Acacetin, which is present in damiana (Turnera diffusa ) and black locust (Robinia pseudoacacia ), has several pharmacologic activities such as antioxidant, anti-inflammatory, and anti-proliferative effects on cancer cells. However, the effect of acacetin on head and neck cancers has not been clearly established. This study aimed to examine the effects of acacetin on cell growth and apoptosis induction in FaDu human pharyngeal carcinoma cells. These were investigated by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay, Live/Dead cell assay, 4′,6-diamidino-2-phenylindole dihydrochloride staining, caspase-3 and caspase-7 activation assay, and immunoblotting in FaDu cells. Acacetin induced FaDu cell death in a dose-dependent manner, with an estimated IC50 value of 41.9 µM, without affecting the viability of L-929 mouse fibroblasts as normal cells. Acacetin treatment resulted in nuclear condensation in the FaDu cells. It promoted the proteolytic cleavage of procaspase-3, -7, -8, and -9 with increasing amounts of the cleaved caspase isoforms in FaDu cells. Acacetin-induced apoptosis in FaDu cells was mediated by the expression of Fas and activation of caspase-8, caspase-3, and poly (ADP-ribose) polymerase. Immunoblotting showed downregulation of the anti-apoptotic mitochondrial proteins Bcl-2 and Bcl-xL, but upregulation of the mitochondria-dependent pro-apoptotic proteins Bax and Badin FaDu cells after acacetin treatment. These findings indicate that acacetin inhibits cell proliferation and induces apoptotic cell death in FaDu human pharyngeal carcinoma cells via both the death receptor-mediated extrinsic apoptotic pathway and the mitochondria-mediated intrinsic apoptotic pathway.

Ficus carica L. (fig ) is one of the first cultivated crops and is as old as humans. This plant has been extensively used as a traditional medicine for treating diseases, such as cough, indigestion, nutritional anemia, and tuberculosis. However, the physiological activity of fig leaves on oral cancer is as yet unknown. In this study, we investigated the anticancer effect of methanol extracts of Ficus carica (MeFC) and the mechanism of cell death in human FaDu hypopharyngeal squamous carcinoma cells. MeFC decreased the viability of oral cancer (FaDu) cells but did not affect the viability of normal (L929) cells, as determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and Live and Dead assay. In addition, MeFC induced apoptosis through the proteolytic cleavage of procaspase-3, -9, poly (ADP-ribose) polymerase (PARP), downregulation of Bcl-2, and upregulation of Bax, as determined by 4′,6-diamidino-2-phenylindole dihydrochloride staining and western blot analysis. Moreover, a concentration of MeFC without cytotoxicity (0.25 mg/mL) significantly suppressed colony formation, a hallmark of cancer development, and completely inhibited the colony formation at 1 mg/mL. Collectively, these results suggest that MeFC exhibits a potent anticancer effect by suppressing the growth of oral cancer cells and colony formation via caspase- and mitochondrial-dependent apoptotic pathways in FaDu human hypopharyngeal squamous carcinoma cells. Therefore, the methanol extract of Ficus carcica leaves provide a natural chemotherapeutic drug for human oral cancer.

The present study was conducted to investigate the effects of feed types (powder vs. pellet) from potato by-product containing glycoalkaloid on rumen fermentation with different rumen pH (low vs. high) using in vitro technique. Rumen fluid was collected from two cannulated Hanwoo heifers before morning feeding (high rumen pH at 6.71) and 3 h after feeding (low rumen pH at 6.20). A 0.5 gram of powder or pellet from potato by-product was incubated in the rumen buffer for 48 h at 39℃. Four replications for each treatment were used along with two blanks. Pellet had higher (p<0.05) the immediately degradable fraction, the fractional degradation rate, and the lag phase than those of powder. However, feed type had no effects on the potentially degradable fraction and the total degradable fraction. In fermentation characteristics, powder had higher (p<0.05) concentrations of ammonia-N, total volatile fatty acid (VFA), and acetate than those of pellet. In addition, a high rumen pH had higher (p<0.05) concentrations of ammonia-N and acetate, but lower (p<0.05) total VFA concentration than those of a low rumen pH. The interaction effects between feed type and rumen pH were found on the total degradable fraction and concentrations of ammonia-N, total VFA, and acetate. The present study concluded that generally feed type had no effects on rumen degradation kinetics, but potato by-product in the powder form is recommended due to produce higher fermentation characteristics than in the pellet form.

Enamel knot (EK)—a signaling center—refers to a transient morphological structure comprising epithelial tissue. EK is believed to regulate tooth development in early organogenesis without its own cellular alterations, including proliferation and differentiation. EKs show a very simple but conserved structure and share functions with teeth of recently evolved vertebrates, suggesting conserved signaling in certain organs, such as functional teeth, through the course of evolution. In this study, we examined the expression patterns of key EK-specific genes including Dusp26 , Fat4, Meis2, Sln , and Zpld1 during mice embryogenesis. Expression patterns of these genes may reveal putative differentiation mechanisms underlying tooth morphogenesis.

The goal of this study was to evaluate effects of various microbial and organic additives on chemical compositions, fermentation indices, and aerobic stability of barley silage. Youngyang barley harvested at 31.5% dry matter (DM), and ensiled into 20 L bucket silo for 0, 1, 3, 7, 48, and 100 d in quadruplicates with four additives following: sterile destilled water at 1% of fresh forage (CON); Lactobacillus plantarum at 1.2 x 105 cfu/g of fresh forage (CL); Lactobacillus buchneri at rate of 1.2 x 105 cfu/g fresh forage (LB); Fermented Persimmon Extract at 1% of fresh forage (FPE); and Essential Oil at 1% of fresh forage (EO). On 100 d of ensiling, CL and FPE silages had lower (p<0.05) DM than CON silage. Additionally, FPE silage had higher (p<0.05) crude protein than CON silage. All silages with additives, except EO, had higher (p<0.05) neutral detergent fiber (NDF) than CON silage. Silage treated with CL, LB, and FPE had lower in vitro DM digestibility than CON silage, and silages treated with LB and FPE had higher in vitro NDF digestibility (IVNDFD) on 100 d of ensiling. The PFE silage produced the highest (p<0.05) lactate during ensiling period, while LB silage produced the highest (p<0.05) acetate. All inoculated silages had higher (p<0.05) LAB count than control silage. Only CL silage had higher (p<0.05) yeast count than control silage, while the other silages were not differ compared to CON silage. The aerobic stability was higher (p<0.05) in LB and FPE silages than in CON silage. In conclusion, FPE could be an alternative additive to increase IVNDFD, fermentation indices, and aerobic stability of barley silage.

In this research, a new medium-entropy alloy with an equiatomic composition of FeCuNi was designed using a phase diagram (CALPHAD) technique. The FeCuNi MEA was produced from pure iron, copper, and nickel powders through mechanical alloying. The alloy powders were consolidated via a high-pressure torsion process to obtain a rigid bulk specimen. Subsequently, annealing treatment at different conditions was conducted on the four turn HPT-processed specimen. The microstructural analysis indicates that an ultrafine-grained microstructure is achieved after post-HPT annealing, and microstructural evolutions at various stages of processing were consistent with the thermodynamic calculations. The results indicate that the post-HPT-annealed microstructure consists of a dual-phase structure with two FCC phases: one rich in Cu and the other rich in Fe and Ni. The kernel average misorientation value decreases with the increase in the annealing time and temperature, indicating the recovery of HPT-induced dislocations.

To attenuate and control the spread of infectious disease, a body of research has been conducted to generate safe vaccines and to continue national-level surveillance. However, understanding on viability and persistence of avian influenza virus (AIV) in infected carcasses, and effective disposal approaches are still limited up to date. Here, using HA test and RT-PCR, we assessed active status of AIV and degradation of viral RNA in collected specimens at different sites and time points. First, AIV infectivity was recovered until day 2, and viral nucleic acids persisted to day 14 and 21 in inorganic and organic samples, respectively, in sealed vials incubated at room temperature. Second, AIV was totally inactivated in all examined specimens, and viral RNA was not detectable at all time points tested at least one month post-infection in AIV-inoculated carcasses buried directly in soil or fiber reinforced plastic (FRP) bin. Lastly, among different burial sites in South Korea, 6 out of 17 sampling sites in Jeonbuk province showed the presence of viral genetic materials, while the rest of the field samples displayed neither the presence of infective AIV nor detectable viral RNA. This study showed a linear relation between time and degradation degree of viral RNA in buried samples suggesting that burial disposal method is effective for the control or at least attenuation of spread of AI infection in infected animals although consistent monitoring is required to verify safety of disposal.

Trifolium pratense leaves (red clover) has been used in Oriental and European folk medicine for the treatment of whooping cough, asthma, and eczema, and is now being used to treat and alleviate the symptoms, such as hot flushes, cardiovascular health effects that occur in postmenopausal women. However, relatively little scientific data is available on the physiological activity of this plant. Therefore, in this study, we investigated the anti-cancer activity of T. pratense leaves using methanol extract of T. pratense leaves (MeTP) on human FaDu hypopharyngeal squamous carcinoma cells. MeTP inhibited the viability of FaDu cells by inducing apoptosis through the cleavage of procaspase- 3, -7, and -9 and poly (adenosine diphosphate ribose-ribose) polymerase (PARP), downregulation of Bcl- 2, and upregulation of Bax, as determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Live & dead assay, 4’6-diamidino-2-phenylindole stain, fluorescence-activated cell sorting analysis, and Western blot analysis. In addition, colony formation was slightly inhibited when FaDu cells were treated with a non-cytotoxic concentration (0.125 mg/mL) of MeTP and almost completely inhibited when cells were treated with 0.25 mg/mL MeTP. Collectively, these results indicate that MeTP induced cell apoptosis via caspase- and mitochondrial-dependent apoptotic pathways, and inhibited colony formation of cancer cells in FaDu human hypopharyngeal squamous carcinoma cells. These findings suggest MeTP should be considered for clinical development as a chemotherapeutic option in oral cancer.

A previous studies depicting origin and sequence variability of the species using DNA barcoding region with the samples collected from Korea showed relatively low sequence variability. Thus, additional markers that reveal higher variability were necessitated to scrutinize population structure in connection with dispersal and invasive dynamics among international populations. Therefore, we sequenced two complete mitochondrial genomes (mitogenomes) of M. pruinosa from the two haplotypes occurring in Korea (H1 and H3). Comparison of the two mitogenomes each with 16,312 and 16,314 bp evidenced that one region located in the A+T-rich region to provide higher number of haplotypes (4 vs. 3), sequence divergence (1.636% vs. 0.636%), and variable sites (7 vs. 3) than those of DNA barcoding region from the screening test using 13 representative individuals. This variable region, in concatenation with the currently available DNA barcoding region might be useful for population genetic analysis of worldwide populations including those of Korea. †These authors contributed equally to this paper.