Identifying intense inflammatory subtype of esophageal squamous cell carcinoma using clustering approach.

OBJECTIVE: To establish a risk-stratification system for predicting the postoperative recurrence of esophageal squamous cell carcinoma, this study aimed to evaluate the prognostic value of clusters based on blood inflammation and coagulation markers and investigate their correlation with serum cytokines and genetic alteration. METHOD: This single-center, retrospective cohort study enrolled 491 patients with esophageal cancer who underwent subtotal esophagectomy between 2004 and 2012. For cluster exploration, nonhierarchical cluster analysis and k-means were applied using serum C-reactive protein, albumin, fibrinogen, and platelet-lymphocyte ratio as variables. Then, multivariate survival analysis was conducted to investigate the association of clusters with recurrence-free survival. To characterize the clusters, serum interleukin-6, interleukin-8, and genetic alteration in primary tumors, the PleSSision-Rapid panel, which can evaluate 160 representative driver genes, was used. RESULTS: Patients were classified into clusters 1, 2, and 3, which included 24 (5%), 161 (33%), and 306 (62%) patients, respectively. Compared with cluster 3, cluster 1 or 2 had significantly worse recurrence-free survival. Based on the multivariable analysis using cluster, pStage, and age as covariates, cluster was an independent prognostic factor for recurrence-free survival (hazard ratio, 1.55; 95% confidence interval, 1.08-2.21; P = 0.02). The percentage of serum interleukin-6 and interleukin-8 levels was the highest in cluster 1, followed by clusters 2 and 3. In 23 patients with available genomic profiles, no significant difference in representative genomic alterations was observed. CONCLUSIONS: Non-biased clustering using inflammation and coagulation markers identified the intense inflammatory subtype, which had an independent prognostic effect on recurrence-free survival.

Successful treatment of postoperative nonobstructive recurrent cholangitis by tract conversion surgery after total pancreatectomy: a case report.

BACKGROUND: Postoperative cholangitis is a complication of biliary reconstruction during hepatobiliary pancreatic surgery. Most cases are associated with anastomotic stenosis, but there are also cases of cholangitis without stenosis, and treatment can be difficult, especially in patients with recurrent symptoms. In this report, we describe a case of repeated nonobstructive cholangitis in a patient after total pancreatectomy, in which a good outcome was obtained after performing tract conversion surgery. CASE PRESENTATION: The patient was a 75-year-old man. He underwent total pancreatectomy for stage IIA cancer of the pancreatic body, hepaticojejunostomy via the posterior colonic route, gastrojejunostomy and Braun anastomosis via the anterior colonic route using the Billroth II method. The patient had a good postoperative course and was receiving adjuvant chemotherapy on an outpatient basis, but he developed his first episode of cholangitis 4 months after surgery. Although conservative treatment with antimicrobial agents was successful, the patient continued to have recurrent biliary cholangitis and was repeatedly admitted and discharged from the hospital. Since stenosis at the anastomosis was suspected, endoscopic observation of the anastomosis was performed using small bowel endoscopy for close examination, but no apparent stenosis was observed. Small bowel imaging indicated a possible influx of contrast medium into the bile duct, and reflux due to food residue was suspected as the cause of cholangitis. Since conservative treatment alone did not suppress the flare-up of symptoms, the decision was made to perform tract conversion surgery for curative purposes. The afferent loop was cut midstream, and jejunojejunostomy was performed downstream. The postoperative course was good, and the patient was discharged on the 10th day after surgery. He is currently an outpatient and has been free of cholangitis symptoms for 4 years without cancer recurrence. CONCLUSIONS: Although the diagnosis of nonobstructive retrograde cholangitis can be difficult, surgical treatment should be considered in patients with recurrent symptoms and refractory treatment.

Elevation of the Prognostic Factor Plasma Fibrinogen Reflects the Immunosuppressive Tumor Microenvironment in Esophageal Squamous Cell Carcinoma.

INTRODUCTION: Despite previous reports on the clinical significance of plasma fibrinogen (FNG) levels as a prognostic indicator of ESCC, its underlying mechanism remains unclear. This study aimed to validate the prognostic impact of plasma FNG levels and clarify its relationship with primary tumors in patients with esophageal squamous cell carcinoma (ESCC). METHODS: The prognostic impact of FNG was evaluated in patients with ESCC who underwent esophagectomy between 2000 and 2019. The RNA sequencing of the primary ESCC site, which was from pre-operative biopsy, was performed, followed by immune profile characterization using an immunogram. Those profiles were assessed via the immunohistochemical staining of tumor-associated macrophages (TAMs) and clinical response to nivolumab. RESULTS: Multivariate analysis identified FNG as a significant prognostic factor in ESCC. The immunogram suggested an immunosuppressive tumor environment in the high-FNG group. Immunostaining with the TAM markers CD163 and CD204, revealed that the high-FNG group had significantly higher number of TAMs compared with the low-FNG group. The immunosuppressive characteristics were clinically validated in patients with metastatic ESCC; those who had elevated FNG levels showed poor response to nivolumab. CONCLUSION: This study successfully validated the prognostic impact of plasma FNG levels in an expanded cohort with ESCC. Accordingly, our findings showed that increased plasma FNG reflects an immunosuppressive tumor microenvironment that facilitates tumor progression and poor responses to nivolumab.

Neutrophil-to-lymphocyte ratio change predicts histological response to and oncological outcome of neoadjuvant chemotherapy for esophageal squamous cell carcinoma.

BACKGROUND: Evaluating tumor response to neoadjuvant chemotherapy (NAC) is important to predict survival and to select the optimal strategy for patients with esophageal cancer. The aim of this study is to investigate the relation between neutrophil-to-lymphocyte ratio (NLR) change after NAC and histological response and oncological outcomes in patients with esophageal cancer. METHODS: This study enrolled 209 patients who underwent NAC and thoracic esophagectomy for esophageal cancer as the primary treatment between 2000 and 2019 in our department. Several predictors of survival including NLR change, which was calculated as post-NAC NLR/pre-NAC NLR, were investigated. We used classification and regression tree (CART) analysis to determine the optimal cutoff values of NLR change for the prediction of histological response. RESULTS: The best cutoff value of NLR change was 0.55 using the CART analysis. Then we divided all patients into two groups; the patients with NLR change below the cutoff were allocated to the NLR reduction group (n = 53), whereas the patients with NLR change above the cutoff were allocated to the no-NLR reduction group (n = 156). NLR change was identified as a significant predictor for histological responder (odds ratio 3.80; 95% confidence interval (CI) 1.69-8.58; p = 0.001) and recurrence-free survival (hazard ratio 0.55; 95% CI 0.33-0.89; p = 0.015) in multivariable analysis. CONCLUSION: The present study demonstrated that NLR change is associated with both histological response to and oncological outcomes of NAC for patients with esophageal cancer. NLR change can help not only to predict survival, but also to facilitate personalized multidisciplinary treatment.

Lymphocyte-to-C-Reactive Protein Ratio as a Novel Marker for Predicting Oncological Outcomes in Patients with Esophageal Cancer.

BACKGROUND: Esophageal cancer has a poor prognosis because of its rapid progression and early and extensive lymph node metastasis. Simple, objective indicators for predicting long-term outcomes are needed to select optimal perioperative treatment and appropriate follow-up for patients with esophageal cancer. The aim of this study is to investigate the relationship between the lymphocyte-to-C-reactive protein ratio (LCR) and the survival of patients with esophageal cancer, by performing time-dependent receiver operating characteristic (ROC) curve analysis. The results were compared to those of traditional inflammation-based markers. METHODS: This study enrolled 495 patients who underwent thoracic esophagectomy for esophageal cancer as the primary treatment between 2000 and 2019 in our department. We investigated the predictability of the LCR for oncological outcomes compared to that of other traditional inflammatory markers. RESULTS: The 3-year overall survival (OS) and recurrence-free survival (RFS) were 72.6% and 57.5%, respectively. Low LCR was significantly associated with higher cancer stage, included depth of invasion (p < 0.001), lymph node metastasis (p < 0.001) and cStage (p < 0.001). The LCR had the highest AUC value (0.675) for predicting OS compared to the other examined inflammatory markers. In multivariate analysis, the LCR (optimal cutoff threshold = 19,000) was identified as a significant predictor of death (hazard ratio, 2.24; 95% confidence interval [CI], 1.61-3.12; p < 0.001) and recurrence (hazard ratio, 1.97; 95%CI, 1.48-2.63; p < 0.001). CONCLUSION: The LCR is novel indicator for oncological outcomes for patients with esophageal cancer and may assist to facilitate personalized multidisciplinary treatments.

The Results of Sentinel Node Mapping for Patients with Clinically Early Staged Gastric Cancer Diagnosed with pT2/deeper Tumors.

BACKGROUND: Sentinel node (SN) mapping based on the SN concept has been applied to early gastric cancer. However, it is still controversial whether or not the oncological safety is ensured in case pathological stage was advanced in these patients. The aim of this study was to investigate the validity of SN mapping in patients with clinically early staged gastric cancer diagnosed with pT2/deeper tumors. METHODS: We retrospectively analyzed 40 patients with a diagnosis of cT1N0 or cT2N0 single-lesion gastric cancer who were shown to have pT2 or deeper tumors after gastrectomy with SN mapping. We adopted a dual-tracer method using a radioactive colloid and blue dye to detect SNs. The diagnostic accuracy and distribution of SNs at each tumor site were analyzed. RESULTS: Of the 40 patients, 24 (60%) were postoperatively diagnosed as pT2, and 16 (40%) as pT3 or T4. SNs were detected in all patients. The false negative rate was 9% (1/11), and in that patient, the non-SN metastasis was observed within the SN basin. Diagnostic accuracy was 98% (39/40). Overall distribution of SNs was similar to that for patients with early gastric cancer. No significant differences in overall and recurrence-free survival were observed between the patients who underwent standard gastrectomy and those who underwent function-preserving gastrectomy, based on the results of SN mapping. CONCLUSIONS: Our results confirmed validity of SN mapping for patients with clinically early staged gastric cancer diagnosed with pT2/deeper tumors after gastrectomy. Closed surveillance without additional surgical treatment is an option for these patients.

Usefulness of Neutrophil to Lymphocyte Ratio at Recurrence for Predicting Long-Term Outcomes in Patients with Recurrent Esophageal Squamous Cell Carcinoma.

BACKGROUND: Although radical esophagectomy with multifield lymph node dissection is a promising treatment to achieve long-term survival for resectable esophageal cancer, survival after postoperative recurrence remains poor. To select the optimal treatment for patients with recurrent esophageal cancer, simple, objective indicators for predicting of long-term outcomes are needed. PATIENTS AND METHODS: We conducted a single-institution, retrospective cohort study between 2004 and 2019, wherein 586 patients underwent transthoracic esophagectomy for primary esophageal squamous cell carcinoma. Of these, 133 patients with postoperative recurrence were included in this analysis. Several predictors of survival after recurrence were investigated. RESULTS: Among all patients, the 1- and 3-year survival rates after recurrence were 48.0% and 23.1%, respectively. On multivariate analysis, the neutrophil to lymphocyte ratio (NLR) at recurrence was identified as a significant predictor of death after recurrence (hazard ratio 1.061; 95% confidence interval 1.002-1.125; p = 0.043). Time-dependent receiver operating characteristics curves showed that the area under the curve value of the NLR at recurrence was superior to the modified Glasgow Prognostic Score at recurrence in all terms. To simulate the clinical decision process, we set the cut-off NLR at recurrence for survival using survival classification and regression tree (CART) and defined the optimal cut-off value as 3.374. CONCLUSIONS: NLR at recurrence was a significant indicator of survival after recurrence in patients with recurrent esophageal cancer. CART analysis was used to determine the optimal cut-off value for the prediction of survival, allowing the NLR to be used clinically to facilitate decision making.

Anisotropic energy transfer in a clay-porphyrin layered system with environment-responsiveness.

The adsorption orientation behavior of tetrakis(1-methylpyridinium-3-yl)porphyrin (m-TMPyP) and tetrakis(1-methylpyridinium-4-yl)porphyrin (p-TMPyP) on the clay monolayer prepared by the Langmuir Blodgett (LB) technique was investigated using the absorption and dichroic spectra obtained on a waveguide. It was revealed that the orientation of m-TMPyP and p-TMPyP on the clay monolayer, that is parallel and tilted with respect to the clay surface, depends on the surrounding environments such as water and N,N-dimethylformamide (DMF). The anisotropic photochemical energy transfer between m-TMPyP as a donor and p-TMPyP as an acceptor in the layered system was investigated in water and in DMF-water (9/1 (v/v)) by a fluorescence observation. As a result, while energy transfer efficiency (ηET) was 60% for the parallel-parallel orientation in water, that was 10% for the tilted-tilted orientation in DMF-water (9/1 (v/v)). The major factor for the change of ηET could be a change of the distance between m-TMPyP and p-TMPyP, and the J value that is a parameter for spectral overlap between energy donor's fluorescence and acceptor's absorption.

Crystallization and preliminary X-ray data analysis of the pXO1 plasmid-partitioning factor TubZ from Bacillus cereus.

TubZ is a structural homologue of tubulin and FtsZ GTPases, which are involved in the type III plasmid-partitioning system. TubZ assembles into polymers in a GTP-dependent manner and drives plasmid segregation as `cytomotive' filaments. In this study, C-terminally truncated TubZ from Bacillus cereus was crystallized in the presence or absence of GDP by the hanging-drop vapour-diffusion method. The crystal of TubZ in complex with GDP belonged to the monoclinic space group P2(1), with unit-cell parameters a=67.05, b=84.49, c=67.66 Å, ß=92.92°, and was non-isomorphous with GDP-bound TubZ previously crystallized in the presence of the slowly hydrolysable GTP analogue GTPγS. TubZ was also crystallized in the free form and the crystal belonged to space group P2(1), with unit-cell parameters a=53.91, b=65.54, c=58.18 Å, ß=106.19°. Data were collected to 1.7 and 2.1 Šresolution for the free and GDP-bound forms, respectively.

Filament formation of the FtsZ/tubulin-like protein TubZ from the Bacillus cereus pXO1 plasmid.

Stable maintenance of low-copy-number plasmids requires partition (par) systems that consist of a nucleotide hydrolase, a DNA-binding protein, and a cis-acting DNA-binding site. The FtsZ/tubulin-like GTPase TubZ was identified as a partitioning factor of the virulence plasmids pBtoxis and pXO1 in Bacillus thuringiensis and Bacillus anthracis, respectively. TubZ exhibits high GTPase activity and assembles into polymers both in vivo and in vitro, and its "treadmilling" movement is required for plasmid stability in the cell. To investigate the molecular mechanism of pXO1 plasmid segregation by TubZ filaments, we determined the crystal structures of Bacillus cereus TubZ in apo-, GDP-, and guanosine 5'-3-O-(thio)triphosphate (GTPγS)-bound forms at resolutions of 2.1, 1.9, and 3.3 Å, respectively. Interestingly, the slowly hydrolyzable GTP analog GTPγS was hydrolyzed to GDP in the crystal. In the post-GTP hydrolysis state, GDP-bound B. cereus TubZ forms a dimer by the head-to-tail association of individual subunits in the asymmetric unit, which is similar to the protofilament formation of FtsZ and B. thuringiensis TubZ. However, the M loop interacts with the nucleotide-binding site of the adjacent subunit and stabilizes the filament structure in a different manner, which indicates that the molecular assembly of the TubZ-related par systems is not stringently conserved. Furthermore, we show that the C-terminal tail of TubZ is required for association with the DNA-binding protein TubR. Using a combination of crystallography, site-directed mutagenesis, and biochemical analysis, our results provide the structural basis of the TubZ polymer that may drive DNA segregation.