MUC1-C integrates PD-L1 induction with repression of immune effectors in non-small-cell lung cancer.

Immunotherapeutic approaches, particularly programmed death 1/programmed death ligand 1 (PD-1/PD-L1) blockade, have improved the treatment of non-small-cell lung cancer (NSCLC), supporting the premise that evasion of immune destruction is of importance for NSCLC progression. However, the signals responsible for upregulation of PD-L1 in NSCLC cells and whether they are integrated with the regulation of other immune-related genes are not known. Mucin 1 (MUC1) is aberrantly overexpressed in NSCLC, activates the nuclear factor-κB (NF-κB) p65→︀ZEB1 pathway and confers a poor prognosis. The present studies demonstrate that MUC1-C activates PD-L1 expression in NSCLC cells. We show that MUC1-C increases NF-κB p65 occupancy on the CD274/PD-L1 promoter and thereby drives CD274 transcription. Moreover, we demonstrate that MUC1-C-induced activation of NF-κB→︀ZEB1 signaling represses the TLR9 (toll-like receptor 9), IFNG, MCP-1 (monocyte chemoattractant protein-1) and GM-CSF genes, and that this signature is associated with decreases in overall survival. In concert with these results, targeting MUC1-C in NSCLC tumors suppresses PD-L1 and induces these effectors of innate and adaptive immunity. These findings support a previously unrecognized central role for MUC1-C in integrating PD-L1 activation with suppression of immune effectors and poor clinical outcome.

MUC1-C activates BMI1 in human cancer cells.

B-cell-specific Moloney murine leukemia virus integration site 1 (BMI1) is a component of the polycomb repressive complex 1 (PRC1) complex that is overexpressed in breast and other cancers, and promotes self-renewal of cancer stem-like cells. The oncogenic mucin 1 (MUC1) C-terminal (MUC1-C) subunit is similarly overexpressed in human carcinoma cells and has been linked to their self-renewal. There is no known relationship between MUC1-C and BMI1 in cancer. The present studies demonstrate that MUC1-C drives BMI1 transcription by a MYC-dependent mechanism in breast and other cancer cells. In addition, we show that MUC1-C blocks miR-200c-mediated downregulation of BMI1 expression. The functional significance of this MUC1-C→︀BMI1 pathway is supported by the demonstration that targeting MUC1-C suppresses BMI1-induced ubiquitylation of H2A and thereby derepresses homeobox HOXC5 and HOXC13 gene expression. Notably, our results further show that MUC1-C binds directly to BMI1 and promotes occupancy of BMI1 on the CDKN2A promoter. In concert with BMI1-induced repression of the p16INK4a tumor suppressor, we found that targeting MUC1-C is associated with induction of p16INK4a expression. In support of these results, analysis of three gene expresssion data sets demonstrated highly significant correlations between MUC1-C and BMI1 in breast cancers. These findings uncover a previously unrecognized role for MUC1-C in driving BMI1 expression and in directly interacting with this stem cell factor, linking MUC1-C with function of the PRC1 in epigenetic gene silencing.

Survival outcomes of 220 consecutive patients with three-staged thoracoscopic esophagectomy.

Patients with thoracic esophageal cancer are often treated by minimally invasive esophagectomy. However, the long-term survival benefits of minimally invasive esophagectomy remain unclear. Two approaches are available for thoracoscopic surgery: one with the patient in the left lateral decubitus position (LLDP), and the other with the patient in the prone position (PP). We investigated the survival benefit of thoracoscopic esophagectomy according to the tumor stage and patient position during the thoracoscopic procedure. We reviewed the records of 220 consecutive patients with esophageal cancer treated from 1998 to 2012. In total, 146 and 74 patients were treated with thoracoscopic esophagectomy in the LLDP and PP, respectively. No patients were initially proposed to be candidates for esophagectomy by thoracotomy during the study period. Data collection was performed with a focus on survival and recurrent disease. Among all the 220 patients, the overall 5-year survival rates were 83.7%, 74.1%, 45.5%, 78.6%, 44.2%, 29.4% and 24.3% in the patients with pStage IA, IB, IIA, IIB, IIIA, IIIB and IIIC disease, respectively. Despite the greater number of dissected mediastinal lymph nodes in the PP procedure, there were no significant differences in the survival curves between the LLDP and PP procedures. The long-term results of thoracoscopic esophagectomy are comparable and acceptable. The PP procedure was not confirmed to offer a superior survival benefit to the LLDP procedure in this retrospective study.

Beamline AR-NW12A: high-throughput beamline for macromolecular crystallography at the Photon Factory.

AR-NW12A is an in-vacuum undulator beamline optimized for high-throughput macromolecular crystallography experiments as one of the five macromolecular crystallography (MX) beamlines at the Photon Factory. This report provides details of the beamline design, covering its optical specifications, hardware set-up, control software, and the latest developments for MX experiments. The experimental environment presents state-of-the-art instrumentation for high-throughput projects with a high-precision goniometer with an adaptable goniometer head, and a UV-light sample visualization system. Combined with an efficient automounting robot modified from the SSRL SAM system, a remote control system enables fully automated and remote-access X-ray diffraction experiments.

Intra-arterial therapy with cisplatin suspension in lipiodol and 5-fluorouracil for hepatocellular carcinoma with portal vein tumour thrombosis.

BACKGROUND: Portal vein tumour thrombosis is a negative prognostic factor for hepatocellular carcinoma (HCC). AIM: To assess the efficacy of cisplatin in lipiodol emulsion combined with 5-fluorouracil (5-FU) for patients with HCC and portal vein tumour thrombosis. METHODS: The study subjects were 51 patients with the above-specified criteria who received injection of cisplatin suspension in lipiodol emulsion followed by intra-arterial infusion of 5-FU. The primary objective was to determine tumour response to the treatment, while the secondary objectives were safety and tolerability. Independent factors for survival were also assessed. RESULTS: Ten patients had complete response and 34 patients had partial response (response rate, 86.3%). The median survival for all 51 patients was 33 months, while that for 10 complete response patients and 21 patients who showed disappearance of HCC following additional therapies was 39 months. The single factor that significantly influenced survival was therapeutic effect. Treatment was well tolerated and severe toxicity was infrequent, with only grade 3 toxicity (thrombocytopenia) in one patient. CONCLUSIONS: The present study demonstrated the efficacy of hepatic arterial infusion chemotherapy using cisplatin-lipiodol emulsion and 5-FU without serious adverse effects in patients with unresectable HCC and portal vein tumour thrombosis.

Enhancement of caffeine-induced locomotor hyperactivity produced by the combination with L-arginine or taurine in mice: Possible involvement of nitric oxide.

Combinations of caffeine with L-arginine or with taurine can enhance the effect of caffeine, but the mechanisms remain elusive. This study was designed to test the hypothesis that stimulant effects of central nervous system nitric oxide (NO) may explain the beneficial effect of caffeine on combinations with amino acid, L-arginine or taurine. Caffeine increased the spontaneous locomotor activity dose-dependently (2-10 mg/kg) in mice. The locomotor activity induced by caffeine at a dose of 2 mg/kg was enhanced by combined administration of L-arginine at a dose of 600 mg/kg, or taurine at a dose of 400 mg/kg, respectively. For both combinations, enhancement was significantly inhibited by pretreatment with N-nitro-L-arginine methyl ester (L-NAME) at a dose of 40 mg/kg. These results suggest that the enhancement induced by combining caffeine with amino acid might be regulated at least in part by NO in the central nervous system.

Treatments of non-parasitic giant hepatic cysts.

This retrospective study presents the results of surgical treatments for large cyst of the liver over 10 cm in diameter in 9 patients diagnosed and treated at Kurume University Hospital. There were 8 women and 1 man, with an average age of 71.6 years. Although the chief complaints were abdominal pain or fullness, 1 had obstructive jaundice due to biliary compression by a large cyst. Cyst size ranged from 10 cm to 27 cm. There are several treatment modalities for giant hepatic cyst, such as cyst resection, unroofing, and sclerotherapy after cyst drainage. Operative procedures in the patients reported here were surgical resection of the liver cyst in 3 patients, unroofing with ethanol sclerotherapy in 1 patient and laparoscopic treatment in 2 patients. Sclerotherapy after percutaneous transhepatic cyst drainage was performed in 3 patients because their general condition was poor. There were no postoperative complications among these patients. Costs varied among the patients and depended mainly on the method of treatment and period of hospital stay. We discuss operative procedures and costs of treatment of each patient and review the literature.

Resectable carcinoma of the pancreatic head developing 7 years and 4 months after distal pancreatectomy for carcinoma of the pancreatic tail.

A 67-year-old woman was referred with an abnormal finding on an abdominal echogram but presented with no symptoms; a pancreatic tail tumor was detected by ultrasonography. Biochemical examinations showed slight elevation of serum carcinoembryonic antigen level. The lesion was resected by tail and body pancreatectomy and her postoperative course was uneventful. Seven years and 4 months after the initial operation, however, her serum level of carbohydrate antigen 19-9 was found to be elevated, and a recurrence of pancreatic cancer was suspected. Examinations revealed a mass in the head of the remnant pancreas. The lesion was radically resected by total remnant pancreatectomy. Histological examinations showed that the initial tumor was a well differentiated tubular adenocarcinoma, while the second tumor was characterized as a moderately differentiated tubular adenocarcinoma. The surgical margins of the distal pancreatectomy specimen were free of atypical cells. Therefore, the position of the second lesion diminished the likelihood that it had developed by intrapancreatic metastasis. This suggests that the second carcinoma in the head of the pancreas may have been a second primary lesion.

Therapeutic effect of clarithromycin for respiratory-tract infections in children caused by Chlamydia pneumoniae. Research Group of Sapporo for Pediatric Chlamydial Infections.

Children infected with Chlamydia pneumoniae sometimes experience lower respiratory tract infections such as pneumonia and bronchitis. Although numerous anti-microbial compounds have been reported to be active against the organism, most of them have not been in a clinical trial in infants and children with C. pneumoniae infection. Clarithromycin has been shown to express anti-chlamydial effects in vitro. In this study, we evaluated the clinical anti-C. pneumoniae properties of clarithromycin in children with mainly lower respiratory tract infection. We administered clarithromycin orally to 21 infants and children at a dose of 10-15 mg/kg/day divided into two or three doses for 4-21 days. Clinical symptoms, roentgenographic and laboratory abnormal findings improved. The overall clinical efficacy rate was 85.7% (18 of 21 cases). Administration of clarithromycin was considered to be a suitable treatment for improving lower respiratory infections in infants and children caused by C. pneumoniae.

Hepatic arterial chemotherapy for liver metastases from colorectal cancer.

We have investigated the complications, toxicities, tumor response and survival rate with hepatic arterial (HA) chemotherapy for liver metastases from colorectal cancer. Forty-three patients with measurable liver metastases and 10 patients after the resection of liver metastases were treated with HA bolus mitomycin C (MMC) or continuous fluorouracil (5-FU) infusion between 1986 and 1994. The catheter- or reservoir-related complications such as catheter induced infection, subcutaneous reservoir pocket infection, or catheter or hepatic artery occlusion occurred in 14 patients (26%) mainly in our early cases. Severe gastritis or a gastroduodenal ulcer developed in 12 patients (23%), in particular after treatment with continuous infusion of 5-FU and when the catheter was placed into the gastroduodenal artery during laparotomy. An ulcer often caused a serious complication such as a duodeno-biliary fistula, penetration into the hepatic artery or duodenal perforation. Also noted were upper gastrointestinal symptoms in 26 patients, hepatic toxicities in 3 patients, and bone marrow depression in 4 patients. Because of these complications and toxicities, HA therapy was discontinued in more than 60% of the cases. The overall tumor response rate was 17%. However, in the cases which were treated by 5-FU continuous infusion for longer than 3 months, the response rate was higher than 60%. There was no significant difference in survival rate between HA therapy and systemic palliative chemotherapy. These results suggest that it is important to prevent gastrointestinal toxicities in order to allow continuation of HA continuous 5-FU infusion therapy which could provide a higher response rate and a better survival for colorectal cancer patients with liver metastases.

Evaluation of imbalance of the vestibulo-spinal reflex by "the circular walking test".

A sensitive method of demonstrating latent imbalance of labyrinthine function was devised. The subject was asked to walk at his or her usual speed with eyes closed in a circle 2 m in diameter. This test was done by 22 patients whose vestibular function as tested by the Romberg, Mann, Fukuda and Babinski-Weil's methods had become normal after an attack of vertigo due to unilateral vestibular disorder. Eighteen patients complained of difficulty in walking while turning toward the affected side, and 11 tended to fall to the inside of the circle. Turning in the opposite direction, however, was performed without any difficulty. It was thought that disorders of the cristospinal reflex became manifest during circular walking toward the affected side. The body is inclined to the inner side of circular walking. Endolymph flows toward the ampulla of the lateral semicircular canal on the same side as turning. This would exert a facilitatory action on extensor neurons and give the ipsilateral leg assistance for supporting the body's weight which shifts to the centre of the circle. Disorders of the cristospinal reflex would cause difficulty in circular walking toward the affected side.

[Influence of liver function on stereotactic microwave tissue coagulation therapy for hepatocellular carcinoma].

We studied 21 cases of hepatocellular carcinoma with liver cirrhosis to investigate the influence of liver function on stereotactic microwave tissue coagulation therapy. The cases were divided into three groups by the operation. The first group received only hepatectomy, the second hepatectomy and coagulation therapy for any remaining tumor, and the third group received only coagulation therapy. In group one, the average value of ICG R15 increased from 26.6% to 33.8%. In group 2, the average value of ICG R15 increased from 18.9% to 32.1%. And in group 3, the initial ICG R15 was 26.1%; after four weeks, the ICG test was 25.4%. These results showed that coagulation therapy alone had less influence on hepatic function. We propose that the coagulation therapy should be selected for cases of HCC with liver cirrhosis which have a poor liver function.

Intracellular DNA and RNA in the course of tumor cell growth.

We have investigated the intracellular levels of the nucleic acids, especially intracellular RNA to determine whether or not they could be used as indicators of cell proliferation kinetics and thereby enable an investigation into the biology of tumor cells. HeLa and COLO 205 were the cell lines used in this experiment, and normal human peripheral lymphocytes were used as the control. Cells were stained with acridine orange and subjected to flow cytometry. Based on the cytograms obtained, changes in the DNA and RNA contents in the cell lines were expressed as a ratio of the mean channel to that of the control. The intracellular RNA concentration was reflected in changes in the mean RNA channel ratio on the DNA and RNA cytograms. An increase in the mean RNA channel ratio represented a logarithmic proliferation of the tumor cells. Changes in the mean RNA channel ratio could be noted in cell samples containing as little as 20% logarithmic growth phase cells. There was a close correlation between the RNA content and the proliferative potential, and an increase in the RNA content was predictive of subsequent cellular proliferation. These results suggested that an increase in the intracellular RNA content on flow cytometry could be used as indicator of tumor proliferative potential, suggesting that this method could be useful for assessing the biological properties of tumor cells.

[Intraoperative real time observation of tumor blood flow in hepatocellular carcinoma by using color Doppler imaging].

To evaluate the tumor blood flow in hepatocellular carcinoma, we have introduced color Doppler imaging during operation. 37 hepatocellular carcinoma patient with 54 nodules were included in this study. Following categories were analyzed, those were detectability of pulsatile blood flow and continuous blood flow signal in the tumor, pulsatile blood flow velocity depend on tumor size and pathological feature. Even the size of the tumor is less than 1 cm in diameter in 70% of those nodules we could detect and analyze the flow signal. This new modality could have a possibility to be a useful diagnostic device for small mass lesions in the liver.

Intraoperative ultrasonography during laparoscopic cholecystectomy.

Intraoperative ultrasonography during laparoscopic cholecystectomy is a new procedure for examining the bile duct. The ultrasonic probe used in this study utilized a 7.5-MHz convex array designed for easy insertion through a 10-mm cannula. This probe was 90 cm long, had a 10-mm diameter, and was equipped with an angulation system controlled by levels. Additionally, this ultrasound system could detect the direction of the blood flow using a color Doppler display. Both laparoscopic ultrasound and laparoscopic cholangiography were attempted in 45 patients. In 41 of the 45 patients, laparoscopic ultrasound was successful in visualizing the first-order hepatic branches and the common bile duct. On the other hand, laparoscopic cholangiography was successful only in 38 of the 45 patients. The color Doppler capability helped to distinguish the bile duct from the portal vein and hepatic artery. Visualization of the bile duct was achievable by laparoscopic ultrasound. Also, the color Doppler system easily identified the bile duct. However, further design improvements in the ultrasound probe are required to establish laparoscopic ultrasound as an effective intraoperative routine procedure.

Factors contributing to labyrinthine ataxia.

Twenty-one normal adults were the subjects of this study. They were instructed to step in the same position with eyes closed for a total of 50 steps under two conditions: i) Immediately after simultaneous irrigation of the right and left auditory canals for 10 s with 30 degrees C water and 44 degrees C water, respectively, and ii) following irrigation of the right auditory canal with 30 degrees C water. The two conditions simulate two different types of imbalance of labyrinthine function: The former physiological and the latter non-physiological. Under the first condition, 33% of the subjects tended to fall to the right side during stepping. Under the second conditions, 86% showed ataxia. The difference was statistically highly significant, p < 0.01. The results indicate that physiological imbalance does not always cause ataxia. A physiological imbalance occurs during active turning. The motion brings about an ampullopetal lymphatic flow in the ipsilateral horizontal semicircular canal and an ampullofugal flow in the contralateral canal, as under the second condition. The former increases the function of the labyrinth, while the latter decreases it. This results in imbalance but no ataxia.

[An experimental study of ethanol injection on VX2 liver cancer].

Ethanol injection therapy for rabbit VX2 cancer of the liver was performed to measure the extent of ethanol infiltration using normal hepatic tissue as control, to observe the serial histological findings after ethanol injection, and to determine the influence of the ethanol on survival days with VX2 cancer of the liver. The necrotic area caused by ethanol showed a significant dose-dependent correlation in the range of 0.1-1.0ml in the normal liver, and a significant correlation in the range of 0.2-1.0ml also in VX2 cancer of the liver. Further, VX2 cancer of the liver showed a significant extension in the number of survival days in the injection group after ethanol injection; 39.5 +/- 5.7 days in the injection group, against 29.3 +/- 2.9 days in the control group. In the successive observation of histological findings after injection, fibrosis started from the 3rd-5th day after ethanol injection and showed acceleration after the 7th day. Eosinophilic infiltration was seen in the VX2 cancer group but not among the controls. Since ethanol causes direct necrosis of the injected region, it is suggested that ethanol injection therapy is a potentially effective method of treating cancer of the liver.

Direct observation of the portal vein interior by intra-operative angioscopy in the dog and man.

The purpose of this study was to develop the technique of intra-operative portal angioscopy using a portal angioscope, and to demonstrate its potential use in the therapy for patients with hepatocellular carcinoma (HCC) bearing a portal thrombus. Portal angioscopes, Olympus BF3C10 and CHFP10 of a diameter of 3.5 mm and 4.8 mm, respectively, were used during operation in five dogs, two patients with hepatic metastasis from colon cancer and three patients with HCC having a portal tumour thrombus. The portal vein and hepatic artery were ligated simultaneously, and the angioscope was immediately introduced under direct vision through a small portal venotomy. Blood in the portal vein was almost fully diverted by infusion of heparinized saline through a channel of the angioscope at a rate of 3 mL/min in dogs and 5 mL/min in patients with hepatic metastasis. Rates of 6 mL/min and 10 mL/min in dogs and patients, respectively, were adequate to clear completely the portal vein of blood. In patients with HCC, portal tumour thrombectomy was performed with a Fogarty balloon catheter by suctioning thrombi through a channel of the fibrescope after visual study of the portal thrombus. Observation and treatment of portal thrombus by angioscopy may become an important part of surgical treatment of HCC with portal invasion, but further technical improvement is desirable before this technique becomes a routine procedure.

[Intra-arterial infusion chemotherapy of liver metastases in colorectal cancer].

We report the complications and the results of intra-arterial infusion chemotherapy (IAIC) for liver metastases in colorectal cancer. Fifteen synchronous liver metastases and 11 metachronous liver metastases were treated from September 1986 to February 1989. Eight cases had H1 (metastases in one lobe only), 11 cases H2 (a few scattered metastases in both lobes) and 7 cases had H3 (numerous metastases in both lobes). Eight cases underwent hepatic resection and 20 cases were treated by IAIC with implantable reservoir. Incidence of complication was decreased from 75% to 17% by the improved technique of hepatic arterial catheter intubation. Median survival was 12 months, 20 months and 8 months, respectively. When used with MMC, the IAIC showed unsatisfactory results. We should therefore investigate possible use of alternative drugs, their optimum dosages, and the regimens for improved chemotherapy.