Functional TCR T cell screening using single-cell droplet microfluidics

Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies

Minimax estimator of regression coefficient in normal distribution under balanced loss function

AbstractThis article investigates linear minimax estimators of regression coefficient in a linear model with an assumption that the underlying distribution is a normal one with a nonnegative definite covariance matrix under a balanced loss function. Some linear minimax estimators of regression coefficient in the class of all estimators are obtained. The result shows that the linear minimax estimators are unique under some conditions

Functional TCR T cell screening using single-cell droplet microfluidics

Adoptive T cell transfer, in particular TCR T cell therapy, holds great promise for cancer immunotherapy with encouraging clinical results. However, finding the right TCR T cell clone is a tedious, time-consuming, and costly process. Thus, there is a critical need for single cell technologies to conduct fast and multiplexed functional analyses followed by recovery of the clone of interest. Here, we use droplet microfluidics for functional screening and real-time monitoring of single TCR T cell activation upon recognition of target tumor cells. Notably, our platform includes a tracking system for each clone as well as a sorting procedure with 100% specificity validated by downstream single cell reverse-transcription PCR and sequencing of TCR chains. Our TCR screening prototype will facilitate immunotherapeutic screening and development of T cell therapies

Comparison of multiple detection methods of antibody for the early diagnosis of pediatric mycoplasma pneumonia

Mycoplasma pneumoniae pneumonia (MPP) is a common disease in infants and young children. MPP can also be a potential cause of healthcare-associated pneumonia. However, it is challenging to make an accurate diagnosis in a timely fashion. Our goal is to determine the assay consistencies of acridinium ester chemiluminescence immunoassay (AECLIA) and gelatin particle agglutination (GPA) test for the detection of Mycoplasma pneumoniae (MP) antibody. In this study, a total of 1404 children with suspected MPP were enrolled. Among them, 130 were diagnosed as MPP positive by mycoplasma culture, and 186 were negative. MP antibodies were detected by AECLIA, enzyme-linked immunosorbent assay (ELISA) and GPA. Consistency rates, differences among assays, and diagnosis performance were compared for the three methods. The independent χ 2 test results of AECLIA and ELISA for the detection of MP-IgG and MP-IgM antibodies were χ 2  = 29.210, P  < 0.001; χ 2  = 9.081, P  = 0.017, respectively, suggesting that the two detection methods are well correlated. Similar analyses were done for the comparison of AECLIA and GPA as well as the comparison of ELISA and GPA. The positive rates of these methods agree with epidemiology data and have good consistency. Thus, AECLIA with a much shorter assay time could be a better option for the screening of MPP

RINT-1 Serves as a Tumor Suppressor and Maintains Golgi Dynamics and Centrosome Integrity for Cell Survival▿

Faithful mitotic partitioning of the Golgi apparatus and the centrosome is critical for proper cell division. Although these two cytoplasmic organelles are probably coordinated during cell division, supporting evidence of this coordination is still largely lacking. Here, we show that the RAD50-interacting protein, RINT-1, is localized at the Golgi apparatus and the centrosome in addition to the endoplasmic reticulum. To examine the biological roles of RINT-1, we found that the homozygous deletion of Rint-1 caused early embryonic lethality at embryonic day 5 (E5) to E6 and the failure of blastocyst outgrowth ex vivo. About 81% of the Rint-1 heterozygotes succumbed to multiple tumor formation with haploinsufficiency during their average life span of 24 months. To pinpoint the cellular function of RINT-1, we found that RINT-1 depletion by RNA interference led to the loss of the pericentriolar positioning and dispersal of the Golgi apparatus and concurrent centrosome amplification during the interphase. Upon mitotic entry, RINT-1-deficient cells exhibited multiple abnormalities, including aberrant Golgi dynamics during early mitosis and defective reassembly at telophase, increased formation of multiple spindle poles, and frequent chromosome missegregation. Mitotic cells often underwent cell death in part due to the overwhelming cellular defects. Taken together, these findings suggest that RINT-1 serves as a novel tumor suppressor essential for maintaining the dynamic integrity of the Golgi apparatus and the centrosome, a prerequisite to their proper coordination during cell division

Strong metal oxide-support interaction in MoO₂/N-doped MCNTs heterostructure for boosting lithium storage performance

The low-rate capability and fast capacity decaying of the molybdenum dioxide anode material have been a bottleneck for lithium-ion batteries (LIBs) due to low carrier transport, drastic volume expansion and inferior reversibility. Furthermore, the lithium-storage mechanism is still controversial at present. Herein, we fabricate a new kind of MoO2 nanoparticles with nitrogen-doped multiwalled carbon nanotubes (MoO2/N-MCNTs) as anode for LIBs. The strong chemical bonding (MoOC) endows MoO2/N-MCNTs a strong metal oxide-support interaction (SMSI), rendering electron/ion transfer and facilitate significant Li+ intercalation pseudocapacitance, which is evidenced by both theoretical computation and detailed experiments. Thus, the MoO2/N-MCNTs exhibits high-rate performance (523.7 mAh/g at 3000 mA g-1) and long durability (507.8 mAh/g at 1000 mA g-1 after 500 cycles). Furthermore, pouch-type full cell composed of MoO2/N-MCNTs anodes and commercial LiNi0.6Co0.2Mn0.2O2 (NCM622) cathodes demonstrate impressive rate performance and cyclic life, which displays an unparalleled energy density of 553.0 Wh kg-1. Ex-situ X-ray absorption spectroscopy (XAS) indicates the enhanced lithium-storage mechanism is originated from a partially irreversible phase transition from Li0.98MoO2 to Li2MoO4 via delithiation. This work not only provides fresh insights into the enhanced lithium-storage mechanism but also proposes new design principles toward efficient LIBs.This work is partly supported by the National Natural Science Foundation of China (Grant No. 11705015, U1832147), Natural Science Foundation of the Jiangsu Higher Education Institutions (23KJA430001)

Hice1, a Novel Microtubule-Associated Protein Required for Maintenance of Spindle Integrity and Chromosomal Stability in Human Cells▿ †

Spindle integrity is critical for efficient mitotic progression and accurate chromosome segregation. Deregulation of spindles often leads to structural and functional aberrations, ultimately promoting segregation errors and aneuploidy, a hallmark of most human cancers. Here we report the characterization of a previously identified human sarcoma antigen (gene located at 19p13.11), Hice1, an evolutionarily nonconserved 46-kDa coiled-coil protein. Hice1 shows distinct cytoplasmic localization and associates with interphase centrosomes and mitotic spindles, preferentially at the spindle pole vicinity. Depletion of Hice1 by RNA interference resulted in abnormal and unstable spindle configurations, mitotic delay at prometaphase and metaphase, and elevated aneuploidy. Conversely, loss of Hice1 had minimal effects on interphase centrosome duplication. We also found that both full-length Hice1 and Hice1-N1, which is composed of 149 amino acids of the N-terminal region, but not the mutant lacking the N-terminal region, exhibited activities of microtubule bundling and stabilization at a near-physiological concentration. Consistently, overexpression of Hice1 rendered microtubule bundles in cells resistant to nocodazole- or cold-treatment-induced depolymerization. These results demonstrate that Hice1 is a novel microtubule-associated protein important for maintaining spindle integrity and chromosomal stability, in part by virtue of its ability to bind, bundle, and stabilize microtubules

Protective Effect of Hericium erinaceus on Alcohol Induced Hepatotoxicity in Mice

We investigated the effects of Hericium erinaceus (HEM) on liver injury induced by acute alcohol administration in mice. Mice received ethanol (5 g/kg BW) by gavage every 12 hrs for a total of 3 doses. HEM (200 mg/kg BW) was gavage before ethanol administration. Subsequent serum alanine aminotransferase (ALT) level, aspartate aminotransaminase (AST) level, Maleic dialdehyde (MDA) level, hepatic total antioxidant status (TAOS), and activated nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) were determined by ELISA and immunohistochemistry, respectively. HEM administration markedly (P<0.05) decreased serum ALT, AST, and MDA levels. The hepatic histopathological observations showed that HEM had a relatively significant role in mice model, which had alcoholic liver damage. In conclusion, we observed that HEM (200 mg/kg BW) supplementation could restrain the hepatic damage caused by acute alcohol exposure

Excellent room temperature catalytic activity for formaldehyde oxidation on a single-atom iron catalyst in a moist atmosphere

For human safety, efficient removal of formaldehyde in indoor environments is essential. However, removing formaldehyde from indoor environment given the temperature and moisture remains a challenge. In this study, a metal-organic framework-based single-atom iron catalyst (FeSA) is proposed as a candidate catalyst for oxidation of formaldehyde. The optimal structure between different coordination environments of FeSA was screened by density functional theory (DFT) calculation. Guided by the theoretical results, FeSA with 5 nitrogen coordination (FeSA-N5-C) was selected and prepared experimentally for evaluation. The activity tests revealed that the removal efficiency of formaldehyde reached 85.5% at 25°C and 75% relative humidity, which is not possible for traditional catalysts. More importantly, moisture boosts catalytic oxidation of formaldehyde to some extent, illustrating that FeSA-N5-C is robust for practical applications. To our knowledge, this is the first report of single-atom catalyst for catalytic oxidation of formaldehyde, opening up a new avenue for design of high activity and strongly water-resistant catalysts