Melia azedarach is commonly used in traditional and folk medicine in Korea and China to treat a variety of diseases including diarrheal, diabetic, rheumatic, and hypertensive disease. The aim of this study was to determine the potential prophylactic and therapeutic effects of Melia azedarach against a broad spectrum of viruses in in vitro cell culture model and the protective effect against different influenza A subtypes in BALB/c mice model. An effective dose of pre-treatment, co-treatment, and post-treatment of Melia azedarach significantly reduced the replication of coxsackievirus, herpes simplex virus, influenza A virus, enterovirus, and bovine rhinovirus in both epithelial and macrophage cell lines. Melia azedarach treatment remarkably promoted the phosphorylation of the key molecules associated with the type-1 interferon and NF-κB signaling pathways. Furthermore, it induced the secretion of type-1 interferon and pro-inflammatory cytokines and the subsequent stimulation of the antiviral state in both epithelial and macrophage cells. Interestingly, oral inoculation of an effective dose of herb extract significantly improved viral clearance in the lungs of BALB/c mice, thus exhibiting protection against several subtypes of influenza A virus. Together with our results indicate that an extracts of Melia azedarach and its components could exhibit a potential natural source of an antiviral drug candidate for a broad spectrum of viruses in animal and humans.

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.

The aminoacyl-tRNA synthetases (ARSs) are ancient house-keeping enzymes that catalyze the ligation of tRNAs to their cognate amino acids in the first step of protein synthesis. During the evolution of higher eukaryotes, cytoplasmic ARSs have undergone significant changes including the addition of new domains that are not part of the enzymatic core. These additional regions have been found to be associated with a broad range of biological functions beyond protein synthesis. The non-translational functions of ARSs appear to be regulated by their presence within a cytoplasmic multi-tRNA synthetase complex (MSC), which is assembled through the appended domains. We recently reported that the MSC member glutamylprolyl- tRNA synthetase (EPRS) promotes antiviral gene expression through its infection-specific phosphorylation and release from the MSC. Here, we conducted transcriptome analysis of influenza A virusinfected cells. We particularly focused on the analysis of chemokine-related gene expression, in combination with chemokine array analysis against virus infection. Moreover, the correlation between chemokine expression pattern and EPRS function in response to different stimuli was assessed. The results showed that viral infection increases interferon-response and pro-inflammatory chemokine expression. In contrast, the level of chemokine expression was suppressed in interferon-γ treated cells. Thus, these results further demonstrate the previously reported stimulus-specific EPRS functions in immune responses.

The belief that honey bee venom (BV) can be used to treat certain immune-related diseases, such as arthritis and rheumatic conditions, goes back to antiquity. A growing number of reports have demonstrated that BV contains at least 18 pharmacologically active components, including phospholipase A2 (PLA2). Recent research has shown that bee venom PLA2 (bvPLA2) induces protective immune responses against several diseases including asthma, Parkinson’s disease, and drug-induced organ inflammation. However, the antiviral properties of bvPLA2 have not been well investigated. Hence, we examined the potential inhibitory effects of bvPLA2 and its possible mechanism of action against a broad panel of pathogenic viruses in vitro. Pre-treatment with bvPLA2 significantly inhibited the replication of vesicular stomatitis virus (VSV), coxsackie virus (H3), enterovirus-71 (EV-71), herpes simplex virus (HSV) and Adenovirus (AdV) dramatically. However, bvPLA2 did not show antiviral activity against Influenza A virus (PR8) and Newcastle disease virus (NDV). Such inhibitory effects were explained by blocking of the attachment of the virus to cells upon bvPLA2 treatment. Additionally, we observed that Heparan sulfate (HS) has an inhibitory effect on the attachment of HSV to the cell surface dose dependently, which was inconsistent with bvPLA2 treatment. These findings suggest that bvPLA2 has an inhibitory effect on the replication of diverse viruses by blocking their attachment to the cell surface and could be a promising source of natural antiviral agents.

Iron is an essential trace element for normal functions of the body. Restriction of iron availability directly limits erythropoiesis. The objective of this experiment was to compare the bioavailability of iron nanoparticles (Fe-NPs) with iron-microparticles (Fe-MPs) in anemic mice. There were four experimental groups, including the normal control group, iron-deficiency anemia (IDA) group, Fe-NPs group, and Fe-MPs group. Animals in the normal group fed on an adequate iron-containing diet (45 ppm Fe). Meanwhile, animals in the other three groups fed on a low Fe diet (4.5 ppm Fe) for seven weeks. Double deionized water was supplied as drinking water ad libitum. After feeding for three weeks with the low Fe diet, animals in the Fe-NPs and Fe-MPs groups received oral administration of Fe-NPs or Fe-MPs at a daily dose of 40 mg/kg for four weeks. The IDA group showed markedly decreased red blood cell (RBC) count, hematocrit (Hct), and hemoglobin (Hb) values compared with the normal group throughout the experimental periods. Treatments with Fe-NPs or Fe-MPs for four weeks resulted in restoration of the decreased RBC count and hematological values similar to normal values. The Fe-NPs group showed faster restoration in values than Fe-MPs with passage of time. The iron contents of the upper small intestine in the Fe-NPs and Fe-MPs groups were higher than in the normal group at weeks 2 and 4. Treatment with Fe-NPs and Fe-MPs resulted in a significant increase in hepatic iron contents and lipid peroxidation, compared with the IDA group with passage of time. The iron contents in liver and ferritin deposits in spleen were identified in the Fe-NPs and Fe-MPs groups, similar to the normal group. These results indicate that oral administration of both Fe-NPs and Fe-MPs can result in recovery from anemia and Fe-NPs is more absorbable and available in the body than Fe-MPs.

Iron nanoparticles (Fe-NPs) have recently been used for cancer diagnosis and therapy for imaging contrast and drug delivery. However, no study on nutritional bioavailability of Fe-NPs in the body has been reported. Ascorbic acid (AA) is known to aid in absorption of iron in the stomach by reducing Fe (III) to Fe (II). In this study, we investigated the bioavailability of Fe-NPs with AA in iron-deficiency-anemic mice in comparison with non-nano iron particles. Iron-deficient anemia was induced by feeding an iron-deficient diet (4.5 mg Fe/kg) and ocular bleeding from retro-orbital venous plexus for four weeks. Normal control mice were given a normal diet (45 mg Fe/ kg). After induction of anemia in mice, anemic mice received daily oral administration of Fe (40 mg/kg B.W.) + AA (5 g/kg B.W) and Fe-NPs (40 mg/kg B.W) + AA (5 g/kg B.W). After sacrifice, liver and spleen tissues were observed by haematoxylin & eosin stain. Amount of trace iron in liver and upper small intestine was investigated using an inductively coupled plasma-atomic emission spectrometer. Red blood cells (RBC), hematocrit (Hct), hemoglobin (Hb), and total iron binding capacity were also measured. The concentrations of iron in the Fe-NPs + AA group were significantly higher in liver and in upper small intestine than that in the Fe + AA group. The values of RBC, Hct, and Hb in the Fe-NPs + AA group were more rapidly increased to normal values compared with the Fe + AA group with increasing time. These results suggest that Fe-NPs in the presence of AA may be more bioavailable than non-nano Fe in Fe-deficient anemic mice.

날치 알과 이의 대체 어란인 열빙어 알 및 청어 알의 안전성과 식품성분 특성에 대하여 살펴보았다. 날치 알의 크기는 열빙어 알 및 청어 알에 비하여 컸다. 날치 알의 수분 함량과 염도는 페루산이 중국산에 비하여 수분의 경우 높으나 염도의 경우 낮았고, 인도네시아산에 비하여는 수분의 경우 낮았으나, 염도의 경우 높았다. 한편, 기타 어란의 수분 함량과 염도는 열빙어 알의 경우 각각 80.4% 및 3.2%, 청어 알의 경우 각각 65.4% 및 20.0%를 나타내었다. pH, 휘발성염기질소, 중금속, 생균수 및 대장균군의 결과에 의하면 이들 5종의 어란의 경우 여러 가지 가공소재로 이용하여도 위생적인 문제는 없으리라 판단 되었다. 어란의 주요 지방산은 날치 알의 경우 16:0(27.8-30.5%), 18:1n-9(7.2-8.0%), 20:5n-3 (5.6-8.2%) 및 22:6n-3(22.0-25.6%)이었고, 열빙어 알 및 청어 알의 경우 이외에도 16:1n-7(6.7 -9.3%)이었다. 어란의 총아미노산 함량은 9.44-10.39 g/100g 범위이었고, 주요 아미노산은 aspartic acid, glutamic acid, leucine 및 lysine이었다. 날치 알의 무기질 함량은 인도네시아산의 아연을 제외 한다면 열빙어 알 및 청어 알의 무기질 함량보다 높았다. 관능 검사 결과에 의하면 열빙어 알과 청어 알 에 비하여 날치 알이 색과 조직감에서는 우수하였으나, 향은 차이가 없었다.

The current study was conducted in order to investigate promotional effects of herbal extracts on hair growth in an animal model of mice. There were four experimental groups, including distilled water (DW) as a negative control (NC), 3% minoxidil (MXD) as a positive control (PC), 50% ethanol (EtOH) as a vehicle control (VC), and herbal extract (HE) as the experimental treatment (E). The HE was extracted with ethanol from plants, including Gardenia, Mentha arvensis, Rosemary, and Lavender. Six-week-old C57BL/6 male mice were shaved with an electric clipper and the test materials were topically treated with 0.2 ml per mouse daily for three weeks. Photographic evaluation of hair re-growth was performed weekly during a period of three weeks. The number of mast cells was counted on the dorsal skin section of mice. The enzymes, alkaline phosphatase (ALP) and γ-glutamyl transpeptidase (γ-GT), were determined using a biochemical autoanalyzer. No clinical signs were observed in any of the experimental groups. As a result of photometric analysis, topical application of HE to dorsal skin for two weeks resulted in significantly faster acceleration of hair regrowth, compared with that of the NC or VC group (P<0.05). The PC and E groups showed a significant decrease in mast cell population, compared to the NC group. Activities of ALP and γ-GT were significantly increased in the PC and E groups, compared to the NC or VC group (P<0.05). Taken together, these results suggest that the herbal extract may have hair-growth promoting activity equal to that of MXD.

To investigate the effect of carnosine on exhaustive exercise, swimming tests were conducted weekly with loads corresponding to 5% of body weight attached to the tails of mice, and the swimming time to exhaustion was measured. Eighty male ICR mice were divided into four groups, to which carnosine was administered at doses of 0 (control), 10, 50, and 250 mg/kg/day, respectively, for a period of four weeks. At the end of swimming exercise challenges, serum biochemistry, oxidative stress enzyme activity, and antioxidant enzyme activity in tissues were determined. Treatment with 250 mg/kg carnosine resulted in a significant increase in swimming times to exhaustion, compared to the control group in the first (P<0.01) and third week (P<0.05). Significantly lower serum lactate levels were observed after the swimming exercise in the carnosine-treated groups (10 and 250 mg/kg), compared with the control (P<0.01). Malondialdehyde levels in the liver (10 and 50 mg/kg carnosine treated groups) and skeletal muscle (250 mg/kg carnosine treated group) were significantly lower, compared with the control (P<0.05). Significantly lower protein carbonyl levels in skeletal muscle were observed in the 50 and 250 mg/kg carnosine treated groups, compared with the control (P<0.01). Superoxide dismutase and glutathione peroxidase activities in skeletal muscle did not differ significantly among the groups. These results indicate that carnosine may improve swimming exercise capacity by attenuating production of lactate and reducing oxidative stress in mice.

To develop new nutraceuticals from mushrooms, nutritional properties of two kinds of Hypsizygus marmoreus fruiting bodies were investigated. Hypsizygus marmoreus (white) contained 21.63% H. marmoreus (brown) contained 27.34% of crude protein, 55.75% of total sugar, and 2.69% crude lipid and 7.77% ash. It also contained 10.04 mg% Na, 19.28 mg% Ca, 9.30 mg% Fe and its calorie value was 356.57 Kcal per 100 g. Free sugar and amino acid content of H. marmoreus (brown) were determined.

This study was carried out to obtain new anti-gout agent from mushrooms. Various extracts from twelve kinds of mushrooms were prepared by water and ethanol extractions and its anti-gout xanthine oxidase inhibitory activity were investigated. Water extract of Flammulina velutipes and Agaricus bisporus fruiting bodies showed high xanthine oxidase inhibitory activity of 70.8% and 60.0%, respectively. However, its inhibitory activity of these ethanol extracts were very low excepet 60.3% of Innotus obliquus. Finally, we selected Flammulina velutipes, showing the highest xanthine oxidase inhibitory activity as a producer of new anti-gout agent.

A level of dietary iron may play a role in colon carcinogenesis. The effect of dietary iron on colon carcinogenesis was investigated in male ICR mice. Five-week old mice were acclimated for one week and fed on iron-normal diet (35 ppm Fe), iron-deficient diet (3 ppm), or iron-overloaded diet (350 ppm Fe) for 8 weeks. Animals received three (0-2^nd weeks after starting experiment) injections of azoxymethane (AOM; 10 mg/kg b.w.) to induce colonic aberrant crypt foci (ACF). There were five experimental groups including normal control without AOM, AOM+iron-normal diet (AOM+NFe), AOM+iron-deficient (AOM+LFe), AOM+ironoverloaded diet (AOM+HFe) groups. The total numbers of ACF and aberrant crypt (AC) were measured in the colonic mucosa after staining with methylene blue. The blood and serum were analyzed with a blood cell differential counter and an automatic serum analyzer. The hepatic iron levels were significantly dependent on the presence of iron in the diets. Iron-deficient diet significantly decreased the several hematological values. The values of glutamic oxaloacetic transaminase (GOT) and glutamic pyruvate transaminase (GPT) were also significantly decreased in iron-overloaded or iron-deficient diet groups, compared with normal iron diet group. Dietary iron-deficiency decreased the numbers of ACF (64.9) and AC (79.8) per colon by 20.6 and 21.8%, respectively, compared with AOM+NFe group (72.4 ACF/colon and 90.3 AC/colon). However, ironoverloaded diet increased ACF (82.9) and AC (96.0) induction by AOM, compared with normal iron diet. These results suggest that dietary iron can affect the colon carcinogenesis in the animal model of mice.

3The goal of this study was to develop high value mushroom foods possessing functionality. The effect of the Pholiota adiposa on the alcohol fermentation and its antihypertensive angiotensin I-converting(ACE) inhibitory activity of Korean traditional rice wine, Yakju was investigated. The ACE inhibitory activity of traditional rice wine was increased about 8% by the addition of 0.1% of the edible mushroom Pholiota adiposa ASI 24012 fruiting bodies into the mash containing 1% Lycii fructus, cooked rice, koji and antihypertensive S. cerevisiae and its Pholiota adiposa PAD-022 which showed anticholesteromia HMGCoA reductase inhibitory activity. Its quality characteristics and stability were investigated during the storage of room temperature and 40℃ for 3 months. It showed very high acceptability and also was very stable at 40℃ for 3 months[This study was supported by a grant from ARPC, 2005-2007].

3The goal of this study was to develop high value mushroom foods possessing functionality. The effect of the Pholiota adiposa on the alcohol fermentation and its antihypertensive angiotensin I-converting(ACE) inhibitory activity of Korean traditional rice wine, Yakju was investigated. The ACE inhibitory activity of traditional rice wine was increased about 8% by the addition of 0.1% of the edible mushroom Pholiota adiposa ASI 24012 fruiting bodies into the mash containing 1% Lycii fructus, cooked rice, koji and antihypertensive S. cerevisiae and its Pholiota adiposa PAD-022 which showed anticholesteromia HMG-CoA reductase inhibitory activity. Its quality characteristics and stability were investigated during the storage of room temperature and 40℃ for 3 months. It showed very high acceptability and also was very stable at 40℃ for 3 months[This study was supported by a grant from ARPC, 2005-2007].

This study was performed to obtain basal data for development of new functional food by using mushroom Pholiota spp.. Among several kinds of extracts, water extracts of Pholiota adiposa ASI 24027 showed the highest solid yield of 58.7%. Crude protein of Pholiota adiposa ASI 24015 and β-glucan contents of Pholiota adiposa ASI 24005 were 25.0% and 0.66%, respectively. K and Mg were also contained plentifully in the fruiting body of Pholiota adiposa ASI 24004 (3,649 mg and 138 mg/100g dried fruiting body, respectively) Amino acid and total fatty acid contents were similar between almost all of Pholiota spp.,

β-Hydroxy-β-methylglutaryl coenzyme A reductase(HMG-CoA reductase, E.C. 1.1.1.34) is known to be rate-limiting enzyme in cholesterol biosynthesis. HMG-CoA reductase inhibitor is very useful in the remedy or precaution of hyperlipidemia. The methanol extracts of Pholiota adiposa ASI 24018 showed the highest HMG-CoA reudctase inhibitory activity of 55.8%. The HMG-CoA reductase inhibitor of Pholiota adiposa ASI 24018 was maximally extracted when it was treated with methanol at 30℃ for 12hrs. After purified the HMG-CoA reductase inhibitor by systematic solvent extraction, LH-20 gel column chromatography and RP-HPLC, finally obtained the HMG-CoA reductase inhibitor with an activity of IC50 6.8ug. Molecular weight of the purified HMG-CoA reductase inhibitor was 765.4Da by LC-Mass, NMR, FT-IR, UV spectrometry.