Phase 1b investigation of the MEK inhibitor binimetinib in patients with advanced or metastatic biliary tract cancer.

Background The MAPK pathway plays a central role in regulation of several cellular processes, and its dysregulation is a hallmark of biliary tract cancer (BTC). Binimetinib (MEK162), a potent, selective oral MEK1/2 inhibitor, was assessed in patients with advanced BTC. Patients and Methods An expansion cohort study in patients who received ≤1 line of therapy for advanced BTC was conducted after determination of the maximum tolerated dose in this Phase 1 trial. Patients received binimetinib 60 mg twice daily. The primary objectives were to characterize the safety profile and pharmacokinetics of binimetinib in advanced BTC. Secondary objectives included assessment of clinical efficacy, changes in weight and lean body mass, and pharmacodynamic effects. Tumor samples were assessed for mutations in relevant genes. Results Twenty-eight patients received binimetinib. Common adverse events (AEs) were mild, with rash (82%) and nausea (54%) being most common. Two patients experienced grade 4 AEs, one generalized edema and the other pulmonary embolism. The pharmacokinetics in this patient population were consistent with those previously reported (Bendell JC et al., Br J Cancer 2017;116:575-583). Twelve patients (43%) experienced stable disease and two had objective responses (1 complete response, 1 partial response) per Response Evaluation Criteria in Solid Tumors and stable metabolic disease by positron emission tomography/computed tomography. Most patients (18/25; 72%) did not have KRAS, BRAF, NRAS, PI3KCA, or PTEN mutations, nor was there correlation between mutation status and response. The average non-fluid weight gain was 1.3% for lean muscle and 4.7% for adipose tissue. Conclusion Binimetinib was well tolerated and showed promising evidence of activity in patients with BTC. Correlative studies suggested the potential for binimetinib to promote muscle gain in patients with BTC.

Baseline tumour measurements predict survival in advanced non-small cell lung cancer.

BACKGROUND: The association between tumour measurements and survival has been studied extensively in early-stage and locally advanced non-small cell lung cancer (NSCLC). We analysed these factors in patients with advanced NSCLC. METHODS: Data were derived from the E4599 trial of paclitaxel-carboplatin±bevacizumab. Associations between the Response Evaluation Criteria in Solid Tumors (RECIST) baseline sum longest diameter (BSLD), response rate, progression-free survival (PFS) and overall survival (OS) were evaluated using univariate and multivariable Cox regression models. RESULTS: A total of 759 of the 850 patients (89%) in the E4599 trial had measurable diseases and were included in this analysis. The median BSLD was 7.5 cm. BSLD predicted OS (hazard ratio (HR) 1.41; P<0.001) and had a trend towards association with PFS (HR 1.14; P=0.08). The median OS was 12.6 months for patients with BSLD <7.5 cm compared with 9.5 months for BSLD ≥ 7.5 cm. This association persisted in a multivariable model controlling multiple prognostic factors, including the presence and sites of extrathoracic disease (HR 1.24; P=0.01). There was no association between BSLD and response rate. CONCLUSION: Tumour measurements are associated with survival in the E4599 trial. If validated in other populations, this parameter may provide important prognostic information to patients and clinicians.

Factors affecting the activity of anammox bacteria during start up in the continuous culture reactor.

Factors affecting cultivation of extremely slow-growing bacteria (anaerobic ammonium oxidiser, doubling time 11 days) were investigated by using upflow anaerobic sludge blanket (UASB) reactors which can maintain high solid retention time. The effects of concentrations of DO, free ammonia (FA), and nitrite on activation of anammox activity were tested during the start-up period. The reactor was inoculated with granular sludge collected from a full-scale UASB reactor used for treating brewery wastewater, and sludge from a piggery wastewater treatment plant and rotating biological contactor treating sewage. Results of continuous operation showed that concentrations of DO, free ammonia (FA) and nitrite in the reactors played a key role in stimulating the anammox activity during start-up period. It is crucial to keep DO below 0.2 ppm, FA below 2 mg/L and nitrite nitrogen below 35 mg/L to cultivate anammox cells in the continuous bioreactor. When the levels of DO, FA and nitrite in the influent were controlled at less than the inhibition levels, the anammox activity increased gradually in the anaerobic condition. Addition of hydrogen sulphide into the reactor enhanced anammox activity in the continuous culture. Through the SEM, TEM and FISH analysis, anammox bacteria were detected in the granular sludge after 3 months of continuous operation.

Modified irinotecan and infusional 5-fluorouracil (mFOLFIRI) in patients with refractory advanced pancreas cancer (APC): a single-institution experience.

Pancreatic adenocarcinoma is the fourth leading cause of cancer death. Recently, MM-398 (nanoliposomal irinotecan) was shown to be associated with significant improvement in outcome measures with acceptable toxicities when combined with 5-fluorouracil (5-FU)/leucovorin (LV) compared to 5-FU/LV alone in patients failing one line of gemcitabine-based therapy. There is a paucity of data evaluating the role of irinotecan in combination with 5FU in advanced pancreas cancer (APC). We performed a retrospective analysis of all patients who received mFOLFIRI (minus bolus 5FU and LV). All patients with metastatic disease who had failed at least one line of gemcitabine-based therapy prior to receiving mFOLFIRI were included in this study. Descriptive statistics were used to assess the continuous variables and adverse events (AEs), and Kaplan-Meier methods were used to calculate the median progression-free survival (PFS) and overall survival (OS). Forty patients were included in this analysis. Patients received 1-5 lines of prior therapy (25 % with more than 3 lines of prior therapy). The mean age at diagnosis was 60, and 98 % had ECOG of 1. The mean CA 19-9 at the start of therapy was 33,169 U/ml. The median PFS was 2.59 months [95 % confidence interval (CI) (1.90, 3.54)], and OS was 4.75 months [95 % CI (3.14, 8.98)]. The most common AEs included fatigue (98 %), neuropathy (83 %), anorexia (68 %), nausea (60 %) and constipation (55 %). Grade 3 toxicities included fatigue (13 %) and rash (3 %). There were no observed grade 4 toxicities. In this single-institution retrospective analysis, mFOLFIRI was found to be both tolerable and relatively effective in a heavily pretreated patient population with APC. Future prospective studies should consider evaluating the role of mFOLFIRI in refractory APC.

Biological properties and expression of mucins in 5-fluorouracil resistant HT29 human colon cancer cells.

We have previously reported that HT29 human colon cancer cells selected by adaptation to 5-fluorouracil (5FU) (HT29-5FU cells) express increased levels of a major intestinal mucin MUC2 mRNA compared with parental HT29 cells. In this study, we examined in detail the changes in synthesis and secretion of mucin that occur in these cells and accompanying changes in the expression of cancer associated mucin related carbohydrate antigens and cell lineage associated biochemical markers. We further investigated their relationship to biological properties of cells. Northern blot analysis revealed a markedly increased level of MUC2 mRNA but no significant change in the mRNA levels of other mucins in HT29-5FU cells compared with parental HT29 cells. Labeling with radiolabeled precursors demonstrated increased synthesis and secretion of mucin glycoproteins by HT29-5FU cells. Immunoblot analysis showed a higher expression of mucin associated carbohydrate antigens such as T, Tn, sialyl Tn, sialyl Lea, sialyl Lex and non-O-acetylated sialic acid concomitant with significant increases in the expression of goblet cell lineage marker, MUC2 apomucin and a panepithelial cell marker, carcinoembryonic antigen. HT29-5FU cells showed significantly higher adhesion to E-selectin and to matrigel and in vitro invasive properties and significantly increased liver colonization capacity in nude mice following splenic vein injection. Nude mouse xenograft tumors produced by HT29-5FU cells showed a greater degree of differentiation, consisting of mucin secreting glands than those produced by parental HT29 cells. These results indicate that predominantly colonic type mucin, MUC2, has been selectively induced in HT29-5FU cells and that altered regulation of mucin genes associated with altered synthesis and secretion of mucin glycoproteins and the degree of differentiation in cancer cells may be responsible for the altered biological properties of these cells.

Phorbol 12-myristate 13-acetate induces alteration in mucin gene expression and biological properties of colon cancer cells.

Phorbol esters such as phorbol 12-myristate 13-acetate (PMA) have been reported to modulate diverse cellular responses through signal transduction pathways including the protein kinase C (PKC) pathway. In the present study, we sought to determine the effect of PMA on mucin gene expression and on the biological properties of a human colon cancer cell line, HM3. The cells were treated for 8 and 24 h with various concentrations of PMA and total RNA was extracted and Northern and slot blot analyses were carried out using MUC2, MUC3 and MUC5AC mucin cDNA probes to assess the steady state levels of mRNA. Spent media were collected and the level of cancer associated carbohydrate antigens (T, Tn, sialyl Tn, sialyl Lex, and sialyl Lea) and matrix-degrading metalloproteinase (MMPs) activity were examined. Trypsinized cells were used for assessing in vitro invasion, motility and adhesion to matrigel. Our results showed that PMA caused upregulation of steady state mRNA levels of MUC2, MUC3 and MUC5AC which was inhibited after treatment with protein synthesis inhibitors. Calphostin C, a highly specific inhibitor of protein kinase C significantly inhibited the PMA induced induction of mRNA levels of MUC2, MUC3, and MUC5AC. The levels of all cancer-associated mucin carbohydrate antigens examined in the media were increased by PMA treatment. PMA also caused an increase in MMPs activity and in in vitro invasion and motility properties, but did not affect adhesion of HM3 cells to matrigel. Thus, PMA caused a significant increase in the expression of all three mucin genes through signaling pathways involving protein kinase C and increased secretion of mucin associated carbohydrate antigens. These changes were associated with increases in MMP activity as well as by increases in the invasive and motility properties of HM3 colon cancer cells. These data suggest that protein kinase C signaling pathways may be involved in mucin gene regulation and in modulating the invasive and metastatic properties of colon cancer cells.

Structural changes in Z-disks of skeletal muscle myofibrils during growth of chicken.

Postnatal structural changes in Z-disks of skeletal muscles of chicken from 2 to 35 wk after hatching were examined to elucidate how Z-disks develop from the embryonic to the mature stage. The mechanical strength of Z-disks of breast muscle (i.e. the ratio of the number of myofibrils to the total number of myofibrils and myofibrillar fragments composed of 1-4 sarcomeres, which were formed under the mechanical forces exerted during homogenization) increased from 66% at 2 wk of age to 75% at 10 wk, and leveled off at 76% at 25 wk. The Z-disks of leg muscle were stronger than those of breast muscle throughout growth. Measurements by electron microscope showed that the width of Z-disks increased during growth from 26 to 33 nm, 48 to 53 nm, and 89 to 101 nm, in white, intermediate, and red muscle fibers, respectively. It was proved that the configuration of Z-filaments, the structural backbone of Z-disks, takes its final shape within 2 wk after hatching; the immature Z-disks are then reinforced by the accumulation of amorphous matrix materials. These results confirm that the maturation of Z-disks is brought about by physical motion in muscle tissue and the accompanying development of tension.

Fluorescence in situ hybridization analysis of nitrifiers in piggery wastewater treatment reactors.

Fluorescence in situ hybridization (FISH) was performed to analyze the nitrifying microbial communities in an activated sludge reactor (ASR) and a fixed biofilm reactor (FBR) for piggery wastewater treatment. Heterotrophic oxidation and nitrification were occurring simultaneously in the ASR and the COD and nitrification efficiencies depend on the loads. In the FBR nitrification efficiency also depends on ammonium load to the reactor and nitrite was accumulated when free ammonia concentration was higher than 0.2 mg NH3-N/L. FISH analysis showed that ammonia-oxidizing bacteria (NSO1225) and denitrifying bacteria (RRP1088) were less abundant than other bacteria (EUB338) in ASR. Further analysis on nitrifying bacteria in the FBR showed that Nitrosomonas species (NSM156) and Nitrospira species (NSR1156) were the dominant ammonia-oxidizing and nitrite-oxidizing bacteria, respectively, in the piggery wastewater nitrification system.

Liberation of phospholipids from Z-disks of chicken skeletal muscle myofibrils by 0.1 mM calcium ions: weakening mechanism for Z-disks during post-mortem aging of meat.

Weakening of the Z-disks of skeletal muscle myofibrils contributes to the tenderization of meat during post-mortem aging. To elucidate the weakening mechanism, we compared Z-disks weakened by post-mortem aging of chicken breast muscle with those of myofibrils treated with a solution containing 0.1 mM CaCl2 and 1 microM calpastatin domain I. In both cases, the Z-disks were weakened with a corresponding liberation of their constituent phospholipids (PLs). The liberation of PLs specific to 0.1 mM calcium ions was minimal at pH 6.5 and maximal at 35 degrees C together with the Z-disk weakening. Binding of calcium ions to PLs in the Z-disks was determined by 45Ca-autoradiography. Acidic PLs were strongly radioactive and neutral PLs were appreciably radioactive. It is very probable that acidic PLs would bind electrostatically to alpha-actinin under physiological conditions, and that this interaction would be broken by the binding of calcium ions at 0.1 mM to PLs, resulting in the partial liberation of PLs from Z-disks. We conclude, therefore, that the liberation of PLs by the binding of 0.1 mM calcium ions was the main cause for Z-disk weakening during the post-mortem aging of chicken.

Adsorption of Bacillus subtilis endo-beta-1,4-glucanase to cellulosic materials.

The intact and the truncated endo-beta-1,4-glucanase expressed in Bacillus megaterium by a B. subtilis gene were purified and its adsorption characteristics to cellulosic materials were investigated. The intact enzyme was purified by affinity towards insoluble cellulose and Mono-Q chromatography. The truncated enzyme was purified by gel filtration and chromatofocusing. The molecular activities of these enzymes towards the soluble substrates were identical. Optimum pH and temperature of these two types of enzyme were same, 5.5 and 60 degrees C, respectively. However, the insoluble substrate, Avicel, was hydrolyzed slowly by the intact endoglucanase while the truncated endoglucanase did not hydrolyze the Avicel. Isoelectric points of the intact and the truncated enzyme were 6.2 and 5.6, respectively. In a Sephadex column the large form of intact endoglucanase was eluted later than the small form of truncated enzyme. Only the intact enzyme was strongly adsorbed to Avicel. The practical maximum adsorption was about 20 mg/g of Avicel at pH 6.0 and 4 degrees C.

Identification of lipids as the main component of skeletal muscle Z-discs.

The existence of lipids in Z-discs of the vertebrate skeletal muscle was suggested by the results of microscopy using rhodamine 6G reagent, a fluorescent probe. After removal of lipids by treatment of myofibrils with 0.5% Trition X-100, intact configurations of Z-filaments composed of alpha-actinin were clearly visible under an electron microscope. These findings indicated that lipids were amorphous-matrix materials of the Z-disc. Lipids were proved to be the main component of Z-discs by their extraction from I-Z-I brushes with a mixture of methanol and chloroform and by analysing them. The total amount of lipids in Z-discs exceeded that of alpha-actinin and varied from 2.4 to 7.1 g per 100 g of myofibrillar proteins depending on the phenotype of skeletal muscle. The sum of the amounts of lipids and alpha-actinin was 4.5 g per 100 g of myofibrillar proteins in chicken pectoralis profundus muscle (fast type), while it was 10.1 g in chicken soleus muscle (slow type). Lipid classes were phospholipids, triacylglycerols, cholesterol and free fatty acids. Since the lipids extracted from I-Z-I brushes were hardly contaminated with those from the sarcoplasmic reticulum and mitochondria, their amounts and classes were considered to be characteristic of Z-discs. The lipids probably cement neighbouring Z-filaments electrostatically, reinforce the Z-disc structure, and play an important role in the force transmission of skeletal muscle myofibrils.

Annual Report of the Central Cancer Registry in Korea-1999: Based on Registered Data from 128 Hospitals.

PURPOSE: The Central Cancer Registry Center in Korea (KCCR) conducted a nationwide hospital-based cancer registry to provide basic statistical data on cancer. MATERIALS AND METHODS: In 1999, 128 hospitals participated in the cancer registry program. All cancer registry data, which was submitted from the participating hospitals by diskettes during the year, were reviewed and analyzed by the committee members who were all Board-qualified clinical oncologists and pathologists. To avoid duplication, every resident registration number was compared by a computer. Cases that had been diagnosed by a histological examination were preferentially chosen for inclusion in this data. RESULTS: Of 94,003 cases that were registered, there was a total of 8,452 (9.0%) duplication cases which were excluded. Of the remaining 85,551 cases, there were 3,231 cases (3.8%) of carcinoma in situ (morphology code/2) which were excluded. A final total of 82,320 cases were analyzed. Of the analyzed cases, 46,908 (57.0%) were males and 35,412 (43.0%) were females. The leading age groups in the order of their relative frequency were those who were 60~64 years of age (15.3%), followed by the 55~59 age group (13.8%). The six leading primary cancer sites in the order of their relative frequency were stomach (20.7%), followed by the bronchus and lung (12.1%), the liver and intrahepatic bile duct (12.0%), the colorectum (9.9%), the breast (6.4%), and then the uterine cervix (5.0%). In males, the five leading primary cancer sites were the stomach (24.2%), the liver and intrahepatic bile duct (16.3%), the bronchus and lung (16.1%), the colorectum (9.7%), and the urinary bladder (3.3%). In females, the stomach (16.2%) was the most common cancer site, followed by the breast (14.7%), the uterine cervix (11.6%), the colorectum (10.2%), and the thyroid (6.8%). Among the 1,077 cases of childhood malignancies, leukemia (35.4%), CNS tumors (16.7%), malignant lymphomas (7.0%), and sympathetic nervous system tumors (6.9%) were the most common cancer types. CONCLUSION: We analyzed and report the KCCR data from 128 nationwide hospitals during 1999.

A Study of the Survival Rate of Childhood Cancer in Korea.

PURPOSE: It is known that the prognosis of childhood cancer is relatively good, however actual representative nationwide data on childhood cancer, particularly of survival rate, are rare. In this study we attempted to establish the overall survival rate of major childhood cancer. MATERIALS AND METHODS: The primary source of data of childhood cancer under 15 years of age were the registry files of the Central Cancer Registry Report (Ministry of Health & Welfare) from 1993 to 1997. The above data was compared to death case data files of the same period obtained from the Korea National Statistical Office using the personal identification code. We calculated the 1, 3, and 5 year survival rates using the life table of SPSS and Kaplan-Meier method and compared the survival rate of disease according to prognostic factors. RESULTS: A total of 6,720 cases of pediatric cancer from the Central Cancer Registry files were computerized and sorted by personal identification (ID) code to extract duplicated cases as well as cases with incomplete data. The final number of cases entered in this study was 4,983. 1) The number of confirmed death cases was 1,448 (29.1%). 2) The disease distribution showed that the most common pediatric cancer was leukemia (1,468/4,983, 29%), followed by brain tumors (503/4,983, 10%), lymphoma (315/4,983, 6%), Wilms tumor (165/4,983, 3%), etc. in order by number of patients. 3) The 5 year survival rate of disease was as follows: overall 62%, acute lymphocytic leukemia 61%, acute non-lymphocytic leukemia 32%, malignant lymphoma 72%, neuroblastoma 47%, medulloblastoma 51%, Astrocytoma 66%, Wilms tumor 83%, etc. CONCLUSION: We analyzed and report the 5 year survival rate of overall childhood cancer and of each of the twelve major childhood cancers from in Korea 1993 to 1997 to provide basic data on childhood cancer statistics.

1991 cancer incidence in Seoul, Korea: results of the Implementation Study of the Seoul Cancer Registry.

This article presents the results of the Implementation Study of the Seoul Cancer Registry, which started in July, 1991 as a population based cancer registry in Seoul, Korea. The completeness and validity of the registered data were evaluated using Mortality/Incidence ratio (M/I ratio), Histologically Verified Cases (HV%), Primary Site Uncertain (PSU%), and Age Unknown (Age UNK%). Owing to the additional active surveillance, the completeness of the data turned out to be fairly acceptable, except for the aged over 75(Mortality/Incidence ratio was over 100%). Eventhough the Seoul cancer registry(SCR) has further way to go in the completeness especially among elderly persons, the validity of SCR data was also acceptable in terms of HV%, PSU%, and Age UNK%. However, PSU% and Age UNK% might need to be further reduced to be comparable with other well established cancer registries. The age standardized incidence rates(ASR) of all cancers between July 1, 1991 and June 30, 1992 were 232.4/100,000 in males and 147.9/100,000 in females. The top five major sites of cancers in Seoul were the stomach, liver, lung, colo-rectum, and bladder in order in males, and the uterine cervix, stomach, breast, colo-rectum, and liver in females. Those 5 cancer sites comprised 68.9% and 64.7% of the total cancer incidence in males and females, respectively.

Subacute toxicity of a Pseudomonas vaccine prepared from outer membrane fraction of Pseudomonas aeruginosa in beagle dogs.

CFC-101, a Pseudomonas vaccine, was administered to beagle dogs by intramuscular injection for 4 weeks (5 days/week) at 0.05, 0.15 and 0.45 mg/kg/d. Clinical signs considered to be related to treatment were restricted to swelling at the injection sites, being apparent 1-2 h after treatment. There was no effect on body weight, food consumption, ophthalmoscopy, electrocardiography, hematology, biochemical and urinary parameters. The histopathological examination revealed treatment-related changes at the injection sites at all dosages, particularly in the hindlimbs where both perivascular and intramuscular aggregations of inflammatory cells were seen. Thus, the only treatment-related changes seen in this study were local reactions to the test substance at the injection sites; furthermore these changes seem to represent a pharmacological response to the test material. Because no evidence of any systemic toxicity was observed at any dosage level, it is concluded that dosages of CFC-101 up to and including 0.45 mg/kg/d were well tolerated over a period of 4 weeks in the beagle dog.

Human immune response to a Pseudomonas aeruginosa outer membrane protein vaccine.

In order to evaluate in humans the safety and immunogenicity of a Pseudomonas aeruginosa vaccine composed of outer membrane proteins (OMPs), CFC-101, we carried out a phase I/IIa clinical trial in healthy male volunteers. Groups of six volunteers were immunized either subcutaneously (s.c.) or intramuscularly (i.m.) with three dosages of the vaccine three times at 7-day intervals. The vaccine was well tolerated by volunteers. Local reactions in the injection sites were generally mild and transient. Significant increases in OMP-specific antibody were observed in both route groups after vaccinations but was higher in the i.m.-immunized group, where vaccination with 0.5 or 1.0 mg doses yielded 100% seroconversion. The specificity of the induced antibodies to P. aeruginosa OMP was demonstrated by western blot analysis and immunoprecipitation assay. An increase in Clq-binding capacity and ability to confer mice protection from lethal challenges with P. aeruginosa indicated the protective efficacy of the elicited antibodies. Based on these data, we concluded that the P. aeruginosa OMP vaccine is safe and effective in humans with an optimal dose of 0.5 and 1.0 mg and that i.m. is the better route than s.c. for this vaccine.