Accurate expression of PD-L1/L2 in lung adenocarcinoma cells: A retrospective study by double immunohistochemistry.

The percentage of programmed death ligand 1 (PD-L1) positivity in cancer cells, named as the tumor proportion score, is considered to be a predictive biomarker for anti-PD-1/PD-L1 therapy in lung cancer. PD-L1 is expressed on not only cancer cells but also on immune cells, including macrophages. Although previous studies related to PD-L1/2 expression in cancer tissues have been generally based on single immunohistochemistry (IHC), in the present study, we attempted to evaluate accurate PD-L1/2 expression in cancer cells in lung adenocarcinoma cells using double IHC to also evaluate macrophages. Of the 231 patients, PD-L1 expression was negative in 169 patients (73.2%), 1%-49% positive in 47 patients (20.3%), and ≥50% positive in 15 patients (6.5%). Interestingly, PD-L1 positivity was decreased when using double IHC compared with the estimation by single IHC. High PD-L1 expression was associated with high-grade cancer cells and in higher stage cancer. PD-L2 was negative in 109 patients (47.2%), 1%-49% positive in 50 patients (21.6%), and ≥50% positive in 72 patients (31.2%). The number of PD-L2-positive patients was increased in cases that had an epidermal growth factor receptor (EGFR) mutation and in lower stage cancer. Thirty-five patients (15.2%) were positive for both PD-L1 and PD-L2, whereas 81 patients (35.1%) were negative for both PD-L1 and PD-L2. Log-rank analysis showed that progression-free survival and overall survival were significantly the longest in the PD-L1-negative and PD-L2-positive groups (P < .0001 and P = .0120). We observed lower PD-L1 or PD-L2 expression in lung adenocarcinoma than previously reported. Double IHC for macrophages may help clinicians to evaluate PD-L1 or PD-L2 expression specifically in cancer cells.

Novel approach to pleurodesis with 50 % glucose for air leakage after lung resection or pneumothorax.

PURPOSE: Pleurodesis is performed in patients demonstrating air leakage after lung resection and in those with pneumothorax who must avoid surgery. However, there have so far been very few reports of pleurodesis with 50 % glucose. We herein examined the feasibility and effectiveness of this novel pleurodesis technique. METHODS: Thirty-five patients after lung resection and 11 pneumothorax patients without surgery were treated with pleurodesis using 50 % glucose. Approximately, 200 mL of 50 % glucose solution was injected into the pleural space and repeated until the air leakage stopped. Cases in which the air leakage did not stop after three injections were considered to be unsuccessful and subsequently treated with conventional pleurodesis using OK-432. RESULTS: Thirty-nine patients were successfully treated with 50 % glucose, although 7 patients required further treatment with OK-432. The unsuccessful group had some pulmonary comorbidities (P < 0.001), and the pleural effusion volume after pleurodesis was less than that in the successful group (P < 0.001). Although the air leakage did not stop in unsuccessful patients, the amount of air leakage markedly decreased. A temporary elevation of the blood sugar level was observed in 20 patients, but no other side effects had appeared. CONCLUSIONS: Pleurodesis with 50 % glucose is an easy, safe, and effective treatment modality. It is therefore considered to be a useful alternative method for pleurodesis.

[Chronic Progressive Pulmonary Aspergillosis after Surgery for Lung Cancer].

Case 1:A 62-year-old man underwent right S9+10 segmentectomy for non-small cell lung cancer. Three years later, pleural thickening appeared in the apex, and gradually developed cystic change. Case 2:A 64-year-old man underwent left upper lobectomy for non-small cell lung cancer. One year after surgery, chest computed tomography (CT) showed cysts in the apex of the lower lobe. The cysts expanded slowly with consolidation. Both cases were diagnosed as chronic progressive pulmonary aspergillosis. Although anti-fungal drugs were administered in both cases, case 1 died of the disease. Chronic progressive pulmonary aspergillosis is a rare late postoperative complication;when CT shows progressive cysts with consolidation in the apex area of the residual lung, frequent check-ups are needed.

A QM/MM study of the L-threonine formation reaction of threonine synthase: implications into the mechanism of the reaction specificity.

Threonine synthase catalyzes the most complex reaction among the pyridoxal-5'-phosphate (PLP)-dependent enzymes. The important step is the addition of a water molecule to the Cß-Cα double bond of the PLP-α-aminocrotonate aldimine intermediate. Transaldimination of this intermediate with Lys61 as a side reaction to form α-ketobutyrate competes with the normal addition reaction. We previously found that the phosphate ion released from the O-phospho-l-homoserine substrate plays a critical role in specifically promoting the normal reaction. In order to elucidate the detailed mechanism of this "product-assisted catalysis", we performed comparative QM/MM calculations with an exhaustive search for the lowest-energy-barrier reaction pathways starting from PLP-α-aminocrotonate aldimine intermediate. Satisfactory agreements with the experiment were obtained for the free energy profile and the UV/vis spectra when the PLP pyridine N1 was unprotonated and the phosphate ion was monoprotonated. Contrary to an earlier proposal, the base that abstracts a proton from the attacking water was the ε-amino group of Lys61 rather than the phosphate ion. Nevertheless, the phosphate ion is important for stabilizing the transition state of the normal transaldimination to form l-threonine by making a hydrogen bond with the hydroxy group of the l-threonine moiety. The absence of this interaction may account for the higher energy barrier of the side reaction, and explains the mechanism of the reaction specificity afforded by the phosphate ion product. Additionally, a new mechanism, in which a proton temporarily resides at the phenolate O3' of PLP, was proposed for the transaldimination process, a prerequisite step for the catalysis of all the PLP enzymes.

Association of a genetic variant of CYP19A1 with multicentric development of lung adenocarcinomas.

BACKGROUND: The detection rate of multiple lung adenocarcinomas, which display multiple ground glass opacity nodules in the peripheral lung, is increasing because of advances in high resolution computed tomography. The genetic backgrounds of multiple nodules and the mechanisms that underlie their multicentric development are unknown. In this study, we examined single nucleotide polymorphisms (SNPs) of the cytochrome P450 19A1 gene to determine if they are associated with multiple adenocarcinomas risk. METHODS: Fifty-one cases of multiple adenocarcinomas with lepidic growth, 62 cases of a single adenocarcinoma with lepidic growth, and 126 control cases were analyzed. Three SNPs were analyzed by using a 5' nuclease assay with TaqMan minor-groove-binder probe. The expression level of CYP19A1 in the noncancerous lung was quantified by real-time reverse transcription polymerase chain reaction (RT-PCR). RESULTS: A minor allele of SNP rs3764221, which is located in the CYP19A1 gene, was significantly associated with multiple adenocarcinomas risk (adjusted odds ratio = 3.06; P = 0.006). Other polymorphisms of CYP19A1 were not significantly associated with the risk of multiple adenocarcinomas. A minor allele of SNP rs3764221 was also associated with a higher level of CYP19A1 messenger RNA expression (P = 0.03). CONCLUSIONS: SNP rs3764221 contributes to the development of multicentric adenocarcinomas in the peripheral lung by causing higher levels of CYP19A1 expression.

Photoluminescence characterization in silicon nanowire fabricated by thermal oxidation of nano-scale Si fin structure.

Low-temperature photoluminescence (PL) spectra of electron-hole systems in Si nanowires (NWs) prepared by thermal oxidization of Si fin structures were studied. Mapping of PL reveals that NWs with uniform width are formed over a large area. Annealing temperature dependence of PL peak intensities was maximized at 400 °C for each NW type, which are consistent with previous reports. Our results confirmed that the micro-PL demonstrated here is one of the important methods for characterizations of the interface defects in Si NWs.

Durable Response of Pembrolizmab for EGFR Mutation-positive Lung Adenocarcinoma with Early Progression to Osimertinib in First-line Treatment.

Osimertinib, a third-generation epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), is the standard first-line treatment for EGFR mutation-positive non-small-cell lung cancer (NSCLC) and demonstrates favorable disease control. Conversely, immune checkpoint inhibitors (ICIs) that target programmed cell death-1/programmed cell death ligands demonstrate a restrictive tumor response. We herein report a patient who achieved a durable response to pembrolizumab following early progression within two months of osimertinib administration for EGFR mutation-positive lung adenocarcinoma. Our findings suggest that treatment with ICIs for patients with EGFR mutation-positive NSCLC experiencing early progression to osimertinib as first-line treatment might represent a viable approach.

Hierarchical approach to evaluating storage requirements for renewable-energy-driven grids.

Energy storage can accelerate the decarbonization of the electrical grid. As useful energy storage technologies are developed, investors and manufacturers want to determine the needs for storage in a wide range of scenarios. In this study, we introduce a strategy for identifying the types of storage that will be most valuable to the grid given specific generation and load profiles. This method estimates the annual minimum number of cycles for each storage, how long each holds the charge, and charging and discharging rates for an idealized system, giving insight into tomorrow's complex systems. We demonstrate the proposed hierarchical approach and quantify how many fewer times wind-driven grids cycle the storage at night compared with solar-driven grids, as well as how winter-dominant wind generation and latitude-tilt solar may reduce the need for seasonal storage. Also, we quantify how higher discharging rates are required for energy storage products that cycle most frequently.

Antigen-antibody interactions and structural flexibility of a femtomolar-affinity antibody.

The femtomolar-affinity mutant antibody (4M5.3) generated by directed evolution is interesting because of the potential of antibody engineering. In this study, the mutant and its wild type (4-4-20) were compared in terms of antigen-antibody interactions and structural flexibility to elucidate the effects of directed evolution. For this purpose, multiple steered molecular dynamics (SMD) simulations were performed. The pulling forces of SMD simulations elucidated the regions that form strong attractive interactions in the binding pocket. Structural analysis in these regions showed two important mutations for improving attractive interactions. First, mutation of Tyr102(H) to Ser (sequence numbering of Protein Data Bank entry 1FLR ) played a role in resolving the steric hindrance on the pathway of the antigen in the binding pocket. Second, mutation of Asp31(H) to His played a role in resolving electrostatic repulsion. Potentials of mean force (PMFs) of both the wild type and the mutant showed landscapes that do not include obvious intermediate states and go directly to the bound state. These landscapes were regarded as funnel-like binding free energy landscapes. Furthermore, the structural flexibility based on the fluctuations of the positions of atoms was analyzed. It was shown that the fluctuations in the positions of the antigen and residues in contact with antigen tend to be smaller in the mutant than in the wild type. This result suggested that structural flexibility decreases as affinity is improved by directed evolution. This suggestion is similar to the relationship between affinity and flexibility for in vivo affinity maturation, which was suggested by Romesberg and co-workers [Jimenez, R., et al. (2003) Proc. Natl. Acad. Sci. U.S.A.100, 92-97]. Consequently, the relationship was found to be applicable up to femotomolar affinity levels.

[Esophageal fish bone migrating to the lung; report of a case].

A 55-year-old female felt a transient chest pain soon after a fish meal. Sixteen days later she presented a local clinic with fever of 38°C. After chest computed tomography( CT) and upper gastrointestinal endoscopy showed fish bone embedded in the wall of the middle portion of the esophagus, she referred to our hospital 22 days after the meal. Thirty days after the meal ,endoscopic removal of the fish bone was challenged bygastroentenologists in vain. Chest CT after the attempt showed migration of the fish bone to the lung adjacent to the right inferior pulmonary vein. The fish bone which stuck out from the lung after division of the pulmonary ligament was removed successfully under thoracoscopic surgery. The patient is well 26 months after the surgery.

Pan-cancer methylome analysis for cancer diagnosis and classification of cancer cell of origin.

The accurate and early diagnosis and classification of cancer origin from either tissue or liquid biopsy is crucial for selecting the appropriate treatment and reducing cancer-related mortality. Here, we established the CAncer Cell-of-Origin (CACO) methylation panel using the methylation data of the 28 types of cancer in The Cancer Genome Atlas (7950 patients and 707 normal controls) as well as healthy whole blood samples (95 subjects). We showed that the CACO methylation panel had high diagnostic potential with high sensitivity and specificity in the discovery (maximum AUC = 0.998) and validation (maximum AUC = 1.000) cohorts. Moreover, we confirmed that the CACO methylation panel could identify the cancer cell type of origin using the methylation profile from liquid as well as tissue biopsy, including primary, metastatic, and multiregional cancer samples and cancer of unknown primary, independent of the methylation analysis platform and specimen preparation method. Together, the CACO methylation panel can be a powerful tool for the classification and diagnosis of cancer.

Peptides derived from human insulin-like growth factor-II mRNA binding protein 3 can induce human leukocyte antigen-A2-restricted cytotoxic T lymphocytes reactive to cancer cells.

Insulin-like growth factor-II mRNA binding protein 3 (IMP-3) is an oncofetal protein expressed in various malignancies including lung cancer. This study aimed to identify immunogenic peptides derived from IMP-3 that can induce tumor-reactive and human leukocyte antigen (HLA)-A2 (A*02:01)-restricted cytotoxic T lymphocytes (CTL) for lung cancer immunotherapy. Forty human IMP-3-derived peptides predicted to bind to HLA-A2 were analyzed to determine their capacity to induce HLA-A2-restricted T cells in HLA-A2.1 (HHD) transgenic mice (Tgm). We found that three IMP-3 peptides primed HLA-A2-restricted CTL in the HLA-A2.1 Tgm. Among them, human CTL lines reactive to IMP-3 (515) NLSSAEVVV(523) were reproducibly established from HLA-A2-positive healthy donors and lung cancer patients. On the other hand, IMP-3 (199) RLLVPTQFV(207) reproducibly induced IMP-3-specific and HLA-A2-restricted CTL from healthy donors, but did not sensitize CTL in the HLA-A2.1 Tgm. Importantly, these two IMP-3 peptide-specific CTL generated from healthy donors and cancer patients effectively killed the cancer cells naturally expressing both IMP-3 and HLA-A2. Cytotoxicity was significantly inhibited by anti-HLA class I and anti-HLA-A2 monoclonal antibodies, but not by the anti-HLA-class II monoclonal antibody. In addition, natural processing of these two epitopes derived from the IMP-3 protein was confirmed by specific killing of HLA-A2-positive IMP-3-transfectants but not the parental IMP-negative cell line by peptide-induced CTL. This suggests that these two IMP-3-derived peptides represent highly immunogenic CTL epitopes that may be attractive targets for lung cancer immunotherapy.

A novel tumor-associated antigen, cell division cycle 45-like can induce cytotoxic T-lymphocytes reactive to tumor cells.

The present study attempted to identify a useful tumor-associated antigen (TAA) for lung cancer immunotherapy and potential immunogenic peptides derived from the TAA. We focused on cell division cycle 45-like (CDC45L), which has a critical role in the initiation and elongation steps of DNA replication, as a novel candidate TAA for immunotherapy based on a genome-wide cDNA microarray analysis of lung cancer. The CDC45L was overexpressed in the majority of lung cancer tissues, but not in the adjacent non-cancerous tissues or in many normal adult tissues. We examined the in vitro and in vivo anti-tumor effects of cytotoxic T-lymphocytes (CTL) specific to CDC45L-derived peptides induced from HLA-A24 (A*24:02)-positive donors. We identified three CDC45L-derived peptides that could reproducibly induce CDC45L-specific and HLA-A24-restricted CTL from both healthy donors and lung cancer patients. The CTL could effectively lyse lung cancer cells that endogenously expressed both CDC45L and HLA-A24. In addition, we found that CDC45L (556) KFLDALISL(564) was eminent in that it induced not only HLA-A24 but also HLA-A2 (A*02:01)-restricted antigen specific CTL. Furthermore, the adoptive transfer of the CDC45L-specific CTL inhibited the growth of human cancer cells engrafted into immunocompromised mice. These results suggest that these three CDC45L-derived peptides are highly immunogenic epitopes and CDC45L is a novel TAA that might be a useful target for lung cancer immunotherapy.

Significant change in electronic structures of heme upon reduction by strong Coulomb repulsion between Fe d electrons.

We report total-energy electronic-structure calculations based on the density functional theory performed on a low-spin heme. We have found that the high-lying occupied and low-lying unoccupied states having Fe d and/or porphyrin pi orbital character are significantly rearranged upon the reduction of the heme. An analysis of these states shows that the remarkable elevation of the Fe d levels takes place due to the strong Coulombic repulsion between accommodated d electrons. Due to a peculiarity of the heme, this elevation could be controlled by lower-lying empty porphyrin pi states, leading to electron transfer from Fe d orbitals to the porphyrin pi ones in order to reduce the Coulomb-energy cost. This self-limiting mechanism provides a natural explanation not only for the present calculated results, but also for general electron delocalization appearing under various physiological conditions, regardless of the types of the hemes.

Sialadenoma papilliferum of the bronchus: rare tracheobronchial tumor of salivary gland type.

Presented herein is the second case of sialadenoma papilliferum of the bronchus in a 75-year-old man. The bronchial tumor had an exophytic papillary lesion protruding into the lumen of the ostial region of the right superior lobe bronchus on chest CT and bronchoscopy. Grossly, the resected tumor was pedunculated, 5 x 3 x 3 mm in size and consisted of both an exophytic papillary lesion protruding into the bronchial lumen and a glandular component in the bronchial submucosa. Histologically, the papillary structures were lined by ciliated or non-ciliated columnar epithelium and metaplastic non-keratinizing stratified squamous epithelium. The submucosal glandular components were well circumscribed beneath and in continuity with the papillary lesion. The glandular components of cysts and duct-like structures were lined by a double-layer epithelium composed of luminal cells and basal cells. Sialadenoma papilliferum is a rarely recognized salivary gland tumor, mostly occurring at intraoral mucosal sites including the hard and soft palate and buccal mucosa. There have been sporadic cases of sialadenoma papilliferum of the esophagus, nasopharynx and nasal cavity. To the authors' knowledge only one case of sialadenoma papilliferum of the bronchus has been reported previously.

Theoretical study of the time-dependent phenomenon of photon-assisted tunneling through a charged quantum dot.

We have studied numerically the time-dependent photon-assisted tunneling (TD-PAT) process for electrons confined in quantum dots (QDs) by employing the finite difference method under the scheme of the TD-density functional theory (DFT). We have found the quasi-dark state (quasi-DS), where the injected electron remains in the QD. By varying the barrier thickness, we have calculated the TD profile of the electron density in a QD, and found the optimal geometry of the lozenge QD. We have also discussed how the charged QD modulates the PAT process.

Influence of edge magnetization and electric fields on zigzag silicene, germanene and stanene nanoribbons.

Using a multi-orbital tight-binding model, we have studied the edge states of zigzag silicene, germanene, and stanene nanoribbons (ZSiNRs, ZGeNRs and ZSnNRs, respectively) in the presence of the Coulomb interaction and a vertical electric field. The resulting edge states have non-linear energy dispersions due to multi-orbital effects, and the nanoribbons show induced magnetization at the edges. Owing to this non-linear dispersion, ZSiNRs, ZGeNRs and ZSnNRs may not provide superior performance in field effect transistors, as has been proposed from single-orbital tight-binding model calculations. We propose an effective low-energy model that describes the edge states of ZSiNRs, ZGeNRs, and ZSnNRs. We demonstrate that the edge states of ZGeNR and ZSnNR show anti-crossing of bands with opposite spins, even if only out-of-plane edge magnetization is present. The ability to tune the spin polarizations of the edge states by applying an electric field points to future opportunities to fabricate silicene, germanene and stanene nanoribbons as spintronics devices.

Quantum effects in a cylindrical carbon-nanotube capacitor.

We have theoretically investigated the electric polarization and capacitance in a nano-scale coaxial-cylindrical capacitor made of a double-walled carbon nanotube, (6,0)@(36,0), using a first-principles density-functional method with the enforced Fermi-energy difference scheme. We show that the distribution of the accumulated charge in the inner tube is quantum-mechanically spilled outward, while that in the outer tube is penetrating inward. Reflecting these charge spills, the electrostatic capacitance of the system is larger than what would be expected from classical theory. Furthermore, the total capacitance of the system is smaller than the electrostatic capacitance, which is another quantum effect showing large contributions from the DOS of the electrodes to the capacitance. We believe that these two quantum effects in the capacitance are important for the successful design of carbon-nanotube devices.

First-principles molecular dynamics study of proton transfer mechanism in bovine cytochrome c oxidase.

Density functional based first-principles molecular dynamics calculations, performed on a model system extracted from the bovine cytochrome c oxidase, have been performed in an attempt to inspect the proton transfer mechanism across a peptide group. Our model system includes the specific Tyr440-Ser441 peptide group involved in a novel proton transfer path and shows that the Y440-S441 enol peptide group [-C(OH) = N-], which is a structural isomer of a keto form [-CO-NH-], is the product of the deprotonation of an imidic acid [-C(OH)-NH-] occurring in the vicinity of the deprotonated aspartic acid residue. For the subsequent enol-to-keto tautomerization, a direct H(+) transfer path in the Y440-S441 peptide group has been identified, in which the transition state takes a distorted four-membered ring structure.

Interstitial fluid pressure of thymic epithelial tumours.

Tumours show an increased interstitial fluid pressure, which correlates with various pathophysiological features. Moreover, interstitial fluid pressure is a prognostic factor for cervical and lung cancer. However, there have been no reports on the usefulness of measuring interstitial fluid pressure in thymic epithelial tumours. Therefore, this study aimed to examine the relationship between interstitial fluid pressure and the clinicopathological characteristics of thymic epithelial tumours. Interstitial fluid pressure was prospectively measured at the centre of the tumour using a 1-Fr Mikro-Tip sensor catheter in 44 patients with thymic epithelial tumours, 40 with thymomas and 4 with thymic carcinomas. Data from these 44 patients were analysed for correlations between interstitial fluid pressure and clinicopathological and demographic factors including sex, age, tumour size, World Health Organization histological subtypes, myasthenia gravis, capsular invasion, mediastinal pleura invasion, lung invasion, pericardium invasion, dissemination, Masaoka-Koga stage, maximal standardized uptake value and recurrence-free survival (RFS). The mean interstitial fluid pressure was 11.3 mmHg; interstitial fluid pressure was significantly correlated with maximal standardized uptake value, lung invasion, dissemination and Masaoka-Koga stage. Low interstitial fluid pressure (≤14 mmHg) showed a tendency for better RFS compared with high interstitial pressure (P = 0.053). Lung invasion, dissemination and Masaoka-Koga stage were correlated with RFS in univariable analysis; lung invasion was selected as an independent prognostic factor in multivariable analysis. On the basis of these results, interstitial fluid pressure of thymic epithelial tumours has been shown to correlate with their clinicopathological features.