Early development of de novo hepatocellular carcinoma after direct-acting agent therapy: Comparison with pegylated interferon-based therapy in chronic hepatitis C patients.

Patients with chronic hepatitis C who achieve a sustained viral response after pegylated interferon therapy have a reduced risk of hepatocellular carcinoma, but the risk after treatment with direct-acting antivirals is unclear. We compared the rates of early development of hepatocellular carcinoma after direct-acting antivirals and after pegylated interferon therapy. We retrospectively analysed 785 patients with chronic hepatitis C who had no history of hepatocellular carcinoma (211 treated with pegylated interferon, 574 with direct-acting antivirals) and were followed up for at least 24 weeks after antiviral treatment. De novo hepatocellular carcinoma developed in 6 of 574 patients receiving direct-acting antivirals and in 1 of 211 patients receiving pegylated interferon. The cumulative incidence of early hepatocellular carcinoma development did not differ between the treatment groups either for the whole cohort (1.05% vs 0.47%, P = .298) or for those patients with Child-Pugh Class A cirrhosis (3.73% vs 2.94%, P = .827). Multivariate analysis indicated that alpha-fetoprotein level >9.5 ng/mL at the time of end-of-treatment response was the only independent risk factor for early development of hepatocellular carcinoma in all patients (P < .0001, hazard ratio 176.174, 95% confidence interval 10.768-2882.473) and in patients treated with direct-acting agents (P < .0001, hazard ratio 128.402, 95% confidence interval 8.417-1958.680). In conclusion, the rate of early development of hepatocellular carcinoma did not differ between patients treated with pegylated interferon and those treated with direct-acting antivirals and was associated with the serum alpha-fetoprotein level at the time of end-of-treatment response.

Combinations of eight key mutations in the X/preC region and genomic activity of hepatitis B virus are associated with hepatocellular carcinoma.

Accumulation of eight key mutations located in the X/preC regions of the hepatitis B virus (HBV) genome (G1613A, C1653T, T1753V, A1762T, G1764A, A1846T, G1896A and G1899A) is a risk marker for the development of hepatocellular carcinoma (HCC). In this study, we analysed the 8 key mutations in 442 serum samples collected from 310 non-HCC and 132 HCC patients to identify the combinations linked to HCC. After the patients were stratified according to the age groups and mutation combinations, clinical parameters were compared between the HCC and the non-HCC groups. Analyses were focused on patient ≥40 years of age infected by HBV genotype C with A1762T and G1764A mutations in the basal core promoter region (BCP double mutation). In patients with ≥6 mutations, the combination of [G1613A + C1653T + A1846T + G1896A] mutations was closely linked to HCC, whereas no specific single or double mutation combination was associated with HCC. In patients with ≤5 mutations, HBeAg and HBV DNA serum titres were lower in the HCC group than those in the non-HCC group. Unlike the number of mutations, no specific combination correlated with advanced clinical stage in HCC. Of the BCP double mutation-based HBV mutant types, combinations of ≥6 mutations that include G1613A + C1653T + A1846T + G1896A, and combinations of ≤5 mutations with reduced HBeAg production, may be more specific indicators of HCC risk than only the number of mutations or any specific combination(s).

Pretreatment HBeAg level and an early decrease in HBeAg level predict virologic response to entecavir treatment for HBeAg-positive chronic hepatitis B.

There are few reports on hepatitis B e antigen (HBeAg) titres during nucleos(t)ide analogues treatment. We investigated the changes in HBeAg levels in patients treated with entecavir and the usefulness of HBeAg quantification for predicting antiviral response. Ninety-five consecutive HBeAg-positive patients treated with entecavir for more than 48 weeks were enrolled. Serum levels of hepatitis B surface antigen (HBsAg), HBeAg and HBV DNA were assessed at 4-week intervals to week 24 and thereafter at 12-week intervals. Virologic response (Y1VR) was defined as an undetectable HBV DNA level at week 48 of therapy. During 48 weeks, HBeAg and HBV DNA level decreased significantly in a biphasic manner and HBsAg level tended to decease. Fifty-three patients (55.8%) attained Y1VR. Pretreatment HBeAg levels were significantly lower in the Y1VR group than in no Y1VR group. At week 4 and 12 of therapy, 25% and 41.4% of patients showed a decrease of HBeAg levels with >0.5 log(10) and >1.0 log(10) from baseline, respectively. These patients achieved more Y1VR than those with less decrease of HBeAg levels (97.7%vs 22.2% and 86.2%vs 29.3%, respectively). HBeAg level at week 12 had higher predictive values for Y1VR than HBV DNA level. Multivariate analysis revealed that a pretreatment HBeAg level of <360 PEIU/mL and the reduction in HBeAg level >1.0 log(10) at week 12 were associated with Y1VR. These results suggest that pretreatment HBeAg level and an early decrease in HBeAg level are useful measurements for predicting one-year virologic response during entecavir treatment.

Direct and simultaneous determination of Co and Cu on a silicon wafer using a chemiluminescence system.

In this work, we developed a drop-type chemiluminescence (CL) system with a partial least squares (PLS) calibration in which the coaxial optical fiber sensing head was developed for sampling and detection to determine Cu(2+) and Co(2+) on a silicon wafer directly. The use of time-resolved signal generation and PLS calibration in addition to CL allowed us to determine the metal ions simultaneously and selectively, based on the kinetic difference of Cu and Co ions in the luminol-H(2)O(2) system. Two component mixtures with a set of 15 wafer fragments were orthogonally calibrated. After prediction test, the method was applied to an intentionally contaminated silicon wafer and validated by inductively coupled plasma-mass spectrometer (ICP-MS) measurement with a HF-HNO(3) scanning solution. The average concentrations of Cu(2+) and Co(2+) of 3.45 (±0.95) × 10(13) and 2.30 (±1.18) × 10(11) atoms per cm(2), respectively, were obtained, which were very close to the ICP-MS results of 3.70 × 10(13) for Cu(2+) and 2.46 × 10(11) atoms per cm(2) for Co(2+). In conclusion, this drop mode CL showed almost more than 10 times better reproducibility than the typical batch mode for the profile measurement. Moreover, the adoption of PLS calibration added the function of selectivity for the simultaneous determination to this CL system, in addition to the direct mapping capability for the solid surface analysis.

Thermoelectric properties of dispersant-free semiconducting single-walled carbon nanotubes sorted by a flavin extraction method.

Semiconducting single-walled carbon nanotubes (s-SWNTs) were extracted from SWNT mixtures using a flavin derivative (FC12). We evaluated the thermoelectric properties of the s-SWNT sheets. Electrical conductivity, power factor and figure of merit values of the sheets were increased by two orders of magnitude after removing FC12 simply by dipping in dichloromethane.

The best candidates for transarterial chemotherapy in patients with hepatocellular carcinoma awaiting liver transplantation: a cohort-based characterization of dropout times.

BACKGROUND: Although transarterial chemotherapy is used to retard tumour progression for hepatocellular carcinoma (HCC) patients awaiting orthotopic liver transplantation (OLT), information regarding the acceptable waiting time and appropriate patient selection for the therapy is lacking. AIM: To examine dropout times and determine the best candidates for pre-transplant transarterial therapy in a cohort study. METHODS: In total, 180 consecutive HCC candidates receiving pre-transplant chemo-lipiodolization were included in the study. RESULTS: Overall, 70 (38.9%) patients dropped off the waiting list during the median follow-up of 19 months. According to the Child-Pugh (C-P) classification, the estimated dropout rates at 1 and 2 years were 17.2% and 44.8% for the C-P A group and 33.4% and 81.3% for the C-P B/C group, respectively. C-P B/C patients experienced more frequent dropouts than C-P A patients (P < 0.001). Risk factor analysis identified C-P classification to be the strongest predictor of dropout (P < 0.001). On multivariate analysis, alpha-fetoprotein (AFP) >100 ng/mL, tumour size >3 cm and multiple nodules remained independently predictive of dropout for C-P A group (all P < 0.05). Candidates with none of these factors were found to be at the lowest risk of dropout, with only a 22.5% dropout rate up to 41 months. CONCLUSIONS: This study suggests that Child-Pugh A patients with one nodule <3 cm and AFP < 100 ng/mL may be the best candidates for pre-transplant chemo-lipiodolization, with the lowest dropout rate. However, comparative studies with other therapeutic options are needed to assess the definitive role of transarterial therapy in this setting.

RAC-LATS1/2 signaling regulates YAP activity by switching between the YAP-binding partners TEAD4 and RUNX3.

The tumor-suppressor RUNX3 has a critical role in a lineage determination, cell cycle arrest and apoptosis. Lozenge (Lz), a Drosophila homolog of mammalian RUNX family members, has integral roles in these processes and specifically in eye cell fate determination. To elucidate the genetic modifiers of Lz/RUNX3, we performed a large-scale functional screen in a fly mutant library. The screen revealed genetic interactions between the Lz, Rac and Hippo pathways. Analysis of interactions among these genes revealed that the defective phenotype resulting from activation of Yki, an end point effector of the Hippo pathway, was suppressed by Lz and enhanced by Rac-Trio. Molecular biological analysis using mammalian homologs reveled that LATS1/2-mediated YAP phosphorylation-facilitated dissociation of the YAP-TEAD4 complex and association of the YAP-RUNX3 complex. When cells were stimulated to proliferate, activated RAC-TRIO signaling inhibited LATS1/2-mediated YAP phosphorylation; consequently, YAP dissociated from RUNX3 and associated with TEAD, thereby replacing the YAP-RUNX3 complex with YAP-TEAD. RUNX3 contributed to both association and dissociation of YAP-TEAD complex, most likely through the formation of the YAP-TEAD-RUNX3 ternary complex. Ectopic expression of RUNX3 in MKN28 gastric cancer cells reduced tumorigenicity, and the tumor-suppressive activity of RUNX3 was associated with its ability to interact with YAP. These results identify a novel regulatory mechanism, mediated by the Hippo and RAC-TRIO pathways, that changes the binding partner of YAP.

Endostatin inhibits endothelial and tumor cellular invasion by blocking the activation and catalytic activity of matrix metalloproteinase.

Here we report the inhibition of cellular invasion by a recombinant mouse endostatin and the possible mechanism of the inhibition. Endostatin significantly reduced endothelial as well as tumor cellular invasion into the reconstituted basement membrane in vitro. Gelatin zymographic analysis revealed that the activation of promatrix metalloproteinase-2 (proMMP-2) that was secreted from endothelial cells was blocked upon endostatin treatment. Studies with recombinant MMPs confirmed that endostatin inhibited proMMP-2 activation, mediated by both membrane-type 1 MMP and 4-aminophenylmercuric acetate. Furthermore, enzymatic assays using a peptide substrate demonstrated that endostatin inhibited the catalytic activities of both MMP-2 and membrane-type 1 MMP. Finally, coimmunoprecipitation experiments revealed that endostatin formed a stable complex with proMMP-2. These novel findings would, at least in part, explain the mechanism of the potent antiangiogenic and antitumor activities of endostatin.

Improvement of polypyrrole nanowire devices by plasmonic space charge generation: high photocurrent and wide spectral response by Ag nanoparticle decoration.

In this study, improvement of the opto-electronic properties of non-single crystallized nanowire devices with space charges generated by localized surface plasmon resonance (LSPR) is demonstrated. The photocurrent and spectral response of single polypyrrole (PPy) nanowire (NW) devices are increased by electrostatically attached Ag nanoparticles (Ag NPs). To take advantage of plasmon-exciton coupling in the photocurrent of the device, 80 nm of Ag NPs (454 nm = λmax) were chosen for matching the maximum absorption with PPy NWs (442 nm = λmax). The photocurrent density is remarkably improved, up to 25.3 times (2530%), by the Ag NP decoration onto the PPy NW (PPyAgNPs NW) under blue light (λ = 425-475 nm) illumination. In addition, the PPyAgNPs NW shows a photocurrent decay time twice that of PPy NW, as well as an improved spectral response of the photocurrent. The improved photocurrent efficiency, decay time, and spectral response resulted from the space charges generated by the LSPR of Ag NPs. Furthermore, the increasing exponent (m) of the photocurrent (JPC ∼ V(m)) and finite-differential time domain (FDTD) simulation straightforwardly indicate relatively large plasmonic space charge generation under blue light illumination. These results prove that the performance of non-single crystallized polymer nanowire devices can also be improved by plasmonic enhancement.

Analysis of matrix metalloproteinase mRNAs expressed in hepatocellular carcinoma cell lines.

To analyze matrix metalloproteinase (MMP) mRNAs that are expressed in hepatocellular carcinoma cell lines, the kinds of MMP mRNAs were surveyed in HepG2 and Hep3B cells and normal liver by a reverse transcription-polymerase chain reaction using two degenerate primer pairs, derived from conserved domains of known MMPs. The level for each MMP mRNA was examined by Northern blot analysis in HepG2 and Hep3B cells, as well as in normal tissues. It was also examined by a reverse transcription-polymerase chain reaction analysis in 8 different hepatocellular carcinoma cell lines. MMP-2, MMP-14, and MMP-15 mRNAs were expressed at elevated levels in most of the cell lines studied, reflecting that these MMPs would play an important role in the invasion and metastasis of hepatocellular carcinoma. MMP-1, MMP-3, MMP-8, MMP-10, MMP-11, and MMP-13 mRNAs were also expressed in some or most of the cell lines. Interestingly, MMP-9 mRNA, as well as its polypeptide, was undetected in all of the cell lines studied. This implies that MMP-9, which was suggested as a tumor marker for hepatocellular carcinoma, would be expressed in stromal cells, rather than tumor cells. These results provide information for the basal levels of MMP mRNAs in various hepatocellular carcinoma cell lines. It will also facilitate study on the transcriptional regulation of each MMP mRNA by oncogenes.

Bacteriology of chronic sinusitis after amoxicillin-clavulanate potassium therapy.

The bacteriology of chronic sinusitis was studied after amoxicillin-clavulanate potassium therapy. Patients with chronic sinusitis were randomly divided into 2 groups. In the study group, 90 patients were given a 2-week course of amoxicillin-clavulanate potassium before functional endoscopic sinus surgery. In the control group, 113 patients did not take any antibiotics within 2 weeks before the surgery. Swab specimens were taken from the maxillary and ethmoid sinuses during surgery and sent for aerobic and anaerobic culture. In the study group, the culture rates of maxillary and ethmoid sinuses were 45.6% and 28.9%, respectively. In the control group, the culture rates of maxillary and ethmoid sinuses were 53.1% and 34.5%, respectively. The culture rates between the control group and the study group were not significantly different, either for the maxillary sinus or the ethmoid sinus. This showed that treatment with amoxicillin-clavulanate potassium did not change the bacteriology of chronic sinusitis.

Effects of Qi-therapy on blood pressure, pain and psychological symptoms in the elderly: a randomized controlled pilot trial.

Recently, we reported that Qi-therapy may be beneficial in reducing negative psychological symptoms and increasing melatonin levels, neutrophil function and natural killer cell cytotoxicity in young subjects. However, there is little scientific evidence of its efficacy in elderly subjects. Therefore, this study was designed to investigate the effects of Qi-therapy on anxiety, depression, fatigue, pain and blood pressure in elderly subjects. Ninety-four elderly subjects were randomly assigned to either Qi-therapy (n=47) or mimic therapy (n=47) groups. Both groups received a 10-min intervention period once using similar procedures. The Qi-therapy group exhibited greater reduction in anxiety, depression, fatigue, pain level and blood pressure compared to the placebo group; the difference in anxiety was significant (P=0.014). These results suggest that even a brief application of Qi-therapy may exert a positive psychological and physiological effect. However, further research is necessary in order to fully understand the long-term impact of Qi-therapy on psychological health and the cardiovascular system.

Comparison of corneal centering in photorefractive keratectomy.

The present study compares three centering methods for excimer laser photorefractive keratectomy (PRK. VISX 20/20) by analyzing the corneal topography. The subjects were grouped according to three different centering methods used in the procedure: an ablation using a light reflex from the patient's cornea pursued by both eyes of the surgeon (Group 1, n = 49); an ablation using a red light reflex from the patient's cornea pursued by the surgeon's left eye only while the right eye remained closed (Group 2, n = 27); an ablation using the patient's center of the pupil pursued by the surgeon's left eye only while the right eye remained closed (Group 3, n = 21). The mean distance from the center of ablation zone to the center of the pupil were; 0.69 +/- 0.45 mm for Group 1, 1.05 +/- 0.48 mm for Group 2 and 0.63 +/- 0.28 mm for Group 3. The degree of deviation in Group 2 was significantly greater than in Group 1 or Group 3. The deviation was greater in the right eyes than the left eyes in Group 2 only. The decentration of the right eye in Group 2 was due to angle Kappa with misalignment of the fixation light and viewing tube containing reticule.

Bacteriology of the maxillary and ethmoid sinuses in chronic sinusitis.

The bacteriology of chronic sinusitis was studied by using swab and mucosal specimens from both the maxillary and ethmoid sinuses. The specimens of the maxillary sinus were taken through translabial antroscopy. The specimens of the ethmoid sinus were taken after removing the ethmoid bulla during functional endoscopic sinus surgery (FESS). Eighty-six samples of each type of specimen were collected. Among the maxillary sinus samples, the culture rate was 60.5 per cent from the swab specimens and 36 per cent from the mucosal specimens. Among the ethmoid sinus samples, the culture rate was 58.1 per cent from the swab specimens and 75.6 per cent from the mucosal. The p-value by the Chi-Square test is higher than 0.01 (p = 0.015). As there were more isolates of Staphylococcus epidermidis from the mucosal specimens, they are not a better choice of specimen for sampling the ethmoid sinus than a swab specimen.

Bacteriology of endoscopically normal maxillary sinuses.

The bacteriology of maxillary sinuses with normal endoscopic findings is reported in this study. When transantral sinoscopy was used to examine the maxillary sinuses, the whole maxillary sinus was inspected with different-angle endoscopes. If no lesion was seen over the whole maxillary sinus mucosa, no secretion existed in the maxillary sinus cavity, and the maxillary sinus ostium was wide open, the maxillary sinus was considered endoscopically normal. The bacteriology of these endoscopically normal maxillary sinuses was studied by passing cotton-tipped sticks through the cannula to collect swab specimens. In some cases, a biopsy forceps was also passed to obtain mucosal specimens. The specimens were sent to the laboratory for aerobic and anaerobic cultures. Between July 1990 and May 1998, 83 swab and 31 mucosal specimens were collected from 69 patients who had not taken any antibiotic within 10 days before endoscopy. The culture rates were 62.3 per cent (35/53) from swab specimens and 57.1 per cent (eight out of 14) from mucosal specimens in patients with the diagnosis of chronic paranasal sinusitis, and were 46.7 per cent (14/30) from swab specimens and 41.2 per cent (seven out of 17) from mucosal specimens in patients without this diagnosis. This study shows that endoscopically normal maxillary sinuses are not sterile.

The effect of excipients on glass transition temperatures for FK906 in the frozen and lyophilized states.

FK906, a tripeptide, has been used as a model compound to study the effect of excipients on the glass transition temperature for peptides and related compounds in the frozen solution and lyophilized states. Three kinds of excipients were chosen for study, low molecular weight sugars, high molecular weight polymers and salts. Our results indicate that the Gordon-Taylor equation can be used to predict the glass transition temperature of FK906 in the presence of sucrose, lactose, trehalose and maltose in the frozen solutions and in lyophilized products. Dextrans having different molecular weights can have different effects on the glass transition temperature of FK906 in the frozen state. Cooling rate can also have an effect on the glass transition temperature for FK906 frozen solutions in the presence of dextran. Finally, salts have a significant effect on the glass transition temperature of FK906 in the frozen state but not in lyophilized FK906.

Some physicochemical properties of FK906: glass transition and relaxation.

FK906, a tripeptide antihypertensive agent, was used as a model compound to study some physicochemical properties of peptides and peptide-related compounds. Our results indicate that lyophilization, grinding and melting or storing the samples under conditions of high relative humidity affect chemical stability and crystallinity of FK906. In a preliminary study of lyophilization process design, an extended phase diagram for FK906 in aqueous solution was constructed, and the effects of cooling rate and and heating rate on the glass transition temperatures of lyophilized FK906 and the FK906 solution were investigated. Following lyophilization, the DSC thermogram of FK906 exhibits an endothermic peak whose area is storage temperature- and time-dependent. The endothermic peak shifts to higher temperatures as the heating rate is increased. Our results indicate that the endothermic peak is related to structural change during incubation of lyophilized FK906.

Dynamic changes of the inflammation-based index predict mortality following chemoembolisation for hepatocellular carcinoma: a prospective study.

BACKGROUND: Transarterial chemoembolisation (TACE) is a standard treatment for unresectable, intermediate stage hepatocellular carcinoma (HCC). Survival after TACE, however, can be highly variable, with no suitable biomarker predicting therapeutic outcome. The inflammation-based index (IBI) has previously been shown to independently predict overall survival (OS) in all stages of HCC. AIM: To explore the prognostic ability of IBI as a predictor of survival after TACE. METHODS: Baseline staging, biochemical and clinicopathological features including IBI were studied in a derivation set of 64 patients undergoing TACE for intermediate stage HCC. Dynamic changes in IBI before and after TACE were studied as predictors of survival using both a univariate and multivariate Cox regression model and further validated in two independent patient cohorts from Korea (n = 76) and Japan (n = 577). RESULTS: Pre-treatment IBI predicted for OS in the derivation set (P = 0.001). Other univariate predictors of OS included radiological response to TACE (P < 0.001), pre-TACE CLIP score (P < 0.01), tumour diameter >5 cm (P = 0.05) and AFP ≥400 (P < 0.001). Normalisation of IBI post-TACE was associated with radiological response by mRECIST criteria and improved OS (P < 0.001). Normalisation of IBI remained a significant multivariate predictor of OS in both the derivation and validation sets (P < 0.001). CONCLUSIONS: Normalisation of IBI after TACE is shown to be an independent predictor of survival and may be integrated into the retreatment criteria for repeat TACE in intermediate stage HCC. IBI and its dynamic changes after treatment are validated as a biomarker allowing the stratification of patients with a significant survival advantage following initial TACE.

Rapid genotyping of Plasmodium vivax Pvs25 and Pv38 genes by using mismatch specific endonuclease.

Mismatch specific endonuclease (MSE) method was used to detect natural polymorphisms in Pvs25 and Pv38 genes of Plasmodium vivax. Eighty seven patients with P. vivax were recruited in the Republic of Korea (ROK). Pvs25 and Pv38 genes were amplified by polymerase chain reaction (PCR), and the PCR amplicons were mixed with reference DNA sequences. Following the denaturation and gradual annealing, the product mixtures were cleaved by the MSE. Heteroduplex types were readily detected by gel electrophoresis, where extra bands with shorter sizes would appear from the cleavage. After MSE cleavage of 657- bp product from Pvs25 mixtures, three genotypes were detected, while Pv38 mixtures with 1220-bp products presented two genotypes in ROK isolates. After the MSE cleavage, the mismatched samples of Pvs25 and Pv38 were completely sequenced, and the results were in complete agreement with the MSE analyses. In conclusion, genotyping of Pvs25 and Pv38 with MSE cleavage could be a potential method for the high-throughput screening of the large field samples.

RUNX family members are covalently modified and regulated by PIAS1-mediated sumoylation.

Transcription factors of the RUNX family (RUNXs), which play pivotal roles in normal development and neoplasia, are regulated by various post-translational modifications. To understand the molecular mechanisms underlying the regulation of RUNXs, we performed a large-scale functional genetic screen of a fly mutant library. The screen identified dPias (the fly ortholog of mammalian PIASs), an E3 ligase for the SUMO (small ubiquitin-like modifier) modification, as a novel genetic modifier of lz (the fly ortholog of mammalian RUNX3). Molecular biological analysis revealed that lz/RUNXs are sumoylated by dPias/PIAS1 at an evolutionarily conserved lysine residue (K372 of lz, K144 of RUNX1, K181 of RUNX2 and K148 of RUNX3). PIAS1-mediated sumoylation inhibited RUNX3 transactivation activity, and this modification was promoted by the AKT1 kinase. Importantly, PIAS1 failed to sumoylate some RUNX1 mutants associated with breast cancer. In nude mice, tumorigenicity was promoted by RUNX3 bearing a mutation in the sumoylation site, but suppressed by wild-type RUNX3. Our results suggest that RUNXs are sumoylated by PIAS1, and that this modification could play a critical role in the regulation of the tumor-suppressive activity of these proteins.