In vivo quantification of programmed death-ligand-1 expression heterogeneity in tumors using fluorescence lifetime imaging.

Cancer patient selection for immunotherapy is often based on programmed death-ligand-1 (PD-L1) expression as a biomarker. PD-L1 expression is currently quantified using immunohistochemistry, which can only provide snapshots of PD-L1 expression status in microscopic regions of ex vivo specimens. In vivo imaging using targeted agents can capture dynamic variations of PD-L1 expression in entire tumors within and across multiple subjects. Towards this goal, several PD-L1 targeted molecular imaging probes have been evaluated in murine models and humans. However, clinical translation of these probes has been limited due to a significant non-specific accumulation of the imaging probes and the inability of conventional imaging modalities to provide quantitative readouts that can be compared across multiple subjects. Here we report that in vivo time-domain (TD) fluorescence imaging can provide quantitative estimates of baseline tumor PD-L1 heterogeneity across untreated mice and variations in PD-L1 expression across mice undergoing clinically relevant anti-PD1 treatment. This approach relies on a significantly longer fluorescence lifetime (FLT) of PD-L1 specific anti-PD-L1 antibody tagged to IRDye 800CW (αPDL1-800) compared to nonspecific αPDL1-800. Leveraging this unique FLT contrast, we show that PD-L1 expression can be quantified across mice both in superficial breast tumors using planar FLT imaging, and in deep-seated liver tumors (>5 mm depth) using the asymptotic TD algorithm for fluorescence tomography. Our results suggest that FLT contrast can accelerate the preclinical investigation and clinical translation of novel molecular imaging probes by providing robust quantitative readouts of receptor expression that can be readily compared across subjects.

Preventing NK cell activation in the damaged liver induced by cabozantinib/PD-1 blockade increases survival in hepatocellular carcinoma models.

The addition of anti-VEGF antibody treatment to immune checkpoint blockade (ICB) has increased the efficacy of immunotherapy in advanced hepatocellular carcinoma (HCC). Despite an initial promise, adding multitargeted kinase inhibitors of VEGFR with ICB has failed to increase survival in HCC. To reveal the mechanisms underlying treatment failure, we studied the effects of cabozantinib/ICB using orthotopic murine HCC models with or without liver damage. We monitored tumor growth and liver function, recorded survival outcomes, and performed immune profiling studies for intra-tumoral and surrounding liver. Cabozantinib/ICB treatment led to tumor regression and significantly improved survival in mice with normal livers. However, consistent with the clinical findings, combination therapy failed to show survival benefits despite similar tumor control when tested in the same models but in mice with liver fibrosis. Moreover, preclinical and clinical data converged, showing that activating immune responses by cabozantinib/ICB treatment induced hepatoxicity. Immune profiling revealed that combination therapy effectively reprogrammed the tumor immune microenvironment and increased NK cell infiltration and activation in the damaged liver tissue. Surprisingly, systemic depletion of NK reduced hepatotoxicity elicited by the combination therapy without compromising its anti-cancer effect, and significantly enhanced the survival benefit even in mice with HCC and underlying liver fibrosis. These findings demonstrate that preventing NK activation allowed for maintaining a favorable therapeutic ratio when combining ICB with cabozantinib in advanced HCC models.

The liver-to-spleen ratio is a risk factor predicting oxygen demand in COVID-19 patients.

Background: We aimed to investigate risk factors predicting oxygen demand in COVID-19 patients. Methods: Patients admitted to Shizuoka General Hospital with COVID-19 from August 2020 to August 2021 were included. First, we divided patients into groups with and without oxygen demand. Then, we compared patients' clinical characteristics and laboratory and radiological findings to determine factors predicting oxygen demand. Results: One hundred seventy patients with COVID-19 (aged 58±15 years, 57 females) were enrolled. Common comorbidities were cardiovascular diseases (47.6%), diabetes mellitus (28.8%), and dyslipidemia (26.5%). Elder age, higher body mass index, cardiovascular diseases, diabetes mellitus, lower lymphocyte count, albumin, hepatic attenuation value, and the liver-to-spleen ratio (L/S), higher D-dimer, aspartate aminotransferase, lactate dehydrogenase, troponin-T, C-reactive protein, KL-6, chest and abdominal circumference, and visceral fat were found in patients with oxygen demand. According to the multivariate logistic regression analysis, L/S, lymphocyte count, D-dimer, and abdominal circumference under the diaphragm were independent risk factors predicting oxygen demand in COVID-19 patients. Conclusions: On admission, L/S, lymphocyte count, D-dimer, and abdominal circumference were predictive factors for oxygen demand. These factors may help in the appropriate triage of COVID-19 patients in the decision to admit them to the hospital.

Stratification of Stage II Colon Cancer Using Recurrence Prediction Value: A Multi-institutional International Retrospective Study.

OBJECTIVE: To create a recurrence prediction value (RPV) of high-risk factor and identify the patients with high risk of cancer recurrence. SUMMARY BACKGROUND DATA: There are several high-risk factors known to lead to poor outcomes. Weighting each high-risk factor based on their association with increased risk of cancer recurrence can provide a more precise understanding of risk of recurrence. METHODS: We performed a multi-institutional international retrospective analysis of patients with Stage II colon cancer patients who underwent surgery from 2010 to 2020. Patient data from a multi-institutional database were used as the Training data, and data from a completely separate international database from two countries were used as the Validation data. The primary endpoint was recurrence-free survival (RFS). RESULTS: A total of 739 patients were included from Training data. To validate the feasibility of RPV, 467 patients were included from Validation data. Training data patients were divided into RPV low (n = 564) and RPV high (n = 175). Multivariate analysis revealed that risk of recurrence was significantly higher in the RPV high than the RPV low (Hazard ratio (HR) 2.628; 95% confidence interval (CI) 1.887-3.660; P < 0.001). Validation data patients were divided into two groups (RPV low, n = 420) and RPV high (n = 47). Multivariate analysis revealed that risk of recurrence was significantly higher in the RPV high than the RPV low (HR 3.053; 95% CI 1.962-4.750; P < 0.001). CONCLUSIONS: RPV can identify Stage II colon cancer patients with high risk of cancer recurrence world-wide.

Cancer cell plasticity and MHC-II-mediated immune tolerance promote breast cancer metastasis to lymph nodes.

Tumor-draining lymph nodes (TDLNs) are important for tumor antigen-specific T cell generation and effective anticancer immune responses. However, TDLNs are often the primary site of metastasis, causing immune suppression and worse outcomes. Through cross-species single-cell RNA-Seq analysis, we identified features defining cancer cell heterogeneity, plasticity, and immune evasion during breast cancer progression and lymph node metastasis (LNM). A subset of cancer cells in the lymph nodes exhibited elevated MHC class II (MHC-II) gene expression in both mice and humans. MHC-II+ cancer cells lacked costimulatory molecule expression, leading to regulatory T cell (Treg) expansion and fewer CD4+ effector T cells in TDLNs. Genetic knockout of MHC-II reduced LNM and Treg expansion, while overexpression of the MHC-II transactivator, Ciita, worsened LNM and caused excessive Treg expansion. These findings demonstrate that cancer cell MHC-II expression promotes metastasis and immune evasion in TDLNs.

Diet-mediated constitutive induction of novel IL-4+ ILC2 cells maintains intestinal homeostasis in mice.

Group 2 innate lymphoid cells (ILC2s) expressing IL-5 and IL-13 are localized at various mucosal tissues and play critical roles in the induction of type 2 inflammation, response to helminth infection, and tissue repair. Here, we reveal a unique ILC2 subset in the mouse intestine that constitutively expresses IL-4 together with GATA3, ST2, KLRG1, IL-17RB, and IL-5. In this subset, IL-4 expression is regulated by mechanisms similar to but distinct from those observed in T cells and is partly affected by IL-25 signaling. Although the absence of the microbiota had marginal effects, feeding mice with a vitamin B1-deficient diet compromised the number of intestinal IL-4+ ILC2s. The decrease in the number of IL-4+ ILC2s caused by the vitamin B1 deficiency was accompanied by a reduction in IL-25-producing tuft cells. Our findings reveal that dietary vitamin B1 plays a critical role in maintaining interaction between tuft cells and IL-4+ ILC2s, a previously uncharacterized immune cell population that may contribute to maintaining intestinal homeostasis.

Out-of-Plane Needle Placements Using 3D Augmented Reality Protractor on Smartphone: An Experimental Phantom Study.

PURPOSE: To evaluate the accuracy of needle placement using a three-dimensional (3D) augmented reality (AR) protractor on smartphones (AR Puncture). MATERIALS AND METHODS: An AR protractor that can be rotated in three directions against the CT plane with angle guidance lines for smartphones was developed. The protractor center can be adjusted to an entry point by manually moving the smartphone with the protractor center fixed at the center of the screen (Fix-On-Screen) or by image tracking with a printed QR code placed at an entry point (QR-Tracking). Needle placement was performed by viewing a target line in the tangent direction with the Bull's eye method. The needle placement errors placed by four operators in six out-of-plane directions in a phantom using a smartphone (iPhone XR, Apple, Cupertino, CA, USA) were compared with two registration methods. RESULTS: No significant difference in the average needle placement error was observed between the Fix-On-Screen and QR-Tracking methods (5.6 ± 1.7 mm vs. 6.1 ± 2.9 mm, p = 0.475). The average procedural time of the Fix-On-Screen method was shorter than that of the QR-Tracking method (71.0 ± 23.9 s vs. 98.4 ± 59.5 s, p = 0.042). CONCLUSION: The accuracies of out-of-plane needle placements using the 3D AR protractor with the two registration methods were equally high, with short procedure times. In clinical use, the Fix-On-Screen registration method would be more convenient because no additional markers are required.

Evaluating the impact of human flow on the spread of infectious diseases.

This study explores infectious disease transmission through contact during daily trips between municipalities. We propose an extended susceptible-infectious-recovered model that considers daily movements on the spatial spread of infectious disease. The current model considers two types of movement: long-term movements such as migration and shorter activities completed within a day. We present analytical results using a next-generation matrix and numerical results using actual human flow data, focusing on the number of days it takes for an outbreak from each region to reach the entire area. Our results suggest that the likelihood of infection depends on the ratio of human flow to population rather than the population per se.

Social dilemma in the excess use of antimicrobials incurring antimicrobial resistance.

The emergence of antimicrobial resistance (AMR) caused by the excess use of antimicrobials has come to be recognized as a global threat to public health. There is a 'tragedy of the commons' type social dilemma behind this excess use of antimicrobials, which should be recognized by all stakeholders. To address this global threat, we thus surveyed eight countries/areas to determine whether people recognize this dilemma and showed that although more than half of the population pays little, if any, attention to it, almost 20% recognize this social dilemma, and 15-30% of those have a positive attitude toward solving that dilemma. We suspect that increasing individual awareness of this social dilemma contributes to decreasing the frequency of AMR emergencies.

Exploring sexual contact networks by analyzing a nationwide commercial-sex review website.

Understanding the structure of human sexual contact networks is vital in a broad range of disciplines, including sociology, biology, public health, and anthropology. However, sexual contact networks are yet to be understood because technical and privacy issues make it difficult to conduct accurate, large-scale surveys. In this study, we surveyed data openly available on one of the largest adult entertainment websites in Japan, where male clients (MCs) can write online customer reviews of female commercial sex workers (FCSWs). In particular, our investigation focused on a type of establishment called "soapland," the only type of sex industry in Japan where sexual intercourse is publicly permitted. Soaplands are scattered throughout Japan, and the study website covers approximately 66% of them. Using such a vast amount of data on a nationwide scale, we clarified the network structure of commercial sex, characterized by small-world, scale-free, and disassortative mating properties. To study geographical characteristics, we compared the resulting network with three different artificially generated networks via the random rewiring of links. Moreover, we considered a simple epidemic model on the resulting network, and investigated whether it would be more effective to provide infection control measures to FCSWs or MCs. We determined that active FCSWs constitute an important pathway of infection propagation in commercial sex networks, but MCs also play an essential role as weak ties.

PMEL p.Leu18del dilutes coat color of Kumamoto sub-breed of Japanese Brown cattle.

BACKGROUND: Coat color is important for registration and maintenance of livestock. Standard coat color of Kumamoto sub-breed of Japanese Brown cattle is solid brown, but individuals with diluted coat color have been observed recently. In this study, we attempted to identify polymorphism(s) responsible for coat color dilution by whole genome analysis. RESULTS: One of the diluted cattle possessed 7302 exonic polymorphisms which could affect genes' function. Among them, 14 polymorphisms in 10 coat color-related genes were assumed to be specific for the diluted cattle. Subsequent genotyping with three diluted cattle and 74 standard cattle elucidated that PMEL p.Leu18del was the causative polymorphism for coat color dilution in this sub-breed. Individuals with del/del type of this polymorphism showed diluted coat color, but coat color of heterozygotes were intermediate with various dilution rates. CONCLUSIONS: Coat color dilution of Kumamoto sub-breed was caused by PMEL p.Leu18del. The causative del allele has been detected in several genetically distant cattle breeds, suggesting that PMEL p.Leu18del can be used as a DNA marker to control cattle coat color.

Basic reproduction number of epidemic models on sparse networks.

The basic reproduction number R_{0} is a standard indicator of infection control in epidemiology. Although R_{0} has been studied extensively for deterministic epidemic models, it has not been formulated accurately for models adopting network structures. Here, we extend a four-compartment model that includes commonly used epidemic models to a Markov process on networks. By examining the Markov process in detail, we derive a canonical formula for R_{0} involving two probability values. Numerical calculations show that the derived formula is a better approximation than the conventional formula when the network is very sparse. We propose this as a standard formula for controlling infections that can only be transmitted through intimate contact, where contacts between individuals can be described as a sparse network.

Optimal strategies and cost-benefit analysis of the [Formula: see text]-player weightlifting game.

The study of cooperation has been extensively studied in game theory. Especially, two-player two-strategy games have been categorized according to their equilibrium strategies and fully analysed. Recently, a grand unified game covering all types of two-player two-strategy games, i.e., the weightlifting game, was proposed. In the present study, we extend this two-player weightlifting game into an [Formula: see text]-player game. We investigate the conditions for pure strategy Nash equilibria and for Pareto optimal strategies, expressed in terms of the success probability and benefit-to-cost ratio of the weightlifting game. We also present a general characterization of [Formula: see text]-player games in terms of the proposed game. In terms of a concrete example, we present diagrams showing how the game category varies depending on the benefit-to-cost ratio. As a general rule, cooperation becomes difficult to achieve as group size increases because the success probability of weightlifting saturates towards unity. The present study provides insights into achieving behavioural cooperation in a large group by means of a cost-benefit analysis.

Increased CD8+ T-cell Infiltration and Efficacy for Multikinase Inhibitors After PD-1 Blockade in Hepatocellular Carcinoma.

Immune checkpoint blockade combined with antiangiogenic therapy induces vascular normalization and antitumor immunity and is efficacious in hepatocellular carcinoma (HCC); but whether and how initial immunotherapy affects the efficacy of subsequent antiangiogenic therapy are unknown. We evaluated a cohort of HCC patients (n = 25) who received the pan-vascular endothelial growth factor receptor multikinase inhibitor sorafenib after initial therapy with an antiprogrammed cell death protein (PD)-1 antibody and found superior outcomes in these patients (12% overall response rate to sorafenib and a median overall survival of 12.1 months). To prove this potential benefit, we examined the impact of an anti-PD-1 antibody on response to subsequent sorafenib treatment in orthotopic models of murine HCC. Prior anti-PD-1 antibody treatment amplified HCC response to sorafenib therapy and increased survival (n = 8-9 mice per group, hazard ratio = 0.28, 95% confidence interval = 0.09 to 0.91; 2-sided P = .04). Anti-PD-1 therapy showed angioprotective effects on HCC vessels to subsequent sorafenib treatment, which enhanced the benefit of this therapy sequence in a CD8+ T-cell-dependent manner. This priming approach using immunotherapy provides an immediately translatable strategy for effective HCC treatment while reducing drug exposure.

Noncontact measurement of puncture needle angle using augmented reality technology in computed tomography-guided biopsy: stereotactic coordinate design and accuracy evaluation.

PURPOSE: This study aims to introduce a new handheld device application for noncontact and real-time measurements of the angle of a biopsy needle using augmented reality (AR) image tracking technology. Furthermore, this study discusses the methods used to optimize the related coordinate design for computed tomography (CT)-guided biopsy procedures. METHODS: An in-house noncontact angle measurement application was developed using AR platform software. This application tracks the position and direction of a printed texture located on the handle of a biopsy needle. The needle direction was factorized into two directions: tilting or rolling. Tilting was defined following the tilting of the CT gantry so that rolling would match the angle measured in CT images. In this study, CT-guided tumor biopsies were performed using a conventional guiding method with a protractor. The true value of needle rolling was measured by CT imaging and was then compared to the rolling measurement provided by the application developed in the current study using a mobile phone. RESULTS: This study enrolled 18 cases of tumor biopsy (five renal tumors, five lung tumors, four retroperitoneal tumors, one soft tissue tumor, one thyroid tumor, one mesentery tumor, and one bone tumor). The measurement accuracy was - 0.2°, which was the average difference between AR and CT, and the measurement precision was 2.0°, which was the standard deviation of the difference between AR and CT measurements. The coefficient of determination (R2) was 0.996. CONCLUSION: The noncontact needle measurement software using AR technology is sufficiently reliable for use in clinical settings. A real-time display of the needle angle that also shows the direction of the CT gantry is expected to enable a simple biopsy needle navigation.

Mixed Reality Needle Guidance Application on Smartglasses Without Pre-procedural CT Image Import with Manually Matching Coordinate Systems.

PURPOSE: To develop and assess the accuracy of a mixed reality (MR) needle guidance application on smartglasses. MATERIALS AND METHODS: An MR needle guidance application on HoloLens2, without pre-procedural CT image reconstruction or import by manually matching the spatial and MR coordinate systems, was developed. First, the accuracy of the target locations in the image overlay at 63 points arranged on a 45 × 35 × 21 cm box and needle angles from 0° to 80°, placed using the MR application, was verified. The needle placement errors from 12 different entry points in a phantom by seven operators (four physicians and three non-physicians) were compared using a linear mixed model between the MR guidance and conventional methods using protractors. RESULTS: The average errors of the target locations and needle angles placed using the MR application were 5.9 ± 2.6 mm and 2.3 ± 1.7°, respectively. The average needle insertion error using the MR guidance was slightly smaller compared to that using the conventional method (8.4 ± 4.0 mm vs. 9.6 ± 5.1 mm, p = 0.091), particularly in the out-of-plane approach (9.6 ± 3.5 mm vs. 12.3 ± 4.6 mm, p = 0.003). The procedural time was longer with MR guidance than with the conventional method (412 ± 134 s vs. 219 ± 66 s, p < 0.001). CONCLUSION: MR needle guidance without pre-procedural CT image import is feasible when matching coordinate systems, and the accuracy of needle insertion is slightly better than that of the conventional method.

Blood eosinophil count and FeNO to predict benralizumab effectiveness in real-life severe asthma patients.

OBJECTIVE: Benralizumab is a promising drug for severe uncontrolled asthma. This study aimed to clarify the effectiveness of benralizumab in a real-life setting. METHODS: Subjects included 24 patients with severe type 2 asthma who received benralizumab between April 2018 and July 2019. Changes in parameters, exacerbation frequency, and oral corticosteroid (OCS) use after 4 and 24 weeks of administration were examined. The parameters included the Global Evaluation of Treatment Effectiveness (GETE) scale, Asthma Control Questionnaire (ACQ), Asthma Control Test (ACT), blood eosinophils, fractional exhaled nitric oxide (FeNO), and spirometry. The response to treatment was defined as follows: for patients with exacerbations or OCS use before treatment initiation, a reduction of ≥50% in exacerbation frequency or OCS use; and for patients without exacerbations or OCS use, an improvement of ≥0.5 in ACQ scores and ≥3 in ACT scores, or of ≥10.38% in FEV1. RESULTS: Twenty-one patients completed the treatment for 24 weeks. Excellent and good GETE scales and ACQ and ACT improvement were found in 67% of the patients at 4 weeks, and the effect continued until 24 weeks. The patients' rate with exacerbations was significantly reduced compared to the previous 24 weeks before administration. In 17 patients receiving OCS, the use could be reduced or quit in 14 patients. Overall, 16 patients (76.2%) met the responder definition and could be predicted by the baseline eosinophil count and FeNO levels with the best cutoff values of 100/µL and 40 ppb, respectively. CONCLUSIONS: Blood eosinophil and FeNO could predict benralizumab effectiveness.

Impact of intraoperative hypocapnia on postoperative complications in laparoscopic surgery for colorectal cancer.

PURPOSE: In laparoscopic surgery (LS) for colorectal cancer (CRC), the relationship between intraoperative end-tidal carbon dioxide concentration (EtCO2) and surgery-related complications remains unexplored. This study assessed the impact of intraoperative EtCO2 on postoperative complications in LS for CRC. METHODS: In total, 909 patients who underwent LS for CRC were enrolled. Hypocapnia and hypercapnia were defined as EtCO2 < 35 mmHg and > 40 mmHg, respectively, and the relationships between hypocapnia or hypercapnia duration and postoperative complications were analyzed. RESULTS: The median (range) durations of hypocapnia and hypercapnia were 2.0 (0-8.3) h and 0.3 (0-5.8) h, respectively. Complications were observed in 208 cases (23.0%), which included 37 (4.1%) instances of anastomotic leakage and 86 (9.5%) of superficial surgical site infection (SSI). While the hypercapnia duration was not associated with the short-term outcomes, prolonged hypocapnia was significantly correlated with complications (p = 0.02), specifically superficial SSI (p = 0.005). Multivariate analyses adjusted for confounding factors confirmed that hypocapnia prolongation was an independent risk factor for postoperative superficial SSI [OR 1.19; 95% confidence interval (Cl) 1.03-1.36, p = 0.01]. CONCLUSION: Intraoperative hypocapnia may be a risk factor for postoperative complications, in particular superficial SSI, in LS for CRC.

Evaluation of abdominal aortic calcification by plain CT predicts anastomotic leakage in laparoscopic surgery for colorectal cancer.

BACKGROUND: Anastomotic leakage is one of the most severe and critical complications of laparoscopic surgery for colorectal cancer. However, definitive preoperative predictors of anastomotic leakage remain elusive. With the ageing of society, the number of colorectal cancer patients with arteriosclerotic disease in Japan is increasing. This study was performed to evaluate the correlation between preoperative arteriosclerosis and anastomotic leakage. METHODS: In total, 98 patients undergoing laparoscopic surgery for colorectal cancer with reconstruction using the double-stapling technique without diversion of the stoma were enrolled in the study. Preoperative assessment of arteriosclerotic disease was performed by abdominal computed tomography. The calcification volume percentage of the aorta between the level of the celiac artery root and aortic bifurcation was calculated using ZIOstation2 software, and the relationship between arteriosclerosis and anastomotic leakage was analysed. RESULTS: Among 98 cases, anastomotic leakage was observed in 16 (16.3%). The median calcification volume percentage (range) was 2.35% (0-40.3%). Age, male sex, hypertension, dissection number, estimated glomerular filtration rate and tumour location were correlated with anastomotic leakage on statistical analysis. Statistical analysis showed that calcification volume percentage was one of the robust risk factors for anastomotic leakage (odds ratio: 1.09, 95% confidence interval: 1.03-1.17, P < 0.01). CONCLUSIONS: Calcification of the abdominal aorta may be a promising predictor of AL after laparoscopic surgery for colorectal cancer reconstruction using the double-stapling technique.