Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.

Animal crude drugs (natural medicines derived from animal organs) have been widely used in various Chinese medicine for the therapeutic effects and for enhancement of immunologic functions. We found the specific identification methods using DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analyses for mitochondrial DNA (mt DNA) in order to discriminate between the animal species and organs as well as the placenta of humans. Species-specific PCR bands of D-loop mt DNA for equine, bovine, porcine, and human were 133 bp, 137 bp, 231 bp, and 240 bp, respectively. Porcine organs were identified using restriction enzyme, HphI cut into two subfragments, 36 bp and 195 bp bands in the heart, spleen, and liver, except for kidney. The heart and liver of porcine were identified using restriction enzyme, SpeI cut into two subfragments, 84 bp and 147 bp bands, except for kidney and spleen. Bovine organs were cut into 68 bp and 69 bp bands in the liver, kidney, and spleen using NalIV, except heart and placenta. Placentas of bovine and humans were easily identified using each primer. Our results suggest that sequencing of mt DNA and its PCR-RFLP methods are useful for identification and discrimination of inter- and intra-specific variations in equine, bovine, porcine, and human by routine analysis.

The effects of chitosan upon the experimentally induced differentiation of MDPC-23 cells, derived from mouse dental papilla cells, were investigated by RT-PCR, observations of cell morphology and Alizaline red-S staining. Chitosan was found to significantly increase and accelerate the expression of ALP mRNA but decrease the ColI transcript levels, as compared with the control, in a time-dependent manner during the differentiation of MDPC-23 cells. Chitosan also significantly downregulated ON mRNA expression and accelerated mineralization in differentiating MDPC-23 cells. These results suggest that chitosan facilitates odontoblast differentiation and mineralization and may have potential clinical applications as a dentin regeneration material.

The spontaneous mutant circling mouse (cir/cir) shows a circling behavior and hearing loss. We produced transgenic mice overexpressing transmembrane inner ear (tmie) gene, the causative gene, for the phenotypic rescue of the circling mouse. Through the continuous breeding with circling mice, the cir/cir homozygous mice carrying the transgene (cir/cir‐gtg) were produced. The rescued cir/cir‐gtg mice were able to swim in the water with proper orientation and did not show any circling behavior like wild type mice. Western blot and immunohistochemical analysis exhibited that the transgenic tmie was expressed in the inner ear. Inner and outer hair cells were recovered in the cochlea and spiral ganglion neurons were also recovered in the rescued mice. Auditory brainstem response (ABR) test demonstrated that the cir/cir‐gtg mice are able to respond to sound. This study demonstrates that tmie transgene can recover the hearing impairment and abnormal behavior in the circling mouse.

Selenoprotein S (SelS) is widely expressed in diverse tissues where it localizes in the plasma membrane and endoplasmic reticulum. We studied the potential function of SelS in erythrocyte differentiation using K562 cells stably over-expressing SelS wild-type (WT) or one of two SelS point mutants, U188S or U188C. We found that in the K562 cells treated with 1μM Ara-C, SelS gradually declined over five days of treatment. On day 4, intracellular ROS levels were higher in cells expressing SelS-WT than in those expressing a SelS mutant. Moreover, the cell cycle patterns in cells expressing SelS-WT or U188C were similar to the controls. The expression and activation of SIRT1 were also reduced during K562 differentiation. Cells expressing SelS-WT showed elevated SIRT1 expression and activation (phosphorylation), as well as higher levels of FoxO3a expression. SIRT1 activation was diminished slightly in cells expressing SelS-WT after treatment with the ROS scavenger NAC (12 mM), but not in those expressing a SelS mutant. After four days of Ara-C treatment, SelS-WT-expressing cells showed elevated transcription of β-globin, y-globin, ε-globin, GATA-1 and zfpm-1, whereas cells expressing a SelS mutant did not. These results suggest that the suppression of SelS acts as a trigger for proerythrocyte differentiation via the ROS-mediated downregulation of SIRT1.

Antioxidants partially ameliorated the detrimental effects of reactive oxygen species (ROS) on sperm characteristics during in vitro storage. The objective of the present study was to investigate the single or synergetic antioxidative effect of curcumin and Vit. E on the characteristics of fresh boar sperm during in vitro storage. The sperm viability in curcumin, Vit. E supplementation and curcumin+Vit. E+H2O2 groups remained over 85.0% in 3 hr incubation period, but in 6 hr incubation period, curcumin+Vit. E+H2O2 groups was sharply dropped than those of curcumin and Vit. E group. The membrane intergrity in all evaluated groups except for H2O2 group did not significantly difference in 3 hr incubation period. The viability in curcumin or Vit. E supplementation were significantly increased than in curcumin+H2O2 and Vit. E+H2O2 group in 6 hr incubation period. The percentage of mitochondrial activity and acrosome intergrity obtained similar trends within same incubation periods irrespective of treatment. The lipid peroxidation of spermatozoal plasma membrane ranged from 11.6∼17.5 nM/l×106 and 14.0∼ 19.0 nM/l×106 in 3 hr and 6 hr incubation periods. In conclusion, curcumin or Vit. E rpplementation alone or cooperatively improved sperm viability index (motility, membrane intergrity, viability and survival rates) and fertility index (mitochondria activity, acrosome intergrity and lipid peroxidation) of fresh boar sperm, indicating that curcumin and Vit. E have a antioxidative properties through its scavenging activity against hydrogen peroxide.

Seoul is one of the most dynamic capital cities in the world of which population is about 10.49 million. The air quality of Seoul subway is a great concern to users and the public authorities, especially the pollution of particulate matters. The metallic particulate matters are generated from the friction of wheels and subway rails, which causes dispersion into the platform and even to the trains. This study attempted to characterize the current filters in subway MVAC system and a newly developed filter which was a type of thin electret fiber bundle. Collection efficiency of the filtration system varies from 20% to 90% depending on filter life and dust particle size. The removal of fine dust with diameter smaller than 1㎛, which could not be well achieved by current filtration system, is very important in the subway air cleaning system. The new electret filter was found to have a high potential for the use in subway MVAC with high collection efficiency, especially for fine mode dust.

14-3-3 proteins are known to play a pivotal role in a diverse array of cellular events such as cell survival, apoptosis, and signal transduction. Numerous 14-3-3 ζ have been cloned and characterized from a host of eukaryotic organisms including human, plants, yeast, fruit fly and silkworm. However, no study on Spodoptera exigua 14-3-3ζ in conjunction with virus infection has so far been reported in insects. It appears that expression of Se14-3-3ζ was decreased starting 24 h post-SeNPV infection as SeNPV titers seemed to increase as evidenced by intense bands of SeNPV IAP3. Interestingly, confocal microscopic analysis revealed that Se14-3-3ζ is expressed at the apical side of the NPV-uninfected gut cells, whereas it was detected mainly in the nucleus of the NPV-infected cells. Thus, despite the biological significance of Se14-3-3ζ in S. exigua in conjunction with molecular interactions between SeNPV and S. exigua is unclear now, our data suggest that Se14-3-3 ζ protein plays a role to protect S. exigua from the infection or inhibit replication of SeNPV.

Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway is known to play a pivotal role in various cellular events and antiviral responses in both vertebrates and insects. In an attempt to elucidate the potential involvement of STAT on S. exigua-SeNPV interactions, the full length cDNA of SeSTAT was cloned from S. exigua. Analysis of temporal expression patterns shows that SeSTAT is expressed in all stages of life cycle such as larvae, pupae, and adult. Spatial expression analysis shows that it is highly expressed in fat body and Malpighian tubule. Interestingly, SeSTAT is induced at 24 h in response to either laminarin or LTA injection in larvae. Electrophoretic mobility shift assay (EMSA) shows that the binding of nuclear extracts from fat body cells immune-challenged with LTA to STAT5 probe was observed. In addition, SeSTAT was nuclear-translocalized in both fat body and gut cells that were challenged with LTA and laminarin, respectively. Finally, gene silencing of SeSTAT shows that SeNPV number appears to be increased. It suggests that SeSTAT may act as a negative regulator against SeNPV in midgut.

The experimental work was conducted to investigate the effects of the application level of composted cattle manure (CCM) on forage productivity using rye-whole crop rice (WCR) double cropping system for 3 years. Main plot was consisted of application level of CCM such as 150%, 200% and control plot. The total DMY of of rye-WCR in treatment of CCM 150% significantly decreased as compared with that of treatment of CF and CCM 200%. However, NDF, ADF and CP content of rye and WCR were hardly influenced by CCM application.

Periodontal ligament (PDL) tissue is a connective tissue that is interposed between the roots of the teeth and the inner wall of the alveolar bone socket. PDL is always exposed to physiologic mechanical force such as masticatory force and PDL cells play important roles during orthodontic tooth movement by synthesizing and secreting different mediators involved in bone remodeling. The Wnt/β-catenin signaling pathway was recently shown to play a significant role in the control of bone formation. In the present study, we applied cyclic tensile stress of 20% elongation to cultured human PDL cells and assessed its impact after six days upon components of the Wnt/β-catenin signaling pathway. RTPCR analysis showed that Wnt1a, Wnt3a, Wnt10b and the Wnt receptor LRP5 were down-regulated, whereas the Wnt inhibitor DKK1 was up-regulated in response to these stress conditions. In contrast, little change was detected in the mRNA expression of Wnt5a, Wnt7b, Fz1, and LRP6. By western blotting we found decreased expression of the β-catenin and p-GSK-3β proteins. Our results thus show that mechanical stress suppresses the canonical Wnt/β-catenin signaling pathway in PDL cells.

The objective of this study was to develop a reliable and reproducible descriptive analysis procedure for Korean style sweet pumpkin gruel (Hobakjuk). The sensory attributes of the sweet pumpkin gruel were developed and defined, the sample preparation method was standardized， and the sensory evaluation procedure for a sample was established. Seven types of sweet pumpkin gruel (five ready-to-eat type vs. two ready-to-heat type) were selected to be analyzed. Panel training and descriptive analysis were carried out with these 7 samples. A total of 12 sensory attributes (2 aroma/odor, 5 taste/flavor, 4 texture/mouthfeel, and 1 aftertaste attributes) were developed to describe the sensory characteristics of the sweet pumpkin gruel. The definition and reference standards for each sensory attribute were determined to clearly understand each attribute. In the main experiment, trained panelists evaluated the sensory characteristics of the 7 gruel samples based on a fifteen-point intensity scale using the developed attributes. The results were statistically analyzed by analysis of variance, principal component analysis, and cluster analysis. The results showed that the 7 sweet pumpkin gruel samples significantly diffieredin their intensities of all attributes except for sweet pumpkin aroma and viscosity. The ready-to-eat style samples were distinctly characterized by their sweet pumpkin aroma and flavor, whereas the ready-to-heat style samples were markedly characterized by their low intensity of gelatinized starch and pumpkin flavor retention.