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.

Association between Decreased SGK1 and Increased Intestinal α-Synuclein in an MPTP Mouse Model of Parkinson's Disease.

Parkinson's disease (PD) is a globally common progressive neurodegenerative disease resulting from the loss of dopaminergic neurons in the brain. Increased α-synuclein (α-syn) is associated with the degeneration of dopaminergic neurons and non-motor symptoms like gastrointestinal disorders. In this study, we investigated the association between serum/glucocorticoid-related kinase 1 (SGK1) and α-syn in the colon of a PD mouse model. SGK1 and α-syn expression patterns were opposite in the surrounding colon tissue, with decreased SGK1 expression and increased α-syn expression in the PD group. Immunofluorescence analyses revealed the colocation of SGK1 and α-syn; the PD group demonstrated weaker SGK1 expression and stronger α-syn expression than the control group. Immunoblotting analysis showed that Na+/K+ pump ATPase α1 expression levels were significantly increased in the PD group. In SW480 cells with SGK1 knockdown using SGK1 siRNA, decreasing SGK1 levels corresponded with significant increases in the expression levels of α-syn and ATPase α1. These results suggest that SGK1 significantly regulates Na+/K+ pump ATPase, influencing the relationship between electrolyte balance and fecal formation in the PD mouse model. Gastrointestinal disorders are some of the major prodromal symptoms of PD. Therefore, modulating SGK1 expression could be an important strategy for controlling PD.

A novel cyclopentyl methyl ether electrolyte solvent with a unique solvation structure for subzero (-40 °C) lithium-ion batteries.

1 M LiFSI in cyclopentyl methyl ether is shown as a novel electrolyte with a unique solvation structure to form a thin robust multilayer solid electrolyte interface with an inorganic LiF-rich inner layer. Aggregates and contact ion pairs are actively formed in the solvation shell and reduced on the graphite anode during lithiation. This EC-free electrolyte provides 86.9% initial efficiency, and 355 mA h g-1 over 350 cycles with an excellent capacity retention of 84% at a 1C rate. An excellent low-temperature performance of 370, 337, and 330 mA h g-1 at 0, -10, and -20 °C, respectively, at a 0.1C rate is recorded. Furthermore, at -40 °C, the graphite half-cell has a capacity of 274 mA h g-1 without electrolyte freezing.

Clinical characteristics and prognosis of elderly patients with colorectal cancer: Comparison between surgical resection and supportive care.

ABSTRACT: Elderly colorectal cancer (CRC) patients tend to avoid standard treatment, especially curative surgical resection, because of concerns about surgical complications or underlying diseases. This study is intended to compare clinical characteristics and prognosis between patients who had undergone surgical resection and received supportive care, and to evaluate the usefulness of surgical treatment in elderly patients.A total of 114 patients aged ≥80 years who were diagnosed with CRC were analyzed retrospectively. Of these patients, 73 patients underwent surgical resection for malignancy and 41 patients received supportive care. Clinicopathological factors and overall survival (OS) rates were compared.The surgical resection group had better Eastern Cooperative Oncology Group performance status, American Society of Anesthesiologists (ASA) physical status, and a lower stage than did the supportive-care group. The 3-year OS rate of the surgical group was significantly higher than that of the supportive-care group (60.7% vs 9.1%, P < .001). In extremely elderly patients (age ≥85 years), the surgical group showed a better 3-year OS rate than did the supportive-care group (73.9% vs 6.3%, P < .001), although Eastern Cooperative Oncology Group performance status and ASA physical status were not different. The post-operative mortality rate was 2.7%. In the analysis of risk factors related to survival, surgical resection was a good prognostic factor.Surgical treatment in elderly CRC patients showed a survival benefit, even in the extremely elderly patients. Surgical resection for CRC in elderly patients can be considered to improve survival.

Inactivation of Bacillus cereus Spores on Stainless Steel by Combined Superheated Steam and UV-C Irradiation Treatment.

Bacillus cereus spore contamination on food contact surfaces is of great concern in the food industry. Thus, in the present study, superheated steam (SHS) was used alone or combined with UV-C irradiation for inactivation of B. cereus spores inoculated on stainless steel coupons. Temperatures higher than 250°C were needed to effectively inactivate B. cereus spores by SHS treatment alone, while a synergistic bactericidal effect resulted from the sequential treatment of SHS before or after UV-C irradiation. The increased dipicolinic acid ratio obtained by the combined treatment had a significant role in the synergistic bactericidal effect. Therefore, the combined treatment of SHS and UV-C could be used effectively to inactivate B. cereus on stainless steel. It is recommended to use hurdle technology with reduced energy consumption to ensure microbiological safety on food contact surfaces.

Inactivation of Staphylococcus aureus Biofilms on Food Contact Surfaces by Superheated Steam Treatment.

The objective of this study was to compare the inactivation efficacy of saturated steam (SS) and superheated steam (SHS) on Staphylococcus aureus biofilms on food contact surfaces, including type 304 stainless steel coupons with No. 4 finish (STS No. 4), type 304 stainless steel coupons with 2B finish (STS 2B), high-density polyethylene (HDPE), and polypropylene (PP). In addition, the effects of the surface characteristics on the inactivation efficacy were evaluated. Biofilms were formed on each food contact coupon surface using a three-strain cocktail of S. aureus. Five-day-old biofilms on STS No. 4, STS 2B, HDPE, and PP coupons were treated with SS at 100°C and SHS at 125 and 150°C for 2, 4, 7, 10, 15, and 20 s. Among all coupon types, SHS was more effective than SS in inactivating the S. aureus biofilms. S. aureus biofilms on steel coupons were more susceptible to most SS and SHS treatments than the biofilms on plastic coupons. S. aureus biofilms on HDPE and PP coupons were reduced by 4.00 and 5.22 log CFU per coupon, respectively, after SS treatment (100°C) for 20 s. SS treatment for 20 s reduced the amount of S. aureus biofilm on STS No. 4 and STS 2B coupons to below the detection limit. With SHS treatment (150°C), S. aureus biofilms on HDPE and PP needed 15 s to be inactivated to below the detection limit, while steel coupons only needed 10 s. The results of this study suggest that SHS treatment has potential as a biofilm control intervention for the food industry.

Small-molecule drug screening identifies drug Ro 31-8220 that reduces toxic phosphorylated tau in Drosophila melanogaster.

The intraneuronal aggregates of hyperphosphorylated and misfolded tau (neurofibrillary tangles, NFTs) cause a stereotypical spatiotemporal Alzheimer's disease (AD) progression that correlates with the severity of the associated cognitive decline. Kinase activity contributes to the balance between neuron survival and cell death. Hyperactivation of kinases including the conventional protein kinase C (PKC) is a defective molecular event accompanying associative memory loss, tau phosphorylation, and progression of AD or related neurodegenerative diseases. Here, we investigated the ability of small therapeutic compounds (a custom library) to improve tau-induced rough-eye phenotype in a Drosophila melanogaster model of frontotemporal dementia. We also assessed the tau phosphorylation in vivo and selected hit compounds. Among the potential hits, we investigated Ro 31-8220, described earlier as a potent PKCα inhibitor. Ro 31-8220 robustly improved the rough-eye phenotype, reduced phosphorylated tau species in vitro and in vivo, reversed tau-induced memory impairment, and improved the fly motor functions. In a human neuroblastoma cell line, Ro 31-8220 reduced the PKC activity and the tau phosphorylation pattern, but we also have to acknowledge the compound's wide range of biological activity. Nevertheless, Ro 31-8220 is a novel therapeutic mitigator of tau-induced neurotoxocity.

Association between hepatic steatosis and the development of hepatocellular carcinoma in patients with chronic hepatitis B.

BACKGROUND/AIMS: Nonalcoholic fatty liver disease (NAFLD) is becoming a worldwide epidemic, and is frequently found in patients with chronic hepatitis B (CHB). We investigated the impact of histologically proven hepatic steatosis on the risk for hepatocellular carcinoma (HCC) in CHB patients without excessive alcohol intake. METHODS: Consecutive CHB patients who underwent liver biopsy from January 2007 to December 2015 were included. The association between hepatic steatosis (≥ 5%) and subsequent HCC risk was analyzed. Inverse probability weighting (IPW) using the propensity score was applied to adjust for differences in patient characteristics, including metabolic factors. RESULTS: Fatty liver was histologically proven in 70 patients (21.8%) among a total of 321 patients. During the median (interquartile range) follow-up of 5.3 (2.9-8.3) years, 17 of 321 patients (5.3%) developed HCC: 8 of 70 patients (11.4%) with fatty liver and 9 of 251 patients (3.6%) without fatty liver. The five-year cumulative incidences of HCC among patients without and with fatty liver were 1.9% and 8.2%, respectively (P=0.004). Coexisting fatty liver was associated with a higher risk for HCC (adjusted hazards ratio [HR], 3.005; 95% confidence interval [CI], 1.122-8.051; P=0.03). After balancing with IPW, HCC incidences were not significantly different between the groups (P=0.19), and the association between fatty liver and HCC was not significant (adjusted HR, 1.709; 95% CI, 0.404-7.228; P=0.47). CONCLUSION: Superimposed NAFLD was associated with a higher HCC risk in CHB patients. However, the association between steatosis per se and HCC risk was not evident after adjustment for metabolic factors.

Xenograft Failure of Pulmonary Valved Conduit Cross-linked with Glutaraldehyde or Not Cross-linked in a Pig to Goat Implantation Model.

BACKGROUND: Biologic valved grafts are important in cardiac surgery, and although several types of graft are currently available, most commercial xenografts tend to cause early disfiguration due to intimal proliferation and calcification. We studied the graft failure patterns on non-fixed and glutaraldehyde-fixed pulmonary xenograft in vivo animal experiment. MATERIALS AND METHODS: Pulmonary valved conduits were obtained from the right ventricular outflow tract of eleven miniature pigs. The grafts were subjected to 2 different preservation methods; with or without glutaraldehyde fixation: glutaraldehyde fixation (n=7) and non-glutaraldehyde fixation (n=4). The processed explanted pulmonary valved grafts of miniature pig were then transplanted into eleven goats. Calcium quantization was achieved in all of the explanted xenograft, hemodynamic, histopathologic and radiologic evaluations were performed in the graft which the transplantation period was over 300 days (n=7). RESULTS: Grafts treated with glutaraldehyde fixation had more calcification and conduit obstruction in mid-term period. Calcium deposition also appeared much higher in the glutaraldehyde treated graft compared to the non-glutaraldehyde treated graft (p<0.05). CONCLUSION: The present study suggests that xenografts prepared using glutaraldehyde fixation alone appeared to have severe calcification compared to the findings of non-glutaraldehyde treated xenografts and to be managed with proper anticalcification treatment and novel preservation methods. This experiment gives the useful basic chemical, histologic data of xenograft failure model with calcification for further animal study.

Do Class III patients have a different growth spurt than the general population?

INTRODUCTION: Understanding the timing and length of the growth spurt of Class III prognathic patients is fundamental to the strategy of interceptive orthopedic orthodontics as well as to the timing of orthognathic surgery. Consequently, this study was undertaken to determine whether there are any significant differences in the stature growth pattern of Class III subjects compared with non-Class III subjects and the general population. METHODS: Twelve-year longitudinal stature growth data were collected for 402 randomly selected adolescents in the general population, 55 Class III mandibular prognathic patients, and 37 non-Class III patients. The growth data were analyzed by using the traditional linear interpolation method and nonlinear growth functions. The 6 stature growth parameters were measured: age at takeoff, stature at takeoff, velocity at takeoff, age at peak height velocity, stature at peak height velocity, and velocity at peak height velocity. Comparisons in the stature growth parameters and 15 cephalometric variables among the general population, Class III subjects, and non-Class III subjects were made with multivariate analysis. RESULTS: Patients with Class III prognathism did not have different growth parameters compared with Class II subjects or the general population. CONCLUSIONS: This study does not allow meaningful conclusions with regard to the relationship of mandibular size and stature growth pattern. The application of nonlinear growth curves vs the traditional linear interpolation method was also discussed.

A more accurate method of predicting soft tissue changes after mandibular setback surgery.

PURPOSE: To propose a more accurate method to predict the soft tissue changes after orthognathic surgery. PATIENTS AND METHODS: The subjects included 69 patients who had undergone surgical correction of Class III mandibular prognathism by mandibular setback. Two multivariate methods of forming prediction equations were examined using 134 predictor and 36 soft tissue response variables: the ordinary least-squares (OLS) and the partial least-squares (PLS) methods. After fitting the equation, the bias and a mean absolute prediction error were calculated. To evaluate the predictive performance of the prediction equations, a 10-fold cross-validation method was used. RESULTS: The multivariate PLS method showed significantly better predictive performance than the conventional OLS method. The bias pattern was more favorable and the absolute prediction accuracy was significantly better with the PLS method than with the OLS method. CONCLUSIONS: The multivariate PLS method was more satisfactory than the conventional OLS method in accurately predicting the soft tissue profile change after Class III mandibular setback surgery.

Changes of the Structural and Biomechanical Properties of the Bovine Pericardium after the Removal of α-Gal Epitopes by Decellularization and α-Galactosidase Treatment.

BACKGROUND: Bovine pericardium is one of the most widely used materials in bioprosthetic heart valves. Immunologic responses have been implicated as potential causes of limited durability of xenogenic valves. This study aimed to determine the effectiveness of decellularization and α-galactosidase (α-gal) to remove major xenoreactive antigens from xenogenic tissues. MATERIALS AND METHODS: Recombinant Bacteroides thetaiotaomicron (B. thetaiotaomicron) α-gal or decellularization, or both were used to remove α-gal from bovine pericardium. It was confirmed by α-gal-bovine serum albumin-based enzyme-linked immunosorbent assay (ELISA), high-performance anion exchange chromatography, flow cytometry, 3,3'-diaminobenzidine-staining, and lectin-based ELISA. The mechanical properties of bovine pericardium after decellularization or α-gal treatment were investigated by tests of tensile-strength, permeability, and compliance. Collagen fiber rearrangement was also evaluated by a 20,000× transmission electron microscope (TEM). RESULTS: Recombinant B. thetaiotaomicron α-gal could effectively remove α-gal from bovine pericardium B. thetaiotaomicron (0.1 U/mL, pH 7.2) while recombinant human α-gal removed it recombinant human α-gal (10 U/mL, pH 5.0). There was no difference in the mechanical properties of fresh and recombinant α-gal-treated bovine pericardium. Furthermore, the TEM findings demonstrated that recombinant α-gal made no difference in the arrangement of collagen fiber bundles with decellularization. CONCLUSION: Recombinant B. thetaiotaomicron α-gal effectively removed α-gal from bovine pericardium with a small amount under physiological conditions compared to human recombinant α-gal, which may alleviate the harmful xenoreactive immunologic responses of α-gal. Recombinant α-gal treatment had no adverse effects on the mechanical properties of bovine pericardium.

Detoxification of Glutaraldehyde Treated Porcine Pericardium Using L-arginine & NABH(4).

BACKGROUND: Calcification is the most frequent cause of clinical failure of bioprosthetic tissues fabricated from GA-fixed porcine valves or bovine pericardium. A multi-factorial approach using different mechanisms was recently developed to reduce the calcification of bioprosthetic tissues. The purpose of the present study was to evaluate the synchronized synergism of using L-arginine and NaBH(4), compared with ethanol and L-lysine, in glutaraldehyde treated porcine pericardium from the standpoint of calcification and tissue elasticity. MATERIALS AND METHODS: Porcine pericardium was fixed at 0.625% GA (7 days at room temperature after 2 days at 4℃). An interim step of ethanol (80%; 1 day at room temperature) or L-lysine (0.1 M; 2 days at 37℃) or L-arginine (0.1 M; 2 days at 37℃) was followed by completion of the GA fixation. A final step of NaBH(4) (0.1 M; 2 days at room temperature) was followed. Their tensile strength, thickness, and thermal stability were measured. Treated pericardia were implanted subcutaneously into three-week-old Sprague-Dawley rats for 8 weeks. Calcium content was assessed by atomic absorption spectroscopy and histology. RESULTS: L-arginine and NaBH(4) pretreatment (1.81±0.39 kgf/5 mm p=0.001, 0.30±0.08 mm p<0.001) significantly increased tensile strength and thickness compared with the control (0.53±0.34 kgf/5 mm, 0.10±0.02 mm). In a thermal stability test, L-arginine and NaBH(4) pretreatment (84.25±1.12℃, p=0.023) caused a significant difference from the control (86.25±0.00℃). L-lysine and NaBH(4) pretreatment (183.8±42.6 ug/mg, p=0.804), and L-arginine and NaBH(4) pretreatment (163.3±27.5 ug/mg, p=0.621) did not significantly inhibit calcification compared to the control (175.5±45.3 ug/mg), but ethanol and NaBH(4) pretreatment did (38.5±37.3 ug/mg, p=0.003). CONCLUSION: The combined pretreatment using L-arginine and NaBH(4) after GA fixation seemed to increase the tensile strength and thickness of porcine pericardium, fixed with GA. Additionally, it seemed to keep thermal stability. However it could not decrease the calcification of porcine pericardium fixed with GA. NaBH(4) pretreatment seemed to decrease the calcification of porcine pericardium fixed with GA, but only with ethanol.

Ethylene-induced opposite redistributions of calcium and auxin are essential components in the development of tomato petiolar epinastic curvature.

Calcium has been suggested as an important mediator of gravity signaling transduction within the root cap statocyte. In a horizontally-placed root, it is redistributed in the direction of the gravity vector (i.e. it moves downward) and its redistribution is closely correlated with auxin downward movement. However, the involvement of calcium in the regulation of ethylene-induced epinasty and auxin movement is not known. In this report, we examined the involvement of calcium in lateral auxin transport during ethylene-induced epinasty in an effort to understand the relationship among calcium, auxin, and ethylene. Ethylene-induced epinasty was further stimulated by exogenously applied Ca2+, the calcium effect being the strongest among divalent cations tested. Pretreatment with NPA, an auxin transport inhibitor, negated the promotive effect of calcium ions on the petiolar epinasty. Ethylene caused redistribution/differential accumulation of 45Ca2+ toward the morphologically lower (abaxial) side of the leaf petioles, an effect opposite to that of 14C-IAA redistribution. Verapamil, a Ca2+ channel blocker, inhibited ethylene-induced epinasty, as well as the redistribution of 14C-IAA and 45Ca2+. When the petiole was inverted in the presence or absence of ethylene, the direction of 45Ca2+ differential accumulation was still toward the morphologically abaxial side of the petiole during epinastic movement regardless of gravitational direction. These results suggest that gravity-insensitive, ethylene-induced Ca2+ redistribution and accumulation toward the abaxial side are closely coupled to the adaxial auxin redistribution/accumulation and, in turn, to the petiolar epinasty.

Discovery of diarylacrylonitriles as a novel series of small molecule sortase A inhibitors.

On the basis of a hit from random screening, a novel class of small-molecule sortase A inhibitors was generated. The primary structure-activity relationship and the minimal structural requirements for potency were established through structural modifications and molecular modeling studies.

Inhibition of the bacterial surface protein anchoring transpeptidase sortase by isoquinoline alkaloids.

The inhibitory activity of Coptis chinensis rhizome-derived material was evaluated against sortase, a bacterial surface protein anchoring transpeptidase, from Staphylococcus aureus ATCC 6538p and compared to that of four commercially available isoquinoline alkaloids. The biologically active constituent of C. chinensis extract was characterized as the isoquinoline alkaloid, berberine chloride, by spectral analysis. The isolate was a potent inhibitor of sortase, with an IC50 value of 8.7 microg/ml and had antibacterial activity against Gram-positive bacteria with a minimum inhibitory concentration (MIC) in the range of 50-400 microg/ml. Among the four isoquinoline alkaloids tested, berberine chloride had strong inhibitory activity. These results indicate that berberine is a possible candidate for the development of a bacterial sortase inhibitor.

Inhibition of sortase, a bacterial surface protein anchoring transpeptidase, by beta-sitosterol-3-O-glucopyranoside from Fritillaria verticillata.

A glucosylsterol, beta-sitosterol-3-O-glucopyranoside, has been isolated as an active principle with sortase inhibitory effect from the bulbs of Fritillaria verticillata by bioassay-guided chromatographic fractionation. The isolate was a potent inhibitor of sortase, with an IC(50) value of 18.3 microg/ml and had antibacterial activity against Bacillus subtilis, Staphylococcus aureus, and Micrococcus leuteus with MIC values of 50, 200, and 400 microg/ml, respectively, indicating that this compound is a possible candidate for the development of a bacterial sortase inhibitor. In addition, sitosterol was found to be inactive upon sortase and bacterial cell growth. These results suggest that the inhibitory potency of beta-sitosterol-3-O-glucopyranoside is sensitively dependent upon the glucopyranoside side chain moiety.