Whole-Slide Imaging, Mutual Information Registration for Multiplex Immunohistochemistry and Immunofluorescence.

Multiplex immunohistochemistry/immunofluorescence (mIHC/mIF) is a developing technology that facilitates the evaluation of multiple, simultaneous protein expressions at single-cell resolution while preserving tissue architecture. These approaches have shown great potential for biomarker discovery, yet many challenges remain. Importantly, streamlined cross-registration of multiplex immunofluorescence images with additional imaging modalities and immunohistochemistry (IHC) can help increase the plex and/or improve the quality of the data generated by potentiating downstream processes such as cell segmentation. To address this problem, a fully automated process was designed to perform a hierarchical, parallelizable, and deformable registration of multiplexed digital whole-slide images (WSIs). We generalized the calculation of mutual information as a registration criterion to an arbitrary number of dimensions, making it well suited for multiplexed imaging. We also used the self-information of a given IF channel as a criterion to select the optimal channels to use for registration. Additionally, as precise labeling of cellular membranes in situ is essential for robust cell segmentation, a pan-membrane immunohistochemical staining method was developed for incorporation into mIF panels or for use as an IHC followed by cross-registration. In this study, we demonstrate this process by registering whole-slide 6-plex/7-color mIF images with whole-slide brightfield mIHC images, including a CD3 and a pan-membrane stain. Our algorithm, WSI, mutual information registration (WSIMIR), performed highly accurate registration allowing the retrospective generation of an 8-plex/9-color, WSI, and outperformed 2 alternative automated methods for cross-registration by Jaccard index and Dice similarity coefficient (WSIMIR vs automated WARPY, P < .01 and P < .01, respectively, vs HALO + transformix, P = .083 and P = .049, respectively). Furthermore, the addition of a pan-membrane IHC stain cross-registered to an mIF panel facilitated improved automated cell segmentation across mIF WSIs, as measured by significantly increased correct detections, Jaccard index (0.78 vs 0.65), and Dice similarity coefficient (0.88 vs 0.79).

Cu2+-Pyropheophorbide a-Cystine Conjugate: Synergistic Photodynamic/Chemodynamic Therapy and Glutathione Depletion Improves the Antitumor Efficacy and Downregulates the Hypoxia-Inducing Factor.

Cancer immune escape, metastasis, recurrence, and multidrug resistance are all associated with hypoxia in the tumor microenvironment (TME). We synthesized a CuPPaCC conjugate for reactive oxygen species (ROS)-mediated cancer therapy. CuPPaCC continuously produced cytotoxic ROS and oxygen through a photo-chemocycloreaction, alleviated hypoxia, and inhibited the expression of a hypoxia-inducing factor (HIF-1α). CuPPaCC was synthesized from pyromania phyllophyllic acid a (PPa), cystine (CC), and copper ions, and its structure was characterized by nuclear magnetic resonance (NMR) and mass spectrometry (MS). The ability of CuPPaCC to produce ROS and oxygen after photodynamic therapy (PDT) in vitro and in vivo was investigated. The ability of CuPPaCC to consume glutathione was investigated. CuPPaCC toxicity (light and dark) in CT26 cells was analyzed by MTT and live/dead cell staining. The anticancer effect of CuPPaCC in vivo was investigated in CT26 Balb/c mice. When stimulated by the TME, CuPPaCC released Cu2+ and PPaCC, and the singlet oxygen yield increased from 34 to 56.5%. The dual ROS-generating mechanism via a Fenton-like reaction/photoreaction and dual glutathione depletion via Cu2+/CC multiplied the antitumor efficacy of CuPPaCC. The photo-chemocycloreaction continued to produce oxygen and maintained high ROS levels even after PDT, significantly alleviating hypoxia in the TME and downregulating the expression of HIF-1α. CuPPaCC thus showed excellent antitumor activity in vitro and in vivo. These results showed that the strategy could be effective in improving the antitumor efficacy of CuPPaCC and could be used as a synergistic regimen for cancer therapy.

Diagnosis of breast implant associated anaplastic large cell lymphoma by analysis of cytokines in peri-implant seromas.

More than 1300 women with breast implants have developed an anaplastic large cell lymphoma (ALCL) in fluid (seroma) around their implant. More often, seromas are due to benign causes, for example, capsule contracture, leakage, or trauma. Our report in American Journal of Hematology identified several cytokines (IL-9, IL-10, IL-13) as significantly elevated only in seromas due to ALCL. We further showed that the most robust biomarker, IL-10, could be detected by a lateral flow assay (similar to COVID detection) within minutes allowing physicians to quickly plan management, eliminate or reduce costly testing and patient time away from family. Early detection of ALCL in seromas before infiltration may avoid need for cytotoxic or immunotherapy and is possibly life-saving.

Exciton-polariton light-emitting diode based on a single ZnO superlattice microwire heterojunction with performance enhanced by Rh nanostructures.

One-dimensional (1D) wirelike superlattice micro/nanostructures have received considerable attention for potential applications due to their versatility and capability for modulating optical and electrical characteristics. In this study, 1D superlattice microwires (MWs), which are made of undoped ZnO and Ga-doped ZnO with periodic and alternating crystalline layers (ZnO/ZnO:Ga), were synthesized individually. Under optical excitation, a series of resonance peaks in the photoluminescence spectrum can be ascribed to polariton emission, which originates from the coupling interaction of the 1D photonic crystal and confined excitons along the wire direction. Using a p-type GaN layer as the hole transport layer, a kind of waveguide light source based on an individual ZnO/ZnO:Ga superlattice MW was proposed and constructed. By analysing the spatially resolved electroluminescence spectra, the observed multipeak was ascribed to exciton-polariton emission with a vacuum Rabi splitting of about 275 meV. Cladding with Rh nanostructures gives rise to appropriate ultraviolet plasmons, and the Rabi splitting energy of our device was enhanced up to 413 meV. The exciton-polariton properties were further examined using angle-resolved electroluminescence measurements. Therefore, individual superlattice MWs can act as optical microresonators to achieve photon-exciton coupling with a large Rabi splitting energy. The experimental results indicate that an individual ZnO/ZnO:Ga superlattice MW can be generally used in developing exciton-polariton luminescence/lasing light sources, particularly for constructing low-threshold/thresholdless lasers toward pragmatic applications.

Plasmonic and catalytic Au NBP@AgPd nanoframes for highly efficient photocatalytic reactions.

Heterogeneous metal nanostructures with excellent plasmonic performance and catalytic activity are urgently needed to realize efficient light-driven catalysis. Herein, we demonstrate the preparation of hollow Au nanobipyramid (NBP)@AgPd nanostructures by employing Au NBP@Ag nanorods as templates. The products could transform from Au NBP@AgPd nanoframes to nanocages, along with the redshift and broadening of the plasmon wavelength. Particularly, the plasmon intensity of these nanostructures remained considerable among the shape evolution process. Based on the selective absorption of CTAB, the Ag atoms on the side surfaces of the Au NBP@Ag nanorods were employed as the sacrificial templates to reduce Pd atoms through galvanic replacement. The reduced Pd and Ag atoms produced through the reduction reaction were preferably co-deposited on the corners and edges at the early stage and later deposited directly on the defect sites of the side facets, as more Ag atoms were released. The discontinued distribution of the Pd atoms gives an opportunity to etch away the Ag atoms in the cores, leading to the formation of hollow Au NBP@AgPd nanostructures after the etching process. It is worth noting that the deposition of the ultrathin AgPd nanoframe had little influence on the plasmonic properties of Au NBPs, as verified by electrodynamic simulations. The Au NBP@AgPd nanoframe showed great photocatalytic activity toward Suzuki coupling reactions under laser irradiation. Taken together, these results suggest that the hot electrons successfully transfer from Au NBP to the AgPd nanoframes to participate in the photocatalytic reactions. This study affords a promising route for the synthesis of anisotropic bimetallic nanostructures with excellent plasmonic performances.

Atractylodin targets GLA to regulate D-mannose metabolism to inhibit osteogenic differentiation of human valve interstitial cells and ameliorate aortic valve calcification.

Atractylodin (ATL) has been reported to exert anti-inflammatory effects. Osteogenic changes induced by inflammation in valve interstitial cells (VICs) play a key role in the development of calcified aortic valve disease (CAVD). This study aimed to investigate the anti-calcification effects of ATL on aortic valves. Human VICs (hVICs) were exposed to osteogenic induction medium (OM) containing ATL to investigate cell viability, osteogenic gene and protein expression, and anti-calcification effects. Gas chromatography-mass spectroscopy (GC-MS) metabolomics analysis was used to detect changes in the metabolites of hVICs stimulated with OM before and after ATL administration. The compound-reaction-enzyme-gene network was used to identify drug targets. Gene interference was used to verify the targets. ApoE-/- mice fed a high-fat (HF) diet were used to evaluate the inhibition of aortic valve calcification by ATL. Treatment with 20 µM ATL in OM prevented calcified nodule accumulation and decreases in the gene and protein expression levels of ALP, RUNX2, and IL-1ß. Differential metabolite analysis showed that D-mannose was highly associated with the anti-calcification effect of ATL. The addition of D-mannose prevented calcified nodule accumulation and inhibited succinate-mediated HIF-1α activation and IL-1ß production. The target of ATL was identified as GLA. Silencing of the GLA gene (si-GLA) reversed the anti-osteogenic differentiation of ATL. In vivo, ATL ameliorated aortic valve calcification by preventing decreases in GLA expression and the up-regulation of IL-1ß expression synchronously. In conclusion, ATL is a potential drug for the treatment of CAVD by targeting GLA to regulate D-mannose metabolism, thereby inhibiting succinate-mediated HIF-1α activation and IL-1ß production.

Atractylenolide-1 affects glycolysis/gluconeogenesis by downregulating the expression of TPI1 and GPI to inhibit the proliferation and invasion of human triple-negative breast cancer cells.

Atractylenolide-1 (AT-1) is a major octanol alkaloid isolated from Atractylodes Rhizoma and is widely used to treat various diseases. However, few reports have addressed the anticancer potential of AT-1, and the underlying molecular mechanisms of its anticancer effects are unclear. This study aimed to assess the effect of AT-1 on triple-negative breast cancer (TNBC) cell proliferation and migration and explore its potential molecular mechanisms. Cell invasion assays confirmed that the number of migrating cells decreased after AT-1 treatment. Colony formation assays showed that AT-1 treatment impaired the ability of MDA-MB-231 cells to form colonies. AT-1 inhibited the expression of p-p38, p-ERK, and p-AKT in MDA-MB-231 cells, significantly downregulated the proliferation of anti-apoptosis-related proteins CDK1, CCND1, and Bcl2, and up-regulated pro-apoptotic proteins Bak, caspase 3, and caspase 9. The gas chromatography-mass spectroscopy results showed that AT-1 downregulated the metabolism-related genes TPI1 and GPI through the glycolysis/gluconeogenesis pathway and inhibited tumor growth in vivo. AT-1 affected glycolysis/gluconeogenesis by downregulating the expression of TPI1 and GPI, inhibiting the proliferation, migration, and invasion of (TNBC) MDA-MB-231 cells and suppressing tumor growth in vivo.

Nutrient status regulates MED19a phase separation for ORESARA1-dependent senescence.

The mediator complex is highly conserved in eukgaryotes and is integral for transcriptional responses. Mediator subunits associate with signal-responsive transcription factors (TF) to activate expression of specific signal-responsive genes. As the key TF of Arabidopsis thaliana senescence, ORESARA1 (ORE1) is required for nitrogen deficiency (-N) induced senescence; however, the mediator subunit that associates with ORE1 remains unknown. Here, we show that Arabidopsis MED19a associates with ORE1 to activate -N senescence-responsive genes. Disordered MED19a forms inducible nuclear condensates under -N that is regulated by decreasing MED19a lysine acetylation. MED19a carboxyl terminus (cMED19a) harbors a mixed-charged intrinsically disordered region (MC-IDR) required for ORE1 interaction and liquid-liquid phase separation (LLPS). Plant and human cMED19 are sufficient to form heterotypic condensates with ORE1. Human cMED19 MC-IDR, but not yeast cMED19 IDR, partially complements med19a suggesting functional conservation in evolutionarily distant eukaryotes. Phylogenetic analysis of eukaryotic cMED19 revealed that the MC-IDR could arise through convergent evolution. Our result of MED19 MC-IDR suggests that plant MED19 is regulated by phase separation during stress responses.

Nonmalignant CD30+ Cells in Contralateral Peri-Implant Capsule of Patient With BIA-ALCL: A Premalignant Step?

CD30 lymphocyte activation antigen and phosphorylated STAT3 (pSTAT3) are consistent markers of tumor cells in breast implant-associated anaplastic large cell lymphoma (BIA-ALCL). We present a case of BIA-ALCL in a breast implant capsule containing clustered tumor cells expressing CD30, pSTAT3, pSTAT6, interleukin 9, and granzyme B tumor cell biomarkers. Remarkably, the contralateral breast contained many scattered large, atypical CD30+ cells surrounded by inflammatory cells, raising a suspicion of bilateral BIA-ALCL, known to occur in some patients. To clarify the diagnosis, immunohistochemistry and multilabel immunofluorescence were performed. Unlike the tumor cells, the atypical CD30+ cells of the contralateral breast lacked pSTAT3, pSTAT6, interleukin 9, and granzyme B, eliminating a diagnosis of bilateral BIA-ALCL. This case highlights the importance of interpreting CD30 staining in the context of other tumor cell biomarkers and histopathology to avoid an incorrect diagnosis of BIA-ALCL. We believe the findings also suggest the possibility of CD30 expression as an early event in the multistep pathogenesis of BIA-ALCL.

[Study on Parametric Release of Ethylene Oxide Sterilization of Medical Devices].

This study briefly introduces the basic theory of sterilization, the characteristics of ethylene oxide sterilization for medical devices and the key factors about sterilization effectiveness, analyzes and compares three methods used in the product release of medical devices sterilized by ethylene oxide: test for sterility, traditional release and parametric release, and focuses on the theoretical basis, feasibility, validation requirements, advantages and disadvantages of parametric release.

Granzyme B Is a Biomarker for Suspicion of Malignant Seromas Around Breast Implants.

BACKGROUND: Granzyme B (GrB) is a serine protease secreted, along with pore-forming perforin, by cytotoxic lymphocytes to mediate apoptosis in target cells. GrB has been detected in tumor cells associated with systemic and breast implant-associated anaplastic large cell lymphoma (BIA-ALCL) but its potential use for detection of early BIA-ALCL has not been fully investigated. OBJECTIVES: Prompted by the increased incidence of BIA-ALCL, the aim of this study was to assess GrB as a new biomarker to detect early disease in malignant seromas and to better understand the nature of the neoplastic cell. METHODS: A Human XL Cytokine Discovery Magnetic Luminex 45-plex Fixed Panel Performance Assay was used to compare cytokine levels in cell culture supernatants of BIA-ALCL and other T-cell lymphomas, as well as malignant and benign seromas surrounding breast implants. Immunohistochemistry was employed to localize GrB to cells in seromas and capsular infiltrates. RESULTS: Differences in GrB concentrations between malignant and benign seromas were significant (P < 0.001). GrB was found in and around apoptotic tumor cells, suggesting that the protease may be involved in tumor cell death. CONCLUSIONS: GrB is a useful marker for early detection of malignant seromas and to identify tumor cells in seromas and capsular infiltrates. Because there is an overlap between the lowest concentrations of soluble GrB in malignant seromas and the highest concentrations of GrB in benign seromas, it is recommended that GrB be used only as part of a panel of biomarkers for the screening and early detection of BIA-ALCL.

Novel stable QTLs identification for berry quality traits based on high-density genetic linkage map construction in table grape.

BACKGROUND: Aroma, berry firmness and berry shape are three main quality traits in table grape production, and also the important target traits in grapevine breeding. However, the information about their genetic mechanisms is limited, which results in low accuracy and efficiency of quality breeding in grapevine. Mapping and isolation of quantitative trait locus (QTLs) based on the construction of genetic linkage map is a powerful approach to decipher the genetic determinants of complex quantitative traits. RESULTS: In the present work, a final integrated map consisting of 3411 SLAF markers on 19 linkage groups (LGs) with an average distance of 0.98 cM between adjacent markers was generated using the specific length amplified fragment sequencing (SLAF-seq) technique. A total of 9 significant stable QTLs for Muscat flavor, berry firmness and berry shape were identified on two linkage groups among the hybrids analyzed over three consecutive years from 2016 to 2018. Notably, new stable QTLs for berry firmness and berry shape were found on LG 8 respectively for the first time. Based on biological function and expression profiles of candidate genes in the major QTL regions, 3 genes (VIT_08s0007g00440, VIT_08s0040g02740 and VIT_08s0040g02350) related to berry firmness and 3 genes (VIT_08s0032g01110, VIT_08s0032g01150 and VIT_08s0105g00200) linked to berry shape were highlighted. Overexpression of VIT_08s0032g01110 in transgenic Arabidopsis plants caused the change of pod shape. CONCLUSIONS: A new high-density genetic map with total 3411 markers was constructed with SLAF-seq technique, and thus enabled the detection of narrow interval QTLs for relevant traits in grapevine. VIT_08s0007g00440, VIT_08s0040g02740 and VIT_08s0040g02350 were found to be related to berry firmness, while VIT_08s0032g01110, VIT_08s0032g01150 and VIT_08s0105g00200 were linked to berry shape.

High performance lasing in a single ZnO microwire using Rh nanocubes.

High-purity and size-controlled Rh nanocubes (RhNCs) with plasmonic responses in the ultraviolet spectrum range were synthesized; the ultraviolet plasmonic features of RhNCs have potential applications in wide bandgap semiconductors and optoelectronic devices because of their optical tunability and stability, as well as the compatibility with neighboring semiconductor micro/nanostructures. In this work, by incorporating RhNCs, the near-band-edge emission of a single ZnO microwire is considerably enhanced. When optically pumped by a fs pulsed laser at room temperature, RhNCs-plasmon enhanced high-performance whispering gallery mode (WGM) lasing characteristics, including lower lasing threshold, higher Q-factor, and lasing output enhancement, can be achieved from a single ZnO microwire covered by RhNCs. To further probe the modulation effect of RhNCs plasmons on the lasing characteristics of the ZnO microwires, time-resolved photoluminescence (TRPL) and electromagnetic simulation analyses were also performed. Based on our results, it can be concluded that size-controlled RhNCs with ultraviolet energy-tunable plasmons have the potential for use in optoelectronic devices requiring stable and high-performance in the short wavelength spectrum band owing to their unique ultraviolet plasmonic features.

B7-h2 is a costimulatory ligand for CD28 in human.

CD28 and CTLA-4 are cell surface cosignaling molecules essential for the control of T cell activation upon the engagement of their ligands B7-1 and B7-2 from antigen-presenting cells. By employing a receptor array assay, we have demonstrated that B7-H2, best known as the ligand of inducible costimulator, was a ligand for CD28 and CTLA-4 in human, whereas these interactions were not conserved in mouse. B7-H2 and B7-1 or B7-2 interacted with CD28 through distinctive domains. B7-H2-CD28 interaction was essential for the costimulation of human T cells' primary responses to allogeneic antigens and memory recall responses. Similar to B7-1 and B7-2, B7-H2 costimulation via CD28 induced survival factor Bcl-xL, downregulated cell cycle inhibitor p27(kip1), and triggered signaling cascade of ERK and AKT kinase-dependent pathways. Our findings warrant re-evaluation of CD28 and CTLA-4's functions previously attributed exclusively to B7-1 and B7-2 and have important implications in therapeutic interventions against human diseases.

Closed-Loop Control of Droplet Transfer in Electron-Beam Freeform Fabrication.

In the process of electron-beam freeform fabrication deposition, the surface of the deposit layer becomes rough because of the instability of the feeding wire and the changing of the thermal diffusion condition. This will make the droplet transfer distance change in the deposition process, and the droplet transfer cannot always be stable in the liquid bridge transfer state. It is easy to form a large droplet or make wire and substrate stick together, which makes the deposition quality worsen or even interrupts the deposition process. The current electron-beam freeform fabrication deposition is mostly open-loop control, so it is urgent to realize the real-time and closed-loop control of the droplet transfer and to make it stable in the liquid bridge transfer state. In this paper, a real-time monitoring method based on machine vision is proposed for the droplet transfer of electron-beam freeform fabrication. The detection accuracy is up to ± 0.08 mm. Based on this method, the measured droplet transfer distance is fed back to the platform control system in real time. This closed-loop control system can stabilize the droplet transfer distance within ± 0.14 mm. In order to improve the detection stability of the whole system, a droplet transfer detection algorithm suitable for this scenario has been written, which improves the adaptability of the droplet transfer distance detection method by means of dilatation/erosion, local minimum value suppression, and image segmentation. This algorithm can resist multiple disturbances, such as spatter, large droplet occlusion and so on.

Comparative Analysis of Cytokines of Tumor Cell Lines, Malignant and Benign Effusions Around Breast Implants.

BACKGROUND: More than 700 women have developed an anaplastic large T cell lymphoma (ALCL) surrounding textured surface breast implants, termed breast implant-associated ALCL (BIA-ALCL). Most patients with BIA-ALCL present with an accumulation of fluid (delayed seroma) around the implant. However, benign seromas without malignant cells complicating scar contracture, implant rupture, trauma, infection, and other causes are more common. For proper patient management and to avoid unnecessary surgery, a simple diagnostic test to identify malignant seromas is desirable. OBJECTIVES: The aim of this study was to develop an ancillary test for the diagnosis of malignant seromas and to gain insight into the nature of the malignant cells and their microenvironment. METHODS: We employed an immunologic assay on only 50 µL of aspirated seroma fluid. The assay measures 13 cytokines simultaneously by flow cytometry. To establish a baseline for clinical studies we measured cytokines secreted by BIA-ALCL and cutaneous ALCL lines. RESULTS: Our study of cell line culture supernatants, and 8 malignant compared with 9 benign seromas indicates that interleukin 9 (IL-9), IL-10, IL-13, IL-22, and/or interferon γ concentrations >1000 pg/mL distinguish malignant seromas from benign seromas. IL-6, known to be a driver of malignant cells, is also elevated in benign seromas and does not distinguish them from malignant seromas. CONCLUSIONS: The cytokine assay introduced in this study can be used together with levels of soluble CD30 to identify malignant seromas. Validation of these findings in a larger prospective patient cohort is warranted. The unique pattern of cytokine expression in malignant effusions surrounding breast implants gives further insight into the pathogenesis and cells of origin of BIA-ALCL.Level of Evidence: 5.

Circulating MicroRNA-30e Predicts Left Ventricular Hypertrophy In Essential Hypertensive Patients.

BACKGROUND: The biomarker for left ventricular hypertrophy (LVH) in patients with essential hypertension (EH) remains an unmet clinical need. The microRNA-30 (miR-30) family has been associated with LVH in cellular and animal studies, but not in a clinical setting. OBJECTIVE: The objective of the study was to investigate the clinical significance of circulating levels of miR-30 family as a biomarker for LVH in EH patients. METHODS: A total of 239 EH patients and 239 healthy controls were enrolled in this study. Circulating levels of miR-30 family members, namely, miR-30a, miR-30b, miR-30c-1, miR-30c-2, miR- 30d, and miR-30e, were evaluated using real-time polymerase chain reaction assays. RESULTS: The circulating miR-30a, miR-30b, and miR-30e were significantly reduced in EH patients in contrast to controls. EH patients with LVH (EH-LVH) had substantially lower circulating miR-30b and miR-30e levels compared to EH patients without LVH (EH-nLVH). Moreover, the expression levels of miR-30b and miR-30e were positively related to LVMI, respectively. Receiver operating curve analysis showed that circulating miR-30e levels distinguished EH patients from controls, and EH-LVH from EH-nLVH patients. Logistic regression analysis identified the circulating miR-30e as a risk factor for LVH in EH patients. CONCLUSION: Circulating miR-30e level can be used as a biomarker in distinguishing EH-LVH from EH-nLVH. A further prospective study is warranted to validate this finding.

Association of PD-1/PD-L axis expression with cytolytic activity, mutational load, and prognosis in melanoma and other solid tumors.

Programmed cell death protein-1 (PD-1)/programmed death ligand-1 (PD-L1) checkpoint blockade has led to remarkable and durable objective responses in a number of different tumor types. A better understanding of factors associated with the PD-1/PD-L axis expression is desirable, as it informs their potential role as prognostic and predictive biomarkers and may suggest rational treatment combinations. In the current study, we analyzed PD-L1, PD-L2, PD-1, and cytolytic activity (CYT) expression, as well as mutational density from melanoma and eight other solid tumor types using The Cancer Genome Atlas database. We found that in some tumor types, PD-L2 expression is more closely linked to Th1/IFNG expression and PD-1 and CD8 signaling than PD-L1 In contrast, mutational load was not correlated with a Th1/IFNG gene signature in any tumor type. PD-L1, PD-L2, PD-1, CYT expression, and mutational density are all positive prognostic features in melanoma, and conditional inference modeling revealed PD-1/CYT expression (i.e., an inflamed tumor microenvironment) as the most impactful feature, followed by mutational density. This study elucidates the highly interdependent nature of these parameters, and also indicates that future biomarkers for anti-PD-1/PD-L1 will benefit from tumor-type-specific, integrated, mRNA, protein, and genomic approaches.

Online Measurement of Deposit Surface in Electron Beam Freeform Fabrication.

In the process of electron beam freeform fabrication (EBF3), due to the continuous change of thermal conditions and variability in wire feeding in the deposition process, geometric deviations are generated in the deposition of each layer. In order to prevent the layer-by-layer accumulation of the deviation, it is necessary to perform online geometry measurement for each deposition layer, based on which the error compensation can be done for the previous deposition layer in the next deposition layer. However, the traditional three-dimensional reconstruction method that employs structured laser cannot meet the requirements of long-term stable operation in the manufacturing process of EBF3. Therefore, this paper proposes a method to measure the deposit surfaces based on the position information of electron beam speckle, in which an electron beam is used to bombard the surface of the deposit to generate the speckle. Based on the structured information of the electron beam in the vacuum chamber, the three-dimensional reconstruction of the surface of the deposited parts is realized without need of additional structured laser sensor. In order to improve the detection accuracy, the detection error is theoretically analyzed and compensated. The absolute error after compensation is smaller than 0.1 mm, and the precision can reach 0.1%, which satisfies the requirements of 3D reconstruction of the deposited parts. An online measurement system is built for the surface of deposited parts in the process of electron beam freeform fabrication, which realizes the online 3D reconstruction of the surface of the deposited layer. In addition, in order to improve the detection stability of the whole system, the image processing algorithm suitable for this scene is designed. The reliability and speed of the algorithm are improved by ROI extraction, threshold segmentation, and expansion corrosion. In addition, the speckle size information can also reflect the thermal conditions of the surface of the deposited parts. Hence, it can be used for online detection of defects such as infusion and voids.

Expression of LAG-3 and efficacy of combination treatment with anti-LAG-3 and anti-PD-1 monoclonal antibodies in glioblastoma.

Like in many tumor types, immunotherapy is currently under investigation to assess its potential efficacy in glioblastoma patients. Trials are under way to assess the efficacy of new immune checkpoint inhibitors including anti-PD-1 or CTLA4. We here investigate the expression and efficacy of a novel immune-checkpoint inhibitor, called LAG-3. We show that LAG-3 is expressed in human glioblastoma samples and in a mouse glioblastoma model we show that knock out or LAG-3 inhibition with a blocking antibody is efficacious against glioblastoma and can be used in combination with other immune checkpoint inhibitors toward complete eradication of the model glioblastoma tumors. From a mechanistic standpoint we show that LAG-3 expression is an early marker of T cell exhaustion and therefore early treatment with LAG-3 blocking antibody is more efficacious than later treatment. These data provide insight and support the design of trials that incorporate LAG-3 in the treatment of glioblastoma.