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This study investigated the effect of electron beam (EB) treatment on the microbial reduction of dried laver (Porphyra tenera) and identified EB-resistant bacteria from the treated dried laver. After EB treatments of 4 kGy and 7 kGy, the numbers of total bacteria and EB-resistant bacteria were measured using tryptic soy agar and mannitol salt agar, respectively. The morphological and biochemical characteristics of each isolated EB-resistant bacteria were investigated and these bacteria were identified. Compared to the control (1.5±0.2)×106 CFU/g, the total bacterial number was significantly decreased to (5.4±0.5)×104 CFU/g and (1.1±0.6)×104 CFU/g after EB treatments of 4 kGy and 7 kGy, respectively. With a higher EB dosage, the number of red colonies was almost same, whereas the number of yellow colonies was significantly decreased to (3.3±1.2)×103 CFU/g and 0 CFU/g for 4 kGy and 7 kGy, respectively. All red and yellow colonies were gram-positive cocci, catalase-positive, and resistant to 3% and 5% NaCl media. From the 16S rDNA sequence analysis, yellow and red colonies were identified as either Micrococcus flavus or M. luteus, with 99% similarity for the yellow colonies, and Deinococcus proteolyticus and D. piscis, with 99% and 97% similarity for the red colonies, respectively.

The objective of this study was to demonstrate the applicability of immunosensors combined with a light microscopic imaging system (LMIS) for the rapid detection of Salmonella in chicken. The reactivity and specificity of purified anti-Salmonella polyclonal antibodies (pAbs) were compared with commercial pAbs using indirect ELISA. After the optimum antibody concentration was determined, the immobilization of antibody was confirmed using an atomic force microscopy (AFM). The immunosensor combined with LMIS method was applied to chicken for Salmonella detection. No significant difference in reactivity was found between the purified and commercial anti-Salmonella pAbs over the entire concentration range (p > 0.05). In addition, the purified anti-Salmonella pAbs showed sufficient specificity for 11 non-Salmonella bacterial strains and reactivity for 10 chicken-associated Salmonella serovars. The optimal antibody concentration for immobilization on the sensor was determined to be 100 μg/mL. The AFM images confirmed the binding of antibody on a sensor by providing irregular and globular surface features. When applied to chicken, the number of detected Salmonella increased quantitatively as the concentration of inoculated Salmonella increased. The limit of detection was 103 cfu/chicken, which was considerably lower than those of other biosensor methods. With the advantage of direct observation and enumeration of Salmonella on the sensors, the immunosensor combined with LMIS method detected Salmonella in chicken in a specific, sensitive, simple, and practical manner.

In this study, the probiotic properties of nine lactic acid bacteria (LAB) were evaluated by using an in silico analysis comprising principal component, heat map, and network analyses. Acid and bile resistances, bile salt hydrolase (BSH) activity, antimicrobial activity, auto- and co-aggregation, and hydrophobicity were analyzed in the strains. All tested LAB exhibited >60% of survival percentage in acid and bile resistances. Five strains of LAB exhibited positive BSH activities and all LAB demonstrated antimicrobial activities against at least one pathogen except L. acidophilus IDCC3302. Following a time-dependent manner, L. acidophilus IDCC3302 and L. plantarum LP-K1791 exhibited the greatest auto- and co-aggregation percentages, respectively. The hydrophobicity of all LAB was satisfied with the minimum requirement of hydrophobicity (40%) as potential probiotics. Based on the in silico analysis, L. casei IDCC3451, L. plantarum LP-K1791, and L. rhamnosus IDCC3201 were selected as the most promising probiotics. This in silico analysis will be useful for the precise selection of probiotics for the development of functional foods and healthy dietary supplements in the food and pharmaceutical industries.

Biosensing systems based on electrochemical detection is popular to the industrial community, because the electronic readout from biosensor assays could replace conventional microarray methods. However, the greatest practical barrier to a wider acceptance of biosensors within the industrial field is due to their associated manufacturing and operating costs. In this study, a single-walled carbon nanotube (SWCNT)-based biosensor was tested for reusability after cleaning. The reusability of the existing biosensing system was examined using diverse cleaning reagents. Successful sensor cleaning without affecting their reusable properties was confirmed using sodium hydroxide (NaOH) solution. A range of NaOH concentrations between 70 and 160 mM was tested and the optimum reusable concentration for the biosensor was determined as 100 mM. A 5-time-reuse of the biosensor was achieved using 100 mM NaOH solution for peanut allergen control. Furthermore, re-adding anti-Yersinia antibodies to the biosensor led to the reaction of bacterial cells of Yersinia enterocolitica with the potential biosensor response. Scanning electron microscopy (SEM) results showed that microorganisms (Y. enterocolitica) and peanut allergen (Ara h1) were washed away from the biosensor chip after washing using NaOH (100 mM) for 2 h.

Malarial infection induces tissue hypoxia in the host through destruction of red blood cells. Tissue hypoxia in malarial infection may increase the activity of HIF1α through an intracellular oxygen-sensing pathway. Activation of HIF1α may also induce vascular endothelial growth factor (VEGF) to trigger angiogenesis. To investigate whether malarial infection actually generates hypoxia-induced angiogenesis, we analyzed severity of hypoxia, the expression of hypoxia-related angiogenic factors, and numbers of blood vessels in various tissues infected with Plasmodium berghei. Infection in mice was performed by intraperitoneal injection of 2×106 parasitized red blood cells. After infection, we studied parasitemia and survival. We analyzed hypoxia, numbers of blood vessels, and expression of hypoxia-related angiogenic factors including VEGF and HIF1α. We used Western blot, immunofluorescence, and immunohistochemistry to analyze various tissues from Plasmodium berghei-infected mice. In malaria-infected mice, parasitemia was increased over the duration of infection and directly associated with mortality rate. Expression of VEGF and HIF1α increased with the parasitemia in various tissues. Additionally, numbers of blood vessels significantly increased in each tissue type of the malaria-infected group compared to the uninfected control group. These results suggest that malarial infection in mice activates hypoxia-induced angiogenesis by stimulation of HIF1α and VEGF in various tissues.

The aim of this study was to isolate and characterize a lytic

The Hippo pathway is involved in intestinal epithelial homeostasis with Wnt, BMP, Notch, and EGF signaling. We investigated the relationship between Hippo and other signaling pathways and the role of MOB kinase activator 1A/1B (MOB1A/B) in intestinal homeostasis. Mice with intestinal epithelial cell (IEC)-specific depletion of MOB1A/B showed hyperproliferation in IECs, defects in secretory lineage differentiation and loss of intestinal stem cells and eventually died at 10–12 days after tamoxifen treatment. In MOB1A/B-depleted IECs, expression of Wnt target genes were downregulated but Bmp2 and Tgfbr2 were transcriptionally activated with enhanced YAP activity. In in vivo and in vitro experiments with several signaling inhibitors, it has been shown that the BMP inhibitor LDN193189 or TGF-β inhibitor SB431542 had effects on partial restoration of the intestinal degenerative phenotype. Treatment with these inhibitors restored differentiation of secretory lineage cells in MOB1A/B-deficient mice, but not ISC pools in the crypt region. These studies reveal that IEC-specific depletion of MOB1A/B induced overexpression of Bmp2 and Tgfbr2 and inhibited Wnt activity, finally leading to loss of ISCs and functional epithelia in the mouse intestine. These results suggest that MOB1A/B has an essential function for intestinal epithelial homeostasis by regulating YAP, Wnt activity, and BMP/TGF-β signaling.

Background: The peanut protein Arachis hypogaea (Ara h) 6 is one of the most serious food allergens that contributes to food-related, life-threatening problems worldwide. The extremely low allergic dose demands for more selective and rapid methods for detecting Ara h 6. Objective: The goal of this study was to develop a single-walled carbon nanotube (SWCNT)-based biosensor for the rapid detection of Ara h 6 in commercial food products. Methods: The detection principle of this biosensor was based on the binding of Ara h 6 to the anti-Ara h 6 antibody (pAb) through 1-pyrenibutanoic acid succinimidyl ester. The resistance difference (ΔR) was calculated via linear sweep voltammetry using a potentiostat. Results: The ΔR increased as the Ara h 6 concentrations increased above the range of 100–107 pg/L. A specificity analysis showed that the anti-Ara h 6 pAb selectively interacted with Ara h 6 molecules in the buffer solution (pH 7.4). Conclusions: This research proposes that an SWCNT-based biosensor in self-assembly with antibodies could be an effective tool for the rapid detection of allergen proteins in food. Highlights: The developed biosensor exhibited higher sensitivity and selectivity. Application studies resulted in precise Ara h 6 detection in peanut-containing processed food.

BackgroundIn a previous study, we found thatTrichinella spiralisexcretory and secretory proteins (ES-P) most likely activate collagen synthesis via TGF-β/Smad signaling, and this event could influence collagen capsule formation.Methodology/Principal FindingsIn order to identify the specific collagen inducing factor, ES-P was fractionated by a Superdex 200 10/300 GL column. We obtained three large fractions, F1, F2, and F3, but only F3 had collagen gene inducing ability. After immunoscreening, 10 collagen inducing factor candidates were identified. Among them, TS 15-1 and TS 15-2 were identical to the putative trypsin ofT. spiralis. The deduced TS 15-1 (M.W. = 72 kDa) had two conserved catalytic motifs, an N-terminal Tryp_SPc domain (TS 15- 1n) and a C-terminal Tryp_SPc domain (TS 15-1c). To determine their collagen inducing ability, recombinant proteins (rTS 15-1n and rTS 15-1c) were produced using the pET-28a expression system. TS 15-1 is highly expressed during the muscle larval stage and has strong antigenicity. We determined that rTS 15-1c could elevate collagen I via activation of the TGF-β1 signaling pathwayin vitroandin vivo.Conclusion/SignificanceIn conclusion, we identified a host collagen inducing factor fromT. spiralisES-P using immunoscreening and demonstrated its molecular characteristics and functions.Author SummaryTrichinella spiraliscan make collagen capsules in host muscle cells during its life cycle, which encapsulates muscle stage larvae. Many investigators have tried to reveal the complex mechanism behind this collagen capsule architecture, and it has been suggested that several serine proteases in excretory-secretory proteins of the parasite are potential collagen capsule inducing factors. In addition, collagen synthesis is activated through the TGF-β/Smad signaling pathway and these events are closely related with protease activated receptor 2 which was activated by various serine proteases. In this study, we isolated and characterized a collagen gene expression inducer fromT. spiralisES-P using immunoscreening and investigated the candidate protein for its usefulness as a wound healing therapeutic agent.

Aminoacyl-tRNA synthetase-interacting multifunctional protein-3 (AIMP3) is a component of the multi-aminoacyl-tRNA synthetase complex and is involved in diverse cellular processes. Given that AIMP3 deficiency causes early embryonic lethality in mice, AIMP3 is expected to play a critical role in early mouse development. To elucidate a functional role of AIMP3 in early mouse development, we induced AIMP3 depletion in mouse embryonic stem cells (mESCs) derived from blastocysts of AIMP3f/f; CreERT2 mice. In the present study, AIMP3 depletion resulted in loss of self-renewal and ability to differentiate to three germ layers in mESCs. AIMP3 depletion led to accumulation of DNA damage by blocking double-strand break repair, in particular homologous recombination. Through microarray analysis, the p53 signaling pathway was identified as being activated in AIMP3-depleted mESCs. Knockdown of p53 rescued loss of stem cell characteristics by AIMP3 depletion in mESCs. These results imply that AIMP3 depletion in mESCs leads to accumulation of DNA damage and p53 transactivation, resulting in loss of stemness. We propose that AIMP3 is involved in maintenance of genome stability and stemness in mESCs.

This study investigated the feasibility of the lytic, tailed

Lung cancer is a fatal disease with a high mortality rate. The perinuclear reorganization of keratin 8 (K8) is an important biochemical phenomenon reflecting changes in the physical properties of metastatic cancer. However, there is not much of information about the regulatory molecules involved in phosphorylation and perinuclear reorganization of K8. In this study, we investigated the role and molecular mechanisms of YdjC chitooligosaccha- ride deacetylase homolog (YDJC) in sphingosylphosphorylcholine (SPC)-induced phosphorylation and reorganization of K8, and migration and invasion (SPC-induced events). SPC induced expression of YDJC in a dose- and time-dependent manner. Gene silencing of YDJC suppressed SPC-induced events. YDJC overexpression induced the SPC-induced events. YDJC deacetylase dominant negative mutant (YDJCD13A) did not induce SPC-induced events. YDJC siRNA reduced ERK activation and overexpression of YDJC induced ERK activation. The siRNA of ERK1 or ERK2 suppressed YDJC-induced phosphorylation and reorganization of K8, and migration and invasion. Co-immunoprecipitation revealed that YDJC binds to CDC16. Interestingly, CDC16 siRNA induced SPC-induced events. Overexpression of CDC16 blocked SPC-induced events. KMPLOT analysis based on public microarray data revealed the poor prognosis of lung cancer patients with high expression of YDJC compared with patients with low expression of YDJC. The collective results indicate that YDJC is involved in SPC-induced events in A549 lung cancer cells by interacting with CDC16. YDJC overexpression might be involved in the progression of lung cancer. These results also suggest that suppression of YDJC or boosting of CDC16 interaction with YDJC might be a novel way to prevent progression of lung cancer.

High-mobility group box 1 (HMGB1) enhances inflammatory reactions by potentiating the activity of pro-inflammatory mediators and suppressing the phagocytosis of apoptotic neutrophils. However, the effects of HMGB1 on phagocytosis induced by pro-resolving mediators, such as resolvins, have not been studied up until this point. In this study, we investigated the effects and underlying mechanism of HMGB1 on resolvin D1-induced phagocytosis of MDA-MB-231 cells, which were selected as a model system based on their phagocytic capability and ease of transfecting them with a plasmid or siRNA in several cancer cell lines. Then we confirmed effects of HMGB1 in THP-1 cells.Resolvin D1 (RvD1) enhanced phagocytosis in MDA-MB-231 and THP-1 cells. HMGB1 suppressed RvD1-induced phagocytosis in MDA-MB.231 and THP-1 cells. HMGB1 dose-dependently induced the expression of 15-hydroxyprostaglandin dehydrogenase (15-PGDH), the inactivating enzyme in pro-resolving lipid mediators such as RvE1 and RvD1. Involvement of 15-PGDH in-HMGB-1-induced suppression of phagocytosis was examined using siRNA of 15-PGDH or 15-PGDH inhibitor, TD23. Surprisingly, the silencing of 15-PGDH increased phagocytotic activity of MDA-MB-231 cells. TD23 also enhanced phagocytosis of MDA-MB-231 and THP-1 cells.In conclusion, the release of HMGB1 during the inflammatory phase induces 15-PGDH expression, which suppresses the phagocytotic activity of macrophages. These processes might be involved in the mechanism that blocks the resolution of inflammation, thereby allowing acute inflammation to progress to chronic inflammation.

In Trichinella spiralis infection, type 2 helper T (Th2) cell-related and regulatory T (Treg) cell-related immune responses are the most important immune events. In order to clarify which Toll-like receptors (TLRs) are closely associated with these responses, we analyzed the expression of mouse TLR genes in the small intestine and muscle tissue during T. spiralis infection. In addition, the expression of several chemokine- and cytokine-encoding genes, which are related to Th2 and Treg cell mediated immune responses, were analyzed in mouse embryonic fibroblasts (MEFs) isolated from myeloid differentiation factor 88 (MyD88)/TIR-associated proteins (TIRAP) and Toll receptor-associated activator of interferons (TRIF) adapter protein deficient and wild type (WT) mice. The results showed significantly increased TLR4 and TLR9 gene expression in the small intestine after 2 weeks of T. spiralis infection. In the muscle, TLR1, TLR2, TLR5, and TLR9 gene expression significantly increased after 4 weeks of infection. Only the expression of the TLR4 and TLR9 genes was significantly elevated in WT MEF cells after treatment with excretory-secretory (ES) proteins. Gene expression for Th2 chemokine genes were highly enhanced by ES proteins in WT MEF cells, while this elevation was slightly reduced in MyD88/TIRAP-/- MEF cells, and quite substantially decreased in TRIF-/- MEF cells. In contrast, IL-10 and TGF-β expression levels were not elevated in MyD88/TIRAP-/- MEF cells. In conclusion, we suggest that TLR4 and TLR9 might be closely linked to Th2 cell and Treg cell mediated immune responses, although additional data are needed to convincingly prove this observation.

Toxicants can perturb cellular homeostasis by modifying phosphorylation-based signaling. In the present study, we examined the effects of cerulein, an inducer of acute pancreatitis, on keratin 8 (K8) phosphorylation. We found that cerulein dose-dependently induced K8 phosphorylation and perinuclear reorganization in PANC-1 cells, thus leading to migration and invasion. The extracellular signal-regulated kinases (ERK) inhibitor U0126 suppressed cerulein-induced phosphorylation of serine 431 and reorganization of K8. Cerulein reduced the expressions of protein phosphatase 2A (PP2A) via ubiqutination and alpha4. PP2A's involvement in K8 phosphorylation of PANC-1 cells was also confirmed by the observation that PP2A gene silencing resulted in K8 phosphorylation and migration of PANC-1 cells. Overall, these results suggest that cerulein induced phosphorylation and reorganization through ERK activation by downregulating PP2A and alpha4, leading to increased migration and invasion of PANC-1 cells. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 2090-2098, 2016.

The histone H3K27 demethylase, UTX, is a known component of the H3K4 methyltransferase MLL complex, but its functional association with H3K4 methylation in human cancers remains largely unknown. Here we demonstrate that UTX loss induces epithelial-mesenchymal transition (EMT)-mediated breast cancer stem cell (CSC) properties by increasing the expression of the SNAIL, ZEB1 and ZEB2 EMT transcription factors (EMT-TFs) and of the transcriptional repressor CDH1. UTX facilitates the epigenetic silencing of EMT-TFs by inducing competition between MLL4 and the H3K4 demethylase LSD1. EMT-TF promoters are occupied by c-Myc and MLL4, and UTX recognizes these proteins, interrupting their transcriptional activation function. UTX decreases H3K4me2 and H3 acetylation at these promoters by forming a transcriptional repressive complex with LSD1, HDAC1 and DNMT1. Taken together, our findings indicate that UTX is a prominent tumour suppressor that functions as a negative regulator of EMT-induced CSC-like properties by epigenetically repressing EMT-TFs.

This study was conducted to elucidate comprehensive sensory profiles of potential natural alternative sweeteners, xylooligosaccharides (XOS), and luo han guo extract (LHGE). Relative sweetness to 5% sucrose was obtained using the two-alternative forced-choice method. Sensory and temporal profiles were identified using descriptive analysis and time-intensity analysis, respectively. XOS and LHGE were 0.25- and 75.76-fold sweeter than 5% sucrose solution, respectively. At equi-sweetness concentrations to 5% sucrose, XOS was characterized by sourish and corn silk flavor. LHGE was distinguished from other sweeteners by its yellowness, honey-like flavor, and licorice flavor. Bitterness and acridness of these sweeteners were not significantly different from those of sucrose. Both sweeteners exhibited a delayed time to peak intensity and persistent sweet aftertaste. Further studies are required to enhance the sweetness profile of XOS and LHGE toward achieving more sucrose-like sweetness impression.

To date, the magnetoelastic (ME) biosensor method has not been used for the detection of methicillin-resistant Staphylococcus aureus (MRSA). In this study, we introduced, for the first time, the use of a lytic phage as a novel bio-recognition element for ME biosensors for the detection of MRSA. Optimal conditions for the binding of lytic phage to the ME sensor platform were determined by evaluating different phage concentrations (108–1012 pfu/ml) and immobilization times (10, 30, 90, 270, 810, and 2430 min). Scanning electron microscopy (SEM) analysis was carried out to examine the phages immobilized on the sensor platforms. The mean free length (MFL) between successive phages was calculated to determine the optimum immobilization time and phage concentration. Different concentrations of bovine serum albumin (BSA) were used to block the spaces between the phages on the sensors. The lytic phage-based ME biosensor method was used to detect MRSA in solution and limit of detection was compared with other biosensor methods. SEM images revealed that the lytic phage immobilized on the sensor platforms had slightly oval heads and relatively long tails. The optimum concentration and immobilization time for efficient binding of the lytic phage to the sensor was determined to be 1011 pfu/ml and 30 min, respectively, with an MFL of (917 ± 0.080) nm. The optimum concentration of the blocking agent, BSA, was determined to be 1 mg/ml, at which the maximum number of MRSA bound on the measurement sensor. Finally, the ME biosensor method was successfully used to detect MRSA with a limit of detection of 3.0 log cfu/ml, which was approximately 2 log lower than that of the surface plasmon resonance method.

Epithelial mesenchymal transition (EMT) is the first step in metastasis and implicated in the phenotype of cancer stem cells. Therefore, understanding and controlling EMT, are essential to the prevention and cure of metastasis. In the present study, we examined, by Western blot, reverse transcription polymerase chain reaction (RT-PCR), and confocal microscopy, the effects of cardamonin (CDN) on transforming growth factor-β1 (TGF-β1)-induced EMT of A549 lung adenocarcinoma cell lines. TGF-β1 induced expression of N-cadherin and decreased expression of E-cadherin. CDN suppressed N-cadherin expression and restored E-cadherin expression. Further, TGF-β1 induced migration and invasion of A549 cancer cells, which was suppressed by CDN. TGF-β1 induced c-Jun N-terminal kinase (JNK) activation during EMT, but CDN blocked it. Protein serine/threonine phosphatase 2A (PP2A) expression in A549 cancer cells was reduced by TGF-β1 but CDN restored it. The overall data suggested that CDN suppresses TGF-β1-induced EMT via PP2A restoration, making it a potential new drug candidate that controls metastasis.

In our previous studies, the recombinant type II macrophage migration inhibitory factor homologue (rAs-MIF) secreted from Anisakis simplex suppressed experimental inflammation mouse model through IL-10 production and CD4(+)CD25(+)Foxp3(+) T-cell recruitment. Also, TLR2 gene expression was significantly increased following rAs-MIF treatment. To know the relation between TLR2 and amelioration mechanisms of rAs-MIF, we induced allergic airway inflammation by ovalbumin and alum with or without rAs-MIF under TLR2 blocking systems [anti-TLR2-specific antibody (α-mTLR2 Ab) treatment and using TLR2 knockout mice]. As a result, the amelioration effects of rAs-MIF in allergic airway inflammation model (diminished inflammation and Th2 response in the lung, increased IL-10 secretion, CD4(+)CD25(+)Foxp3(+) T-cell recruitment) were diminished under two of the TLR2 blocking model. The expression of TLR2 on the surface of lung epithelial cell was significantly elevated by rAs-MIF treatment or Pam3CSK (TLR2-specific agonist) treatment, but they might have some competition effect on the elevation of TLR2 expression. In addition, the elevation of IL-10 gene expression by rAs-MIF treatment was significantly inhibited by α-mTLR2 Ab or Pam3CSK pretreatment. In conclusion, anti-inflammatory effects of the rAs-MIF on OVA-induced allergic airway inflammation might be closely related to TLR2.

본 논문은 미국 남부지역 특화작물인 머스커다인 포도의 부분별 추출물의 항산화능 및 항균활성을 구명하기 위한 목적으로, Higgins, Jumbo, Noble 3종의 머스커다인 품종을 대상으로 하여 포도의 과피/과육부분과 종자부분을 추출하여 실험에 사용하였다. 각 추출물에 대하여, 항산화능은 총페놀함량 및 라디칼 소거능(Scavenging activity 및 $EC_{50}$ )을, 항균활성은 E.coli K12에 대한 최소생육저해농도(minimum inhibitory concentration, MIC)를 조사하였다. 종자 추출물의 총페놀함량은 231.24~294.81 mg/mL GAE로 과피/과육 추출물에 비하여 높은 함량을 보였다. 또한 Higgins품종의 종자 추출물이 가장 우수한 라디칼소거능( $EC_{50}=0.026mg/mL$ )을 나타내, 종자추출물이 과피/과육추출물보다 우수한 항산화능을 보유하였다. 반면, E.coli K12에 대한 항균활성은 Higgins 품종의 과피/과육추출물이 40 mg/mL의 MIC를 보임으로써 가장 우수하였다. 따라서, 본 연구는 머스커다인 포도 추출물이 천연 유래의 항산화 및 항균 기능성을 보유한 소재로 활용될 수 있는 잠재적 가치를 제시하였다. 【The objective of this research was to investigate the antioxidant and antimicrobial activities of muscadine grape extracts. Three different cultivars of muscadine grapes including Higgings, Jumbo, and Noble were selected. The skin/pulp and seed parts of three selected muscadine grape cultivars were used for extraction. The total phenolic contents of muscadine grape extracts were expressed as gallic acid equivalents (GAE). The antioxidant activity of muscadine grape extracts were determined by scavenging activity of diphenylpicrylhydrazyl (DPPH) radical and expressed as effective concentration ( $EC_{50}$ ), which represented the concentration of the extract exhibiting 50% DPPH radical scavenging. The antimicrobial activity against E. coli K12 was determined and expressed as the minimum inhibition concentration (MIC). The seed extracts exhibited greater total phenolic contents than the skin/pulp extracts, ranging from 231.24 to 294.81 mg/mL GAE. The seed extracts exhibited greater antioxidant activities than the skin/pulp extracts ( $EC_{50}$ of Higgins seed extract=0.026 mg/mL). However, the skin/pulp extracts exhibited greater antimicrobial activities than the seed extracts, exhibiting the minimum inhibition concentration (MIC) in Higgins skin/pulp extract (MIC=4.0 mg/mL). This research indicated that the seed part and skin/pulp parts of the muscadine grapes possessed antioxidant activity and antimicrobial activity, respectively. Therefore, it was concluded that muscadine grapes possess the potential to be utilized as functional foods or nutraceuticals.】

C).Afteroptimizationofincubationtimeandtemperature,spinachleaveswere spiked with serial concentration of MRSA and the attachment of MRSA was confirmed using SEM. The phage-immobilizedsensorandcontrol(devoidofthelyticphage)sensorwereplacedonthesurfaceoftheleavesandtheresonantfrequencyshiftsofbothsensors were compared. The optimal incubation time and temperature for contact of the phage-immobilized ME biosensor with theMRSA were determined to be ≤ 30 min and ≤ 22

Simulations of the dust cycle and its interactions with the changing Earth system are hindered by the empirical nature of dust emission parameterizations in weather and climate models. Here we take a step towards improving dust cycle simulations by using a combination of theory and numerical simulations to derive a physically based dust emission parameterization. Our parameterization is straightforward to implement into large-scale models, as it depends only on the wind friction velocity and the soil's threshold friction velocity. Moreover, it accounts for two processes missing from most existing parameterizations: a soil's increased ability to produce dust under saltation bombardment as it becomes more erodible, and the increased scaling of the dust flux with wind speed as a soil becomes less erodible. Our treatment of both these processes is supported by a compilation of quality-controlled vertical dust flux measurements. Furthermore, our scheme reproduces this measurement compilation with substantially less error than the existing dust flux parameterizations we were able to compare against. A critical insight from both our theory and the measurement compilation is that dust fluxes are substantially more sensitive to the soil's threshold friction velocity than most current schemes account for.

Alpinia katsumadai is known to suppress thromboxane A 2 (TXA 2 ) receptor agonist-induced scratching in mice. The specific components of A. katsumadai responsible for these biological effects, however, are not known. In the present study, we investigated whether cardamonin (CDN), one of major principles of A. katsumadai , has suppressive effects on TXA 2 -induced scratching in mice. Scratching induced by U46619 (the TXA 2 receptor agonist) at a dose of 10 nmol/site was shown to be suppressed by CDN (0.1 nmol–0.5 nmol/site). Suppression of the U46619-induced scratching response by CDN was found to be unrelated to competition with the ligand at the TXA 2 receptor, since CDN did not suppress [ 3 H] SQ29548 (the TXA 2 receptor antagonist) binding to TXA 2 receptor. TXA 2 receptor expression in A549, HaCaT, and SH-SY5Y cell lines was examined and determined to be significant in the A549 and SH-SY5Y cell lines. Further, binding of high molecular G protein G h /transglutaminase-2 (G h /Tgase-2) to TXA 2 receptor was confirmed in the A549 and SH-SY5Y cells by co-immunoprecipitation. CDN suppressed the binding of TXA 2 receptor with G h /Tgase-2, which also acts as a G protein involved in TXA 2 signaling. These results suggested that CDN suppresses TXA 2 receptor agonist-induced scratching by suppressing TXA 2 signaling, specifically via blocking of the binding of G h /Tgase-2 to TXA 2 receptor.

The high mortality rates associated with cancer reflect the metastatic spread of tumor cells from the site of their origin. Metastasis, in fact, is the cause of 90% of cancer deaths. Therefore, considerable effort is being made to inhibit metastasis. In the present study, we screened ketotifen for anti-migratory and anti-invasive activities against MDA-MB-231 breast cancer and HT-1080 fibrosarcoma cancer cells. Cancer cell migration and invasion were measured using multi-well chambers. Additionally, western blots were used to examine the effects of ketotifen on the expressions of CDC42, Rho, Rac, and matrix metalloproteinase 9 (MMP-9). The results showed that ketotifen dose-dependently suppressed the migration and invasion of MDA-MB-231 and HT-1080 cells. Ketotifen also suppressed the expressions of CDC42, Rac, and Rho, which, significantly, are involved in MDA-MB-231 and HT-1080 cancer cell migration. Moreover, ketotifen suppressed the expression and activity of MMP-9, which is involved in degradation of the extracellular matrix leading to invasion. The overall data suggested that ketotifen suppresses the migration and invasion of MDA-MB-231 and HT-1080 cancer cells via inhibition of CDC42, Rac, Rho, and MMP-9 expression.

To investigate the spatial characteristics of seasonality in phytoplankton productivity over the entire East/Japan Sea (EJS), a wavelet analysis was applied to SeaWiFS chlorophyll-a (chl-a) data over the period from 1998 to 2007. The wavelet analysis revealed that a 6-month period of change in chl-a concentration, explained by spring and fall blooms, was persistent throughout EJS during the ten years examined. The time series seasonality could be further classified into three patterns: (1) the highest variance peak at the 6-month period, (2) double variance peaks with similar values at the periods of 6- and 12-month, and (3) the highest variance at the 12-month period. A spatial map of the ratios between variances at the 6- and 12-month periods illustrated geographical distribution of the chl-a seasonality patterns. The Japan Basin and Ulleung Warm Eddy area were characterized by pattern 1 associated with clear spring and fall blooms. The region near the sub-polar front had seasonality pattern 2 characterized by two similar, weak blooms, and the area of East Koran Bay and the Yamato Basin are distinguished by pattern 3 with strong spring bloom, but irregular and unnoticeable fall bloom due to relatively high chl-a concentrations in wintertime. Distinct regional distribution of the seasonality patterns may suggest that phytoplankton productivity and its seasonal cycles respond not only to basin-scale or mesoscale climate forcing but also to changes in local physical properties characterizing each region.

Anisakis (Anisakidae) is one of the most important causes of helminth-induced allergic reactions and elicits clinical responses that include urticaria, rhinitis, bronco-constriction, cough, and/or gastrointestinal symptoms. More than 13 reactive allergens have been identified in the serum of Anisakis allergy patients, but the allergenicity of only a few of these have been evaluated in vivo using a mouse model. To evaluate the allergenicity of two important allergens, Ani s 1 and Ani s 9, we induced experimental allergic airway inflammation in a mouse model by repeated intranasal administration of the allergens. Both recombinant proteins (rAni s 1 and rAni s 9) elicited increased airway hyperresponsivity, airway infiltration by inflammatory cells (especially eosinophils), bronchial epithelial cell hyperplasia, all of which are characteristic of allergic airway inflammation. These allergens significantly increased the levels of Th2-related cytokines (IL-4, IL-5, IL-13, and IL-25) and Th17 related cytokines (IL-6 and IL-17) in both splenocytes and airway (except IL-17 in airway by rAni s 9). OVA-specific IgE and total IgE were increased in rAni s 1 and rAni s 9 treated mice as compared with controls treated with OVA alone. In addition, these two allergens induced gene expression of thymic stromal lymphopoietin (TSLP) and IL-25 (initiators of the Th2 response), as well as CXCL1 (initiator of the Th17 response) in mouse lung epithelial cells. In conclusion, repeated intranasal treatments with rAni s 1 and rAni s 9 induced airway inflammation in mice by elevating of Th2 and Th17 responses in the lung.

Introduction: Trichinella spiralis establishes a chronic infection in skeletal muscle by developing nurse cells within muscle fibers. During symbiosis in host, changes in the muscle fibers and inflammation may affect muscle function. Methods: We investigated muscle strength and inflammation in T. spiralis-infected mice during 1 to 48 weeks after infection. Results: Muscle strength decreased compared to that in uninfected control mice during the late infection stage. Additionally, inflammatory related cytokines increased significantly during early stage of infection and then rapidly decreased. In pathological study, nuclear infiltration maintained from the early infection stage to chronic infection stage. Moreover, vacuoles and eosinophil infiltration were observed in infected muscle in chronic stage. Conclusion: These results suggest that infection by T. spiralis significantly affects muscle function was continuously being weakness because vacuoles formation and maintained nucleus and eosinophil infiltration during chronic phase of T. spiralis infection.

The potential application of surface-enhanced Raman spectroscopy (SERS) was investigated to detect Salmonella Typhimurium and Escherichia coli. The silver colloidal suspension with optimum optical characteristics was obtained for SERS detection. The SERS spectra were collected the wavelengths from 500 to 1200 cm-1. The characteristic SERS spectra were obtained from both S. Typhimurium and E. coli; however the observed SERS peak profiles for both pathogens exhibited similar peak profiles. Therefore, the results suggest that even though SERS possesses the potential capability to detect pathogens rapidly, further study is required to resolve the ambiguous spectra profiles for S. Typhimurium and E. coli.

The aerosol characteristics between haze episode and Asian dust event were identified in January and March, 2013 in Gwang-ju of Korea to investigate the metal elements, ionic concentrations and carbonaceous particles of and . In the haze episode, the concentrations were increased 1~3.2 times of ionic species and 1.6~2.7 of metal elements. Especially, the concentration of , and consists of 50 percent in ionic species during haze episode that was higher than Asian dust event. This suggests that secondary aerosols from anthropogenic air pollution were mainly contributed by haze episode. During the Asian dust event, increase of metal concentrations was higher than haze episode because of remarkable increase of Ti, K and Fe originated from soil. The concentrations of carbonaceous particles were increased 2.5 times during haze episode, and 2.4 times of OC and 2.1 times of EC during Asian dust event in . However, these aerosol mass concentration does not affect the OC/EC ratio. The average equivalence ratios of cations/anions in were 0.99 in haze episodes and 0.94 during non-event day. The neutralization factor of was higher than that of . Futhermore, aerosol was aged due to atmospheric stagnation that might be affected by the haze episode.

Autophagy is a self-digestion process in which intracellular structures are degraded in response to stress. Notably, prolonged autophagy leads to cell death. In this study, we investigated whether CX-4945, an orally available protein kinase 2 (CK2) inhibitor, induces autophagic cell death in human cervical cancer-derived HeLa cells and in human prostate cancer-derived LNCaP cells. CX-4945 treatment of both cell lines resulted in the formation of autophagosomes, in the conversion of microtubule-associated protein 1 light chain 3 (LC3), and in down-regulation of the Akt-mammalian target of rapamycin (mTOR)-p70 ribosomal protein S6 kinase (S6K) signaling cascade. Thus, pharmacologic inhibition of CK2 by CX-4945 induced autophagic cell death in human cancer cells by down-regulating Akt-mTOR-S6K. These results suggest that autophagy-inducing agents have potential as anti-cancer drugs.

United States. National Aeronautics and Space Administration (Upper Atmospheric Research Program Grant NNX11AF17G)

Statistical trajectory analysis has been widely used to identify potential source regions for chemically and radiatively important chemical species in the atmosphere. The most widely used method is a statistical source-receptor model developed by Stohl (1996), of which the underlying principle is that elevated concentrations at an observation site are proportion- ally related to both the average concentrations on a specific grid cell where the observed air mass has been passing over and the residence time staying over that grid cell. Thus, the method can compute a residence-time-weighted mean concentration for each grid cell by superimpos- ing the back trajectory domain on the grid matrix. The concentration on a grid cell could be used as a proxy for potential source strength of corresponding species. This technical note describes the statistical trajectory approach and introduces its application to estimate potential source regions of CO2 enhancements observed at Korean Global Atmosphere Watch Observa- tory in Anmyeon-do. Back trajectories are calculated using HYSPLIT 4 model based on wind fields provided by NCEP GDAS. The identified CO2 potential source regions responsible for the pollution events observed at Anmyeon-do in 2010 were mainly Beijing area and the North- ern China where Haerbin, Shenyang and Changchun mega cities are located. This is consistent with bottom-up emission information. In spite of inherent uncertainties of this method in esti- mating sharp spatial gradients within the vicinity of the emission hot spots, this study suggests that the statistical trajectory analysis can be a useful tool for identifying anthropogenic poten- tial source regions for major GHGs.

Objective The small molecule halofuginone has been shown to inhibit fibrosis, angiogenesis, and tumor progression. This study was undertaken to evaluate the effects of halofuginone in preventing autoimmune arthritis in mice. Methods The effects of halofuginone on joint diseases were assessed by clinical scoring and histologic analysis. Protein expression levels were confirmed by immunohistochemistry, enzyme-linked immunosorbent assay, flow cytometry, and/or Western blotting. The expression levels of messenger RNA (mRNA) for various molecules were determined by real-time polymerase chain reaction (PCR). Proliferation of osteoclast precursors was assessed by bromodeoxyuridine uptake. Osteoclast differentiation and activity were determined by quantifying tartrate-resistant acid phosphatase (TRAP)–positive multinucleated cells and area of resorbed bone. Results Treatment with halofuginone suppressed the development of autoimmune arthritis and reciprocally regulated Th17 cells and FoxP3+ Treg cells. These effects of halofuginone on Th17 differentiation involved increased signaling of ERK and reduction of STAT-3 and NF-ATc1 expression. Furthermore, halofuginone induced the expression of indoleamine 2,3-dioxygenase (IDO) in dendritic cells, leading to reduced production of Th17 cells. In addition, halofuginone prevented the formation and activity of osteoclasts through suppression of transcription factors, such as activator protein 1 and NF-ATc1, and inhibited cell cycle arrest by the committed osteoclast precursors via expression of Ccnd1 encoding cyclin D1. Conclusion Taken together, our results suggest that halofuginone is a promising therapeutic agent for the treatment of Th17 cell–mediated inflammatory diseases and bone diseases.

Recently, we reported that Alpinia katsumadai (AK) has anti-nociceptive activity in vivo and that cardamonin (CDN) from AK suppresses the activity and expression of transglutaminase-2 (Tgase-2). However, it remains unknown whether CDN contributes to the anti-nociceptive activities of AK in vivo. We examined the anti-inflammatory effects of CDN in MG63 osteoblast-like cells and Raw264.7 macrophage-like cells treated with interleukin-1β treatment. CDN suppressed the expression of Tgase-2, cyclooxygenase-2 (COX-2), and p65 (nuclear factor-κB) in a concentration-dependent manner, and restored the expression of IκB in MG63 and Raw264.7 cells. However, CDN did not inhibit the activity of COX-2. Gene silencing of Tgase-2 reduced the COX-2 expression in MG63 cells. Phenylbenzoquinone (PBQ)-induced writhing, carrageenan-induced hyperalgesia, and rota-rod test were used to evaluate the anti-nociceptive activity in vivo. CDN (3-30 mg/kg, orally administered) significantly inhibited PBQ-induced writhing. CDN also produced a significant, dose-dependent increase in the withdrawal response latencies in carrageenan-induced hyperalgesia. The effects of CDN on PBQ-induced writhing were not caused by impaired motor functions. These results suggest that CDN might be helpful in controlling the pain from inflammatory diseases.

Skin hyperpigmentation is one of the most common skin disorders caused by abnormal melanogenesis. The mechanism and key factors at play are not fully understood. Previous reports have indicated that cystamine (CTM) inhibits melanin synthesis, though its molecular mechanism in melanogenesis remains unclear. In the present study, we investigated the effect of CTM on melanin production using ELISA reader and the expression of proteins involved in melanogenesis by Western blotting, and examined the involvement of transglutaminase-2 (Tgase-2) in SK-MEL-2 human melanoma cells by gene silencing. In the results, CTM dose-dependently suppressed melanin production and dendrite extension in α-MSH-induced melanogenesis of SK-MEL-2 human melanoma cells. CTM also suppressed α-MSH-induced chemotactic migration as well as the expressions of melanogenesis factors TRP-1, TRP-2 and MITF in α-MSH-treated SK-MEL-2 cells. Meanwhile, gene silencing of Tgase-2 suppressed dendrite extension and the expressions of TRP-1 and TRP-2 in α-MSH-treated SK-MEL-2 cells. Overall, these findings suggested that CTM suppresses α-MSH-induced melanogenesis via Tgase-2 inhibition and that therefore, Tgase-2 might be a new target in hyperpigmentation disorder therapy.

The stiffness of cancer cells is attributable to intermediate filaments such as keratin. Perinuclear reorganization via phosphorylation of specific serine residue in keratin is implicated in the deformability of metastatic cancer cells including the human pancreatic carcinoma cell line (PANC-1). 12-O-Tetradecanoylphorbol-13-acetate (TPA) is a potent tumor promoter and protein kinase C (PKC) activator. However, its effects on phosphorylation and reorganization of keratin 8 (K8) are not well known. Therefore, we examined the underlying mechanism and effect of TPA on K8 phosphorylation and reorganization. TPA induced phosphorylation and reorganization of K8 and transglutaminase-2 (Tgase-2) expression in a time- and dose-dependent manner in PANC-1 cells. These effects peaked after 45 min and 100 nM of TPA treatment. We next investigated, using cystamine (CTM), Tgase inhibitor, and Tgase-2 gene silencing, Tgase-2’s possible involvement in TPA-induced K8 phosphorylation and reorganization. We found that Tgase-2 gene silencing inhibited K8 phosphorylation and reorganization in PANC-1 cells. Tgase-2 gene silencing, we additionally discovered, suppressed TPA-induced migration of PANC-1 cells and Tgase-2 overexpression induced migration of PANC-1 cells. Overall, these results suggested that TPA induced K8 phosphorylation and reorganization via Tgase-2 expression in PANC-1 cells.

Sea cucumber extracts have potent biological effects, including anti-viral, anti-cancer, antibacterial, anti-oxidant, and anti-inflammation effects. To understand their anti-asthma effects, we induced allergic airway inflammation in mice after 7 oral administrations of the extract. The hyper-responsiveness value in mice with ovalbumin (OVA)-alum-induced asthma after oral injection of sea cucumber extracts was significantly lower than that in the OVA-alum-induced asthma group. In addition, the number of eosinophils in the lungs of asthma-induced mice pre-treated with sea cucumber extract was significantly decreased compared to that of PBS pre-treated mice. Additionally, CD4[Formula: see text]CD25[Formula: see text]Foxp3[Formula: see text]T (regulatory T; Treg) cells significantly increased in mesenteric lymph nodes after 7 administrations of the extract. These results suggest that sea cucumber extract can ameliorate allergic airway inflammation via Treg cell activation and recruitment to the lung.

Background Although indoleamine 2,3-dioxygenase (IDO)-mediated immune suppression of mesenchymal stem cells (MSCs) has been revealed in septic and tumor microenvironments, the role of IDO in suppressing allergic airway inflammation by MSCs is not well documented. We evaluated the effects of adipose-derived stem cells (ASCs) on allergic inflammation in IDO-knockout (KO) asthmatic mice or asthmatic mice treated with ASCs derived from IDO-KO mice. Methods and Findings ASCs were injected intravenously in wild-type (WT) and IDO-KO asthmatic mice. Furthermore, asthmatic mice were injected with ASCs derived from IDO-KO mice. We investigated the immunomodulatory effects of ASCs between WT and IDO-KO mice or IDO-KO ASCs in asthmatic mice. In asthmatic mice, ASCs significantly reduced airway hyperresponsiveness, the number of total inflammatory cells and eosinophils in bronchoalveolar lavage fluid (BALF), eosinophilic inflammation, goblet hyperplasia, and serum concentrations of total and allergen-specific IgE and IgG1. ASCs significantly inhibited Th2 cytokines, such as interleukin (IL)-4, IL-5, and IL-13, and enhanced Th1 cytokine (interferon-γ) and regulatory cytokines (IL-10, TGF-β) in BALF and lung draining lymph nodes (LLNs). ASCs led to significant increases in regulatory T-cells (Tregs) and IL-10+ T cell populations in LLNs. However, the immunosuppressive effects of ASCs did not significantly differ between WT and IDO-KO mice. Moreover, ASCs derived from IDO-KO mice showed immunosuppressive effects in allergic airway inflammation. Conclusions IDO did not play a pivotal role in the suppression of allergic airway inflammation through ASCs, suggesting that it is not the major regulator responsible for suppressing allergic airway inflammation.

Purpose: This study evaluates the efficacy of a Self-Care Reinforcement Program (SCRP) based on the Selection Optimization Compensation (SOC) model, in socially vulnerable elderly women with metabolic syndrome. Methods: This study adopts a pretest-posttest nonequivalent control group design. The participants were 64 socially vulnerable elderly Korean women with metabolic syndrome (experimental group: 31, control group: 33). Participants’ body composition analysis, nutrient intake, risk factors of metabolic syndrome, depressive symptoms, and social network were measured. Data were analyzed with an independent t-test; statistical significance levels were set at p

Epithelial-mesenchymal-transition (EMT) is a key event for tumor cells to initiate metastasis which lead to switching of E-cadherin to N-cadherin. Resolvins are known to promote the resolution of inflammation and phagocytosis of macrophages. However, the role of resolvins in EMT of cancer is not known. Therefore, we examined the effects of resolvins on transforming growth factor, beta 1 (TGF-β1)-induced EMT. Expression of E-cadherin and N-cadherin in A549 lung cancer cells was evaluated by Western blot and confocal microscopy. Involvement of lipoxin A4 receptor/formyl peptide receptor 2 (ALX/FPR2) was examined by gene silencing. TGF-β1 induced expression of N-cadherin in A549 lung cancer cells, and resolvin D1 and D2 inhibited the expression of N-cadherin at low concentrations (1-100 nM). Resolvin D1 and D2 also suppressed the expression of zinc finger E-box binding homeobox 1 (ZEB1). The effects of resolvin D1 and D2 were confirmed in other lung cancer cell lines such as H838, H1299, and H1703. Resolvin D1 and D2 did not affect the proliferation of A549 lung cancer cells. Resolvin D1 and D2 also suppressed the TGF-β1-induced morphological change. Resolvin D1 and D2 also inhibited the TGF-β1-induced migration and invasion of A549 cells. Resolvin D1 is known to act via ALX/FPR2 and GPR32. Thus, we examined the involvement of ALX/FPR2 and GPR32 in the suppressive effects of resolvin D1 on TGF-β1-induced EMT of A549 cells. Gene silencing of ALX/FPR2 and GPR32 blocked the action of resolvin D1. Overexpression of ALX/FPR2 or GPR32 increased the effects of resolvin D1. These results suggest that resolvin D1 inhibited TGF-β1-induced EMT via ALX/FPR2 and GPR32 by reducing the expression of ZEB1.

To determine whether pinworm infections and head lice infestations spread among children in orphanages, 117 children from 4 orphanages in Busan-si and Ulsan-si, Korea, were examined for enterobiasis and head lice infestation between January and February 2014. The overall rate of Enterobius vermicularis egg positivity was 0.85%, whereas none of the children had head lice infestations. The rate of pinworm infection was much lower among the orphanage children compared to the rates observed in previous studies among kindergarten and primary school students. Moreover, the risk factors for enterobiasis were less frequent among these subjects than previously reported. The personal hygiene and health of the orphanage children were supervised by a regular, employed nurse through a health education program. In conclusion, pinworm infection was efficiently controlled among the children in orphanages, and this might be related to good personal hygiene practices in Korea.

The ascarids, Toxocara canis and Toxascaris leonina, are probably the most common gastrointestinal helminths encountered in dogs. In order to understand biological differences of 2 ascarids, we analyzed gene expression profiles of female adults of T. canis and T. leonina using CLC Genomics Workbench, and the results were compared with those of free-living nematode Caenorhabditis elegans. A total of 2,880 and 7,949 ESTs were collected from T. leonina and T. canis, respectively. The length of ESTs ranged from 106 to 4,637 bp with an average insert size of 820 bp. Overall, our results showed that most functional gene annotations of 2 ascarids were quite similar to each other in 3 major categories, i.e., cellular component, biological process, and molecular function. Although some different transcript expression categories were found, the distance was short and it was not enough to explain their different lifestyles. However, we found distinguished transcript differences between ascarid parasites and free-living nematodes. Understanding evolutionary genetic changes might be helpful for studies of the lifestyle and evolution of parasites.

To determine alteration of immune responses during visceral larva migrans (VLM) caused by Toxascaris leonina at several time points, we experimentally infected mice with embryonated eggs of T. leonina and measured T-helper (Th) cell-related serial cytokine production after infection. At day 5 post infection (PI), most larvae were detected from the lungs, spleen, intestine, and muscle. Expression of thymic stromal lymphopoietin (TSLP) and CCL11 (eotaxin) showed a significant increase in most infected organs, except the intestine. However, expression of the CXCL1 (Gro-α) gene was most highly enhanced in the intestine at day 14 PI. Th1-related cytokine secretion of splenocytes showed increases at day 28 PI, and the level showed a decrease at day 42 PI. Th2-related cytokine secretion of splenocytes also showed an increase after infection; in particular, IL-5 level showed a significant increase at day 14 PI, and the level showed a decrease at day 28 PI. However, levels of Th17-related cytokines, IL-6 and IL-17A, showed gradual increases until day 42 PI. In conclusion, Th1, Th2, and Th17-related cytokine production might be important in immune responses against T. leonina VLM in experimental mice.