculated R2 values of the pseudo-first-order (SCWB-P = 0.998-0.999; SCWB-A = 0.850-0.921) and pseudo-second-order kinetic models (SCWB-P = 0.988-0.998; SCWB-A = 0.935-0.966) are evident that the physisorption mainly controlled the adsorption of radioactive species toward SCWB-P and the chemisorption played a crucial role in their adsorptive removal with SCWB-A. From the calculated intra-particle diffusion, isotherm, thermodynamic parameters, it can be concluded that the intra-particle diffusion and monolayer adsorption primarily governed the adsorption of radioactive species using SCWB-P and SCWB-A, and their adsorption processes occurred spontaneously and endothermically. The dominant adsorption mechanism of spent coffee waste biochars was changed from physisorption (ΔH° of SCWB-P = 21.6-29.8 kJ/mol) to chemisorption (ΔH° of SCWB-A = 42.4-81.3 kJ/mol) through alkaline chemical activation. The distinctive M-OH peak in the O1s XPS spectra of SCWB-A directly corresponding to the decrease in the abundances of O-containing functional groups confirms again that the enrichment of O-containing functional groups markedly facilitated the adsorption removal of radioactive species by chemisorption occurred at the inner and outer surfaces of spent coffee waste biochars.

Antigen-independent activation enhances the efficacy of 4-1BB-costimulated CD22 CAR T cells.

While CD19-directed chimeric antigen receptor (CAR) T cells can induce remission in patients with B cell acute lymphoblastic leukemia (ALL), a large subset relapse with CD19- disease. Like CD19, CD22 is broadly expressed by B-lineage cells and thus serves as an alternative immunotherapy target in ALL. Here we present the composite outcomes of two pilot clinical trials ( NCT02588456 and NCT02650414 ) of T cells bearing a 4-1BB-based, CD22-targeting CAR in patients with relapsed or refractory ALL. The primary end point of these studies was to assess safety, and the secondary end point was antileukemic efficacy. We observed unexpectedly low response rates, prompting us to perform detailed interrogation of the responsible CAR biology. We found that shortening of the amino acid linker connecting the variable heavy and light chains of the CAR antigen-binding domain drove receptor homodimerization and antigen-independent signaling. In contrast to CD28-based CARs, autonomously signaling 4-1BB-based CARs demonstrated enhanced immune synapse formation, activation of pro-inflammatory genes and superior effector function. We validated this association between autonomous signaling and enhanced function in several CAR constructs and, on the basis of these observations, designed a new short-linker CD22 single-chain variable fragment for clinical evaluation. Our findings both suggest that tonic 4-1BB-based signaling is beneficial to CAR function and demonstrate the utility of bedside-to-bench-to-bedside translation in the design and implementation of CAR T cell therapies.