A total of 35 phosphate solubilizing bacterial strains were isolated from waste bed of Agaricus bisporus in Buyeo-Gun, Chungchugnam-do province and screened for the production of indole acetic acid(IAA). The best IAA producing strain was identified as Pantoea rodasii using 16S rRNA analysis. In addition to the IAA production, this strain could act as an efficient phosphate solubilizer (1100 μg ml-1 after 5 days of incubation) also. The selected strain was cultured under different conditions in order to assess the optimum conditions for maximum IAA production. The nutrient broth (NB) medium was recorded as the best medium, where the maximum IAA production (229 μg ml-1) was recorded at the start of stationary phase (12 hours after inoculation) of the bacteria growth. The performance of the strain was found to be maximum at the temperature of 30°C followed by 25°C. IAA production was found to be increased with increasing tryptophan concentration (from 0.1 to 0.6%), however beyond this limit, a slight reduction in IAA production was observed. The strains’ ability to produce IAA was further confirmed by extraction of crude IAA and subsequent TLC analysis. A specific spot from the extracted IAA preparation was found corresponding with the standard spot of IAA with same Rf value. The results of HPLC analysis conducted in identifying and quantifying the IAA production more precisely, are in agreement with the results of the assessment done with colorimetric method. As revealed by the results of the pot experiment, the isolated strain could significantly enhance the growth (as measured by shoot and root growth) of mung bean plants compared to that of non-inoculated plants. Therefore it can be concluded that the present strain, Pantoea rodasii has great potential to be used as bio-inoculants.

Sweetpotato whitefly (Bemisia tabaci), especially Q biotype, has been recognized one of the most destructive insect pests worldwide because of increased resistance to some insecticide groups requiring alternative strategies for its control. We studied the conidia production of entomopathogenic fungus Isaria javanica Pf04, which had been reported high virulence isolate against Q biotype of B. tabaci, using grain. Brown rice was most suitable for conidia mass production of the isolate of I. javanica. Conidia was produced high at 25 ~ 27.5℃. The isolate produced more spores when conidia suspension directly inoculated onto media than two-phase fermentation. When concentration of inoculum was high spore production was high, but increasing rate of conidia production was highest at low inoculum concentration (1×105 conidia/ml) as 6,700 times increase compared with 20 times increase at high inoculum concentration (1×108 conidia/ml). These results indicated that the isolate can produce more conidia with cheap agricultural product and can develop as a microbial pesticide to control sweetpotato whitefly.

In the past four years, outbreaks of acute respiratory diseases associated with canine influenza H3N2 viruses in dogs and cats have been reported in South Korea and China. For prevention of disease from spread of the disease and for administration of timely medical treatments, including countermeasures for quarantine, use of a rapid and highly sensitive detection method are important to detection of the causative viruses. This study was conducted in order to develop a real time RT-PCR for the H3N2 subtype. It was based on primers targeting the highly homologous sequences of matrix, hemagglutinin, and neuraminidase genes. The detection limit of real time RT-PCR was 10 copies/ul with matrix and hemagglutinin genes, and 1 copy with neuraminidase genes, respectively. This real time RT-PCR was as sensitive as virus isolation in 52 clinical samples. The detection system developed in this study might provide more rapid and highly sensitive results than commercial rapid kits based on immunochromatographic assay.

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.

Toxicities of 10 insecticides were examined against late third instars of Culex tritaeniorhynchus using the direct-contact mortality bioassay. Six geospatially distant field mosquitoes were collected from Chuncheon-si (designated CC-CT), Hwaseong (HS-CT), Seosan (SS-CT) Jeonju (JJ-CT), Daegu (DG-CT), and Busan (BS-CT) in the Republic of Korea (ROK). Marked regional variations of insecticide susceptibility were observed. Field populations of SS-CT, JJ-CT and DG-CT from agricultural areas showed higher to extremely higher insecticide susceptibility to pyrethroids than those of CC-CT, HS-CT and BS-CT strains from none agricultural areas. Extremely high to low levels of susceptibility were measured: bifenthrin, susceptible ratio (SR) = 2.7–896.3; β-cyfluthrin, SR = 1.8–633.3; α-cypermethrin, SR = 1.2–1,051.9; deltamethrin, SR = 1.3–711.1; permethrin, SR = 1.5–1,053.4 etofenprox, SR = 2.2–29.3; chlorfenapyr, SR = 5.1–103.6; chlorpyrifos, SR = 2.3–337.0; fenitrothion, SR = 2.0–142.3 and fenthion, SR = 1.4–186.2. Culex tritaeniorhynchus populations from rice paddies had been under heavy selection pressure due to the agricultural insecticides and that’s why the mosquito species demonstrated high resistance to pyrethroids which were used for a long time to control agricultural pests in the localities. These results indicate that careful selection and rotational use of these insecticides mayresult in continued satisfactory control against field populations of Japanese encephalitis vector mosquitoes.

Both iron-deficient and zinc-sufficient diets have been known to be associated with a decreased risk of colon cancer. We investigated that effect of dietary zinc on the formation of colonic aberrant crypt foci (ACF) induced by azoxymethane (AOM) followed by dextran sodium sulfate in iron-deficient mice. Five-week old ICR mice were acclimated for 1 week and fed on iron-deficient diet (4.50 ppm iron) with three different zinc levels (0.01, 0.1, and 1.0 ppm) for 12 weeks. The total number of aberrant crypt (AC) and ACF was measured in the colonic mucosa after methylene blue staining. The total ACF numbers of low Zn (LZn), medium Zn (MZn) and high Zn (HZn) diet groups were 10.00 ± 2.67, 8.78 ± 3.12, and 7.96 ± 2.44, respectively and there were no significant differences among the groups. However, the total AC numbers of HZn (27.07 ± 3.88) and MZn (26.39 ± 5.59) diet groups were significantly low compared to LZn (22.57 ± 5.09) diet group (p<0.01). Cytosolic SOD activity was the highest in LZn diet group. But thiobarbituric acid-reactive substances level in liver was also the highest in LZn diet group compared to other groups. There is no difference in cell proliferation in mucous membrane among the groups, while apoptotic positive cells were increased in the HZn diet group. The high zinc diet exhibited decreased β-catenin-stained areas on the mucous membrane of colon compared to the LZn or MZn diet group. These findings indicate that dietary zinc might exert a modulating effect on development of ACF/AC in the mice with iron-deficient status.

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.

In this study, two duplex real-time PCR approach with melting curve analysis is presented for the detection of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus, which are important food-borne bacterial pathogens usually present in fresh and/or minimally processed vegetables. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the β-glucuronidase (uidA, E. coli), thermonuclease (nuc, S. aureus), hemolycin (hly, L. monocytogenes) and tetrathionate reductase (ttr, Salmonella spp.) genes. Melting curve analysis using a SYBR Green I real-time PCR approach showed characteristic Tm values demonstrating the specific and efficient amplification of the four pathogens; 80.6 ± 0.9 ℃,86.9 ± 0.5 ℃, 80.4 ± 0.6 ℃ and 88.1 ± 0.11 ℃ for S. aureus, E. coli O157:H7, L. monocytogenes and Salmonella spp.,respectively. For all the pathogens, the two duplex, real-time PCR was equally sensitive to uniplex real-time PCR,using same amounts of purified DNA, and allowed detection of 10 genome equivalents. When our established duplex real-time PCR assay was applied to artificially inoculated fresh lettuce, the detection limit was 10³ CFU/g for each of these pathogens without enrichment. The results from this study showed that the developed duplex real-time PCR with melting curve analysis is promising as a rapid and cost-effective test method for improving food safety.

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.

This study is intended to examine the motor skill learning and treadmill exercise on motor performance and synaptic plasticity in the cerebellar injured rats by harmaline. Experiment groups were divided into four groups and assigned 15 rats to each group. GroupⅠ was a normal control group(induced by saline); GroupⅡ was a experimental control group(cerebellar injured by harmaline); GroupⅢ was a group of motor skill learning after cerebellar injured by harmaline; GroupⅣ was a group of treadmill exercise after cerebellar injured by harmaline. In motor performance test, the outcome of groupⅡ was significantly lower than the groupⅢ, Ⅳ(especially groupⅢ)(p<.001). In histological finding, the experimental groups were destroy of dendrities and nucleus of cerebellar neurons. GroupⅢ, Ⅳ were decreased in degeneration of cerebellar neurons(especially groupⅢ). In immunohistochemistric response of synaptophysin in cerebellar cortex, experimental groups were decreased than groupⅠ. GroupⅢ's expression of synaptophysin was more increased than groupⅡ, Ⅳ. In electron microscopy finding, the experimental groups were degenerated of Purkinje cell. These result suggest that improved motor performance by motor skill learning after harmaline induced is associated with dynamically altered expression of synaptophysin in cerebellar cortex and that is related with synaptic plasticity.

본 연구에서는 azoxymethane (AOM)과 dextran sodium sulfate (DSS)로 유도된 대장 발암과정에 대한 셀레늄의 방어 효과를 조사하였다. 셀레늄 결핍(0.02 ppm Se), 정상(0.1 ppm Se), 과다(0.5 ppm Se)사료를 12주간 식이로 급여하여 혈액검사와 대장암 발생의 초기단계인 aberrant crypt foci (ACF)수를 측정했으며, 암 발생율을 조사하였다. ICP-AES 를 사용하여 간의 셀레늄 농도를 측정하였으며, 또한 셀레늄포함 항산화효소인 glutathione peroxidase (GPx) 활성을 알아보았다. 또한 TUNEL assay와 PCNA, β-catenin에 대한 면역조직 염색을 수행하였다. ACF 수 및 종양 발생률에 있어서, 셀레늄과다사료를 급여한 군이 정상셀레늄사료를 급여한 군보다 낮았으며, 셀레늄결핍사료를 급여한 군은 오히려 ACF 수 및 종양 발생률이 높았다. GPx 활성은 셀레늄의 섭취가 과다한 군에서 높게 나타났으며, 이 때, TUNEL 에서 apoptotic positive cell이 증가하는 것을 확인했다. 또 한 셀레늄의 섭취가 과다한 군에서 PCNA와 β-catenin의 발현이 감소됨을 볼 수 있었다. 본 마우스 모델실험에서 셀레늄은 여러 기전에 의해 대장암 발생을 억제할 수 있을 것으로 사료된다.

Wrinkles are an outward sign of cutaneous aging appearing preferentially on ultraviolet B (UVB)-exposed areas. The anti-wrinkle effects of herbal extracts were investigated in an animal model. Female albino hairless mice (HR/ICR) were randomly allocated to the control group (non-irradiated vehicle), positive control group (UVB irradiated-vehicle), and two herbal extract mixture groups (HE-1 and HE-2). HE-1 included Glycyrrhizae radix, Rhei Rhizoma, Cornus officinalis, and Sesami semeni, and HE-2 included Swertia pseudo-chinensis, Sophora flavescens, Scutellaria baicalensis, and Salvia miltiorrhiza. The herbal extract mixtures were pre-treated dorsally with 0.2 ml per individual five times per week for four weeks prior to the start of UVB irradiation. At the fifth week, the animals were exposed to UVB irradiation for a subsequent eight weeks, three times per week. The intensity of irradiation showed a gradual increase, from 30 mJ/cm 2 to 240 mJ/cm2 (1 MED: 60 mJ/cm2 ). Dorsal skin samples were stained with H&E in order to examine the epidermal thickness. In addition, Masson-Trichrome staining was performed for determination of the amount of collagen fiber. Treatments with HE-1&2 resulted in an increase in the amount of collagen fiber, a better appearance, and fewer wrinkles, compared with the positive control. As determined by hydroxyproline assay, treatments with HE-1&2 led to a significant increase in the amount of collagen, compared with the positive control group (p<0.05). Chronic UVB irradiation to skin of hairless mice resulted in an increase in expression of matrix metalloproteinase-1 (MMP-1), however, treatments with HE-1&2 tended to decrease the expression of MMP-1. These results indicate that the herbal extracts used in this study have a preventive effect on UVB-induced wrinkle formation in a hairless mouse model, due in part to inhibition of MMP-1 expression and increment of collagen amount.

Iron catalyzes the production of free radicals, which can be related to a variety of pathological events, such as cancer and aging. The effect of dietary iron was investigated on formation of colonic aberrant crypt foci (ACF) induced by azoxymethane in male F344 rats. Animals were fed three different diets, including iron-deficient (3 ppm Fe), iron-normal (35 ppm Fe), and iron-overloaded (350 ppm Fe) diets for eight weeks. During the first and second weeks of the experiment, animals received two subcutaneous injections of azoxymethane (AOM, 15 mg/kg body weight) to induce ACF. After staining with methylene blue, the total numbers of ACF and aberrant crypts (AC) were counted on the colonic mucosa. Analysis of blood and serum was performed using a blood cell differential counter and an automatic serum analyzer. Iron-deficient diets induced a significant decrease in red blood cell counts and the values of hemoglobin concentration, hematocrit, mean corpuscular hemoglobin, and mean corpuscular volume, while an iron-overloaded diet did not affect these values. The iron-overloaded diet induced an increase in deposits of iron in the liver of rats, as determined by the ICP method and Perl’s staining. The numbers of ACF per colon showed a slight increase in iron-overloaded or iron-deficient rats, without statistical significance, compared to iron-normal rats. The number of total AC per colon in iron-overloaded rats was significantly higher than that in iron-normal rats (p<0.05). The number of large ACF (≥ 4 AC per ACF) in iron-overloaded rats was also significantly higher than that in iron-normal rats (p<0.05). These results suggest that dietary iron intake may play an important role in colon carcinogenesis in humans and animals.

We report for the first time the occurrence of DWV-infected bumble bees (Bombus ignitus). For the present study, the detection of DWV virus from the female and male bumble bee was investigated in the same colony. The Deformed wing virus (DWV) of honeybee (Apis mellifera) is closely associated with characteristic wing deformities, abdominal bloating, paralysis, and rapid mortality of emerging adult bees. Using specific RT-PCR protocols for the detection of DWV followed by sequencing of the PCR products we could demonstrate that the bumble bees were indeed infected with DWV. The virus was detected from Bombus ignitus, and its partial DWV gene was cloned and sequenced. The partial DWV gene encoding the polyprotein is 711-nt of 235 amino acid residues. The deduced nucleotide sequence of the polyprotein partial gene of DWV showed 96.9%, 96.2%, 96.8%, and 96.5% homology to other structure polyprotein partial gene of DWV from insects, respectively. Phylogenetic analysis further conformed that the deduced nucleotide sequence of the polyprotein partial gene of DWV divided to the outside tree. We describe the first time that presence of Deformed wing virus(DWV) from bumble bee(Bombus terrestris) in korea using RT-PCR.

Although hair disorders are not life threatening, a lot of people who suffer hair loss and/or hair thinning is increasing in accordance with changes in lifestyle and nutritional balance. The aim of this study was to examine the effects of herbal extracts on hair regrowth in C3H/HeJ mice. There were 6 experimental groups including distilled water (D.W.), 50% ethanol (a vehicle control), 3% minoxidil (a positive control), and 3 kinds of herbal extracts mixtures (C, D & E). The test compounds included followings; C : Glycyrrhizae radix, Rhei Rhizoma, Cornus officinalis and Sesami semeni, D : Viticus fructus, Pulsatilla chinensis, Gardenia fructus and Artemisiae argyi herb, E : Swertia pseudo-chinensis, Sophora flavescens Scutellaria baicalensis and Salvia miltiorrhiza. The animals were shaved with an electric clipper. The test compounds were daily treated to dorsal skin with 0.2 ml per mouse for 3 weeks. The topical application of the E test solution accelerated hair regrowth after 10 days faster than that of the positive and vehicle controls. The activities of alkaline phosphatase (ALP) and γ-glutamyl transpeptidase significantly were increased in all the treatment groups after 3 weeks, compared with D.W. group. Especially, the E test solution notably increased ALP activity compared with positive or vehicle control group. Epidermal growth factor (EGF) and vascular endothelial growth factor (VEGF) were increased in all the treatment groups after 3 weeks compared with D.W. group. These results suggest that the herbal extracts have hair regrowth effect by increasing enzyme activities and growth factors and it can be useful for treatment for alopecia in humans.