Search

Supplementary Data from CCR4 Blockade Depletes Regulatory T Cells and Prolongs Survival in a Canine Model of Bladder Cancer

Regulatory T-cell (Treg) infiltration can be targeted as a cancer immunotherapy. Here, we describe therapeutic efficacy of this strategy in a canine model of bladder cancer. We used dogs with naturally occurring bladder cancer to study the molecular mechanism of Treg infiltration into bladder cancer tissues and the effect of anti-Treg treatment. Tumor-infiltrating Tregs were evaluated by immunohistochemistry, and their association with prognosis was examined in dogs with bladder cancer. The molecular mechanism of Treg infiltration was explored by RNA sequencing and protein analyses. Murine xenograft experiments and canine studies were used to explore the therapeutic potential of anti-Treg treatment for bladder cancer. We found that tumor-infiltrating Tregs were associated with poor prognosis in dogs bearing spontaneous bladder cancer. Treg infiltration was caused by interaction between the tumor-producing chemokine CCL17 and the receptor CCR4 expressed on Tregs. CCR4 blockade inhibited tumor growth and Treg infiltration into the tissues in a xenograft mouse model. Dogs with spontaneous bladder cancer responded to anti-CCR4 treatment with improved survival and low incidence of clinically relevant toxicities. In human patients with bladder cancer, immunohistochemistry showed that tumor-infiltrating Tregs expressed CCR4. Thus, anti-CCR4 treatment may be a rational approach to test in clinical trials for human patients with bladder cancer.

Supplementary Table from CCR4 Blockade Depletes Regulatory T Cells and Prolongs Survival in a Canine Model of Bladder Cancer

Supplementary Figure from CCR4 Blockade Depletes Regulatory T Cells and Prolongs Survival in a Canine Model of Bladder Cancer

Supplementary Tables S1-S13. Supplementary Table S1. Clinical information and the summary of experiments performed on each sample. Supplementary Table S2. List of genes and regions captured for targeted deep sequencing. Supplementary Table S3. dbSNP ID list for which target baits were designed. Supplementary Table S4. Non-silent mutations detected in a validation cohort by targeted deep sequencing. Supplementary Table S5. Validated non-silent somatic mutations by PCR-based deep sequencing. Supplementary Table S6. Validated silent/non-silent somatic mutations used for clonal analysis of multiple samples from PBL001. Supplementary Table S7. Significantly hypo-/hyper-methylated gene sets in PBL compared to normal pancreas. Supplementary Table S8. Fusion genes detected by whole transcriptome sequencing in PBL. Supplementary Table S9. Genes with a significant differential expression between PBL and normal pancreatic samples. Supplementary Table S10. Significant gene set enrichment in comparison between PBL and normal pancreas. Supplementary Table S11. List of differentially expressed genes in each gene cluster identified by hierarchical clustering. Supplementary Table S12. Top 20 significantly enriched pathways in each gene cluster. Supplementary Table S13. Gene coefficients of PC1 and PC2.

Pancreatoblastoma is a rare pediatric pancreatic malignancy for which the molecular pathogenesis is not understood. In this study, we report the findings of an integrated multiomics study of whole-exome and RNA sequencing as well as genome-wide copy number and methylation analyses of ten pancreatoblastoma cases. The pancreatoblastoma genome was characterized by a high frequency of aberrant activation of the Wnt signaling pathway, either via somatic mutations of CTNNB1 (90%) and copy-neutral loss of heterozygosity (CN-LOH) of APC (10%). In addition, imprinting dysregulation of IGF2 as a consequence of CN-LOH (80%), gain of paternal allele (10%), and gain of methylation (10%) was universally detected. At the transcriptome level, pancreatoblastoma exhibited an expression profile characteristic of early pancreas progenitor-like cells along with upregulation of the R-spondin/LGR5/RNF43 module. Our results offer a comprehensive description of the molecular basis for pancreatoblastoma and highlight rational therapeutic targets for its treatment.Significance: Molecular genetic analysis of a rare untreatable pediatric tumor reveals Wnt/IGF2 aberrations and features of early pancreas progenitor-like cells, suggesting cellular origins and rational strategies for therapeutic targeting. Cancer Res; 78(4); 865–76. ©2017 AACR.

Supplementary Figures S1-S10. Supplementary Figure S1. Depths and coverages of targeted deep sequencing. Supplementary Figure S2. Sequencing coverages and numbers of mutations detected by WES. Supplementary Figure S3. Tumor subclonal populations in multiple samples from PBL001 estimated by PyClone. Supplementary Figure S4. Mutational signatures of primary and metastatic PBL. Supplementary Figure S5. SNP array-based copy number analysis of PBL. Supplementary Figure S6. Targeted deep sequencing based allele-specific copy number analysis of chromosome 11. Supplementary Figure S7. Bisulfite Sanger sequencing of ICR1 on chromosome 11p15.5. Supplementary Figure S8. Relative expression levels of IGF2 in CN-LOH-negative samples. Supplementary Figure S9. Aberrant activation of Wnt signaling pathway in PBL009. Supplementary Figure S10. Global DNA methylation profile of PBL.

Dyskeratosis congenita (DKC) is a rare, multisystem, bone marrow failure disease characterized by abnormalities such as in the skin, mucosa, nervous system, and lungs. Here we report a rare case of presumed DKC causing total retinal detachment in the right eye and severe peripheral retinal vascular occlusion in the left eye. A 3-year-old boy was presented with vitreous hemorrhage and total retinal detachment in the right eye and was scheduled to undergo vitreous surgery in the right eye and detailed ophthalmologic examination of the left eye under general anesthesia. Since a systemic examination revealed anemia and marked thrombocytopenia, he underwent a detailed pediatric examination. Although genetic testing revealed no significant pathologic mutations, the presence of shortened telomere length and other clinical findings suggested the possibility of DKC. His right eye had severe proliferative vitreoretinopathy, and retinal reattachment was not achieved with vitreous surgery, thus resulting in phthisis bulbi. The left eye showed a wide retinal avascular area in the temporal retina, retinal neovascularization, and hard exudates on fluorescein fundus angiography and was treated with laser photocoagulation using a binocular indirect ophthalmoscopic photocoagulator. Following laser surgery, the new blood vessels regressed, and the visual acuity was maintained at 1.0. The findings in this rare case indicate that DKC can cause severe retinal vascular occlusion, thus leading to vitreous hemorrhage and retinal detachment. Therefore, early detection with fundus examination and early treatment with photocoagulation are important.

In this study, the effect of reed vegetation on evapotranspiration (ET) and treatment performance was estimated, and a commonly used meteorological estimate of potential evapotranspiration (PET) was compared with ET with vertical subsurface flow constructed wetlands (VSSFs) in the treatment of the high salinity of landfill leachates. The experimental approaches consisted of three runs: Run A was a poor reed vegetation bed, Run B was a dense reed vegetation bed, and Run C was a bed without reeds. The results of this study are as follows: The salinities of the leachate inflow in Run A and Run B were 15.8 ± 1.9g Cl-/L and 15.5 ± 2.8g Cl-/L, respectively. The average ETs of Run A, Run B and Run C were 4.2mm/d, 7.4mm/d and 3mm/d, respectively. The annual ET rates of Run A, Run B and Run C were 1535mm, 2702mm and 1101mm, respectively. On the other hand, those of PET estimated on the basis of the Hamon equation of 2017and 2018 were 741mm and 791mm, respectively. The PET rate was much less compared to the ET rate in the dense vegetation bed. It was necessary to consider site-specific factors such as growth of plants in the evaluation of the water budget. The water loss by evapotranspiration in Run B was much more than those in Run A and Run C. Although the removal rates calculated from the concentration between inflow and outflow did not differ between the dense vegetation bed and the poor vegetation bed, the load reduction rates calculated from the water budget differed between dense vegetation and poor vegetation.

Tweakable block ciphers (TBCs) have proven highly useful to boost the security guarantees of authentication schemes. In 2017, Cogliati et al. proposed two MACs combining TBC and universal hash functions: a nonce-based MAC called NaT and a deterministic MAC called HaT. While both constructions provide high security, their properties are complementary: NaT is almost fully secure when nonces are respected (i.e., n-bit security, where n is the block size of the TBC, and no security degradation in terms of the number of MAC queries when nonces are unique), while its security degrades gracefully to the birthday bound (n/2 bits) when nonces are misused. HaT has n-bit security and can be used naturally as a nonce-based MAC when a message contains a nonce. However, it does not have full security even if nonces are unique.This work proposes two highly secure and efficient MACs to fill the gap: NaT2 and eHaT. Both provide (almost) full security if nonces are unique and more than n/2-bit security when nonces can repeat. Based on NaT and HaT, we aim at achieving these properties in a modular approach. Our first proposal, Nonce-as-Tweak2 (NaT2), is the sum of two NaT instances. Our second proposal, enhanced Hash-as-Tweak (eHaT), extends HaT by adding the output of an additional nonce-depending call to the TBC and prepending nonce to the message. Despite the conceptual simplicity, the security proofs are involved. For NaT2 in particular, we rely on the recent proof framework for Double-block Hash-then-Sum by Kim et al. from Eurocrypt 2020.

Bosutinib is a second-generation tyrosine kinase inhibitor indicated for treatment of chronic myeloid leukemia (CML) in adult patients. The safety and efficacy of bosutinib in patients younger than 18 years of age have not been established. We here report the case of a 4-year-old male with CML who was treated with bosutinib during coordination of human leukocyte antigen–matched unrelated bone-marrow transplantation because of insufficient responses to imatinib and dasatinib. The patient achieved a complete cytogenetic response immediately after starting bosutinib at 180 mg/day (290 mg/m2/day). Because toxicity was tolerable, the dose was increased to 200 mg/day (330 mg/m2/day). A complete cytogenetic response was maintained, but a major molecular response was not achieved 6 months after initiation of treatment with bosutinib. At steady state, maximum plasma concentration, minimum plasma concentration, and area under the plasma concentration-time curve were 89.2 ng/mL, 16.7 ng/mL, and 1017.4 ng·hr/mL, respectively, at 290 mg/m2/day; and 141.1 ng/mL, 18.9 ng/mL, and 1278.5 ng·hr/mL, respectively, at 330 mg/m2/day. To the best of our knowledge, this is the first case report to show the pharmacokinetics of bosutinib with efficacy and safety in a pediatric patient with CML. This rare case in a very young child with CML can also be valuable reference for clinical practice.

Introduction Chronic rhinosinusitis (CRS) is commonly classified based on the presence or absence of nasal polyps (NPs). Eosinophil infiltration is observed in NPs of patients in Western countries. In contrast, in East Asian countries, including Japan, CRS with NPs (CRSwNP) is subdivided based on the presence (eosinophilic CRS [ECRS]) or absence (non-eosinophilic CRS [NECRS]) of eosinophils in NPs. However, detailed analyses of other immune cells, such as lymphocytes, in NPs have not been performed. Therefore, clarification of the types of cells that infiltrate NPs is important to understand CRS pathogenesis. Objectives We analyzed the lymphocytes that infiltrate the paranasal sinus mucosa of ECRS and NECRS patients. Methods Eighteen patients with CRSwNP participated in this study, out of whom 6 were NECRS patients, and 12 were ECRS patients. The mucosa specimens, collected from patients during sinus surgeries, were subjected to collagenase treatment to prepare single cell suspensions. Then, mononuclear cells were isolated, and CD4+ T, CD8+ T, and CD20+ B-cell populations were examined using flow cytometry. Results In both NECRS and ECRS patients, CD8+ T-cells were dominant over CD4+ T-cells. Notably, CD4+ T-cell/B-cell ratio, but not CD8+ T-cell/B-cell or CD4+ T-cell/CD8+ T-cell ratios, was significantly higher in ECRS patients than in NECRS patients. Conclusion The CD4+ T-cell/B-cell ratio can be used as a potential indicator to differentiate between ECRS and NECRS.

The effect of genetic variation on second malignant neoplasms (SMNs) remains unclear. First, we identified the pathogenic germline variants in cancer-predisposing genes among 15 children with SMNs after childhood leukemia/lymphoma using whole-exome sequencing. Because the prevalence was low, we focused on the association between SMNs and NUDT15 in primary acute lymphoblastic leukemia (ALL) cases. NUDT15 is one of the 6-mercaptopurine (6-MP) metabolic genes, and its variants are common in East Asian individuals. The prevalence of NUDT15 hypomorphic variants was higher in patients with SMNs (n = 14; 42.9%) than in the general population in the gnomAD database (19.7%; P = .042). In the validation study with a cohort of 438 unselected patients with ALL, the cumulative incidence of SMNs was significantly higher among those with (3.0%; 95% confidence interval [CI], 0.6% to 9.4%) than among those without NUDT15 variants (0.3%; 95% CI, 0.0% to 1.5%; P = .045). The 6-MP dose administered to patients with ALL with a NUDT15 variant was higher than that given to those without SMNs (P = .045). The 6-MP–related mutational signature was observed in SMN specimens after 6-MP exposure. In cells exposed to 6-MP, a higher level of 6-MP induced DNA damage in NUDT15-knockdown induced pluripotent stem cells. Our study indicates that NUDT15 variants may confer a risk of SMNs after treatment with 6-MP in patients with ALL.

Objective Continuous positive airway pressure (CPAP) is the mainstay therapy for patients with obstructive sleep apnea (OSA) however compliance with CPAP is variable. Nasal ailments, such as nasal congestion are frequently mentioned as a cause for CPAP non-compliance, and potentially could be addressed prior to CPAP initiation, however, no specific criteria or recommendations for the evaluation and management of these patients exist. The aim of this retrospective study is to evaluate the effects of nasal anatomic features and disease on adherence to CPAP therapy for patients with OSA and determine the indications for pre-CPAP nasal treatment by using data obtained at clinical examination. Methods In total, 711 adult patients with initial diagnosis of OSA and an apnea–hypopnea index of ≥20 who were amenable to CPAP were included. We analyzed nasal parameters, past history of nasal disease, subjective symptoms, and disease severity in addition to whether CPAP therapy had been initiated, rate of CPAP therapy use (initial and 1 year), treatment continuation rate at 2 months and 1 year, and nasal treatments for all patients. Results CPAP therapy was initiated in 543 of 711 patients. Nasal resistance was significantly higher in patients who discontinued therapy soon after CPAP initiation. Nasal disease and nasal parameters were not found to be predictors of treatment adherence at 1 year. Allergic rhinitis, moderate to severe nasal congestion at bedtime, slight or extensive sinus opacification, and a high nasal septum deviation score were found to be independent predictors of nasal treatment, while strong awareness of nasal congestion, a past history of sinusitis, and a total nasal resistance (supine position) of ≥0.35 Pa/cm3/s were independent predictors of surgical treatment. Conclusion Long-term CPAP therapy adherence in patients with OSA can be predicted from initial CPAP adherence. Nasal disease and nasal parameters are important factors for early CPAP therapy discontinuation and should be adequately treated before therapy initiation to ensure long-term adherence. Indications for pre-CPAP nasal treatment and nasal surgery for patients with OSA can be predicted from the data obtained at the first examination, and these patients should be treated differently from those without OSA.

This paper presents a new incremental parallelizable message authentication code (MAC) scheme adaptable to lightweight block ciphers for memory integrity verification. The highlight of the proposed scheme is to achieve both incremental update capability and sufficient security bound with lightweight block ciphers, which is a novel feature. We extend the conventional parallelizable MAC to realize the incremental update capability while keeping the original security bound. We prove that a comparable security bound can be obtained even if this change is incorporated. We also present a hardware architecture for the proposed MAC scheme with lightweight block ciphers and demonstrate the effectiveness through FPGA implementation. The evaluation results indicate that the proposed MAC hardware achieves 3.4 times improvement in the latency-area product for the tag update compared with the conventional MAC.

In recent years, with the progress of data analysis methods utilizing artificial intelligence (AI) technology, concepts of smart cities collecting data from IoT devices and creating values by analyzing it have been proposed. However, making sure that the data is not tampered with is of the utmost importance. One way to do this is to utilize blockchain technology to record and trace the history of the data. Park and Kim proposed ensuring the trustworthiness of the data by utilizing an IoT device with a trusted execution environment (TEE). Also, Guan et al. proposed authenticating an IoT device and mediating data using a TEE. For the authentication, they use the physically unclonable function of the IoT device. Usually, IoT devices suffer from the lack of resources necessary for creating transactions for the blockchain ledger. In this paper, we present a secure protocol in which a TEE acts as a proxy to the IoT devices and creates the necessary transactions for the blockchain. We use an authenticated encryption method on the data transmission between the IoT device and TEE to authenticate the device and ensure the integrity and confidentiality of the data generated by the IoT devices.

Podocyte injury is an independent risk factor for the progression of renal diseases. Semaphorin3A (SEMA3A), expressed in podocytes and tubular cells in the mammalian adult kidneys, has been reported to regulate diverse biological functions and be associated with renal diseases. Here, we investigated pathological roles of SEMA3A signaling on podocyte injury using a doxorubicin (Dox)-induced mouse model and examined the therapeutic effect of SEMA3A-inhibitor (SEMA3A-I). We demonstrated that Dox caused massive albuminuria and podocyte apoptosis as well as an increase of SEMA3A expression in podocytes, all of which were ameliorated with SEMA3A-I treatment. In addition, c-Jun N-terminal kinase (JNK), known as a downstream of SEMA3A signaling, was activated in Dox-injected mouse podocytes while SEMA3A-I treatment partially blocked the activation. In vitro, SEMA3A-I protected against Dox-induced podocyte apoptosis and recombinant SEMA3A caused podocyte apoptosis with activation of JNK signaling. JNK inhibitor, SP600125, attenuated SEMA3A-induced podocyte apoptosis, indicating that the JNK pathway would be involved in SEMA3A-induced podocyte apoptosis. Furthermore, the analysis of human data revealed a positive correlation between levels of urinary SEMA3A and protein, suggesting that SEMA3A is associated with podocyte injury. In conclusion, SEMA3A has essential roles in podocyte injury and it would be the therapeutic target for protecting from podocyte injury.

Human papillomavirus (HPV) is a common sexually transmitted infection worldwide, which spreads via contact with infected genital, anal, and oral/pharyngeal areas (oral sex) owing to diverse manners of sexual intercourse. In this study, we devised an oral HPV detection method using mouthwash waste fluids that causes less psychological resistance to visiting the outpatient otolaryngology departments. We successfully detected only the specific unique reverse sequencing probe (using pyro-genotyping) and identified the nine genotypes of HPV targeted for vaccination by pyrosequencing the mouthwash waste fluids of non-head and neck cancer patient volunteers (n = 52). A relatively large number (11/52) of mouthwash waste fluids tested positive for HPV (21.2%, genotype 6, n = 1, 11, n = 1, 16, n = 1, and 18, n = 8). These results surpassed the sensitivity observed testing the same specimens using the conventional method (1/52, 1.9%). Our method (pyro-genotyping) was developed using nine HPV genotypes targeted for vaccination and the results were highly sensitive compared to those of the conventional method. This less expensive, high-throughput, and simple method can be used for detecting oral HPV infection with fewer socio-psychological barriers.

Introduction Chronic rhinosinusitis (CRS) is commonly classified based on the presence or absence of nasal polyps (NPs). Eosinophil infiltration is observed in NPs of patients in Western countries. In contrast, in East Asian countries, including Japan, CRS with NPs (CRSwNP) is subdivided based on the presence (eosinophilic CRS [ECRS]) or absence (non-eosinophilic CRS [NECRS]) of eosinophils in NPs. However, detailed analyses of other immune cells, such as lymphocytes, in NPs have not been performed. Therefore, clarification of the types of cells that infiltrate NPs is important to understand CRS pathogenesis. Objectives We analyzed the lymphocytes that infiltrate the paranasal sinus mucosa of ECRS and NECRS patients. Methods Eighteen patients with CRSwNP participated in this study, out of whom 6 were NECRS patients, and 12 were ECRS patients. The mucosa specimens, collected from patients during sinus surgeries, were subjected to collagenase treatment to prepare single cell suspensions. Then, mononuclear cells were isolated, and CD4 + T, CD8 + T, and CD20 + B-cell populations were examined using flow cytometry. Results In both NECRS and ECRS patients, CD8 + T-cells were dominant over CD4 + T-cells. Notably, CD4 + T-cell/B-cell ratio, but not CD8 + T-cell/B-cell or CD4 + T-cell/CD8 + T-cell ratios, was significantly higher in ECRS patients than in NECRS patients. Conclusion The CD4 + T-cell/B-cell ratio can be used as a potential indicator to differentiate between ECRS and NECRS.

Objectives This study investigated morphological variations of lamina papyracea, the structure that should be carefully considered when opening posterior ethmoid sinus during endoscopic sinus surgery, to avoid injury. Study design This study employed axial, coronal, and sagittal computed tomography. Methods Using computed tomography images of 228 face-sides, various anatomical parameters were determined: distances of the anterior and posterior ethmoid arteries from the skull base, and from the third lamella; changes in the angles of the lamina papyracea at the anterior and posterior ethmoid sinuses; and presence or absence of supraorbital ethmoid cell (SECs), Onodi cell, and Haller cell. The relationship between the distances which indicate the point of maximum projection by the lamina papyracea among third lamina and posterior ethmoid artery into the posterior ethmoid sinus and these anatomical factors were analyzed statistically. Results The projection distance of lamina papyracea into the posterior ethmoid sinus was -2.6 mm to 3.4 mm, and in 41.2% of cases, projection in the direction of the nasal cavity was greater than that of the lamina papyracea at the anterior ethmoid sinus. This distance increased with increasing distance of the maximum projection point from the skull base and increasing floating distances of the anterior and posterior ethmoid arteries. The number of subjects with large projection distances was increased among those with floating posterior ethmoid arteries. In addition, subjects with SECs had significantly greater projection distances. Conclusions Particular care should be taken to avoid injury to the lamina papyracea when opening the posterior ethmoid sinus in subjects with floating anterior or posterior ethmoid arteries, and/or SEC. Level of evidence 4 Laryngoscope, 131:E19-E25, 2021.

Authenticated encryption (AE) is a symmetric-key encryption function that provides confidentiality and authenticity of a message. One of the evaluation criteria for AE is state size, which is memory size needed for encryption. State size is especially important when cryptosystem is implemented in constrained devices, while trivial reduction by using a small primitive is not generally acceptable as it leads to a degraded security.