Engineering of potent CAR NK cells using non-viral Sleeping Beauty transposition from minimalistic DNA vectors.

Natural killer (NK) cells have high intrinsic cytotoxic capacity, and clinical trials have demonstrated their safety and efficacy for adoptive cancer therapy. Expression of chimeric antigen receptors (CARs) enhances NK cell target specificity, with these cells applicable as off-the-shelf products generated from allogeneic donors. Here, we present for the first time an innovative approach for CAR NK cell engineering employing a non-viral Sleeping Beauty (SB) transposon/transposase-based system and minimized DNA vectors termed minicircles. SB-modified peripheral blood-derived primary NK cells displayed high and stable CAR expression and more frequent vector integration into genomic safe harbors than lentiviral vectors. Importantly, SB-generated CAR NK cells demonstrated enhanced cytotoxicity compared with non-transfected NK cells. A strong antileukemic potential was confirmed using established acute lymphocytic leukemia cells and patient-derived primary acute B cell leukemia and lymphoma samples as targets in vitro and in vivo in a xenograft leukemia mouse model. Our data suggest that the SB-transposon system is an efficient, safe, and cost-effective approach to non-viral engineering of highly functional CAR NK cells, which may be suitable for cancer immunotherapy of leukemia as well as many other malignancies.

Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.

Recombinant adeno-associated virus (rAAV) vector gene delivery systems have demonstrated great promise in clinical trials but continue to face durability and dose-related challenges. Unlike rAAV gene therapy, integrating gene addition approaches can provide curative expression in mitotically active cells and pediatric populations. We explored a novel in vivo delivery approach based on an engineered transposase, Sleeping Beauty (SB100X), delivered as an mRNA within a lipid nanoparticle (LNP), in combination with an rAAV-delivered transposable transgene. This combinatorial approach achieved correction of ornithine transcarbamylase deficiency in the neonatal Spfash mouse model following a single delivery to dividing hepatocytes in the newborn liver. Correction remained stable into adulthood, while a conventional rAAV approach resulted in a return to the disease state. In non-human primates, integration by transposition, mediated by this technology, improved gene expression 10-fold over conventional rAAV-mediated gene transfer while requiring 5-fold less vector. Additionally, integration site analysis confirmed a random profile while specifically targeting TA dinucleotides across the genome. Together, these findings demonstrate that transposable elements can improve rAAV-delivered therapies by lowering the vector dose requirement and associated toxicity while expanding target cell types.

Acrylates in artificial nails-Results of product analyses and glove penetration studies.

BACKGROUND: Artificial nail materials are mixtures that are prone to contain several sensitizing (meth)acrylates. It is not known whether the listing of (meth)acrylates is correct in these products' packages. Protective gloves suited for nail work are needed. OBJECTIVES: To analyse (meth)acrylates in gel nail and acrylic nail products chemically and to compare the results with the information in the product labels, and to study penetration of artificial nail materials through selected disposable gloves. METHODS: We analysed 31 gel nail products and 6 acrylic nail products for their (meth)acrylate content by gas chromatography-mass spectrometry (GC-MS). We tested the penetration of two nail products through three disposable gloves: nitrile rubber, neoprene rubber and polyvinyl chloride (PVC). RESULTS: Altogether 32/37 products contained (meth)acrylates. In all of them, there was discrepancy between the listed (meth)acrylates and those discovered in the analysis. The commonest (meth)acrylates were hydroxyethyl methacrylate (HEMA, 20/37 samples) and hydroxypropyl methacrylate (HPMA, 9/37 samples), but many of the product packages failed to declare them. Isobornyl acrylate (IBA) was discovered in nine gel nail products. The neoprene glove could withstand nail gel for 20 min and thin nitrile glove and PVC glove for 5 min. Acrylic nail liquid penetrated through disposable gloves quickly. CONCLUSIONS: Labelling of artificial nail products was notably incorrect on most products. Requirements for product labelling must be updated so that the risk of sensitization associated with artificial nail products is clearly indicated. Disposable gloves can probably be used short-term in gel nail work, whereas disposable gloves do not protect the user from acrylic nail liquids.

Natural variation in Beauty Mark is associated with UV-based geographical adaptation in Gossypium species.

BACKGROUND: Anthocyanins, a class of specialized metabolites that are ubiquitous among plant species, have attracted a great deal of attention from plant biologists due to their chemical diversity. They confer purple, pink, and blue colors that attract pollinators, protect plants from ultraviolet (UV) radiation, and scavenge reactive oxygen species (ROS) to facilitate plant survival during abiotic stress. In a previous study, we identified Beauty Mark (BM) in Gossypium barbadense as an activator of the anthocyanin biosynthesis pathway; this gene also directly led to the formation of a pollinator-attracting purple spot. RESULTS: Here, we found that a single nucleotide polymorphism (SNP) (C/T) within the BM coding sequence was responsible for variations in this trait. Transient expression assays of BM from G. barbadense and G. hirsutum in Nicotiana benthamiana using luciferase reporter gene also suggested that SNPs in the coding sequence could be responsible for the absent beauty mark phenotype observed in G. hirsutum. We next demonstrated that the beauty mark and UV floral patterns are associated phenotypes and that UV exposure resulted in increased ROS generation in floral tissues; BM thus contributed to ROS scavenging in G. barbadense and wild cotton plants with flowers containing the beauty mark. Furthermore, a nucleotide diversity analysis and Tajima's D Test suggested that there have been strong selective sweeps in the GhBM locus during G. hirsutum domestication. CONCLUSIONS: Taken together, these results suggest that cotton species differ in their approaches to absorbing or reflecting UV light and thus exhibit variations in floral anthocyanin biosynthesis to scavenge reactive ROS; furthermore, these traits are related to the geographic distribution of cotton species.

The role of category valence in prototype preference.

People prefer prototypical stimuli over atypical stimuli. The dominant explanation for this prototype preference effect is that prototypical stimuli are processed more fluently. However, a more recent account proposes that prototypes are more strongly associated with their category's valence, leading to a reversed prototype preference effect for negative categories. One critical but untested assumption of this category-valence account is that no prototype preference should emerge for entirely neutral categories. We tested this prediction by conditioning categories of dot patterns positively, negatively, or neutrally. In line with previous findings on the category-valence account, prototype preference reversed for negatively conditioned categories. However, prototype preference was similarly strong for positive and neutral categories. These findings imply that prototype preferences do not only reflect a transfer of category valence to exemplars. Instead, the results suggest that prototype preference is a multi-process phenomenon arising from the activated category valence and a fluency-based process. We discuss further implications for theories on fluency and prototype preference.

The Ideal Proportion of the Auricle Exposure via a Morphometric Analysis in Asian Women.

BACKGROUND: Ears are an important aesthetic feature that is vital to the overall attractiveness of the face. Although there have been many studies on the aesthetics of the auricle, there is currently a lack of consensus on the ideal proportion of auricle exposure for Asian women in frontal view. OBJECTIVES: This study aimed to investigate ideal proportion of auricle exposure in Asian women. METHODS: An observational study was carried out on the photographs of 84 women on the list of the 100 most beautiful faces in Asia (published by TCC Asia in 2020). The proportion of the distance between the outer canthus and the outermost point of auricle to the distance between the inner canthus and the outermost point of auricle was calculated as the auricle exposure proportion. Evaluators were asked to rank a set of photographs of the volunteer with varying auricle exposure proportions from most attractive to least attractive. RESULTS: Measurements of the photographs of the 84 women showed a mean ear exposure proportion of 0.600. With 487 questionnaire responses received, the proportion of auricle exposure that the evaluators considered most attractive was 0.600. People with aesthetic experience considered 0.625 the most attractive proportion, while the general group considered 0.600 the most attractive. CONCLUSIONS: The ideal proportion of the auricle exposure for Asian women is in the range of 0.60-0.625, which may help surgeons reconstruct aesthetically pleasing ears. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Exploring the Ideal of Breast Beauty: A Morphometric Analysis Based on the Opinions of the Brazilian Population.

Background The quest to identify parameters deemed optimal for breast aesthetics holds paramount significance in both aesthetic and reconstructive surgery. The definition of ideal breast beauty, however, is subject to cultural variations and diverse standards, necessitating a detailed understanding. Objective This study delves into the perspectives of the Brazilian population regarding the aesthetics of the female breast. Methods A selection of images portraying variations in the distribution of upper and lower pole ratios across four distinct proportion models (35:65, 45:55, 50:50, and 55:45) was presented to participants. Through a meticulously crafted questionnaire administered via Google Forms, respondents were tasked with identifying the images they deemed most beautiful and attractive. Comprehensive demographic data of the study participants were gathered and subsequently correlated with their group-specific opinions. Results A total of 3744 questionnaire responses were collected 228 of which were from plastic surgeons/residents. The average age was 35 years, with 55.3% being women. Notably, breasts featuring a ratio of 45:55 emerged as the consensus choice for the epitome of beauty, garnering preference from 73% of the general population, 68% of women, 78% of men, and an equal 78% of plastic surgeons. This preference was consistent across all regions of the country. Conclusion Breasts with a ratio of 45:55 were universally deemed the most aesthetically pleasing. Similarly, breasts with a 50:50 ratio of breast tissue emerged as the next preferred choice. These insights contribute valuable perspectives on aesthetic considerations in breast surgery within the unique cultural context of Brazil.Level of Evidence IV This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Anthropometry and Current Aesthetic Concept of the Lower Third of the Face and Lips in Caucasian Adult Population: A Systematic Review and Meta-Analysis.

BACKGROUND: The lack of standardization of the norms and aesthetic concept of the lower third of the face and lips creates challenges in reconstructive surgery and aesthetic procedures. A large-scale, anthropometric measurements summary poses an alternative for establishing universal patterns. METHODS: A systematic review was conducted by searching PubMed, Embase, Google Scholar and Web of Science using keywords: lip, face, aesthetics, beauty, anthropometry and Caucasian. Seventy records were included in the review and checked in detail in terms of available data. RESULTS: The study contains a meta-analysis of twelve parameters with sufficient quantitative data-lip width, nose width, facial width, lower third of the face height, midline mandible height, total height of the upper and lower lip, upper and lower vermilion height, cutaneous upper lip height and nasolabial and mentolabial angle. CONCLUSIONS: The review provides a detailed database of primary anthropometric studies of lips and perioral regions of the healthy Caucasian population. The attractiveness of the region is focused on uniformity of proportions. Notably, results acquired with different methods of measurement are not interchangeable. Despite many published anthropometric studies, systems for conducting the measurements and reporting the results are not sufficiently unified to quantitatively assess meticulous key aesthetic clinical parameters. LEVEL OF EVIDENCE I: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine Ratings, please refer to Table of Contents or online Instructions to Authors www.springer.com/00266 .

A Systematic Review on the Reinforcement Loop in Aesthetic Medicine and Surgery: The Interplay of Social Media, Self-Perception, and Repeat Procedures.

INTRODUCTION: This systematic review delves into the impact of social media on self-perception and the escalating interest in clinical aesthetic procedures, proposing that social media significantly influences beauty standards and increases demand for aesthetic enhancements. METHOD: Following PRISMA guidelines, a mixed-method analysis of 34 articles was conducted, sourced from various databases, focusing on social media's psychological effects on clinical aesthetics decisions. The studies encompassed a broad spectrum, including qualitative, quantitative, and mixed methodologies, reflecting diverse geographical and cultural perspectives. RESULTS: The quantitative analysis demonstrated a strong positive correlation between social media usage and the consideration of aesthetic procedures (r=0.45, p<0.001), indicating a significant impact. Specific findings included a large effect size (Cohen's d=0.8) for the relationship between time spent on social media and the desire for aesthetic enhancements. Individuals spending more than 3 hours per day on social media platforms were twice as likely to consider aesthetic procedures compared to those with less usage, with a 95% confidence interval indicating robustness in these findings. CONCLUSION: Confirming the reinforcing effect of social media on aesthetic decision-making, this study highlights the complex interplay between digital media exposure, altered self-perception, and the increased inclination towards aesthetic procedures. It suggests a critical need for practitioners to carefully navigate the digital influence on patient's desires, reinforcing the significance of understanding psychological motivations and societal pressures in clinical aesthetics. This comprehensive analysis offers pivotal insights for clinical practice and ongoing research into social media's role in contemporary beauty standards. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

A Gaze into the Eyebrow Standards of Asian Versus Caucasian Women.

BACKGROUND: Eyebrow position affects perceived facial expression and youthfulness, and its modification is a key component of facial rejuvenation. OBJECTIVE: This investigation aimed to assess the preferred vertical eyebrow position, apex location, and eyebrow shape in Caucasian and Asian individuals and to analyze gaze patterns during aesthetic judgment using eye-tracking technology. MATERIALS AND METHODS: The study included 76 Asian and Caucasian volunteers with no medical background. Eye movements were captured with a Tobii Pro Nano eye-tracker. Participants viewed AI-generated images of Caucasian and Asian females with varied eyebrow positions (ratios 1:1.3 to 1:2.5), shapes (angles 8° to 20°), and apex positions. Aesthetic preferences were rated on a 5-point Likert scale. Eye-tracking metrics and aesthetic ratings were statistically analyzed using ANOVA and bivariate correlations. RESULTS: Both genders across ethnicities preferred a moderate eyebrow position ratio of 1:1.6. For eyebrow shapes, a 12° angle received the highest ratings, while extremes were less favored, indicating a preference for moderately curved eyebrows. The most appealing apex position was above the lateral canthus for Asians, and halfway between the lateral limbus and lateral canthus for Caucasians. Eye-tracking revealed longer fixations on unattractive features, suggesting more complex cognitive processing, while attractive features were processed more efficiently. CONCLUSION: The study revealed that aesthetic preferences for eyebrow features are influenced by both ethnic background and gender, with a general preference toward moderately curved eyebrows and subtle variations in preferred positions. These findings suggest a need for culturally sensitive approaches in facial aesthetic procedures and highlight the potential of eye-tracking technology to enhance surgical planning. Surgeons are advised to adopt a conservative, patient-centered approach when modifying eyebrow features, considering individual and cultural aesthetics to maximize patient satisfaction. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Comparative transcriptome and metabolome analyses revealed quality difference between beauty tea processed through indoor withering and outdoor solar withering.

BACKGROUND: Withering is the first processing procedure of beauty tea, and there are few reports on the impact of withering methods on the quality of beauty tea and its regulatory mechanisms. RESULTS: Through comparison of fresh tea leaves (FT) with the leaves after indoor natural withering for 18 h (IWT-18) and outdoor solar withering for 6 h (OWT-6), which were collected at the end of the two withering processes, 17 282 and 13 984 differentially expressed genes (DEGs) were respectively screened and 267 and 154 differential metabolites (DMs) were respectively identified. The coexpression network revealed that a large number of DEGs and DMs were enriched in phenylpropanoid, flavonoid, and adenosine triphosphate binding cassette (ABC) transporter pathways, and the number of DMs and DEGs in IWT-18 versus FT exceeded that in OWT-6 versus FT. Both withering methods promoted a significant increase in content of phenylalanine and upregulation of ß-glucoside expression in the phenylpropanoid metabolism pathway. Five theaflavin-type proanthocyanidins in the flavonoid synthesis pathway were more significantly accumulated in FT versus IWT-18 than in FT versus OWT-6. Meanwhile, both withering methods can affect the ABC transporter pathway to promote the accumulation of amino acids and their derivatives, but different withering methods affect different ABC transporter families. Outdoor withering with more severe abiotic stress has a greater impact on the ABCG family, whereas indoor withering has a more significant effect on the ABCC family. Sensory evaluation results showed that the dry tea of IWT-18 was slightly better than that of OWT-6 because of the longer withering time and more thorough substance transformation. CONCLUSION: In conclusion, the formation of honey flavor in beauty tea may be closely related to the DEGs and DMs in these three pathways. Our research provides theoretical data support for further revealing the mechanism of quality formation during the withering process of beauty tea. © 2023 Society of Chemical Industry.

Cognitive entailments among "the true, the good, the beautiful": a mainland Chinese sample.

Philosophers and cognitive scientists have long debated about the entailments among "the true, the good, the beautiful" (TGB hereafter). In the current article, we directly probed mainland Chinese subjects' cognitive entailment among TGB. Using 1-7 (Experiment 1) and 1-6 (Experiment 2) Likert scales, we convergently observed that mainland Chinese subjects tend to think that the beautiful is not the true, and that the good is the beautiful. Additionally, Experiment 1 also revealed that mainland Chinese subjects tend to think that the true is not the beautiful. Some of these results may reflect anthropological universals, and some others may reflect cultural specifics. Experiment 3 revealed that the most popular translation of TGB in Chinese into English is rather "the true, the kind, the beautiful", suggesting that the three concepts mapped to TGB in Chinese is not one-to-one mapped to the three concepts mapped to TGB in English. Therefore, caution should be exercised when making cross-linguistic or cross-cultural comparisons about TGB in the future.

Higher orthorexia tendency among female fashion models: an empirical international study.

PURPOSE: Female fashion models are more at risk for developing eating disorders than non-models due to the intense occupational pressure they face. The present study focuses on assessing whether female models are more prone to report orthorexia nervosa signs and symptoms than non-models. METHODS: Female fashion models (n = 179, mean age: 25.9 SD = 4.40 years) and an age adjusted control group (n = 261, mean age: 25.0 SD = 4.97 years) were selected by snowball sampling. Participants filled out an online survey containing anthropometric questions and the 18-item Eating Habits Questionnaire. RESULTS: According to BMI, fashion models were underweight (mean BMI = 18.1 SD = 1.68) while control participants' BMI was in the normal range (mean = 22.1 SD = 4.23, p < 0.001). On all three of Eating Habits Questionnaire subscales fashion models showed significantly higher average value (Knowledge subscale: M = 2.42 among models versus M = 2.08 in the control group, p < 0.01, Cohen's d = 0.52; Problems subscale: M = 1.93 among models versus M = 2.61 in the control group, p < 0.01, Cohen's d = 0.49; Feelings subscale: M = 3.20 among models versus M = 2.96 in the control group, p < 0.01, Cohen's d = 0.38). Orthorexic tendencies were reported by 35.1% of the models versus 20.2% of controls. CONCLUSION: Fashion models are at risk for the development of eating disorders. Even though not yet included in the DSM-5, the assessment of orthorexia nervosa among fashion models seems to be important. It is suggested to take appropriate measures to prevent the spread of disordered eating habits among models as they can lead to the development of anorexia nervosa or bulimia nervosa. LEVEL OF EVIDENCE: Level III, well-designed cohort study.

Beauty work or beauty care? Women's perceptions of appearance in the second half of life.

The prevailing cultural emphasis on women's attractiveness being tied to youth raises questions about how women perceive their appearance in the second half of life. The current qualitative study addresses this issue by posing two questions: how do women over fifty perceive and describe changes in their appearance? And how do they cope with these changes? Five focus group meetings with 19 Israeli women aged 54-76 were held to examine the issue. The meetings were structured around viewing three film clips starring older characters, encouraging the reporting of attitudes and perceptions in response to the clips. Based on a latent thematic analysis, the findings led to the conceptualization of a five-attitude model in response to physical changes in women's appearance: Grief-over the loss of youth and attractiveness; Resentment-over gendered media representations and cultural norms; Avoidance-distancing from one's aged appearance; Care-maintaining grooming routines; and Acceptance-coming to terms with the changes in appearance. Differences in responses between women were interpreted as reflecting a distinction between internal and external locus of control. Those with an external locus of control internalized the judgmental gaze of others, thus, reporting a greater sense of loss. Those with an internal locus of control were better able to accept themselves and focus on grooming rather than conforming to an imagined ideal. Results suggest that the distinction between understanding women's relationship with their appearance in terms of beauty work or beauty care may depend on the woman's locus of control.

Beauty in experiment: A qualitative analysis of aesthetic experiences in scientific practice.

A growing literature in philosophy of science focuses on the role of aesthetics in scientific practice, with the experiment recently recognized for its aesthetic value. However, the literature on aesthetics in experimentation grows out of case studies from the history of science, leaving open the question as to how contemporary scientists experience aesthetics in their experimental work. In this paper we offer the first qualitative, empirical analysis of aesthetic experiences regarding experimental practice, drawing from in-depth interviews with 215 scientists in four countries. We identify six categories of aesthetic experience we find in experimentation, their function, and new questions emerging from our study.

The neural determinants of abstract beauty.

We have enquired into the neural activity which correlates with the experience of beauty aroused by abstract paintings consisting of arbitrary assemblies of lines and colours. During the brain imaging experiments, subjects rated abstract paintings according to aesthetic appeal. There was low agreement on the aesthetic classification of these paintings among participants. Univariate analyses revealed higher activity with higher declared aesthetic appeal in both the visual areas and the medial frontal cortex. Additionally, representational similarity analysis (RSA) revealed that the experience of beauty correlated with decodable patterns of activity in visual areas. These results are broadly similar to those obtained in previous studies on facial beauty. With abstract art, it was the involvement of visual areas implicated in the processing of lines and colours while with faces it was of visual areas implicated in the processing of faces. Both categories of aesthetic experience correlated with increased activity in medial frontal cortex. We conclude that the sensory areas participate in the selection of stimuli according to aesthetic appeal and that it is the co-operative activity between the sensory areas and the medial frontal cortex that is the basis for the experience of abstract visual beauty. Further, this co-operation is enabled by "experience dependent" functional connections, in the sense that currently the existence and high specificity of these connections can only be demonstrated during certain experiences.

Sleeping Beauty kit sets provide rapid and accessible generation of artificial antigen-presenting cells for natural killer cell expansion.

Artificial antigen-presenting cells (aAPCs) offer a cost effective and convenient tool for the expansion of chimeric antigen receptor (CAR)-bearing T cells and NK cells. aAPCs are particularly useful because of their ability to efficiently expand low-frequency antigen-reactive lymphocytes in bulk cultures. Commonly derived from the leukemic cell line K562, these aAPCs lack most major histocompatibility complex expression and are therefore useful for NK cell expansion without triggering allogeneic T-cell proliferation. To combat difficulties in accessing existing aAPC lines, while circumventing the iterative lentiviral gene transfers with antibody-mediated sorting required for the isolation of stable aAPC clones, we developed a single-step technique using Sleeping Beauty (SB)-based vectors with antibiotic selection options. Our SB vectors contain options of two to three genes encoding costimulatory molecules, membrane-bound cytokines as well as the presence of antibiotic-resistance genes that allow for stable transposition-based transfection of feeder cells. Transfection of K562 with SB vectors described in this study allows for the surface expression of CD86, 4-1BBL, membrane-bound (mb) interleukin (IL)-15 and mbIL-21 after simultaneous transposition and antibiotic selection using only two antibiotics. aAPCs successfully expanded NK cells to high purity (80-95%). Expanded NK cells could be further engineered by lentiviral CAR transduction. The multivector kit set is publicly available and will allow convenient and reproducible in-house production of effective aAPCs for the in vitro expansion of primary cells.

Targeted delivery of a PD-1-blocking scFv by CD133-specific CAR-T cells using nonviral Sleeping Beauty transposition shows enhanced antitumour efficacy for advanced hepatocellular carcinoma.

BACKGROUND: CD133 is considered a marker for cancer stem cells (CSCs) in several types of tumours, including hepatocellular carcinoma (HCC). Chimeric antigen receptor-specific T (CAR-T) cells targeting CD133-positive CSCs have emerged as a tool for the clinical treatment of HCC, but immunogenicity, the high cost of clinical-grade recombinant viral vectors and potential insertional mutagenesis limit their clinical application. METHODS: CD133-specific CAR-T cells secreting PD-1 blocking scFv (CD133 CAR-T and PD-1 s cells) were constructed using a sleeping beauty transposon system from minicircle technology, and the antitumour efficacy of CD133 CAR-T and PD-1 s cells was analysed in vitro and in vivo. RESULTS: A univariate analysis showed that CD133 expression in male patients at the late stage (II and III) was significantly associated with worse progression-free survival (PFS) (P = 0.0057) and overall survival (OS) (P = 0.015), and a multivariate analysis showed a trend toward worse OS (P = 0.041). Male patients with advanced HCC exhibited an approximately 20-fold higher PD-L1 combined positive score (CPS) compared with those with HCC at an early stage. We successfully generated CD133 CAR-T and PD-1 s cells that could secrete PD-1 blocking scFv based on a sleeping beauty system involving minicircle vectors. CD133 CAR-T and PD-1 s cells exhibited significant antitumour activity against HCC in vitro and in xenograft mouse models. Thus, CD133 CAR-T and PD-1 s cells may be a therapeutically tractable strategy for targeting CD133-positive CSCs in male patients with advanced HCC. CONCLUSIONS: Our study provides a nonviral strategy for constructing CAR-T cells that could also secrete checkpoint blockade inhibitors based on a Sleeping Beauty system from minicircle vectors and revealed a potential benefit of this strategy for male patients with advanced HCC and high CD133 expression (median immunohistochemistry score > 2.284).

Universal allogeneic CAR T cells engineered with Sleeping Beauty transposons and CRISPR-CAS9 for cancer immunotherapy.

Allogeneic CD19-specific chimeric antigen receptor (CAR) T cells with inactivated donor T cell receptor (TCR) expression can be used as an "off-the-shelf" therapeutic modality for lymphoid malignancies, thus offering an attractive alternative to autologous, patient-derived T cells. Current approaches for T cell engineering mainly rely on the use of viral vectors. Here, we optimized and validated a non-viral genetic modification platform based on Sleeping Beauty (SB) transposons delivered with minicircles to express CD19-28z.CAR and CRISPR-Cas9 ribonucleoparticles to inactivate allogeneic TCRs. Efficient TCR gene disruption was achieved with minimal cytotoxicity and with attainment of robust and stable CD19-28z.CAR expression. The CAR T cells were responsive to CD19+ tumor cells with antitumor activities that induced complete tumor remission in NALM6 tumor-bearing mice while significantly reducing TCR alloreactivity and GvHD development. Single CAR signaling induced the similar T cell signaling signatures in TCR-disrupted CAR T cells and control CAR T cells. In contrast, TCR disruption inhibited T cell signaling/protein phosphorylation compared with the control CAR T cells during dual CAR/TCR signaling. This non-viral SB transposon-CRISPR-Cas9 combination strategy serves as an alternative for generating next-generation CD19-specific CAR T while reducing GvHD risk and easing potential manufacturing constraints intrinsic to viral vectors.

Face inversion effect on perceived cuteness of infant faces.

Research has demonstrated that attractiveness evaluations of adult faces were less accurate when faces were inverted than upright. It remains unknown, however, whether a similar effect applies to perceived cuteness of infants, which is assumed to be based on elemental facial features called the "baby schema." In this research, we studied the face inversion effect on perceived cuteness of infant faces in a rating task and a two-alternative forced-choice (2AFC) task. We also examined beauty as a control dimension. Although the rating task revealed no inversion effect, the 2AFC task showed poorer discrimination performance with inverted faces than with upright faces in both evaluations. These results indicate that infant cuteness and beauty dimensions are correlated well with each other, and their perception not only relies on elemental features that are not strongly affected by inversion but is also affected by holistic facial configurations when a detailed comparison is required.