Miracle-dz/UniFAR-FTUs huggingface dataset:
The Generation of CAR-Transfected Natural Killer T Cells for the Immunotherapy of Melanoma

Natural killer T (NKT) cells represent a cell subpopulation that combines characteristics of natural killer (NK) cells and T cells. Through their endogenous T-cell receptors (TCRs), they reveal a pronounced intrinsic anti-tumor activity. Thus, a NKT cell transfected with a chimeric antigen receptor (CAR), which recognizes a tumor-specific surface antigen, could attack tumor cells antigen-specifically via the CAR and additionally through its endogenous TCR. NKT cells were isolated from peripheral blood mononuclear cells (PBMCs), expanded, and electroporated with mRNA encoding a chondroitin sulfate proteoglycan 4 (CSPG4)-specific CAR. The CAR expression on NKT cells and their in vitro functionality were analyzed. A transfection efficiency of more than 80% was achieved. Upon stimulation with melanoma cells, CAR-NKT cells produced cytokines antigen-specifically. Compared with conventional CAR-T cells, cytokine secretion of CAR-NKT cells was generally lower. Specific cytotoxicity, however, was similar with CAR-NKT cells showing a trend towards improved cytotoxicity. Additionally, CAR-NKT cells could kill target cells through their endogenous TCRs. In summary, it is feasible to generate CAR-NKT cells by using mRNA electroporation. Their CAR-mediated cytotoxicity is at least equal to that of conventional CAR-T cells, while their intrinsic cytotoxic activity is maintained. Thus, CAR-NKT cells may represent a valuable alternative to conventional CAR-T cells for cancer immunotherapy.
Heterologous Expression of a Novel Zoysia japonica C2H2 Zinc Finger Gene, ZjZFN1, Improved Salt Tolerance in Arabidopsis

Growing evidence indicates that some grass species are more tolerant to various abiotic and biotic stresses than many crops. Zinc finger proteins play important roles in plant abiotic and biotic stresses. Although genes coding for these proteins have been cloned and identified in various plants, their function and underlying transcriptional mechanisms in the halophyte Zoysia japonica are barely known. In the present study, ZjZFN1 was isolated from Z. japonica using RACE method. Quantitative real time PCR results revealed that the expression of ZjZFN1 was much higher in leaf than in root and stem tissues, and induced by salt, cold or ABA treatment. The subcellular localization assay demonstrated that ZjZFN1 was localized to the nucleus. Expression of the ZjZFN1 in Arabidopsis thaliana improved seed germination and enhanced plant adaption to salinity stress with improved percentage of green cotyledons and growth status under salinity stress. Physiological and transcriptional analyses suggested that ZjZFN1 might, at least in part, influence reactive oxygen species accumulation and regulate the transcription of salinity responsive genes. Furthermore, RNA-sequencing analysis of ZjZFN1-overexpressing plants revealed that ZjZFN1 may serve as a transcriptional activator in the regulation of stress responsive pathways, including phenylalanine metabolism, α-linolenic acid metabolism and phenylpropanoid biosynthesis pathways. Taken together, these results provide evidence that ZjZFN1 is a potential key player in plants’ tolerance to salt stress, and it could be a valuable gene in Z. japonica breeding projects.
The Core Cyber-Defense Knowledge, Skills, and Abilities That Cybersecurity Students Should Learn in School

Our cybersecurity workforce needs surpass our ability to meet them. These needs could be mitigated by developing relevant curricula that prioritize the knowledge, skills, and abilities (KSAs) most important to cybersecurity jobs. To identify the KSAs needed for performing cybersecurity jobs, we administered survey interviews to 44 cyber professionals at the premier hacker conferences Black Hat 2016 and DEF CON 24. Questions concerned 32 KSAs related to cyber defense. Participants rated how important each KSA was to their job and indicated where they had learned that KSA. Fifteen of these KSAs were rated as being of higher-than-neutral importance. Participants also answered open-ended questions meant to uncover additional KSAs that are important to cyber-defense work. Overall, the data suggest that KSAs related to networks, vulnerabilities, programming, and interpersonal communication should be prioritized in cybersecurity curricula.
60-GHz Power Amplifier in 45-nm SOI-CMOS Using Stacked Transformer-Based Parallel Power Combiner

This letter presents the design of a 60-GHz power amplifier (PA) using a new stacked transformer (STF)-based parallel combiner. It begins by highlighting the main limitation of the conventional millimeter-wave (mm-wave) transformer-based parallel combiners, namely, the output parasitic transmission lines. Then, a new STF parallel combiner based on three overlaying coils is proposed. Analyses revealed significant enhancements in terms of self-inductance, quality factors, and coupling factors, which lead to enhanced combining efficiency in the proposed combiner. Subsequently, a two-stage mm-wave PA is implemented in 45-nm silicon-on-insulator CMOS technology. The fabricated PA demonstrates a 3-dB bandwidth equal to 12.5 GHz (50–62.5 GHz). The output power at 1-dB compression/saturation ( ${\mathrm{ OP}}_{\text {1 dB}}/P_{\text {sat}}$ ) and the corresponding power-added efficiencies are recorded as 16.2/18.5 dBm and 18.7/25.5%, respectively.
Bioinstructive Naringin‐Loaded Micelles for Guiding Stem Cell Osteodifferentiation

Naringin is a naturally occurring flavanone with recognized neuroprotective, cardioprotective, anti‐inflammatory, and antiosteoporotic properties. Herein, the delivery of Naringin‐loaded methoxy‐poly(ethylene glycol)–maleimide–thiol–poly(l‐lactide) (mPEGMSPLA) diblock polymeric micelles to human adipose‐derived stem cells (hASCs) with the aim to augment its pro‐osteogenic effect in these cells is reported for the first time. The synthesis of the diblock copolymer is performed via Michael‐type addition reaction between hydrophilic methoxy‐poly(ethylene glycol)–maleimide (mPEGMAL) and hydrophobic thiol–poly(l‐lactide) (PLASH) and confirmed by 1H NMR and attenuated total reflectance Fourier transformed infrared (ATR‐FTIR) spectroscopy. The resulting mPEGMSPLA copolymer self‐assembles into monodispersed polymeric micelles (≈84.4 ± 2 nm) and presents a high Naringin encapsulation efficiency (87.8 ± 4%), with a sustained release profile at physiological pH. Alongside, in vitro data reveal that upon internalization into hASC 2D cultures, Naringin nanomicellar formulations attain a higher pro‐osteogenic effect than that of free drug. Notably, these bioactive carriers also induce superior osteopontin expression and increase matrix mineralization in these cells over free drug administration. Overall, such findings support for the first time the use of polymeric nanomicelles for Naringin delivery into hASCs as a valid approach for modulating stem cell osteogenic differentiation.
RNF169 limits 53BP1 deposition at DSBs to stimulate single-strand annealing repair

Significance 53BP1 restrains DNA end resection, and its dosage imbalance upsets DNA double-strand break (DSB) repair pathway choice. Here, by monitoring 53BP1 distribution on DSB-flanking chromatin, we have established a dose-dependent role of the RING finger protein RNF169 in limiting 53BP1 DSB deposition. Moreover, we found that forced expression of RNF169 overcomes 53BP1 activity and stimulates mutagenic DSB repair via the single-strand annealing pathway. Our findings suggest that aberrant expression of RNF169 may represent a deleterious factor in DSB repair control and in maintenance of genome stability. Unrestrained 53BP1 activity at DNA double-strand breaks (DSBs) hampers DNA end resection and upsets DSB repair pathway choice. RNF169 acts as a molecular rheostat to limit 53BP1 deposition at DSBs, but how this fine balance translates to DSB repair control remains undefined. In striking contrast to 53BP1, ChIP analyses of AsiSI-induced DSBs unveiled that RNF169 exhibits robust accumulation at DNA end-proximal regions and preferentially targets resected, RPA-bound DSBs. Accordingly, we found that RNF169 promotes CtIP-dependent DSB resection and favors homology-mediated DSB repair, and further showed that RNF169 dose-dependently stimulates single-strand annealing repair, in part, by alleviating the 53BP1-imposed barrier to DSB end resection. Our results highlight the interplay of RNF169 with 53BP1 in fine-tuning choice of DSB repair pathways.
A Token Based Algorithm to Distributed Computation in Sensor Networks

We consider distributed algorithms for data aggregation and function computation in sensor networks. The algorithms perform pairwise computations along edges of an underlying communication graph. A token is associated with each sensor node, which acts as a transmission permit. Nodes with active tokens have transmission permits; they generate messages at a constant rate and send each message to a randomly selected neighbor. By using different strategies to control the transmission permits we can obtain tradeoffs between message and time complexity. Gossip corresponds to the case when all nodes have permits all the time. We study algorithms where permits are revoked after transmission and restored upon reception. Examples of such algorithms include Simple-Random Walk(SRW), Coalescent-Random-Walk(CRW) and Controlled Flooding(CFLD) and their hybrid variants. SRW has a single node permit, which is passed on in the network. CRW, initially initially has a permit for each node but these permits are revoked gradually. The final result for SRW and CRW resides at a single(or few) random node(s) making a direct comparison with GOSSIP difficult. A hybrid two-phase algorithm switching from CRW to CFLD at a suitable pre-determined time can be employed to achieve consensus. We show that such hybrid variants achieve significant gains in both message and time complexity. The per-node message complexity for n-node graphs, such as 2D mesh, torii, and Random geometric graphs, scales as $O(polylog(n))$ and the corresponding time complexity scales as O(n). The reduced per-node message complexity leads to reduced energy utilization in sensor networks.
Susceptibility to audio signals during autonomous driving

We investigate how susceptible human drivers are to auditory signals in three situations: when stationary, when driving, or when being driven by an autonomous vehicle. Previous research has shown that human susceptibility is reduced when driving compared to when being stationary. However, it is not known how susceptible humans are under autonomous driving conditions. At the same time, good susceptibility is crucial under autonomous driving conditions, as such systems might use auditory signals to communicate a transition of control from the automated vehicle to the human driver. We measured susceptibility using a three-stimulus auditory oddball paradigm while participants experienced three driving conditions: stationary, autonomous, or driving. We studied susceptibility through the frontal P3 (fP3) Electroencephalography Event-Related Potential response (EEG ERP response). Results show that the fP3 component is reduced in autonomous compared to stationary conditions, but not as strongly as when participants drove themselves. In addition, the fP3 component is further reduced when the oddball task does not require a response (i.e., in a passive condition, versus active). The implication is that, even in a relatively simple autonomous driving scenario, people’s susceptibility of auditory signals is not as high as would be beneficial for responding to auditory stimuli.
The Positive Relationships of Playfulness With Indicators of Health, Activity, and Physical Fitness

Adult playfulness is a personality trait that enables people to frame or reframe everyday situations in such a way that they experience them as entertaining, intellectually stimulating, or personally interesting. Earlier research supports the notion that playfulness is associated with the pursuit of an active way of life. While playful children are typically described as being active, only limited knowledge exists on whether playfulness in adults is also associated with physical activity. Additionally, existing literature has not considered different facets of playfulness, but only global playfulness. Therefore, we employed a multifaceted model that allows distinguishing among Other-directed, Lighthearted, Intellectual, and Whimsical playfulness. For narrowing this gap in the literature, we conducted two studies addressing the associations of playfulness with health, activity, and fitness. The main aim of Study 1 was a comparison of self-ratings (N = 529) and ratings from knowledgeable others (N = 141). We tested the association of self- and peer-reported playfulness with self- and peer-reported physical activity, fitness, and health behaviors. There was a good convergence of playfulness among self- and peer-ratings (between r = 0.46 and 0.55, all p < 0.001). Data show that both self- and peer-ratings are differentially associated with physical activity, fitness, and health behaviors. For example, self-rated playfulness shared 3% of the variance with self-rated physical fitness and 14% with the pursuit of an active way of life. Study 2 provides data on the association between self-rated playfulness and objective measures of physical fitness (i.e., hand and forearm strength, lower body muscular strength and endurance, cardio-respiratory fitness, back and leg flexibility, and hand and finger dexterity) using a sample of N = 67 adults. Self-rated playfulness was associated with lower baseline and activity (climbing stairs) heart rate and faster recovery heart rate (correlation coefficients were between −0.19 and −0.24 for global playfulness). Overall, Study 2 supported the findings of Study 1 by showing positive associations of playfulness with objective indicators of physical fitness (primarily cardio-respiratory fitness). The findings represent a starting point for future studies on the relationships between playfulness, and health, activity, and physical fitness.
Emodin induces apoptosis in human hepatocellular carcinoma HepaRG cells via the mitochondrial caspase-dependent pathway

Emodin-induced hepatotoxicity in vivo and in vitro has been gaining increasing attention. However, the exact molecular pathways underlying these effects remain poorly clarified. The aim of the present study was to evaluate the cytotoxic effect of emodin on HepaRG cells and to define the underlying mechanism. The results demonstrated that emodin evidently inhibited HepaRG cell growth in a dose- and time-dependent manner by blocking cell cycle progression in the S and G2/M phase and by inducing apoptosis. Emodin treatment also resulted in generation of reactive oxygen species (ROS), which abrogated mitochondrial membrane potential (MMP). The above effects were all suppressed by antioxidants, such as N-acetylcysteine (NAC). Further studies by western blot analysis howed that emodin upregulated p53, p21, Bax, cyclin E, cleaved caspase-3, 8 and 9, and cleaved poly(ADP-ribose)polymerase (PARP). However, the protein expression of Bcl-2, cyclin A and CDK2 was downregulated. Taken together, our results suggest that emodin induces apoptosis via the mitochondrial apoptosis pathway through cell cycle arrest and ROS generation in HepaRG cells.
Hypoxic preconditioned bone mesenchymal stem cells ameliorate spinal cord injury in rats via improved survival and migration

The unique hypoxic inflammatory microenvironment observed in the spinal cord following spinal cord injury (SCI) limits the survival and efficacy of transplanted bone mesenchymal stem cells (BMSCs). The aim of the present study was to determine whether hypoxic preconditioning (HP) increased the therapeutic effects of BMSC on SCI. BMSCs were pretreated with cobalt chloride (CoCl2) in vitro, and the proliferative apoptotic and migratory abilities of these hypoxic BMSCs (H-BMSCs) were assessed. BMSCs and H-BMSCs derived from green fluorescent protein (GFP) rats were transplanted into SCI rats in vivo. The neurological function, histopathology, inflammation, and number and migration of transplanted cells were examined. HP significantly enhanced BMSC migration (increased hypoxia inducible factor 1α and C-X-C motif chemokine receptor 4 expression) and tolerance to apoptotic conditions (decreased caspase-3 and increased B-cell lymphoma 2 expression) in vitro. In vivo, H-BMSC transplantation significantly improved neurological function, decreased spinal cord damage and suppressed the inflammatory response associated with microglial activation. The number of GFP-positive cells in the SCI core and peripheral region of H-BMSC animals was increased compared with that in those of BMSC animals, suggesting that HP may increase the survival and migratory abilities of BMSCs and highlights their therapeutic potential for SCI.
The Vitamin K Metabolome in Chronic Kidney Disease

The purpose of this review is to summarize the research to date on the impact of chronic kidney disease (CKD) on the vitamin K metabolome. Vitamin K-dependent proteins contribute to cardiovascular disease (CVD) prevention via the prevention of ectopic mineralization. Sub-clinical vitamin K deficiency is common in CKD patients, and evidence suggests that it may contribute to the CVD burden in this population. Research from animal models suggests that CKD alters tissue measures of the two predominant forms of vitamin K: KI and MK-4. The expression and/or activity of enzymes that regulate the recycling of vitamin K and the carboxylation of vitamin K-dependent proteins also appear to be altered in CKD. Evidence suggests that statins, a common pharmaceutical prescribed to CKD patients to prevent cardiovascular events, may impact the metabolism of vitamin K and therefore contribute to its relative inefficiency at preventing CVD in this population as kidney disease progresses. Human research on the tissue vitamin K metabolome in CKD patients is lacking.
Spectroscopic ruler for measuring active-site distortions based on Raman optical activity of a hydrogen out-of-plane vibration

Significance Many biological cofactors, including light-absorbing chromophores in photoreceptors, are modulated upon insertion into a protein. The steric contribution can cause structural distortions in the cofactor, and such effects are considered to be crucial for biological function. For example, the out-of-plane distortion of chromophores is a key factor in controlling their absorption spectra. In spite of the functional importance, such structural distortions are difficult to measure experimentally. In this study, we used a unique capability of Raman optical activity (ROA) to address the above-mentioned issue. This study applied ROA spectroscopy to a photoreceptor protein and indicates that a hydrogen out-of-plane ROA band provides a spectroscopic ruler for the out-of-plane distortion of the chromophore that is embedded in a protein environment. Photoactive yellow protein (PYP), from the phototrophic bacterium Halorhodospira halophila, is a small water-soluble photoreceptor protein and contains p-coumaric acid (pCA) as a chromophore. PYP has been an attractive model for studying the physical chemistry of protein active sites. Here, we explore how Raman optical activity (ROA) can be used to extract quantitative information on distortions of the pCA chromophore at the active site in PYP. We use 13C8-pCA to assign an intense signal at 826 cm−1 in the ROA spectrum of PYP to a hydrogen out-of-plane vibration of the ethylenic moiety of the chromophore. Quantum-chemical calculations based on density functional theory demonstrate that the sign of this ROA band reports the direction of the distortion in the dihedral angle about the ethylenic C=C bond, while its amplitude is proportional to the dihedral angle. These results document the ability of ROA to quantify structural deformations of a cofactor molecule embedded in a protein moiety.
Alzheimer's disease and type 2 diabetes mellitus are distinct diseases with potential overlapping metabolic dysfunction upstream of observed cognitive decline

Alzheimer's disease (AD) and type 2 diabetes mellitus (T2DM) are highly prevalent aging‐related diseases associated with significant morbidity and mortality. Some findings in human and animal models have linked T2DM to AD‐type dementia. Despite epidemiological associations between the T2DM and cognitive impairment, the interrelational mechanisms are unclear. The preponderance of evidence in longitudinal studies with autopsy confirmation have indicated that vascular mechanisms, rather than classic AD‐type pathologies, underlie the cognitive decline often seen in self‐reported T2DM. T2DM is associated with cardiovascular and cerebrovascular disease (CVD), and is associated with increased risk of infarcts and small vessel disease in the brain and other organs. Neuropathological examinations of post‐mortem brains demonstrated evidence of cerebrovascular disease and little to no correlation between T2DM and β‐amyloid deposits or neurofibrillary tangles. Nevertheless, the mechanisms upstream of early AD‐specific pathology remain obscure. In this regard, there may indeed be overlap between the pathologic mechanisms of T2DM/“metabolic syndrome,” and AD. More specifically, cerebral insulin processing, glucose metabolism, mitochondrial function, and/or lipid metabolism could be altered in patients in early AD and directly influence symptomatology and/or neuropathology.
Prostanoid EP2 Receptors Are Up-Regulated in Human Pulmonary Arterial Hypertension: A Key Anti-Proliferative Target for Treprostinil in Smooth Muscle Cells

Prostacyclins are extensively used to treat pulmonary arterial hypertension (PAH), a life-threatening disease involving the progressive thickening of small pulmonary arteries. Although these agents are considered to act therapeutically via the prostanoid IP receptor, treprostinil is the only prostacyclin mimetic that potently binds to the prostanoid EP2 receptor, the role of which is unknown in PAH. We hypothesised that EP2 receptors contribute to the anti-proliferative effects of treprostinil in human pulmonary arterial smooth muscle cells (PASMCs), contrasting with selexipag, a non-prostanoid selective IP agonist. Human PASMCs from PAH patients were used to assess prostanoid receptor expression, cell proliferation, and cyclic adenosine monophosphate (cAMP) levels following the addition of agonists, antagonists or EP2 receptor small interfering RNAs (siRNAs). Immunohistochemical staining was performed in lung sections from control and PAH patients. We demonstrate using selective IP (RO1138452) and EP2 (PF-04418948) antagonists that the anti-proliferative actions of treprostinil depend largely on EP2 receptors rather than IP receptors, unlike MRE-269 (selexipag-active metabolite). Likewise, EP2 receptor knockdown selectively reduced the functional responses to treprostinil but not MRE-269. Furthermore, EP2 receptor levels were enhanced in human PASMCs and in lung sections from PAH patients compared to controls. Thus, EP2 receptors represent a novel therapeutic target for treprostinil, highlighting key pharmacological differences between prostacyclin mimetics used in PAH.