Maintaining CD4/CD8 ratio and Th1-CTL subsets of chimeric antigen receptor (CAR)-T cells in serum-free culture conditions

Chimeric antigen receptor (CAR) T cells therapy is a promising strategy that significantly controlled the progress of cancer diseases. CAR-T cells could kill cancer cells through cellular immune response; therefore, CD8+ cytotoxic T cells are critical for CAR-T cell therapy. However, recent papers reported that CD4+ T helper cells were important for the response and maintenance of CAR-T cells in vivo. Here, we developed a serum-free CAR-T cell preparation process that maintained the T cell population and controlled the T cell subsets. The CD4+ and CD8+ T cell population in CAR-T cells were maintained at averagely 59.4 % and 34.6%, and the major T cell subsets were Th1 cells and cytotoxic T lymphocytes (CTLs), implying the potentially high cellular immune response. To verifying whether the prepared CAR-T cells were exhausted, the expression of several immune checkpoint markers was determined. Of interest, only less than 20% of CAR-T cells at endpoint were PD-1+ or CTLA4+, but more than 40% of CAR-T cells at the endpoint were TIM-3+, implying most CAR-T cells were not exhausted. These CAR-T cells produced more than 1 ng/mL of IFN-γ in the response to the antigen. Altogether, CAR-T cells could be prepared in our serum-free process in the controlling of T cell subsets, leading to potential high therapeutic potency. Please click Additional Files below to see the full abstract

Persistent Ballistic Entanglement Spreading with Optimal Control in Quantum Spin Chains

Entanglement propagation provides a key routine to understand quantum many-body dynamics in and out of equilibrium. In this work, we uncover that the ``variational entanglement-enhancing'' field (VEEF) robustly induces a persistent ballistic spreading of entanglement in quantum spin chains. The VEEF is time dependent, and is optimally controlled to maximize the bipartite entanglement entropy (EE) of the final state. Such a linear growth persists till the EE reaches the genuine saturation $\tilde{S} = - \log_{2} 2^{-\frac{N}{2}}=\frac{N}{2}$ with $N$ the total number of spins. The EE satisfies $S(t) = v t$ for the time $t \leq \frac{N}{2v}$, with $v$ the velocity. These results are in sharp contrast with the behaviors without VEEF, where the EE generally approaches a sub-saturation known as the Page value $\tilde{S}_{P} =\tilde{S} - \frac{1}{2\ln{2}}$ in the long-time limit, and the entanglement growth deviates from being linear before the Page value is reached. The dependence between the velocity and interactions is explored, with $v \simeq 2.76$, $4.98$, and $5.75$ for the spin chains with Ising, XY, and Heisenberg interactions, respectively. We further show that the nonlinear growth of EE emerges with the presence of long-range interactions.Comment: 5 pages, 4 figure

In vitro high expansion of chimeric antigen receptor (CAR)-T cells in serum-free process conditions

Manufacturing process is an important and complex factor for preparing chimeric antigen receptor (CAR) T cells for therapy. Although serum was widely applied in the culture or expansion of T cells, the quality of serum could be varied from batch to batch, leading to the variation of T cell expansion and quality. In addition, the safety of pathogens from serum and Chemistry, Manufacturing, and Control (CMC) were required to be considered. To overcome the disadvantages of serum application in T cell culture, serum-free and xeno-free culture conditions were required. We intended to develop a rapid serum-free culture condition for the expansion of immune T cells ex vivo. In our optimized serum-free condition, CAR-T cells could be expanded to about 100-200 times to the initial cell number after 6-day culture and the cell viability of all specimens was above 98%. Of interest, the percentage of CAR+ population in all specimens was increases, and the T cell pollutions could be maintained at averagely about 35-40% of CD8+ T cells and averagely about 50-55% of CD4+ T cells after culture. Taken together, our conditions could be applied in the expansion of CAR-T cells for cell therapy to support the minimum requirement of blood or cell samples from patients and to maintain the T cell population. Please click Additional Files below to see the full abstract

Delayed Airway Obstruction after Internal Jugular Venous Catheterization in a Patient with Anticoagulant Therapy

Delayed onset of neck hematoma following central venous catheterization without arterial puncture is uncommon. Herein, we present a patient who developed a delayed neck hematoma after repeated attempts at right internal jugular venous puncture and subsequent enoxaparin administration. Progressive airway obstruction occurred on the third day after surgery. Ultrasound examination revealed diffuse hematoma of the right neck, and fibreoptic examination of the airway revealed pharyngeal edema. After emergent surgical removal of the hematoma, the patient was extubated uneventfully

The Nexus of Sensory Loss, Cognitive Impairment, and Functional Decline in Older Adults: A Scoping Review

BACKGROUND AND OBJECTIVES: The prevalence of cognitive impairment and sensory loss in hearing or vision increases with age. Based on the Information Processing Model, cognitive impairment coupled with sensory loss may exacerbate disability in late life. Yet this issue has not been systematically studied. The purpose of this scoping review was to examine the literature that studied the relationship between cognitive impairment, sensory loss, and activities of daily living in older adults. RESEARCH DESIGN AND METHODS: Two reviewers independently screened 1,410 studies identified from 5 electronic databases (Medline, EMBASE, PsycINFO, CINHAL, and Web of Science). The search was completed in June 2020. A study was eligible if it included measurements of cognitive function, vision or hearing, and activities of daily living. Additionally, the data analyses must address how cognitive impairment and sensory loss are related to the performance of activities of daily living. RESULTS: The final review included 15 studies. Findings show an additive effect of cognitive impairment and sensory loss on the activities of daily living. Cognitive impairment or vision loss independently relates to the decline in activities of daily living. Hearing loss relates to the decline only when the loss is severe, or if the daily task is hearing - specific. DISCUSSION AND IMPLICATIONS: Older adults with coexisting sensory loss and cognitive impairment have the highest risk or prevalence of disability, comparing to cognitive impairment or sensory loss alone. This finding highlights the importance of developing interventions to reduce the risk of disability for older adults experiencing multiple impairments

Anti-Neuroinflammatory Effects of the Calcium Channel Blocker Nicardipine on Microglial Cells: Implications for Neuroprotection

Background/Objective Nicardipine is a calcium channel blocker that has been widely used to control blood pressure in severe hypertension following events such as ischemic stroke, traumatic brain injury, and intracerebral hemorrhage. However, accumulating evidence suggests that inflammatory processes in the central nervous system that are mediated by microglial activation play important roles in neurodegeneration, and the effect of nicardipine on microglial activation remains unresolved. Methodology/Principal Findings In the present study, using murine BV-2 microglia, we demonstrated that nicardipine significantly inhibits microglia-related neuroinflammatory responses. Treatment with nicardipine inhibited microglial cell migration. Nicardipine also significantly inhibited LPS plus IFN-γ-induced release of nitric oxide (NO), and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Furthermore, nicardipine also inhibited microglial activation by peptidoglycan, the major component of the Gram-positive bacterium cell wall. Notably, nicardipine also showed significant anti-neuroinflammatory effects on microglial activation in mice in vivo. Conclusion/Significance The present study is the first to report a novel inhibitory role of nicardipine on neuroinflammation and provides a new candidate agent for the development of therapies for inflammation-related neurodegenerative diseases

Activation of PI3-Kinase Is Required for AMPA Receptor Insertion during LTP of mEPSCs in Cultured Hippocampal Neurons

AbstractHippocampal CA1 homosynaptic long-term potentiation (LTP) is expressed specifically at activated synapses. Increased insertion of postsynaptic α-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid receptors (AMPARs) appears to be crucial for CA1 LTP. However, the mechanism underlying AMPAR insertion during LTP remains largely unknown. We now report that phosphatidylinositol 3-kinase (PI3K) is complexed with AMPARs at synapses and activated by selective stimulation of synaptic N-methyl-D-aspartate (NMDA) receptors. Activation of the AMPAR-associated PI3K is required for the increased cell surface expression of AMPARs and LTP. Thus, our results strongly suggest that the AMPAR-PI3K complex may constitute a critical molecular signal responsible for AMPAR insertion at activated CA1 synapses during LTP, and consequently, this lipid kinase may serve to determine the polarity of NMDA receptor-dependent synaptic plasticity

High-resolution spatial and genomic characterization of coral-associated microbial aggregates in the coral Stylophora pistillata

Bacteria commonly form aggregates in a range of coral species [termed coral-associated microbial aggregates (CAMAs)], although these structures remain poorly characterized despite extensive efforts studying the coral microbiome. Here, we comprehensively characterize CAMAs associated with Stylophora pistillata and quantify their cell abundance. Our analysis reveals that multiple Endozoicomonas phylotypes coexist inside a single CAMA. Nanoscale secondary ion mass spectrometry imaging revealed that the Endozoicomonas cells were enriched with phosphorus, with the elemental compositions of CAMAs different from coral tissues and endosymbiotic Symbiodiniaceae, highlighting a role in sequestering and cycling phosphate between coral holobiont partners. Consensus metagenome--assembled genomes of the two dominant Endozoicomonas phylotypes confirmed their metabolic potential for polyphosphate accumulation along with genomic signatures including type VI secretion systems allowing host association. Our findings provide unprecedented insights into Endozoicomonas-dominated CAMAs and the first direct physiological and genomic linked evidence of their biological role in the coral holobiont

Use and effectiveness of dapagliflozin in patients with type 2 diabetes mellitus: a multicenter retrospective study in Taiwan

Aims/Introduction To investigate the clinical outcomes of patients with type 2 diabetes mellitus (T2DM) who initiated dapagliflozin in real-world practice in Taiwan. Materials and Methods In this multicenter retrospective study, adult patients with T2DM who initiated dapagliflozin after May 1st 2016 either as add-on or switch therapy were included. Changes in clinical and laboratory parameters were evaluated at 3 and 6 months. Baseline factors associated with dapagliflozin response in glycated hemoglobin (HbA1c) were analyzed by univariate and multivariate logistic regression. Results A total of 1,960 patients were eligible. At 6 months, significant changes were observed: HbA1c by −0.73% (95% confidence interval [CI] −0.80, −0.67), body weight was -1.61 kg (95% CI −1.79, −1.42), and systolic/diastolic blood pressure by −3.6/−1.4 mmHg. Add-on dapagliflozin showed significantly greater HbA1c reduction (−0.82%) than switched therapy (−0.66%) (p = 0.002). The proportion of patients achieving HbA1c <7% target increased from 6% at baseline to 19% at Month 6. Almost 80% of patients experienced at least 1% reduction in HbA1c, and 65% of patients showed both weight loss and reduction in HbA1c. Around 37% of patients had at least 3% weight loss. Multivariate logistic regression analysis indicated patients with higher baseline HbA1c and those who initiated dapagliflozin as add-on therapy were associated with a greater reduction in HbA1c. Conclusions In this real-world study with the highest patient number of Chinese population to date, the use of dapagliflozin was associated with significant improvement in glycemic control, body weight, and blood pressure in patients with T2DM. Initiating dapagliflozin as add-on therapy showed better glycemic control than as switch therapy