Immunohistochemical and mutational analysis of FLASH in gastric carcinomas

FLASH was initially identified as a pro-apoptotic protein that transmits an apoptosis signal during death receptor-induced apoptosis. Additionally, diverse biologic roles of FLASH, including TNF-induced NF-kappaB activation, cell-cycle progression and cell division, have been identified. Although such functions are important in cancer pathogenesis, little is known about the alterations of FLASH gene and FLASH protein expression in human cancers. In this study, we analyzed the expression of FLASH protein in 60 gastric adenocarcinomas by immunohistochemistry. We furthermore analyzed mutation of FLASH in exon 8, where two polyadenine tracts ((A)8 and (A)9) are present, by single-strand conformation polymorphism (SSCP) assay in 184 gastric adenocarcinomas. By immunohistochemistry, FLASH protein expression in cancer cells was detected positively in 42 gastric carcinoma tissues (70%), whereas its expression in epithelial cells of normal gastric mucosa was shown as no or very weak intensity. Mutational analysis detected one FLASH mutation in the gastric carcinomas (0.5%). The increased expression of FLASH in the malignant gastric epithelial cells compared to the normal mucosal epithelial cells suggests that FLASH expression may play a role in gastric tumorigenesis. Also, the data suggest that somatic mutation of FLASH is a rare event in gastric carcinomas.     

Human placenta-derived feeders support prolonged undifferentiated propagation of a human embryonic stem cell line, SNUhES3: comparison with human bone marrow-derived feeders

Co-culture of human embryonic stem (ES) cells on mouse fibroblast feeders is the commonly used method for in vitro expansion of human ES cells in an undifferentiated state. However, it has potential risks of pathogen transmission from animals; thus, human cell-derived feeders have been employed to minimize this problem. In this study, we compared human placenta-derived feeders with bone marrow to demonstrate its effectiveness as feeders for in vitro long-term culture of human ES cells. We cultured a human ES cell line, SNUhES3, on human placenta-derived mesenchymal stem cell feeders and compared their culture efficiency with human bone marrow-derived feeders and control group (mouse fibroblast feeders, STO). The mean number of human ES cell colonies was 166 +/- 35 in the placenta feeders; this was significantly higher than bone marrow-derived feeders (87 +/- 16, p < 0.05). We could propagate the culture of SNUhES3 on the placenta feeders past the 50th week similar to control group. During the culture, the maintenance of undifferentiated state of SNUhES3 was demonstrated by the expression of SSEA-4, TRA-1-81, TRA-1-60, and Oct-4. However, we failed to propagate the culture of human ES cells on the human bone marrow-derived feeders past the 5th week. The efficiency of embryoid body formation was similar between placenta and control group, indicating the preservation of differentiation ability. Thus, placenta-derived feeders are more efficient for the long-term in vitro culture of human ES cells than bone marrow-derived feeders suggesting the possible role of placenta as a source for human cell-derived feeders.     

Practical recombinant hybrid mussel bioadhesive fp-151

Mussel adhesive proteins (MAPs) have received increased attention as potential environmentally friendly adhesives under aqueous conditions and in medicine. However, attempts to produce functional recombinant MAPs (mainly foot protein type 1, fp-1) by several expression systems have failed. Even though we previously reported a functional expression of recombinant foot protein type 5 (fp-5) with significant adhesive ability in Escherichia coli, its practical use was limited by several problems such as low production yield, low purification yield, and high levels of post-purification insolubility. Here, to overcome these limitations, we designed and constructed the novel type of hybrid mussel bioadhesive fp-151, a fusion protein comprising six fp-1 decapeptide repeats at each fp-5 terminus. Using micro- and bulk-scale characterization and mammalian cell-adhesion analyses, we demonstrate that fp-151 has the potential to be a practical bioadhesive with strong adhesive ability, a simple purification process ( approximately 1g-purified protein per 1l-pilot-scale fed-batch bioreactor culture), proper manipulation properties ( approximately 330g/l solubility), and high biocompatibility.     

Molecular cloning and characterization of WD40-repeat protein from Clonorchis sinensis

WD40-repeat proteins have four to eight repeating units flanked by Gly-His (GH) and Trp-Asp (WD) at both termini and folds into a β-propeller. A polypeptide deduced from a Clonorchis sinensis cDNA clone analyzed to have seven WD40-repeats and predicted to form a β-propeller (CsWD1). The CsWD1 protein was expressed stage-specifically in the metacercariae and localized in the tegumental syncytium. The CsWD1 protein is suggested to serve as a platform for interacting partner proteins in the tegumental syncytium of C. sinensis metacercariae.     

Rai1 deficiency in mice causes learning impairment and motor dysfunction, whereas Rai1 heterozygous mice display minimal behavioral phenotypes

Smith-Magenis syndrome (SMS) is associated with an approximately 3.7 Mb common deletion in 17p11.2 and characterized by its craniofacial and neurobehavioral abnormalities. The reciprocal duplication leads to dup(17)(p11.2p11.2) associated with the Potocki-Lupski syndrome (PLS), a neurological disorder whose features include autism. Retinoic acid induced 1 (RAI1) appears to be responsible for the majority of clinical features in both SMS and PLS. Mouse models of these syndromes harboring an approximately 2 Mb chromosome engineered deletion and duplication, respectively, displayed abnormal locomotor activity and/or learning deficits. To determine the contribution of RAI1 in the neurobehavioral traits in SMS, we performed a battery of behavioral tests on Rai1 mutant mice and the Df(11)17-1/+ mice that have a small deletion of approximately 590 kb. The mice with the small deletion were hypoactive like the large deletion mice and they also showed learning deficits. The Rai1+/- mice exhibited normal locomotor activity. However, they had an abnormal electroencephalogram with overt seizure observed in a subset of mice. The few surviving Rai1-/- mice displayed more severe neurobehavioral abnormalities including hind limb clasping, overt seizures, motor impairment and context- and tone-dependant learning deficits. X-gal staining of the Rai1+/- mice suggests that Rai1 is predominantly expressed in neurons of the hippocampus and the cerebellum. Our results suggest that Rai1 is a critical gene in the central nervous system functioning in a dosage sensitive manner and that the neurobehavioral phenotype is modified by regulator(s) in the approximately 590 kb genomic interval, wherein the major modifier affecting the craniofacial penetrance resides.     

Age, sex, and comorbidities were considered in comparing reference data for health-related quality of life in the general and cancer populations

OBJECTIVES: The purpose of this study was to provide reference data for health-related quality of life (HRQOL) in the general Korean population so that the data could be compared with those of cancer patients. Reference data enable more detailed insights into treatments for and care of cancer patients. STUDY DESIGN AND SETTING: We constructed a questionnaire that included the European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire C30, LC13, STO22, and BR23, the Hospital Anxiety and Depression Scale, and the McGill Quality-of-Life questionnaire and administered a population-based, cross-sectional survey to 1,000 persons. RESULTS: Men reported better functioning and existential well-being, but women reported more physical symptoms, anxiety, and depression. Most scores of functioning and well-being scales decreased and most physical symptoms, anxiety, and depression increased with increasing age. Increasing the number of comorbidities had a negative effect on all functions and most symptom scales. CONCLUSION: Our findings suggest that age, sex, and comorbidities must always be considered when comparing HRQOL data from the general population with those from cancer patients.     

Novel brd4 inhibitors with a unique scaffold exhibit antitumor effects

Since bromodomain containing 4 (brd4) has been considered as a prominent cancer target, numerous attempts have been made to develop potent brd4 bromodomain inhibitors. The present study provided a novel chemical scaffold which inhibited brd4 activity. Mid-throughput screening against brd4 bromodomain was performed using alpha-screen and homogeneous time-resolved fluorescence assays. Furthermore, cell cytotoxicity and xenograft assays were performed to examine if the compound was effective both in vitro and in vivo. As a result, it was revealed that compounds having naphthalene-1,4-dione scaffold inhibited the binding of bromodomain to acetylated histone. The compounds with naphthalene-1,4-dione had cytotoxic effects against the Ty82 cell line, a NUT midline carcinoma cell line, whose proliferation is dependent on brd4 activity. A10, one of the compounds with naphthalene-1,4-dione scaffold, also exhibited tumor growth inhibition effects in the xenograft assay. In addition, the compounds exhibited cytotoxic effects against gastric cancer cell lines which were resistant to I-BET-762, a BET bromodomain inhibitor. In conclusion, the novel scaffold to suppress brd4 activity was effective against cancer cells both in vitro and in vivo.     

Novel lbeta-methylcarbapenems having cyclic sulfonamide moieties: synthesis and evaluation of in vitro antibacterial activity

The synthesis of a new series of 1beta-methylcarbapenems having cyclic sulfonamide moieties is described. Their in vitro antibacterial activities against both Gram-positive and Gram-negative bacteria were tested and the effect of substituent on the pyrrolidine ring was investigated. A particular compound (IIIi) having 2-methyl-[1,2,6]thiadiazinan-1,1-dioxide moiety showed the most potent antibacterial activity.     

Investigation of the air kerma response of spherical ionization chambers for unfolded pulse height distributions of (60)Co and (137)Cs using the EGS4 Monte Carlo code

Data required for the determination of the absolute air kerma rate for (60)Co and (137)Cs gamma-rays using spherical cavity chambers were calculated using the EGS4 Monte Carlo system. Mass energy-absorption coefficient ratio and the stopping power ratio were calculated for a 10 cm(3) primary standard graphite-walled ionization chamber from the unfolded energy pulse height distributions of (60)Co and (137)Cs sources. Wall correction factors and non-uniformity correction factors for two graphite and one air equivalent plastic walled ionization chambers were also calculated with EGS4 code. The wall correction factors were compared with those determined by an experimental extrapolation method. To check the accuracy of the calculations the results were compared with those obtained from other primary standard laboratories such as NIST and NRCC. For a 10 cm(3) graphite ionization chamber, the mass energy-absorption coefficient ratios were 0.99917 for (60)Co and 1.0004 for (137)Cs. The values differed by 0.02-0.05 % for (60)Co and 0.11 % for (137)Cs from those of two laboratories. The stopping power ratios were 0.99984 for (60)Co and 1.0087 for (137)Cs. Comparison with NIST values showed differences of 0.06 % for (60)Co and 0.04 % for (137)Cs. The wall correction factors were obtained and they were different by 0.6-1.1 % for (60)Co and (137)Cs compared to the experimental linear extrapolation method. These values were compared with Monte Carlo derived values from other laboratories. The non-uniformity correction factors were also calculated and they differed from unity, the traditional value used in most standard national metrology laboratories.     

Scoring system for risk stratification of viral reactivation during prophylactic antiviral treatment in Korean patients with hepatitis B undergoing anticancer chemotherapy: A multicenter study

Prophylactic antiviral therapy is recommended for hepatitis B virus (HBV)‐infected patients with malignancies who are undergoing systemic chemotherapy. In the current study, we aimed to develop a risk scoring system to guide the selection of prophylactic antiviral agents. In this retrospective analysis, we included consecutive chronic hepatitis B patients who received antiviral prophylaxis for chemotherapy of solid or hematologic malignancies at three large‐volume hospitals in Korea. The primary endpoint was HBV reactivation. The inverse probability treatment weighting method was used to minimize selection bias in terms of antiviral assignments. A total of 419 patients were enrolled: 129 patients received lamivudine (LAM), 216 received telbivudine (LdT), and 74 received entecavir (ETV), respectively. Of these, 36 patients developed on‐treatment HBV reactivation (LAM, 17; LdT, 18; ETV, 1). Multivariate analysis identified three independent predictors for reactivation: hepatitis B e‐antigen positivity, HBV DNA level, and type of malignancy. Accordingly, a risk scoring system was developed wherein one point was assigned for each of the risk factors. HBV reactivation occurred more frequently in the high‐risk group (score ≥ 2) than in the low‐risk group (hazards ratio, 14.17; P < 0.001). ETV exhibited superior prophylactic efficacy over LdT or LAM in the high‐risk group, whereas no significant difference was noted in the low‐risk group. The prognostic scoring system was useful for risk stratification of chemotherapy‐related HBV reactivation. High genetic barrier agents appear to be vital for high‐risk patients, whereas cost‐effectiveness may be more relevant for low‐risk patients.