The purpose of this study was to characterize equine heat-shock protein (Hsp) genes and analyze their expression pattern in various horse tissues and blood leukocytes after exercise. In a previous study, RNA sequencing of blood and skeletal muscles of thoroughbreds before and after exercise was performed using differently expressed gene (DEG) analysis. Three Hsp genes (HspH1, Hsp90 and Hsp70) were selected by DEG analysis and were found to be differentially expressed in either blood or muscle. To validate and extend previous observations on these genes, we performed RT-PCR analyses of horse tissue as well as real-time qPCR analyses of blood leukocytes after exercise. mRNA expression of these Hsp genes was found to be ubiquitous in the analyzed tissues (including thyroid, colon, skeletal muscle, cecum, kidney, spinal cord, heart, and lung). In addition, Hsp mRNA expression of these genes in extracted whole blood increased after 120 minutes of exercise compared to the baseline condition. These results are in agreement with the results of human and other experimental animals, suggesting that regulatory mechanisms that are responsible for upregulation of Hsp gene transcription may be conserved among species. Further investigations to correlate Hsp gene expression patterns with athletic performance or recovery processes after exercise are warranted.

We investigated the effects of extracts from Rubus coreanus (RC) and Artemisia princeps var. orientalis (AP) on DNA damage response in ultraviolet B (UVB)-exposed HaCaT cells. Cell activity upon treatment for 24 h with RC or AP alone was similar to or greater than that of the nontreated control. When UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, cell activity increased in a concentration-dependent manner. Nuclear fragmentation analysis showed that postincubation with RC or AP decreased UVB-induced apoptosis by about 20% and 15%, respectively, of that in cells postincubated with growth medium. When UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, the level of cyclobutane pyrimidine dimer decreased in a concentration- dependent manner. Western blot analysis showed that treatment of cells not exposed to UVB with RC or AP alone did not significantly change the levels of phospho-p53 and GADD45 protein. Interestingly, when UVB-exposed cells were postincubated for 24 h in medium containing RC or AP, phospho-p53 and GADD45 levels decreased in a concentration dependent manner. Our results suggest that RC and AP extract assist the survival of UVB-exposed cells in parallel with a decrease in levels of UVB-induced DNA damage and damage-response proteins, such as p53 and GADD45.

The kinetic properties and isozyme expression of lactate dehydrogenase (EC 1.1.1.27; LDH) in tissues from Rana catesbeiana I and II collected from February (I) and August (II) were studied. LDH activities, A4 isozyme, and LDH/citrate synthase (EC 4.1.3.7; CS) were high in skeletal muscle from R. catesbeiana I, and LDH isozyme increased in several tissues of R. catesbeiana II. In particular, LDH activities were high in heart and brain tissues from R. catesbeiana II. LDH eye-specific C isozyme, detected by native polyacrylamide gel electrophoresis after immunoprecipitation, was expressed in eye tissue and was more similar to the than isozyme. LDH isozyme was purified by oxamate-linked affinity chromatography, and the molecular weight of subunit A was 32.0 kDa. In R. catesbeiana II, levels of , , and tolerance to lactate of LDH were high in all tissues, and of LDH in heart and brain tissue was highly detected. Purified isozyme and LDH in eye tissue were highly tolerate compared to others. The value was highly measured compared to . The degree of inhibition by 10 mM of pyruvate on LDH activities in tissues from R. catesbeiana I and II was more pronounced as the ratio of subunit B increased. As a result, characteristic expression of LDH eye-specific C was found in R. catesbeiana. Anaerobic metabolism seemed to predominate as the LDH of skeletal muscle from I showed higher activity. It also appeared that R. catesbeiana II adapted well to incremental increases in LDH B, becoming tolerant to the lactate of LDH in tissues.

Alginates are linear acidic polysaccharides composed with (1-4)-linked -L-guluronic acid and -Dmannuronic acid. Alginate can be degraded by diverse alginate lyases, which cleave the alginate using a -elimination reaction and produce unsaturated uronate oligomers. A gene for a polyMG-specific alginate lyase possessing a novel structure was previously identified and cloned from Stenotrophomonas maltophilia KJ-2. Homology modeling of KJ-2 polyMG-specific alginate lyase showed it belongs to the PL6 family, whereas three Azotobacter vinelandii polyMG lyases belong to the PL7 family of polysaccharide lyases. From -NMR spectra data, KJ-2 polyMG lyase preferably degraded the M-(1-4)-G glycosidic bond than the G-(1-4)-M glycosidic bond. Seventeen mutants were made by site-directed mutagenesis, and alginate lyase activity was analyzed. Lys220Ala, Arg241Ala, Arg241Lys, and Arg265Ala lost alginate lyase activity completely. Arg155Ala, Gly303Glu, and Tyr304Phe also lost the activity by 60.7-80.1%. These results show that Arg155, Lys220, Arg241, Arg265, Gly303, and Tyr304 are important residues for catalytic activity and substrate binding.

Cortex ulmi pumilae, the cortex of Ulmus davidiana var. japonica, has been used in traditional folk medicine for its anti-inflammatory effect. Although its various bioactivities such as anti-inflammatory, anti-microbial, and anti-cancer, have been reported, the anti-adipogenic activity of cortex ulmi pumilae remains unclarified. In the present study, we investigated the effect of cortex ulmi pumilae extract on adipocyte differentiation in 3T3-L1 preadipocytes. Treatment with cortex ulmi pumilae extract significantly reduced the formation of lipid droplets and triglyceride content in a dose-dependent manner; this is associated with an inhibition of the adipogenic transcription factors, CCAAT/enhancer binding protein (), , and peroxisome proliferator-activated receptor (). In addition, cortex ulmi pumilae extract treatment during the early stage of adipogenesis showed more efficient anti-adipogenic activity than treatment during other stages of adipogenesis. Cortex ulmi pumilae extract also inhibited cell proliferation and induced G1 arrest of 3T3-L1 cells in the early stage of adipogenesis. This was associated with upregulated expression of Cdk inhibitor p21 and downregulated expression of cyclin E and phospho-Rb, indicating that cortex ulmi pumilae extract blocks mitotic clonal expansion by cell cycle regulation. Taken together, these results suggest that cortex ulmi pumilae extract possesses anti-adipogenic activity through the inhibition of adipocyte differentiation by blocking mitotic clonal expansion.

The objective of this study was to elucidate the effect of light-emitting diode treatment on early growth and inorganic elements in leaf lettuce (Lactuca sativa L. `Oak Leaf`). In changes to leaf morphology, shoot elongation and hypocotyl length showed poor growth under red light irradiation, while red+blue light irradiation induced shorter plant height and more leaves, resulting in increased fresh weight. With respect to Hunter`s color and SPAD values, lettuce seedlings grown under red+ blue and fluorescent light irradiation had a higher value but showed no other changes to SPAD values. Interestingly, redness in relative chlorophyll content was 1.4 times higher under red+blue light irradiation. Inorganic element (N, Ca, Mg, and Fe) and ascorbic acid concentrations increased in lettuce plants grown under LED light irradiation compared to those of lettuce grown under fluorescent light, which showed a higher P content. In conclusion, red+blue light irradiation, which stimulates growth and higher nutrient uptake in leaf lettuce, could be employed in containers equipped with LEDs.

Ghost cells have been recognized as eliciting humoral and cell-mediated immune responses and have also been predicted to play a role as an immune adjuvant. In this study, we used the intramuscular (IM) route to inject BALB/c mice with four vaccine groups constructed from Salmonella typhimurium ghost (STG) cells originating from different virulent strains and complex STG groups instead of heat-labile toxin (LT)-B, a type of adjuvant. Although the complex STG groups exhibited a response after a short delay, the groups showed final total IgG levels similar to those of the LT-B group, which encodes LT-B from pMMP300. The IgG1 response to the 3339 group was the highest response at 6 weeks, whereas IgG2a responses to the 3339 and JOL389 groups were higher at 6 and 8 weeks compared to those of the LT-B group. The response of vaginal sIgA to the LT-B group was generally higher than that of the other groups, whereas fecal sIgA to the LT-B group exhibited lower responses. Protection to virulent S. typhimurium in all groups was above 80%, which was similar to the LT-B group. Taken together, we suggest that STG complex groups can be used as an immune adjuvant instead of LT-B.

We screened Paenibacillus sp. strains from Jeotgal, a Korean salted and fermented fish product, for use as a novel probiotic. Among these Paenibacillus sp. isolates, BCNU 5016 was a typical Paenibacillus sp. strain that showed gram-positive, gelatinase-negative, and urease negative activity. On the basis of 16S rDNA sequence comparisons, BCNU 5016 was most closely related phylogenetically to P. polymyxa. When Paenibacillus sp. BCNU 5016 was subjected to the acid tolerance test, this strain showed 91.89% survival after 3 h culture at pH 2.5. Paenibacillus sp. BCNU 5016 also showed excellent bile acid tolerance. Furthermore, its auto-aggregation, coaggregation, and hydrophobic capacities suggest that BCNU 5016 had the capacity to adhere well to the intestinal tract. We conclude that Paenibacillus sp. BCNU 5016 has excellent potential as a probiotic.

Optimal conditions for extraction of bakkenolide B from Petasites japonicus leaves were determined by using response surface methodology. A second-order Box-Behnken design representing three extraction temperatures (80, 100, ), three extraction times (30, 45, 60 min), and three solvent pH`s (5, 7, 9) was executed. The efficiency of the extraction conditions was defined using the -hexosamidase assay by comparing both the bakkenolide B content and its anti-allergic activity expressed as extract inhibition on degranulation. The response surface plot described for the bakkenolide B content showed that the maximum content was predicted as 121.6 with extraction conditions of , 46.6 min, and pH 7.7. Extraction temperature and time were important factors in determining bakkenolide B content. Using regression analysis, correlation between the inhibition effect of mast cell degranulation and bakkenolide B content was found to be low.

Estrogen can promote or inhibit cellular proliferation depending on tissue cell types and physiological condition and acts through the signal transduction pathways mediated primarily by estrogen receptors. This study examined the effects of fulvestrant (Ful), a well-known antagonist for the estrogen receptor, on the action of -estradiol (E2) with respect to the proliferation and apoptosis of Chinese hamster ovarian (CHO) cells. We used different concentrations of E2, Ful, and E2 plus Ful during different treatment durations. Treatment with 15-40 E2 significantly inhibited proliferation in a time-dependent manner, although it had no influence in concentrations up to 1 . Interestingly, Ful at 10-40 also inhibited cellular proliferation in both a concentration- and time-dependent manner. In addition, Ful enhanced rather than decreased the inhibitory effect on cellular proliferation by E2 in combined treatment for 10 days. Thus, Ful does not appear to have an antagonistic effect on estrogen`s anti-proliferative action in CHO cells. In TUNEL assays to confirm DNA fragmentation by E2 and/or Ful, CHO cells treated with 20 E2 showed a TUNEL-positive reaction in most DAPI-stained nuclei, and cells treated with either 40 Ful or 40 Ful plus 20 E2 also exhibited a TUNEL-positive reaction but at a lower rate compared to the E2-treated cells. These results indicate that Ful does not have an antagonistic effect on estrogen`s anti-proliferative action in CHO cells, suggesting that the anti-proliferative and apoptosis-related mechanism(s) through DNA fragmentation by E2 and Ful may be mediated by different signal transduction pathways.

We identified SNPs (single nucleotide polymorphisms) for endothelial nitric oxide synthase (eNOS) genes in the Korean genome. eNOS is present in the vascular endothelium, platelets, and several other cell types that continuously produce modest amounts of NO. Endothelium-derived NO plays a key role in the regulation of vascular tone, and the impaired effects of NO on the cardiovascular system appear to be responsible for coronary atherosclerosis and thrombosis. In recent studies, a missense variant within exon 7 of the eNOS gene in patients with coronary spastic angina-GAG to GAT substitution, which results in the replacement of glutamic acid by aspartic acid (Glu298Asp [G894T])-has been identified and is known to be significantly associated with coronary spasm. We prepared PCR primers based on sequences in Genbank. Primers were prepared for normal and SNPs separately, as reported for other Asian countries, such as G894T. Their sequences were different only at the 3` ends so that primer extension could only by possible when base pairs between templates and primers matched. We also employed ARMS (Amplification Refractory Mutation System) technology to improve the specificity of the PCR reaction. In conclusion, we were able to demonstrate the eNOS G894A polymorphism in Korean gemone. This study should facilitate research on the cause of myocardial infarction and development on further therapy at the genetic level.

Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of and . This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of and/or , for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

Essential oils extracted or purified from various plants have shown various beneficial effects. Seed parts of Schizandra chinensis Baillon (Schisandrae Semen) have been used as a traditional medicine for thousands of years in parts of Asia, including Korea, China, and Japan. However, the pharmacological mechanisms of essential oils purified from S. fructus (S. chinensis Baillon) remain largely unresolved. The aim of this study was to investigate the safety of Schisandrae Semen essential oil (SSeo) by a single- dose toxicity study in mice. SSeo was orally administered at a dose of 5,000 mg/kg in ICR mice. All animals were sacrificed after 14 days of treatment. After a single administration, mortality, clinical signs, body weight changes, and gross pathological findings were observed for 14 days. We also measured parameters of organ weight, clinical chemistry, and hematology. No toxicological change related to the test substance or mortality was observed after administration of a single oral dose of SSeo. There were no adverse effects on clinical signs, body weight, or organ weight and no gross pathological findings in any treatment group. The clinical chemistry and hematological parameters were within the normal ranges except total bilirubin. Therefore, the approximate lethal dose for oral administration of SSeo in mice was considered to be over 5,000 mg/kg. The results on the single-dose toxicity of SSeo indicate that it is not possible to reach oral dose levels related to death or dose levels with any harmful side effects.

During the life cycle of plants, water deficit leads to an adverse effect on its growth and development. To increase the productivity of crops, overcoming such drought stress is one of the most important issues in the field of plant study. Among plant hormones, the phytohormone, abscisic acid (ABA) plays a crucial role in eliciting resistance to drought stress as well as in multiple developmental processes, such as seed germination, stomatal closure, and seedling growth. Therefore, further understanding of the ABA-mediated signal transduction pathway in plants is an effective strategy to generate drought-tolerant plants. Posttranslational modification, such as phosphorylation and ubiquitination, is an efficient mechanism for plants to acquire quick adaptation against environmental stress conditions since this process directly affects pre-existing signaling components by modulating protein activity and stability. Here, recent reports on ABA signaling are reviewed, especially focusing on ABA transport, perception, signaling, and posttranslational modification in ABA-mediated cellular responses. Also, we present future prospects on how the control of such a mechanism can be applied to generate useful agricultural crops.

In a variety of eukaryotic cells, autophagy sequesters a portion of the cytoplasm and targets it to a lytic compartment for degradation in bulk. Autophagy is a dynamic process for degrading cytoplasmic cargoes with various degrees of selectivity, and its activity is tightly regulated in a nutrient- and development-dependent manner. Autophagy research has drawn much attention since autophagy not only is an interesting cell biological phenomenon but also has great potential for medical and agricultural applications. For example, autophagy is associated with cancers and neurodegenerative diseases in human and mammalian cells and is also suggested in remobilization of nutrients during the senescence of plant leaves. In this general review, we describe genetic components of the core autophagic machinery conserved among yeast, animals, and plants and briefly explain how these components are responsible for major steps in plant autophagy. We discuss four common features of autophagic processes: (i) autophagy as a degradation pathway, (ii) the concept of flux in autophagy research, (iii) dependency on developmental and nutritional cues, and (iv) diversity of autophagy, focusing on selective types of autophagy. We also summarize cell biological and physiological functions of plant autophagy. Our intention is to provide a quick guide to autophagy for those who are new to autophagy research.