Unusual Dependence of the Diamond Growth Rate on the Methane Concentration in the Hot Filament Chemical Vapor Deposition Process.

Although the growth rate of diamond increased with increasing methane concentration at the filament temperature of 2100 °C during a hot filament chemical vapor deposition (HFCVD), it decreased with increasing methane concentration from 1% CH4 -99% H2 to 3% CH4 -97% H2 at 1900 °C. We investigated this unusual dependence of the growth rate on the methane concentration, which might give insight into the growth mechanism of a diamond. One possibility would be that the high methane concentration increases the non-diamond phase, which is then etched faster by atomic hydrogen, resulting in a decrease in the growth rate with increasing methane concentration. At 3% CH4 -97% H2, the graphite was coated on the hot filament both at 1900 °C and 2100 °C. The graphite coating on the filament decreased the number of electrons emitted from the hot filament. The electron emission at 3% CH4 -97% H2 was 13 times less than that at 1% CH4 -99% H2 at the filament temperature of 1900 °C. The lower number of electrons at 3% CH4 -97% H2 was attributed to the formation of the non-diamond phase, which etched faster than diamond, resulting in a lower growth rate.

Comparison of diamond nanoparticles captured on the floating and grounded membranes in the hot filament chemical vapor deposition process.

Negatively charged diamond nanoparticles are known to be generated in the gas phase of the hot filament chemical vapor deposition (HFCVD) process. However, the structures of these nanoparticles remain unknown. Also, the effect of charging on the stability of nanodiamond structures has not been studied experimentally. Here, by installing a capturing apparatus in an HFCVD reactor, we succeeded in capturing nanoparticles on the floating and grounded SiO, carbon, and graphene membranes of a copper transmission electron microscope grid during HFCVD. We examined the effect of charge on the crystal structure of nanodiamonds captured for 10 s under various conditions and identified four carbon allotropes, which are i-carbon, hexagonal diamond, n-diamond, and cubic diamond, by analyzing 150 d-spacings of ∼100 nanoparticles for each membrane. Nanoparticles captured on the floating membrane consisted mainly of cubic diamond and n-diamond, whereas those captured on the grounded membrane consisted mainly of i-carbon. Diamond particles deposited for 8 h on the floating silicon (Si) substrate exhibited an octahedron shape with well-developed facets, and a high-intensity 1332 cm-1 Raman peak, whereas diamond particles deposited on the grounded Si substrate showed a spherical shape partially covered with crystalline facets with a broad G-band Raman peak. These results indicate that charging stabilizes the diamond structure.

Various Allotropes of Diamond Nanoparticles Generated in the Gas Phase during Hot Filament Chemical Vapor Deposition.

Diamond nanoparticles have been synthesized using various methods. Nanodiamonds generated in the gas phase were captured on the membrane of a transmission electron microscope grid during a hot filament chemical vapor deposition (HFCVD) diamond process. In total, ~600 nanoparticles, which were captured for 10 s in six conditions of the capture temperatures of 900 °C, 600 °C and 300 °C and the gas mixtures of 1% CH4-99% H2 and 3% CH4-97% H2, were analyzed for phase identification using high-resolution transmission electron microscopy and fast Fourier transformation. Hexagonal diamond, i-carbon, n-diamond, and cubic diamond were identified. The observation of two or more carbon allotropes captured on the same membrane suggested their coexistence in the gas phase during HFCVD. The crystal structure of carbon allotropes was related to the size of the nanodiamond. The crystal structure of the nanoparticles affected the crystal structure of diamond deposited for 8 h. Confirmation of various carbon allotropes provides new insight into the nanodiamond synthesis in the gas phase and the growth mechanism of HFCVD diamond.

Profiling of biomarkers for the exposure of polycyclic aromatic hydrocarbons: lamin-A/C isoform 3, poly[ADP-ribose] polymerase 1, and mitochondria copy number are identified as universal biomarkers.

This study investigated the profiling of polycyclic aromatic hydrocarbon- (PAH-) induced genotoxicity in cell lines and zebrafish. Each type of cells displayed different proportionality of apoptosis. Mitochondrial DNA (mtDNA) copy number was dramatically elevated after 5-day treatment of fluoranthene and pyrene. The notable deregulated proteins for PAHs exposure were displayed as follows: lamin-A/C isoform 3 and annexin A1 for benzopyrene; lamin-A/C isoform 3 and DNA topoisomerase 2-alpha for pentacene; poly[ADP-ribose] polymerase 1 (PARP-1) for fluoranthene; and talin-1 and DNA topoisomerase 2-alpha for pyrene. Among them, lamin-A/C isoform 3 and PARP-1 were further confirmed using mRNA and protein expression study. Obvious morphological abnormalities including curved backbone and cardiomegaly in zebrafish were observed in the 54 hpf with more than 400 nM of benzopyrene. In conclusion, the change of mitochondrial genome (increased mtDNA copy number) was closely associated with PAH exposure in cell lines and mesenchymal stem cells. Lamin-A/C isoform 3, talin-1, and annexin A1 were identified as universal biomarkers for PAHs exposure. Zebrafish, specifically at embryo stage, showed suitable in vivo model for monitoring PAHs exposure to hematopoietic tissue and other organs.

Over-expression of Her-2 in colorectal cancer tissue, but not in serum, constitutes an independent worse prognostic factor.

BACKGROUND: Currently, conflicting information exists regarding Her-2 over-expression and its clinicopathological implications in colorectal cancer (CRC). This study was undertaken to determine Her-2 over-expression in both serum and tumor tissue of CRC patients, and to assess its clinicopathological and targeted therapeutic implications. METHODS: Ninety five CRC patients and sixty healthy controls were prospectively enrolled. Her-2 expression status in serum and CRC tissue were examined by chemiluminescent immunoassay and immunohistochemical staining, respectively. The results were confirmed using fluorescent in situ hybridization. Clinicopathological parameters were analyzed according to Her-2 expression status. RESULTS: Serum Her-2 levels were found to be increased in CRC patients as compared to those of healthy controls. However, serum Her-2 levels were not found to be significantly associated with prognostic parameters. Her-2 expression analysis of CRC tissues revealed Her-2 over-expression in 23 patients (25%), i.e., 13 patients (14%) showed moderate over-expression and 10 patients (11%) showed strong over-expression. The overall survival of patients negative for Her-2 expression was significantly better than that of patients positive for Her-2 expression (P=0.018). The disease-free survival of patients with Her-2 over-expression was significantly shorter than that of patients with no Her-2 expression (P=0.021). CONCLUSIONS: Her-2 over-expression in CRC tissue, but not in serum, acts as a significant independent worse prognostic factor. Assessment of Her-2 expression status may be valuable for the targeted therapeutic management of CRC.

Diagnostic and prognostic value of mitochondrial DNA minisatellites after stem cell transplantation.

Mitochondrial DNA has been used to investigate phylogenetic relationships and pathophysiologic roles in aging, degenerative diseases, and cancer. We investigated the prognostic usefulness of mitochondrial DNA minisatellite (mtMS) markers compared with nuclear short tandem repeat markers by evaluating the laboratory performance and clinical value of these markers in a large sample of patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) with various simulated conditions in vitro and in serial follow-up samples. We examined the value of mtMS markers as a prognostic indicator in 100 patients with various hematologic disorders undergoing allo-HSCT, including 35 patients with longitudinal follow-up for 55 months. The mtMS markers showed high sensitivity and accuracy for the quantitative detection of chimerism compared with nuclear short tandem repeat markers, particularly in unrelated transplantation and under inappropriate sampling conditions. Longitudinal follow-up after allo-HSCT disclosed that chimerism precisely reflected the status of engraftment or relapse during the clinicopathological course. Moreover, changes in mtMS markers in recipients before allo-HSCT were associated with clinical outcomes. Our data indicate that mtMS markers have multiple functions in monitoring mixed chimerism and predicting prognosis after allo-HSCT.

Usefulness of plasma epigenetic changes of five major genes involved in the pathogenesis of colorectal cancer.

PURPOSE: The purpose of present study was to investigate the methylation status of the promoter region in five genes (mothers against decapentaplegic homolog 4, fragile histidine triad protein, death-associated protein kinase 1, adenomatous polyposis coli (APC), and E-cadherin), which are known to be involved in the pathogenesis of colorectal cancer (CRC) and its clinicopathological significance. METHODS: The study subjects were 60 CRC patients, 40 patients with adenomatous colorectal polyp and 60 healthy control individuals. We further enrolled a total of 16 patients (two patients with Crohn's disease, two patients with ulcerative colitis, one patient with serrated adenoma, and 11 patients with colorectal cancer). The methylation states of the five genes were determined in peripheral blood plasma using methylation-specific polymerase chain reaction single-strand conformation polymorphism analysis. RESULTS: This study showed the most sensitive epigenetic markers, E-cadherin (60 %), followed by APC (57 %), for detecting CRC. E-cadherin and APC had similar specificities and amplified 84 and 86 %, respectively, of CRC patients compared to non-CRC patients. Additionally, APC was the only marker to be significantly increased (OR = 6.67, 95 % CI = 1.19-23.4, P = 0.045) and the most sensitive (57 %) and specific (89 %) marker in stage I CRC. Though we have not examined the paired cancer tissues and plasma, there was relatively high concordant rate (60-80 %) in our limited number of colorectal cancer patients. CONCLUSIONS: Five genes, promoter methylation, in plasma were statistically significant risk factors in CRC patients. In this study, E-cad and APC genes may be particularly useful epigenetic biomarkers in plasma for the detection of CRC. Additionally, APC may able to identify early potential CRC.

Frequent occurrence of mitochondrial DNA mutations in Barrett's metaplasia without the presence of dysplasia.

BACKGROUND: Barrett's esophagus (BE) is one of the most common premalignant lesions and can progress to esophageal adenocarcinoma (EA). The numerous molecular events may play a role in the neoplastic transformation of Barrett's mucosa such as the change of DNA ploidy, p53 mutation and alteration of adhesion molecules. However, the molecular mechanism of the progression of BE to EA remains unclear and most studies of mitochondrial DNA (mtDNA) mutations in BE have performed on BE with the presence of dysplasia. METHODS/FINDINGS: Thus, the current study is to investigate new molecular events (Barrett's esophageal tissue-specific-mtDNA alterations/instabilities) in mitochondrial genome and causative factors for their alterations using the corresponding adjacent normal mucosal tissue (NT) and tissue (BT) from 34 patients having Barrett's metaplasia without the presence of dysplasia. Eighteen patients (53%) exhibited mtDNA mutations which were not found in adjacent NT. mtDNA copy number was about 3 times higher in BT than in adjacent NT. The activity of the mitochondrial respiratory chain enzyme complexes in tissues from Barrett's metaplasia without the presence of dysplasia was impaired. Reactive oxygen species (ROS) level in BT was significantly higher than those in corresponding samples. CONCLUSION/SIGNIFICANCE: High ROS level in BT may contribute to the development of mtDNA mutations, which may play a crucial role in disease progression and tumorigenesis in BE.

High-frequency minisatellite instability of the mitochondrial genome in colorectal cancer tissue associated with clinicopathological values.

Most studies of mitochondrial DNA (mtDNA) mutations in colorectal cancer have used case-control and case-database comparisons without searching their clinical relevance. This study was to investigate colorectal cancer tissue-specific mtDNA mutations from 54 matched colorectal cancer and adjacent normal tissues and then to evaluate their clinical values. This study focused on analyzing control region including mtDNA minisatellites and coding regions. Cancer tissue-specific mtDNA mutations were found in over half of the patients (59%). The patterns of mtDNA mutations were substitution only (13%), mtDNA minisatellite instability (mtMSI) (20%) and both mutations combined (26%). mtMSI in colorectal cancer was mainly occurred in the 303 polyC (35%) and 16184 poly C (19%) minisatellite. mtDNA copy number and hydrogen peroxide level were significantly increased in colorectal cancer tissue. The amount of mtDNA large deletions was significantly decreased in colorectal cancer tissue compared with those from matched normal mucosa (p = 0.03). The activity of the mitochondrial respiratory chain enzyme complexes I, II and III in colorectal cancer tissues was impaired. mtDNA haplogroup B4 might be closely associated with colorectal cancer risk. The patient group harboring cancer tissue-specific mtDNA mutations showed larger tumor sizes (p = 0.005) and more advanced TNM stages (p = 0.002). Thus, mtDNA mutations in colorectal cancer might be implicated in risk factors that induce poor outcomes and tumorigenesis.

Mitochondrial DNA copy number and hnRNP A2/B1 protein: biomarkers for direct exposure of benzene.

The present study was performed to identify biomarkers for exposure of benzene in blood cells and hematopoietic tissues. Peripheral mononuclear cells, hematopoietic stem cells, and leukemia cell lines were cultured in RPMI 1640 media with the addition of 0, 1, and 10 mM of benzene. Hydrogen peroxide was measured using an enzyme immunoassay. Mitochondrial mass, membrane potential, and mitochondrial DNA (mtDNA) copy number were measured using MitoTracker Green/Red probes, and real-time polymerase chain reaction. In addition, two-dimensional gel electrophoresis and mass spectrometry matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) technology were performed to identify protein markers. The mitochondrial contents and membrane potentials were dramatically increased after three weeks of direct benzene exposure. The hydrogen peroxide level increased significantly after two weeks of treatment with benzene (4.4 ± 1.9 µM/mg protein) compared to the non-benzene treatment group (1.2 ± 1.0; p = 0.001). The mtDNA copy number gradually increased after exposure to benzene. Numerous protein markers showed significant aberrant expression after exposure to benzene. Among them, the heterogeneous nuclear ribonucleoprotein (hnRNP) A2/B1 was markedly decreased after exposure to benzene. Thus, increased mitochondrial mass, mtDNA copy number, and the hnRNP A2/B1 protein were biomarkers for benzene-related toxicity and hematotoxicity.

Stabilization/Solidification of radioactive molten salt waste via gel-route pretreatment.

The volatilization of radionuclides during the stabilization/solidification of radioactive wastes at high temperatures is one of the major problems to be considered in choosing suitable wasteforms, process, material systems, etc. This paper reports a novel method to convert volatile wastes into nonvolatile compounds via a sol-gel process, which is different from the conventional method using metal-alkoxides and organic solvents. The material system was designed with sodium silicate (Si) as a gelling agent, phosphoric acid (P) as a catalyst/stabilizer, aluminum nitrate (Al) as a property promoter, and H20 as a solvent. A novel structural model for the chemical conversion of molten salt waste, named RPRM (Reaction Product in Reaction Module), was established, and the waste could be solidified with glass matrix via a simple procedure. The leached fraction of Cs and Sr by a PCT leaching method was 0.72% and 0.014%, respectively. In conclusion, the RPRM model isto converttargetwastes into stable and manageable products, not to obtain a specific crystalline product for each radionuclide. This paper suggested a new stabilization/solidification method for salt wastes by establishing the gel-forming material system and showing a practical example, not a new synthesis method of stable crystalline phase. This process, named "gel-route stabilization/solidification (GRSS)", will be a prospective alternative with stable chemical process on the immobilization of salt wastes and various mixed radioactive waste for final disposal.