The effects of lactic acid bacteria (LAB) mixtures on low moisture Italian ryegrass (IRG) silage fermentation was evaluated in field conditions. The experiment was categorized into two groups: Un-inoculated (Control) and Inoculated with LAB mixture for four storage periods (45, 90, 180, and 365 days, respectively). Silage inoculated with the LAB mixture had the lowest pH with highest lactic acid production than the control from beginning at 45-365 days at all moistures. Higher LAB counts were observed in inoculated silages than the control silages at whole experimental periods. It is a key reason for the rapid acidification and higher lactic acid production in silages during the storage periods. Overall results suggest that an adding of LAB mixture had positive effects on the increasing aerobic stability of silage and preserved its quality for an extended duration.

Acute myocardial infarction (AMI) is considered the major cause of mortality in the world. Tremendous animal studies are performed to develop novel therapeutics, and this study aimed to induce porcine myocardial infarction model by using polyethylene terephthalate (PET). Coronary guidewire was placed in left anterior descending artery (LAD). The balloon angioplasty catheter was inserted at the back of the PET. The balloon catheter was carefully pushed forward, until the balloon marker was located in mid-LAD. Coronary angiography was performed pre- and post-occlusion at 28 days by C-arm. Histologic analysis of heart tissue was performed 28 days after inducing AMI. Thirty three pigs were anesthetized and underwent percutaneous coronary catheterization. All pigs were successfully embolized in mid-LAD by PET. Fifteen pigs died due to ventricular fibrillation during post-anesthetic recovery time, and overall experiment mortality was 45.5%. In 2,3,5- triphenyl tetrazolium chloride staining, gross finding of the ischemic heart lesion showed firm and white area of infarction associated with the apex and left ventricular posterior wall. Infarct on H&E-stained sections demonstrated a region without myocytes and rich with cardiomyocyte with atypical nuclei. Successful induction of AMI by using PET may provide the pathophysiological information of ischemic heart disease and improvement of therapy development for AMI.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

Autophagy is an important self-eating process to eliminate damaged or unused organelles. We identified nine autophagy-related genes (Atg) including AaAtg-1, -3, -4b, -4d, -5, -6, -8, -12 and -13 from the Asian tiger mosquito, Aedes albopictus. Developmental expression patterns indicate that mRNA levels of AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 were highly expressed in egg, whereas expression of AaAtg8 was high in 1stand3rdinstarlarvalstages. TissuespecificexpressionofthesegenesindicatesthatAaAtg1 was highly expressed in thorax and midgut in blood-fed adult female mosquitoes (BF), and head and thorax in sugar fed adult female mosquitoes (SF). Transcript level of AaAtg3 was high in thorax in BF, but head, thorax and Malpighian tubules in SF. AaAtg4b, -4d mRNA levels were significantly high in Malpighian tubules in BF, and head in SF, respectively. AaAtg-5 and -6 transcripts were highly expressed in head in BF, and expression of AaAtg-8 was high in Malpighian tubules in BF. Levels of AaAtg-12 and -13 mRNAs were significantly high in head and midgut in BF. Induction patterns of AaAtg genes against pathogens showed that AaAtg-1, -3, -4b, -8, -12 and -13 were strongly induced at 6 h-post injection of S. aureus, and mRNA levels of AaAtg-1, -3 and -13 were significantly induced by E. coli challenge after 3 h-post injection in SF abdominal carcass. In SF midgut, AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 transcripts were drastically induced at 9 h-injection of E. coli and S. aureus, while expression of AaAtg-8 was highly induced by S. aureus and C. albicans at 9 h-post injection. Each AaAtg gene was slightly induced by E. coli, S. aureus or C. albicans at different time points in abdominal carcass in BF. Interestingly, AaAtg-8 was not induced by microbial challenge. While eight other Atg genes except AaAtg-8 were highly influenced by S. aureus at 6 and 9 h-post injection, E. coli at 3 h-post-treatment, and 3, 6, and 9 h-post inoculation. In the future, we will characterize the functional roles of autophagy during mosquito-microbes interaction.

It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

Host defense against pathogen invasion highly relies on immune defense machinery that is controlled by the nuclear factor-κB (NF-κB) of transcription factors. The Toll pathway are well known as an insect innate immune mechanism to protect host itself from invaded pathogens. Basically, in the edible insect, Tenebrio molitor, the Toll pathway is primarily activated by polymeric Lys-type peptidoglycans (PGNs), and components of fungal cell walls, β-1,3-glucan. Based on the current studies, the tremendous study has been focused on recognition and subsequent activation of spätzle in haemolymph, hence, there is a grave gap for intracellular event. Herein, in order to understand intracellular event of Toll signaling pathway, the Dorsal gene were identified. Moreover, domain analyses of TmDorsal2 gene indicate that there are two major domains such as Rel homology domain (RHD), ig-like, plexins, and transcription factors (IPT) domains. Based on the achieved results, TmDorsal2 mRNA was highly expressed in 1-day old pupa. Furthermore, TmDorsal2 was highly expressed in Malpighian tubules and fat body in last instar larvae (LL), and likewise mainly expressed in Malpighian tubules during adult 5-day old period, also the lowest expression of TmDorsal2 was observed in gonads. Moreover, TmDorsal2 mRNA levels after infection with E. coli appreciably went up at 6 and 9h time points. To investigate the effects of TmDorsal2 RNAi on larval susceptibility against various pathogens namely E. coli, S. aureus or C.albicans, dsRNA of TmDorsal2 has been synthesized the larvae dissected after 24h. As a result, TmAttacin1a, 1b and 2, TmDefencine1 and 2, TmTenecin1, 2, 3 and 4, TmCecropin2, TmColeoptericin1 and 2, Thaumatin-like protein 1 and 2 markedly reduced in the gut after injecting all mentioned microbes. In contrast, TmTenecin 2, Thaumatin-like protein 1 and 2 strikingly increased after microbe injection in the fat body. Interestingly, the most AMPs gene expression in whole body experimental case were upregulated. On the horizon, we will investigate effects of TmDorsal1 RNAi on larval susceptibility against various pathogens. Taken together, our studies may aid to understand insect innate immunity.

ICT 기반의 해충 이미지 전송 시스템은 담배거세미나방과 파밤나방을 예찰하고 방제하는데 필요한 효과적인 유인 해충 전송수단이다. 시설 온실내 담배거세미나방과 파밤나방을 유인하기 위하여 스마트 트랩을 이용하였다. 스마트트랩은 나방류 해충유인을 위해 성페로몬 4종 장착하였으며 유인된 해충은 특수 카메라를 이용하여 이미지 촬영이 가능하였다. 또한, 해충유인 발광트랩 처리구는 황색광을 사용한 발광 트랩을 사용하였다. 또한 대조구로 기존에 널리 사용되는 델타트랩은 한 종류의 성페로몬을 장착하여 유인력 검정에 이용하였다. 성페로몬 한 종류만 사용한 델타트랩 처리는 담배거세미나방 페로몬 루어를 사용하였고, 4종류의 성페로몬을 설치한 스마트 트랩 처리구는 담배거세미나방, 뒷날개흰나방, 파밤나방, 왕담배나방용 성페로몬 루어를 사용하였다. 실험은 2018년 6월 1일부터 8월 31일까지 함안 시설원예연구소의 13m2의 소형비닐하우스에서 수행하였으며 공시작물은 파프리카 ‘시로코’ 품종과 토마토 ‘데프니스’ 품종을 사용하였다. 나방류 해충의 유인력 검정결과 파프리카와 토마토를 식재한 온실 모두 성페로몬 루어 한 종류를 설치한 델타트랩보다 유인발광 트랩 처리구에서 높은 나방류 유인율을 나타내었다. 시험기간 유인발광 트랩의 경우 유인 포획된 총 나방수는 파프리카 온실에서 평균 97마리, 토마토 온실에서 75마리가 유인 포획되어 가장 높았다. 그러나, 성페로몬 1종 및 4종을 설치한 델타트랩 및 스마트트랩처리구는 두 개의 온실 모두 평균 5마리 이하의 유인율을 나타내었다. 따라서, 시설내 나방류 예찰 및 유인 방제하기 위해서는 기존 성페로몬만 사용하기 보다는 유인발광 트랩을 함께 설치하는 것이 나방류 유인포획에 효과적이라고 판단되었다.

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

Childhood obesity causes a higher risk of obesity, premature death and disability in adulthood. In addition, obese children experience an increased risk of respiratory problems, hypertension, cardiovascular disease, insulin resistance and psychological effects. This study aimed to investigate how an exercise intervention affects health-related physical fitness and inflammatory-related blood factors in obese children after. We hypothesized that there would be positive effects on serum levels of insulin-like growth factor-1 (IGF-1), connecting peptide(C-peptide) and resistin, as well as in muscle and cardiovascular-related physical capacities, after an exercise intervention in obese children. Thirty-seven obese children haveperformed health-related fitness tests and provided blood samples for the analysis of changes in circulating biomarkers, both before and after an 8-week exercise intervention, which includes stretching, aerobic exercise, resistance exercise and sports games. The results indicate that exercise training beneficially affects body compositions, especially percentage body fat and muscle mass, without influencing to body weight and height. The results of the physical fitness tests show that muscle and cardiovascular capacity were increased in obese children in response to exercise training. Simultaneously, the exercise training decreased circulating levels of C-peptide, which equated to a “large” effect size. Although there were no significant effects on the levels of IGF-1 and resistin, they show a “small” effect size. Therefore, our findings suggest that the exercise intervention have beneficial effects on body composition and physical fitness levels in obese children, whichmight be associated with the decline in circulating C-peptide.

Myosin is considered as the vital motor protein in vertebrates and invertebrates. Our present study was conducted to decipher the occurrence of myosin in dog fish (Squalus mitsukurii). We isolated one clone containing 979 bp cDNA sequence, which consisted of a complete coding sequence of 453 bp and a deduced amino acid sequence of 150 amino acids from the open reading frame with molecular weight, isoelectric point and aliphatic index are 16.72 Kda, 4.49 and 78.00, respectively. It contained 428 bp long 3' UTR with single potential polyadenylation signals (AATAAA). The predicted EF CA2+ binding domains were identified in residue 6-41, 83-118 and 133-150. A BLAST search indicates this protein exhibits a strong similarity to whale shark (Rhincodon typus) MLC3 (91% identical) and also house mouse (Mus musculus) MLC isoform 3f (81% identical). Phylogenetic analysis revealed that this protein is a MLC 3 isoform like protein. This protein also demonstrates highly conserved region with other myosin proteins. Homology modeling of S. mitsukuri was performed using crystal structure of Gallus gallus skeletal muscle myosin II based on high similarity. Reverse transcription-polymerase chain reaction (PCR), quantitative PCR results exhibits dogfish myosin protein is highly expressed in muscle tissue.

Low temperature cracking on asphalt material is one of the serious distresses for asphalt pavement built in northern U.S., Europe and Canada. Thermal stress is a key factor for measuring (and estimating) the resistant capability of asphalt pavement against low temperature cracking. For this reason, many road agencies have recognized thermal stress as a crucial parameter for evaluating the low temperature performance of asphalt pavement materials. Thermal stress is conventionally computed through two steps. First, the relaxation modulus E(t) is generated thorough the conversion of the experimental creep compliance data D(t). Then thermal stress (T℃) is numerically estimated solving convolution integral. In this paper, a one-step approach to the calculation of thermal stress is proposed. This method is based on Laplace transformation. Thermal stress and corresponding critical cracking temperature obtained with single- and double-step procedure on a set of three mixtures are graphically and statistically compared. It is observed that the application of Laplace transformation provides reliable computation results of thermal stress compared to the conventional computation approach.

The objectives of the present study were to examine the antioxidant activity of fractions with different isoelectric points from salmon enzymatic hydrolysates and obtain peptide fractions of sufficient amounts with higher antioxidant peptide fraction, which could be applied to the food and animal model systems. The salmon enzymatic hydrolysates were fractionated on the basis of the amphoteric nature of sample peptides by preparative isoelectric focusing without toxic solvents and reagents, which is termed autofocusing. Acidic and basic fractions showed higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity than the other fractions. The basic fractions showed higher hydroxyl (OH) radical scavenging activity. The weak acidic and weak basic fractions showed higher oxygen radical absorbance capacity (ORAC) values than the acidic and basic fractions. The acidic fraction showed higher metal chelating activity than other fractions. The acidic fraction suppressed lipid oxidation in the cooked patties to a greater extent than other fractions. These results suggest that peptides fractions from salmon enzymatic hydrolysate are effective antioxidant, and that autofocusing could be useful to increase antioxidant activity for application to food and animal model systems.

Salvia miltiorrhiza (Danshen) is perennial plant and commonly used in traditional Chinese medicine to treat numerous diseases like cardiovascular and cerebrovascular diseases, coronary heart disease, myocardial infarction, stroke and some viral diseases. Nevertheless, no study has been conducted on the antiviral properties of crude extract of Salvia miltiorrhiza against Influenza virus. In an attempt to identify new potential anti-influenza virus agents, 200 natural oriental herbal medicines were screened and we found that Salvia miltiorrhiza has a potential anti-influenza effect. Therefore, in this study, we investigated the protective effect of aqueous extract from Salvia miltiorrhiza against divergent influenza A subtypes using murine model of influenza A infection. Effective dose of aqueous extracts of Salvia miltiorrhiza in BALB/c mice displayed higher survival rate and lower lung viral titers when challenged with lethal doses of influenza A subtypes ({A/Aquatic bird/Korea/ W81/2005(H5N2)}, {A/PR/8/34(H1N1)}, {A/Aquatic bird/Korea/W44/2005(H7N3)} and {A/Chicken/ Korea/116/2004(H9N2)}). In vivo results exhibited that Salvia miltiorrhiza induced prophylactic effect in BALB/c mice against Influenza virus by disrupting viral replication or preventing viral infection by creating an antiviral state in the lungs. Taken together, the use of aqueous extracts of Salvia miltiorrhiza as an orally active antiviral agent, will be potential candidates for prophylactic treatments against Influenza A virus for humans and animals.