Development of strain-specific PCR primers based on a DNA probe Fu12 for the identification of fusobacterium nucleatum subsp. nucleatum ATCC 25586T.

The objective of this study was to assess the strain-specificity of a DNA probe, Fu12, for Fusobacterium nucleatum subsp. nucleatum ATCC 25586T (F. nucleatum ATCC 25586T), and to develop sets of strain-specific polymerase chain reaction (PCR) primers. Strain-specificity was tested against 16 strains of F. nucleatum and 3 strains of distinct Fusobacterium species. Southern blot hybridization revealed that the Fu12 reacted exclusively with the HindIII-digested genomic DNA of F. nucleatum ATCC 25586T. The results of PCR revealed that three pairs of PCR primers, based on the nucleotide sequence of Fu12, generated the strain-specific amplicons from F. nucleatum ATCC 25586T. These results suggest that the DNA probe Fu12 and the three pairs of PCR primers could be useful in the identification of F. nucleatum ATCC 25586T, especially with regard to the determination of the authenticity of the strain.

Karyotype analysis of a Korean cucumber cultivar (Cucumis sativus L. cv. Winter Long) using C-banding and bicolor fluorescence in situ hybridization.

An intensive karyotype analysis of a Korean cucumber cultivar (Cucumis sativus L. cv. Winter Long) was carried out with three different methods. These included Feulgen staining, Giemsa C-banding, and fluorescence in situ hybridization (FISH). The mitotic chromosomes of the cucumber (2n = 2x = 14) were characterized, based on the length and arm ratio values. A C-banding analysis showed dark stains on the centromeric, telomeric, and intercalary regions of the chromosomes, except that chromosome 2 had a heavy staining in the long arm. Bicolor FISH, using 45S and 5S rDNA probes, provided additional information to identify cucumber chromosomes. The signals for 45S rDNA were detected on the pericentromeric regions of chromosomes 1, 2, and 4. The signals for 5S rDNA were on the short arm of chromosome 5. Similar band patterns (as the C-banding) were observed when the chromosomes were counter-stained with 4',6-diamidino-2-phenyoindole (DAPI). The data implied that the karyotype of the Korean cucumber cultivar is peculiar and different from previous reports.

Capsaicin-induced apoptosis of FaDu human pharyngeal squamous carcinoma cells.

PURPOSE: To investigate the anti-tumor effect of capsaicin on human pharyngeal squamous carcinoma cells (FaDu). MATERIALS AND METHODS: The expression of apoptosis/cell cycle-related proteins (or genes) was examined by reverse transcriptase- polymerase chain reaction, western blotting and ELISA methods, while the apoptotic cell population, cell morphology and DNA fragmentation levels were assessed using flow cytometry, fluorescence microscopy and agarose gel electrophoresis. RESULTS: Capsaicin was found to inhibit the growth and proliferation of FaDu cells in a dose- and time-dependent manner. Apoptotic cell death was confirmed by observing increases in nuclear condensation, nuclear DNA fragmentation and sub-G1 DNA content. The observed increase in cytosolic cytochrome c, activation of caspase 3 and PARP (p85) levels following capsaicin treatment indicated that the apoptotic response was mitochondrial pathway-dependent. Gene/protein expression analysis of Bcl-2, Bad and Bax further revealed decreased anti-apoptotic Bcl-2 protein and increased pro-apoptotic Bad/Bax expression. Furthermore, capsaicin suppressed the cell cycle progression at the G1/S phase in FaDu cells by decreasing the expression of the regulators of cyclin B1 and D1, as well as cyclin-dependent protein kinases cdk-1, cdk-2 and cdk-4. CONCLUSION: Our current data show that capsaicin induces apoptosis in FaDu cells and this response is associated with mitochondrial pathways, possibly by mediating cell cycle arrest at G1/S.

Changes in mechanical properties of seven light-cured composite resins after thermal cycling.

OBJECTIVE: To examine the changes of the mechanical properties of 7 different light-cured composite resins after thermal cycling and the correlations between these properties. METHODS: Seven different light-cured composite resins, including 2 microfilled composites (A110:AH and ESTELITE :ET), 3 microhybrid composites (AELITE:AT, Z250:ZS, and CharmFil plus:CP), and 2 nanohybrid composites (Z350:ZH and Grandio:GD), were prepared into test specimens with a diameter of 12 mm and a thickness of 1.0 mm. The specimens were stored in distilled water at 37 degrees celsius; for 24 h prior to 1 000 thermal cycles of 5 degrees celsius; for 15 s and 55 degrees celsius; for 15 s. The biaxial flexural strength (δ(f)) was tested using the ball-on-three-ball method at a crosshead speed of 0.5 mm/min (ISO4049). The fracture surface was observed under scanning electron microscope (SEM), and the remaining specimens underwent Knoop hardness test with a 50-g loading for 10 s. RESULTS: The highest and lowest Weibull modulus was observed in AH (18.752) and AT (5.290) group, respectively. The highest and lowest biaxial flexural strength was observed in ZS (158.2 MPa) and ET (54.0 MPa) groups, respectively. The δ(f) of the tested materials decreased in the order of microhybrid composite, nanohybrid composite, and microfiller composite, and the δ(f) showed no significant difference between the composites with a similar filler (P>0.05). The fracture number was positively correlated to the strength of the material. The Knoop hardness numbers (H) was the highest in GD group (110.81∓14.77 kg/mm(2)) and the lowest in AH group (42.81∓1.91 kg/mm(2)). SEM showed that the interface region of the matrix and the filler was vulnerable to crack formation. CONCLUSION: The nanohybrid composite resins better suit clinical applications than microhybrid composites. The applicability of Knoop hardness test in hardness measurement of the composite resins needs to be further demonstrated.