Assessment of the albumin-bilirubin grade as a prognostic factor in patients with non-small-cell lung cancer receiving anti-PD-1-based therapy.

INTRODUCTION: The albumin-bilirubin (ALBI) grade is a novel indicator of the liver function. Some studies showed that the ALBI grade was a prognostic and predictive biomarker for the efficacy of chemotherapy in cancer patients. The association between the ALBI grade and outcomes in patients with non-small-cell lung cancer (NSCLC) treated with cancer immunotherapy, however, is poorly understood. METHODS: We retrospectively enrolled 452 patients with advanced or recurrent NSCLC who received anti-programmed cell death protein 1 (PD-1)-based therapy between 2016 and 2019 at three medical centers in Japan. The ALBI score was calculated from albumin and bilirubin measured at the time of treatment initiation and was stratified into three categories, ALBI grade 1-3, with reference to previous reports. We examined the clinical impact of the ALBI grade on the outcomes of NSCLC patients receiving anti-PD-1-based therapy using Kaplan-Meier survival curve analysis with log-rank test and Cox proportional hazards regression analysis. RESULTS: The classifications of the 452 patients were as follows: grade 1, n = 158 (35.0%); grade 2, n = 271 (60.0%); and grade 3, n = 23 (5.0%). Kaplan-Meier survival curve analysis showed that the ALBI grade was significantly associated with progression-free survival and overall survival. Moreover, Cox regression analysis revealed that the ALBI grade was an independent prognostic factor for progression-free survival and overall survival. CONCLUSION: The ALBI grade was an independent prognostic factor for survival in patients with advanced or recurrent NSCLC who receive anti-PD-1-based therapy. These findings should be validated in a prospective study with a larger sample size.

Exposure to an atomic bomb explosion is a risk factor for in-hospital death after esophagectomy to treat esophageal cancer.

Esophagectomy, one of the most invasive of all gastrointestinal operations, is associated with a high frequency of postoperative complications and in-hospital mortality. The purpose of the present study was to determine whether exposure to the atomic bomb explosion at Hiroshima in 1945 might be a preoperative risk factor for in-hospital mortality after esophagectomy in esophageal cancer patients. We thus reviewed the outcomes of esophagectomy in 31 atomic bomb survivors with esophageal cancer and 96 controls (also with cancer but without atomic bomb exposure). We compared the incidences of postoperative complications and in-hospital mortality. Of the clinicopathological features studied, mean patient age was significantly higher in atomic bomb survivors than in controls. Of the postoperative complications noted, atomic bomb survivors experienced a longer mean period of endotracheal intubation and higher incidences of severe pulmonary complications, severe anastomotic leakage, and surgical site infection. The factors associated with in-hospital mortality were exposure to the atomic bomb explosion, pulmonary comorbidities, and electrocardiographic abnormalities. Multivariate analysis revealed that exposure to the atomic bomb explosion was an independent significant preoperative risk factor for in-hospital mortality. Exposure to the atomic bomb explosion is thus a preoperative risk factor for in-hospital death after esophagectomy to treat esophageal cancer.

Improvement of lentiviral vector-mediated gene transduction by genetic engineering of the structural protein Pr55 Gag.

The lentiviral vector is a promising tool for human gene therapy because of its ability to transduce genes into many cell types. However, one of the technical problems associated with the lentiviral vector is that lentiviral titers in current production systems are relatively low compared with the other viral vectors. In this study, we provide genetic evidence that the attachment of heterologous myristoylation (myr) signals on the amino-terminus of human immunodeficiency virus type 1 Pr55(Gag) (Gag) can increase the viral yield up to 10-fold, leading to the enhancement of gene transduction in many cell lines. The myr signal Gag constructs behaved similarly to the wild-type Gag in targeting to detergent-resistant membrane compartments, Vps4-dependence for viral budding, and virion morphology. However, the myr signal Gag constructs showed improved oligomerization efficiency as measured by bioluminescence resonance energy transfer in living cells, contributing to increased viral production and efficient activation of the viral protease responsible for virion maturation. The genetically modified Gag represents the next generation lentiviral vector, and should contribute to the success of many lentiviral vector applications.

Antitumor activity of a phenoxazine compound, 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one against human B cell and T cell lymphoblastoid cell lines: induction of mixed types of cell death, apoptosis, and necrosis.

PURPOSE: We studied the antitumor activity of 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one (Phx), which was synthesized by the reactions of 2-amino-5-methylphenol with bovine hemoglobin, on human B cell lymphoblastoid cell lines, P3HR-1 and Raji derived from African Burkitt's lymphoma, and the human T cell lymphoblastoid cell line Molt-4. We also studied whether Phx might cause apoptosis and necrosis in these cells. METHODS: We evaluated cell viability and apoptosis and necrosis of the cells in the presence of Phx, by using agarose gel electrophoresis, flow cytometry, and fluorescence microscopy. RESULTS: Phx suppressed the viability of P3HR-1, Raji, and Molt-4 cells, though the suppression patterns were different, i.e., Phx suppressed the viability of P3HR-1, Raji, and Molt-4 cells at higher concentrations, while the drug enhanced the viability of Raji cells, but not those of P3HR-1 and Molt-4 cells at lower concentrations. To investigate which type of cell death - apoptosis or necrosis - is induced by Phx, induction of DNA ladder, phosphatidylserine externalization, and propidium iodide-permeable cells were examined in Phx-treated cells. Although Phx did not induce DNA ladder formation, it induced the phosphatidylserine externalization and propidium iodide-permeable cells, suggesting that Phx caused a mixed type of cell death, both apoptosis and necrosis. The population of early stage apoptotic cells was dominant in Raji cells, and that of the late stage apoptotic/necrotic cells was dominant in Molt-4 cells after 72-h treatment with Phx. The population of the early stage apoptotic cells and the late stage apoptotic/necrotic cells was almost equal in P3HR-1 cells in the presence of Phx, though the population of both types of cells increased with time. The nuclear morphological analysis of Phx-treated Raji, P3HR-1, and Molt-4 cells also showed that Phx induces apoptosis. CONCLUSIONS: The present results suggest that Phx shows antitumor activity against human B cell-derived and T cell-derived lymphoblastoid cell lines, in vitro, causing apoptosis and necrosis.

Lung cancer with p53 expression and a solitary metastasis to the stomach: a case report.

Lung cancer with a solitary metastasis to the stomach occurred in a 65-year-old man, surgically treated for gastric metastasis was followed by pulmonary resection. The gastric metastasis accompanied by upper gastrointestinal hemorrhage. After total gastrectomy to control this hemorrhage, a left lower lobectomy with a partial resection of the lingular segment and combined resection of the chest wall were done. Histopathological features of both the primary tumor in the left lower lobe and the gastric tumor were poorly differentiated adenocarcinoma, and showed the same immunoreactivities of p53 protein, carcinoembryonic antigen and keratin. These results indicate that the gastric tumor was a metastasis originated from the lung cancer.

Phosphatidylserine induces apoptosis in adherent cells.

Phosphatidylserine (PS) is exposed on the outer leaflet of the plasma membrane in apoptotic cell death. However, the roles of PS in apoptotic signaling are still unclear. In this study, we found that exogenous PS, but not other phospholipids, induced cell death in adherent cells, but not in suspension culture. The cell death exhibited typical features of apoptosis such as cell shrinkage, nuclear fragmentation and abnormal chromatin condensation. When PS was added to CHO-K1 cells in monolayer culture, they began to show changes in cell shape and actin cytoskeleton and protein kinase C (PKC) activity, followed by cell detachment, caspase activation, cleavage of focal adhesion kinase (FAK) and finally loss of viability. These results suggested that PS causes apoptosis through actin disorganization, cell detachment and cleavage of FAK.

Zinc induces mixed types of cell death, necrosis, and apoptosis, in molt-4 cells.

To investigate the mode of zinc-induced cell death, the associated morphological changes, and biological events were examined in zinc-treated Molt-4 cells. Fluorescence microscope observations with double staining of zinc-treated cells with Hoechst 33342 and propidium iodide (PI) indicated that the metal induced both necrosis and apoptosis. To confirm this, cells were stained with both PI and FITC-labeled annexin V, which binds phosphatidylserine, and then analyzed by flow cytometry. The results also confirmed that zinc induces mixed types of cell death, necrosis and apoptosis, and that the former induction occurs earlier and at a greater frequency. Hallmarks of apoptosis such as abnormal chromosome condensation and release of cytochrome c, as well as the appearance of annexin-positive cells, appeared along with the expression of mitochondrial membrane protein 7A6. However, zinc did not induce increases in caspase-3 like protease and caspase-8 activities, and caused slightly hypodiploid cells. Furthermore, the induction of cell death and annexin-positive cells was not blocked by the caspase inhibitors Ac-YVAD-CHO and Ac-DEVD-CHO. These results indicate that zinc induces both necrosis and apoptosis, without caspase-3 activation.

Evaluation of p53 alterations in occult lymph node metastases.

BACKGROUND AND OBJECTIVES: This study was designed to evaluate p53 alterations in occult lymph node metastases. METHODS: We examined 41 patients with stage I non-small-cell lung cancer. We investigated p53 gene mutation by polymerase chain reaction and single-strand conformation polymorphism analysis of exons 5-8, p53 protein accumulation by immunostaining with monoclonal antibody DO-7, and detection of tumor cells in lymph nodes by immunohistochemistry with monoclonal antibodies to cytokeratin (CK). RESULTS: p53 gene mutation was detected in 34% of tumors and nuclear p53 accumulation in 46%. CK-positive cells in the hilar and mediastinal region lymph nodes were detected in 43.9% of patients and 29.3%, respectively. Of the 14 cases with p53 mutation and the 19 cases with p53 accumulation, 12 and 15 had micrometastases in the hilar or mediastinal lymph nodes, respectively. However, p53 alterations were not significantly associated with occult lymph node metastases. In cases with occult lymph node metastases, the 5-year survival was 81. 9% for the p53 wild-type group and 45.8% for the p53 mutation group. CONCLUSIONS: p53 alterations are not correlated with occult lymph node metastases, while p53 gene mutation is considered to be an unfavorable prognostic marker in patients with occult lymph node metastases. J. Surg. Oncol. 2000;73:143-147.

Duodenal metastasis from large cell carcinoma of the lung: report of a case.

Duodenal metastasis from primary lung cancer is extremely rare. It rarely shows any symptoms, and the prognosis for this condition is poor. We herein describe the case of a 46-year-old woman with primary lung cancer who underwent a left upper lobectomy. Severe anemia was observed about 20 days after lobectomy. Gastroduodenoscopy showed duodenal metastasis. Simultaneously, brain metastasis was also detected using magnetic resonance imaging. The patient underwent a local resection of the duodenum and a tumor resection of the brain. Postoperative irradiation of the brain metastases and systemic chemotherapy of the lung metastases were performed, and complete remission occurred. However, abdominal lymph node metastasis recurred, and the patient died 1 year after the lobectomy.

Decreased thymosin beta4 in apoptosis induced by a variety of antitumor drugs.

As many antitumor drugs can kill tumors through the induction of apoptosis, the effect of these drugs presumably would be enhanced if they were used in combination with other drugs that interact with apoptotic processes. To clarify the biological events involved in the induction of apoptosis, we examined changes in the proteins associated with induction of apoptosis by antitumor drugs. When Molt-4 cells were exposed to the antitumor drugs etoposide, meso-2,3-bis(3,5-dioxopiperazine-1-yl)butane (ICRF-193), and neocarzinostatin, they exhibited apoptotic cell death as determined by flow cytometry using fluorescein isothiocyanate (FITC)-labeled annexin V staining of phosphatidylserine on membranes and detection of hypodiploid cells. Following the induction of apoptosis, a low molecular weight protein that was identified to be thymosin beta4 by HPLC analysis was commonly decreased, and the morphology of actin filaments changed into clump formations. These results suggest that decreased thymosin beta4 is involved in the induction of apoptosis by antitumor drugs.

Induction of necrosis by zinc in prostate carcinoma cells and identification of proteins increased in association with this induction.

Zinc exhibits inhibitory effects on apoptosis, and a deficiency in this metal generally causes this type of cell death to occur. In the present study, we found that exposure to zinc results in necrosis of prostate carcinoma cells. When zinc acetate was added to LNCaP or PC-3 cells in monolayer culture, they began to detach from the culture dishes, and viability was lost after 4-8 h. Most of the cell death was found to be due to necrosis as determined by double staining with fluorescein-isothiocyanate-labeled annexin V and ethidium bromide, and by detection of hypodiploid cells. Associated with the induction of necrosis was an increase in low molecular-mass proteins, identified by HPLC analysis to be thymosin beta10, parathymosin and GAGE in LNCaP cells, and thymosin beta4, parathymosin and metallothionein in PC-3. The time course of the increase of thymosin beta10 in LNCaP cells and thymosin beta4 in PC-3 cells was consistent with that of appearance of cell detachment and dead cells. These results indicate that zinc can induce necrosis and suggest that production of proteins including beta-thymosins is involved in induction of processes leading to cell detachment.

[Decortication in chronic thoracic empyemas--a report of three cases].

Three cases of chronic thoracic empyema treated by decortication are reported with special reference to the indications for surgery. The first patient was a 68-year-old man who had right chronic thoracic empyema with a bronchopleural fistula. He underwent open thoracostomy, and decortication was performed after 8 months. The second patient was a 74-year-old man who had right chronic empyema without bronchopleural fistula. Open thoracostomy was also performed and decortication was done after 2 months. Postoperative pulmonary function was significantly improved in both patients. The third patient was a 66-year-old man who had left chronic empyema with a bronchopleural fistula. He underwent open thoracostomy and left lower lobectomy, and then decortication and the omental pedicle flap method were performed after 4 months. All three patients are still doing well currently. It is concluded that decortication significantly improves pulmonary function in properly selected patients, and that computed tomography is helpful for assessing the re-expansion ability of the collapsed lung.

Effects of ICRF-193, a catalytic inhibitor of DNA topoisomerase II, on sister chromatid exchange.

To investigate whether mammalian DNA topoisomerase II is directly involved in recombination events, the effects of ICRF-193, a specific catalytic inhibitor on sister chromatid exchange (SCE), were examined in MR-6 cells. ICRF-193 only slightly elevated SCE formation after 3 or 44 h treatments, while VP-16, a cleavable complex forming type of topoisomerase II inhibitor, caused significant enhancement. ICRF-193 had no effect on N-methyl-N'-nitro-N-nitrosoguanidine-induced SCE formation. It would thus appear that the inhibition of topoisomerase II does not affect recombinational repair, and that topoisomerase II inhibitors such as VP-16 and 4'-(9-acridinyl amino) methane sulfon-m-anisidide induce SCE through production of DNA strand breaks, rather than by inhibiting the enzyme activity.

Aggressive pulmonary metastasectomies for synovial sarcoma.

A 51-year-old woman underwent a bilateral wedge resection of lung metastases through a median sternotomy 10 months after an initial operation for synovial sarcoma of the left lower extremity. Since then, five right and two left posterolateral thoracotomies have been performed over a 6-year period. The patient is presently doing well 7 years after the initial operation of the left lower extremity without any evidence of recurrence.

Kinetic analysis of recurrence and survival after potentially curative resection of nonsmall cell lung cancer.

BACKGROUND: About two-thirds of the patients with nonsmall cell lung cancer (NSCLC) who undergo a potentially curative resection eventually suffer from recurrent disease. However, it has yet to be elucidated as to how survival after recurrence is influenced by different variables, including timing, type of recurrence, or other clinicopathological features. There have been few studies concentrating on the kinetics of growth of occult micrometastatic tumor cells that eventually manifest as tumor recurrence. METHODS: We retrospectively reviewed the charts of 197 patients who developed recurrence after a potentially curative resection for NSCLC. RESULTS: The median disease-free interval was a little over 1 year (395 days), as was the median postrecurrence survival-383 days. We created a model for the kinetics of recurrence by assuming that: (1) a tumor of 10(9) cells is the usual limit of detection, (2) patients generally die before the tumor reaches 10(12) cells, and (3) it takes 1 year for average lung cancer cells to show a 10-fold increase. The model indicated that as much as 10(9) tumor cells should have been present immediately after the operation. Alternatively, the residual tumor cells should have an accelerated growth after the surgery. CONCLUSIONS: These models indicate the importance of developing a sensitive detection method for occult metastatic cells and to understand the tumor dormancy mechanism.

Antitumor activity of 2-amino-4,4 alpha-dihydro-4 alpha, 7-dimethyl-3H-phenoxazine-3-one, a novel phenoxazine derivative produced by the reaction of 2-amino-5-methylphenol with bovine hemolysate.

2-Amino-4,4 alpha-dihydro-4 alpha,7-dimethyl-3H-phenoxazine-3-one (Phx) was synthesized by the reaction of 2-amino-5-methyl-phenol with bovine hemolysates. Since Phx is a phenoxazine derivative like actinomycin D, which exerts a strong anti-tumor effect by intercalating DNA, we examined the effects of Phx on cell proliferation and cell cycle progression in human epidermoid carcinoma cells (KB cells). Phx inhibited the proliferation of Kb cells in a dose-dependent manner. When KB cells were incubated for 9 h with medium containing 50 microM Phx, a transient accumulation of cells in S and G2/M phase was observed and at 24 h many of cells had lower DNA content. Although Phx had antitumor activity, the drug did not intercalate DNA, showing a different mode of action from actinomycin D.

Prognostic value and clinicopathological correlation of thrombomodulin in squamous cell carcinoma of the human lung.

Aggregation of host platelets by circulating tumor cells is believed to play an important role in the metastatic process. Because thrombomodulin (TM) is one of the major mediators of the activation of the anticoagulant protein C by thrombin, we examined 136 primary tumor tissues and 45 metastatic lymph node tissues of lung squamous cell carcinomas for TM expression using immunohistochemical methods. The number of tumors with positive TM staining was less in metastatic tumors (44%) than in primary tumors (74%) (P < 0.01). Of various clinicopathological factors, better differentiation and lower N stage were significantly associated with TM expression. A loss of TM expression was associated with a shortened survival in 113 patients who underwent complete resection of the lung tumor (P < 0.01). In this group, TM expression and tumor-node-metastasis staging were independent significant determinants for survival, determined using Cox's multivariate survival analysis. Since TM is apparently associated with tumor progression and differentiation, this correlation may serve as a prognostic indicator in squamous cell carcinoma of the lung.

Video-assisted thoracoscopic surgery of bullous and bleb disorders of the lung using endoscopic stapling device.

The video-assisted thoracic surgical (VATS) approach appears to be a viable alternative to thoracotomy when surgical management of bullous and bleb disorders of the lung is required. Fifty patients with giant bullae (n = 6) and spontaneous pneumothoraces (n = 44) were recently treated by our group using the VATS approach and endoscopic stapling devices. Of the 50 patients, 47 were managed completely by the VATS approach, including six giant bullae that were asymptomatic in five and infectious in one and 41 pneumothoraces, of which 16 were first episode and 25 with recurrent pneumothorax. Median operating times for the bullous and bleb excisions were 147.8 and 45.9 min, respectively (p < 0.01), and median chest tube durations were 5.2 and 1.2 days, respectively (p < 0.05). There was no mortality, and significant morbidity was limited to prolonged air leak in more than 5 days in three patients and postoperative atelectasis in two patients. Median hospital stays of patients with bullous excision was 11.3 days compared with 4.7 days of those with bleb excision. We conclude that the VATS treatment is a safe, effective procedure in patients with bullous and bleb disorders of the lung even in asymptomatic giant bullae or spontaneous pneumothoraces with the first episode. The advantages of the VATS approach for these diseases are ease of operation, less pain, early mobility, and superior cosmetic results.

Characteristics of non-small cell lung cancer 3 cm or less in diameter.

We retrospectively investigated 308 cases of non-small cell lung cancer of < or = 3 cm diameter. There were 204 adenocarcinomas, 78 squamous cell carcinomas, 15 large cell carcinomas, and 11 other carcinomas. According to TNM staging, there were one case stage 0, 208 stage I, 22 stage II, 49 stage IIIA, 15 stage IIIB, and 13 cases stage IV. T1 disease was seen in 262 cases, T2 in 19, T3 in 10, T4 in 16, and Tis in 1. N0 disease was seen in 217 cases, N1 in 30, N2 in 60, and N3 in 1. The 5-year survival rate of all cases was 63%. There were statistically significant differences among T status (T1 vs. T3, T4), N status (N0 vs. N1, N2), and M status (M0 vs. M1) (P < 0.01). The 5-year survival rates of cases with adenocarcinoma and squamous cell carcinoma were 60% and 64%, respectively. In 204 cases of adenocarcinoma, T3 disease was found in one case, T4 disease in 15 (7%), and nodal involvement (N1 + N2) was present in 69 (34%). In 78 cases of squamous cell carcinoma T3 was seen in 6 (8%), T4 in 1, and nodal involvement in 14 (18%). The incidence of T3 disease, T4, and N(+) varied significantly according to histology (P < 0.05). Our investigation suggested that cases of small-sized lung cancer were often at an advanced stage at detection, and that the spread of disease differed according to histology. The patient with small-sized lung cancer should be offered a standard operation regardless of histology.

The close relationship between growth factors and the nucleolar organizer regions in adenocarcinoma of the lung.

We examined immunohistochemically 111 cases of primary adenocarcinoma of the lung, for transforming growth factor alpha (TGF alpha) or epidermal growth factor (EGF), and argyrophilic nucleolar organizer regions (AgNORs). The presence of more than 75% positive cells for both growth factors was designated as a high-GF, while all others were considered to be a low-GF. If AgNORs counts were more than 5.00, it was considered to be a high-AgNORs group, while less than 5.00 was designated as a low-AgNORs group. In our 111 examined specimens, there were 51 (46%) cases of high-GF, and 64 (58%) with high AgNORs. The 5-year survival rates of the patients with a high-GF and low-GF were 34% and 57% (P < 0.05) respectively, while those with high-AgNORs and low-AgNORs were 21% and 81% (P < 0.001), respectively. In the cases of high-AgNORs, the 5-year survival rates of the patients with high-GF and low-GF were 0% and 36% (P < 0.05), respectively. However, in the cases of low-AgNORs, the 5-year survival rates of the patients with high-GF and low-GF were 83% and 79%, respectively. These data suggest that growth factors might be related to the biological malignancy of tumours with a high cell proliferation.