Augmenting Immunotherapy Impact by Lowering Tumor TNF Cytotoxicity Threshold.

New opportunities are needed to increase immune checkpoint blockade (ICB) benefit. Whereas the interferon (IFN)γ pathway harbors both ICB resistance factors and therapeutic opportunities, this has not been systematically investigated for IFNγ-independent signaling routes. A genome-wide CRISPR/Cas9 screen to sensitize IFNγ receptor-deficient tumor cells to CD8 T cell elimination uncovered several hits mapping to the tumor necrosis factor (TNF) pathway. Clinically, we show that TNF antitumor activity is only limited in tumors at baseline and in ICB non-responders, correlating with its low abundance. Taking advantage of the genetic screen, we demonstrate that ablation of the top hit, TRAF2, lowers the TNF cytotoxicity threshold in tumors by redirecting TNF signaling to favor RIPK1-dependent apoptosis. TRAF2 loss greatly enhanced the therapeutic potential of pharmacologic inhibition of its interaction partner cIAP, another screen hit, thereby cooperating with ICB. Our results suggest that selective reduction of the TNF cytotoxicity threshold increases the susceptibility of tumors to immunotherapy.

Anti-tumour immunity induces aberrant peptide presentation in melanoma.

Extensive tumour inflammation, which is reflected by high levels of infiltrating T cells and interferon-γ (IFNγ) signalling, improves the response of patients with melanoma to checkpoint immunotherapy1,2. Many tumours, however, escape by activating cellular pathways that lead to immunosuppression. One such mechanism is the production of tryptophan metabolites along the kynurenine pathway by the enzyme indoleamine 2,3-dioxygenase 1 (IDO1), which is induced by IFNγ3-5. However, clinical trials using inhibition of IDO1 in combination with blockade of the PD1 pathway in patients with melanoma did not improve the efficacy of treatment compared to PD1 pathway blockade alone6,7, pointing to an incomplete understanding of the role of IDO1 and the consequent degradation of tryptophan in mRNA translation and cancer progression. Here we used ribosome profiling in melanoma cells to investigate the effects of prolonged IFNγ treatment on mRNA translation. Notably, we observed accumulations of ribosomes downstream of tryptophan codons, along with their expected stalling at the tryptophan codon. This suggested that ribosomes bypass tryptophan codons in the absence of tryptophan. A detailed examination of these tryptophan-associated accumulations of ribosomes-which we term 'W-bumps'-showed that they were characterized by ribosomal frameshifting events. Consistently, reporter assays combined with proteomic and immunopeptidomic analyses demonstrated the induction of ribosomal frameshifting, and the generation and presentation of aberrant trans-frame peptides at the cell surface after treatment with IFNγ. Priming of naive T cells from healthy donors with aberrant peptides induced peptide-specific T cells. Together, our results suggest that IDO1-mediated depletion of tryptophan, which is induced by IFNγ, has a role in the immune recognition of melanoma cells by contributing to diversification of the peptidome landscape.

Computational and functional analyses of T2D GWAS SNPs for transcription factor binding.

Genome-wide association studies (GWASs) have successfully identified numerous non-coding genetic variants for type 2 diabetes (T2D), but the functional roles underlying these non-coding variants remain largely unknown. The effects of T2D GWAS lead SNPs on transcriptional factors binding motifs were firstly analyzed via JASPAR, followed by functional validations including dual-luciferase reporter assays, biotin-based DNA pull-down assays, real-time quantitative PCR, and western blotting. The results showed that GWAS SNP rs4430796 conferred T allele specific transcriptional enhancer activity via a PAX6 binding element, and upregulated the expression of HNF1B. GWAS SNP rs4607103 showed a bidirectional modulation of ADAMTS9-AS2 and ADAMTS9 by TCF7L2 in a T allele-specific manner. GWAS SNP rs849135 conferred C allele-specific bidirectional transcriptional enhancer activity via a CREB1 binding element. Our findings have uncovered the functional mechanisms of three T2D GWAS SNPs via affecting the binding of transcription factors, providing new insights into the genetics and molecular pathogenesis of T2D.

Unconditional Steady-State Entanglement in Macroscopic Hybrid Systems by Coherent Noise Cancellation.

The generation of entanglement between disparate physical objects is a key ingredient in the field of quantum technologies, since they can have different functionalities in a quantum network. Here we propose and analyze a generic approach to steady-state entanglement generation between two oscillators with different temperatures and decoherence properties coupled in cascade to a common unidirectional light field. The scheme is based on a combination of coherent noise cancellation and dynamical cooling techniques for two oscillators with effective masses of opposite signs, such as quasispin and motional degrees of freedom, respectively. The interference effect provided by the cascaded setup can be tuned to implement additional noise cancellation leading to improved entanglement even in the presence of a hot thermal environment. The unconditional entanglement generation is advantageous since it provides a ready-to-use quantum resource. Remarkably, by comparing to the conditional entanglement achievable in the dynamically stable regime, we find our unconditional scheme to deliver a virtually identical performance when operated optimally.

Hepatocellular Carcinoma: Retrospective Evaluation of the Correlation Between Gadobenate Dimeglumine-Enhanced Magnetic Resonance Imaging and Pathologic Grade.

OBJECTIVE: The aim of this study was to evaluate the usefulness of gadobenate dimeglumine-enhanced magnetic resonance imaging in characterizing the grade of hepatocellular carcinoma (HCC) using the signal intensity (SI) of the erector spinae as internal reference. MATERIALS AND METHODS: Clinical data of 40 patients (a total of 44 lesions) confirmed by pathology for HCC were retrospectively reviewed. Gadobenate dimeglumine-enhanced magnetic resonance imaging was performed in all patients, and SI of lesions (SIles), liver parenchyma around the lesions (SIhep), erector spinae (SImus) and standard deviation of SI of the surrounding noise (SDnoi) on nonenhanced T2WI, nonenhanced T1WI, and contrast-enhanced T1WI (in both arterial and hepatobiliary phase [AP and HBP]) were measured, respectively. Contrast-to-noise ratio (CNR) were separately defined as CNR1 ([SIles - SIhep]/SDnoi) and CNR2 ([SIles - SImus]/SDnoi). Statistical analyses were performed using one-way analysis of variance, least significant difference test, logistic regression analysis, Spearman rank correlation, and receiver operating characteristic curves analysis. RESULTS: Forty-four HCCs, including 3 well-differentiated HCCs, 26 moderately differentiated HCCs, and 15 poorly differentiated (PD) HCCs, were confirmed. On logistic regression analysis, only CNR2 in the HBP was predictor of PD HCCs (P = 0.015, odds ratio = 1.040). The size of lesions, CNR1 in the AP, CNR2 in the AP, and CNR2 in the HBP, showed significant correlations with the degree of differentiation (correlation coefficients = -0.371, 0.435, 0.503, and 0.512, respectively; P = 0.013, 0.003, 0.001, and 0.000, respectively). Contrast-to-noise ratio 2 in the HBP with the cutoff of less than 4.56 could distinguish moderately differentiated HCCs from PD HCC with the sensitivity and specificity of 84.6% and 60.0%, respectively. CONCLUSIONS: Relatively low arterial enhancement and low CNR2 value in the HBP are predictive for poor histological grade of HCCs.

Influence of Access Cavities on Maxillary Central Incisor Fracture Resistance: Finite Element Study.

INTRODUCTION AND AIMS: Altering the position and orientation of the root canal access cavity passway, or modifying the reduction of dentin volume, can influence the strength of dentition. This study aimed to compare the effects of different access cavities on the biomechanical performances of maxillary central incisors with a finite element analysis. METHODS: Based on the micro-computed tomography (CT) scan of a maxillary central incisor, the finite element models of the intact tooth and teeth with 4 access cavity designs: conservative incisal access cavity, incisal access cavity, conservative access cavity, and traditional access cavity were generated. Simulated occlusal forces were applied at the incisal edge of the incisor in the finite element analysis procedure. RESULTS: The maximum von Mises stress and maximum principal stress in the cervical area are highest in the traditional access cavity group, followed by the conservative access cavity group, incisal access cavity group, and conservative incisal access cavity group. CONCLUSION: The conservative access cavities minimise the extent of dentin removal from the cervical region, protecting the mechanical behaviour of the incisor. Moving the access cavity entry point to the incisal edge also improves the fracture resistance of the incisor. CLINICAL RELEVANCE: This study's findings would help clinicians select the most appropriate endodontics access cavity method when performing the root canal on maxillary central incisors.

Arthroscopic and open approaches for autologous matrix-induced chondrogenesis repair of the knee have similar results: a meta-analysis.

BACKGROUND: Cartilage defects are debilitating injuries that can reduce quality of life in patients. However, the poor regenerative properties of cartilage mean that cartilage repair remains challenging, and many methods have arisen to address that. Autologous matrix-induced chondrogenesis (AMIC®) is a popular technique to manage cartilage defects. Recent advances have allowed AMIC® to be done arthroscopically, instead of a mini-open arthrotomy approach. This systematic review and meta-analysis aims to investigate whether the arthroscopic approach to AMIC® provides better clinical outcomes than does the mini-open approach, in hopes of delineating a gold standard in cartilage repair. METHODS: With reference to the Preferred Reporting Items for Systematic Reviews and Meta-analyses guidelines, a systematic search of the following databases (PubMed, Embase, Scopus, and Cochrane Library) was performed on 26th October 2022 using a combination of the following search terms: "autologous matrix induced", "chondrogenesis", and "knee". A total of 390 studies were identified, of which, 24 studies were included in our final analysis. RESULTS: The arthroscopic approach achieves lower Visual Analogue Scale for pain scores. The International Knee documentation Committee) score and Knee Injury and Osteoarthritis Outcome Score were comparable between arthroscopic and open approaches. The open approach achieves a higher Magnetic Resonance Observation of Cartilage Repair Tissue score. Incidence of reported postoperative complications of revision surgery and knee stiffness was higher for the open approach than for the arthroscopic approach, whereas deep vein thrombosis was higher in the arthroscopic approach. CONCLUSION: The AMIC® repair outcomes indicate that the arthroscopic approach does not hold a distinct advantage over the open approach. The choice of approach should consider surgeon expertise, location of lesion, and patient-specific factors. LEVEL OF EVIDENCE: Systematic review and meta-analysis; Level III.

The osteoporosis susceptibility SNP rs188303909 at 2q14.2 regulates EN1 expression by modulating DNA methylation and E2F6 binding.

EN1 encodes a homeodomain-containing transcription factor and is a determinant of bone density and fracture. Previous powerful genome-wide association studies (GWASs) have identified multiple single-nucleotide polymorphisms (SNPs) near EN1 at 2q14.2 locus for osteoporosis, but the causal SNPs and functional mechanisms underlying these associations are poorly understood. The target genes regulated by the transcription factor EN1 are also unclear. In this study, we identified rs188303909, a functional CpG-SNP, as a causal SNP for osteoporosis at 2q14.2 through the integration of functional and epigenomic analyses. Functional experiments demonstrated that unmethylated rs188303909 acted as a strong allele-specific distal enhancer to regulate EN1 expression by modifying the binding of transcription factor E2F6, but rs188303909 methylation attenuated the active effect of E2F6 on EN1 expression. Importantly, transcription factor EN1 could differentially bind osteoporosis GWAS lead SNPs rs4869739-T and rs4355801-G to upregulate CCDC170 and COLEC10 expression, thus promoting bone formation. Our study provided a mechanistic insight into expression regulation of the osteoporosis susceptibility gene EN1, which could be a potential therapeutic target for osteoporosis precision medicine. KEY MESSAGES: CpG-SNP rs188303909 is a causal SNP at the osteoporosis susceptibility locus 2q14.2. Rs188303909 distally regulates EN1 expression by modulating DNA methylation and E2F6 binding. EN1 upregulates CCDC170 and COLEC10 expression through osteoporosis GWAS lead SNPs rs4869739 and rs4355801.

An adverse tumor-protective effect of IDO1 inhibition.

By restoring tryptophan, indoleamine 2,3-dioxygenase 1 (IDO1) inhibitors aim to reactivate anti-tumor T cells. However, a phase III trial assessing their clinical benefit failed, prompting us to revisit the role of IDO1 in tumor cells under T cell attack. We show here that IDO1 inhibition leads to an adverse protection of melanoma cells to T cell-derived interferon-gamma (IFNγ). RNA sequencing and ribosome profiling shows that IFNγ shuts down general protein translation, which is reversed by IDO1 inhibition. Impaired translation is accompanied by an amino acid deprivation-dependent stress response driving activating transcription factor-4 (ATF4)high/microphtalmia-associated transcription factor (MITF)low transcriptomic signatures, also in patient melanomas. Single-cell sequencing analysis reveals that MITF downregulation upon immune checkpoint blockade treatment predicts improved patient outcome. Conversely, MITF restoration in cultured melanoma cells causes T cell resistance. These results highlight the critical role of tryptophan and MITF in the melanoma response to T cell-derived IFNγ and uncover an unexpected negative consequence of IDO1 inhibition.

Feather Damage Monitoring System Using RGB-Depth-Thermal Model for Chickens.

Feather damage is a continuous health and welfare challenge among laying hens. Infrared thermography is a tool that can evaluate the changes in the surface temperature, derived from an inflammatory process that would make it possible to objectively determine the depth of the damage to the dermis. Therefore, the objective of this article was to develop an approach to feather damage assessment based on visible light and infrared thermography. Fusing information obtained from these two bands can highlight their strengths, which is more evident in the assessment of feather damage. A novel pipeline was proposed to reconstruct the RGB-Depth-Thermal maps of the chicken using binocular color cameras and a thermal infrared camera. The process of stereo matching based on binocular color images allowed for a depth image to be obtained. Then, a heterogeneous image registration method was presented to achieve image alignment between thermal infrared and color images so that the thermal infrared image was also aligned with the depth image. The chicken image was segmented from the background using a deep learning-based network based on the color and depth images. Four kinds of images, namely, color, depth, thermal and mask, were utilized as inputs to reconstruct the 3D model of a chicken with RGB-Depth-Thermal maps. The depth of feather damage can be better assessed with the proposed model compared to the 2D thermal infrared image or color image during both day and night, which provided a reference for further research in poultry farming.

Loss-induced nonreciprocity.

Nonreciprocity is important in both optical information processing and topological photonics studies. Conventional principles for realizing nonreciprocity rely on magnetic fields, spatiotemporal modulation, or nonlinearity. Here we propose a generic principle for generating nonreciprocity by taking advantage of energy loss, which is usually regarded as harmful. The loss in a resonance mode induces a phase lag, which is independent of the energy transmission direction. When multichannel lossy resonance modes are combined, the resulting interference gives rise to nonreciprocity, with different coupling strengths for the forward and backward directions, and unidirectional energy transmission. This study opens a new avenue for the design of nonreciprocal devices without stringent requirements.

Influence of post-traumatic status on health-related quality of life among survivors 10 years after the Wenchuan earthquake in China.

BACKGROUND: China is a country with frequent earthquakes. Beichuan was the epicenter of what was later called the Wenchuan earthquake, which caused nearly 80% of the houses in the area to collapse, with 8,605 people killed and 9,693 people missing. The aim of our study was to explore the prevalence of post-traumatic stress disorder (PTSD) and its influence on health-related quality of life (HRQOL) among the survivors 10 years after the 2008 Wenchuan earthquake in China. METHODS: A total of 610 survivors from Leigu town in Beichuan County were investigated by a systematic sampling method. Post-traumatic status, HRQOL, and demographic sources were measured by the PTSD Checklist Civilian Version (PCL-C), 36-item Health Survey Short Form (SF-36), and self-questionnaire, respectively. RESULTS: Ten years after the Wenchuan earthquake, the prevalence of PTSD for survivors was 1.6%. There were significant negative correlations between survivors' SF-36 scores and the scores of PCL-C. Higher scores in post-traumatic status were associated with a higher rate of poor physical HRQOL, which was lower than the mean score [adjusted odds ratio (OR) 1/4 0.96 per SD increase, P 1/4 0.001] and mental HRQOL (adjusted OR 1/4 0.93 per SD increase, P 1/4 0.001). The independent contribution of post-traumatic status to the risk for poor physical and mental HRQOL was 4.9% and 18.7% respectively. CONCLUSIONS: As time has elapsed, the incidence of PTSD has gradually declined after the Wenchuan earthquake. Post-traumatic status was found to influence the health related quality of life of survivors.

The osteoporosis risk variant rs9820407 at 3p22.1 acts as an allele-specific enhancer to regulate CTNNB1 expression by long-range chromatin loop formation.

Previous powerful genome-wide association studies (GWASs) and whole-genome sequencing have identified multiple single-nucleotide polymorphisms (SNPs) located over 69 kb upstream of CTNNB1 at 3p22.1 locus associated with osteoporosis. The CTNNB1 gene encodes ß-catenin that is an integral part of adherens junctions and the primary mediator of the canonical Wnt signaling pathway. The causal variants and underlying molecular mechanisms of the osteoporosis susceptibility locus 3p22.1 remains unknown. Through comprehensive computational analyses, including expression quantitative trait locus (eQTL), high-throughput chromatin interaction (Hi-C), epigenomic and functional annotation, four enhancer SNPs (rs9820407, rs9878224, rs454690 and rs9832204) were prioritized as potential causal SNPs at 3p22.1 for osteoporosis. Rs9820407 displayed the strongest enhancer activity in dual-luciferase assays. Specifically, the minor rs9820407-A can preferentially bind transcription factor FOXC1, elevate the enhancer activity and increase CTNNB1 expression. The architectural protein CTCF was presumably involved in long-range chromatin interaction between rs9820407 and CTNNB1. Our study provided a mechanistic insight into how noncoding enhancer SNP rs9820407 distally regulates CTNNB1 expression and modulates osteoporosis risk.

Nondestructive monitoring of polyphenols and caffeine during green tea processing using Vis-NIR spectroscopy.

Increasing consumption of green tea is attributed to the beneficial effects of its constituents, especially polyphenols, on human health, which can be varied during leaf processing. Processing technology has the most important effect on green tea quality. This study investigated the system dynamics of eight catechins, gallic acid, and caffeine in the processing of two varieties of tea, from fresh leaves to finished tea. It was found that complex biochemical changes can occur through hydrolysis under different humidity and heating conditions during the tea processing. This process had a significant effect on catechin composition in the finished tea. The potential application of visible and near-infrared (Vis-NIR) spectroscopy for fast monitoring polyphenol and caffeine contents in tea leaves during the processing procedure has been investigated. It was found that a combination of PCA (principal component analysis) and Vis-NIR spectroscopy can successfully classify the two varieties of tea samples and the five tea processing procedures, while quantitative determination of the constituents was realized by combined regression analysis and Vis-NIR spectra. Furthermore, successive projections algorithm (SPA) was proposed to extract and optimize spectral variables that reflected the molecular characteristics of the constituents for the development of determination models. Modeling results showed that the models had good predictability and robustness based on the extracted spectral characteristics. The coefficients of determination for all calibration sets and prediction sets were higher than 0.862 and 0.834, respectively, which indicated high capability of Vis-NIR spectroscopy for the determination of the constituents during the leaf processing. Meanwhile, this analytical method could quickly monitor quality characteristics and provide feedback for real-time controlling of tea processing machines. Furthermore, the study on complex biochemical changes that occurred during the tea processing would provide a theoretical basis for improving the content of quality components and effective controlling processes.

Parallel in vivo and in vitro melanoma RNAi dropout screens reveal synthetic lethality between hypoxia and DNA damage response inhibition.

To identify factors preferentially necessary for driving tumor expansion, we performed parallel in vitro and in vivo negative-selection short hairpin RNA (shRNA) screens. Melanoma cells harboring shRNAs targeting several DNA damage response (DDR) kinases had a greater selective disadvantage in vivo than in vitro, indicating an essential contribution of these factors during tumor expansion. In growing tumors, DDR kinases were activated following hypoxia. Correspondingly, depletion or pharmacologic inhibition of DDR kinases was toxic to melanoma cells, including those that were resistant to BRAF inhibitor, and this could be enhanced by angiogenesis blockade. These results reveal that hypoxia sensitizes melanomas to targeted inhibition of the DDR and illustrate the utility of in vivo shRNA dropout screens for the identification of pharmacologically tractable targets.