Bifidobacterium longum IM55 and Lactobacillus plantarum IM76 alleviate allergic rhinitis in mice by restoring Th2/Treg imbalance and gut microbiota disturbance.

This study aimed to examine whether probiotics, which suppressed the differentiation of splenic T cells into type 2 helper T (Th2) cells and induced into regulatory T cells in vitro, alleviate allergic rhinitis (AR) and gut microbiota disturbance. We isolated Bifidobacterium longum IM55 and Lactobacillus plantarum IM76 from human faecal microbiota and kimchi, respectively, and examined their effects on ovalbumin (OVA)-induced AR and gut microbiota disturbance in mice. Treatment with IM55, IM76, or their probiotic mixture (PM) significantly reduced OVA-induced allergic nasal symptoms and blood immunoglobulin E (IgE) levels in mice. These also reduced OVA-induced interleukin (IL)-4 and IL-5 levels in nasal tissues and bronchoalveolar lavage fluid (BALF) but increased OVA-suppressed IL-10 levels. Treatment with IM55, IM76, or PM reduced OVA-induced increase in the populations of mast cells, eosinophils, and Th2 cells and increased OVA-suppressed population of regulatory T cells in the BALF. Treatment with IM55, IM76, or PM also inhibited OVA-induced expression of IL-5 in lung and colon tissues and restored OVA-disturbed composition of gut microbiota Proteobacteria, Bacteroidetes, and Actinobacteria. These results suggest that IM55 and IM67 can alleviate AR by restoring Th2/Treg imbalance and gut microbiota disturbance.

Effect of high advanced-collagen tripeptide on wound healing and skin recovery after fractional photothermolysis treatment.

BACKGROUND: Collagens have long been used in pharmaceuticals and food supplements for the improvement of skin. AIM: We evaluated the efficacy of high advanced-collagen tripeptide (HACP) on wound healing and skin recovery. METHODS: Using an in vitro model, we performed HaCaT cell migration assays and collagen gel contraction assays using HACP concentrations of 1, 10 and 100 µg/mL. In this pilot study, eight healthy volunteers were randomly divided into two groups. Both the control and experimental groups received fractional photothermolysis treatment, but in the experimental group, four subjects received 3 g/day of oral collagen peptide (CP) for 4 weeks. To assess transepidermal water loss in each patient before and after the treatment, we used a Corneometer and a Cutometer, and we also assessed the patient's Erythema Index. RESULTS: The cell migration assay showed that HACP enhanced wound closure, but not in a dose-dependent manner. The collagen gel contraction assay showed increased contractility when patients were treated with 100 µg/mL HACP, but the results were not significantly different from those of controls. We found that post-laser erythema resolved faster in the experimental group than in the control group (P < 0.05). In addition, the recovery of skin hydration after fractional laser treatment was greater in the experimental group than in the control group by day 3 (P < 0.05), and the experimental group showed significantly improved post-treatment skin elasticity compared with the controls by day 14 (P < 0.05). CONCLUSIONS: Collagen tripeptide treatment appears to be an effective and conservative therapy for cutaneous wound healing and skin recovery after fractional photothermolysis treatment.

First Report of Fusarium succisae Causing Flower Rot on Thread-leaf Coreopsis.

In July 2010, flower rot of thread-leaf coreopsis (Coreopsis verticillata) was found in a garden in the Icheon City, Korea. The disease affected about 20 to 50% of a 100 m2 area. The disease was characterized by the appearance of pinkish mycelia on the stigmata and inflorescences of flowers. In some cases, flowers failed to bloom or turned brown before opening fully. Fragments (each 5 × 5 mm) of the symptomatic tissue were surface-sterilized with 1% NaOCl for 1 min, and then rinsed twice in sterilized distilled water. The tissue pieces were placed on water agar (WA) and incubated at 25°C for 4 to 6 days. Twenty-two isolates of Fusarium species were obtained from the diseased flowers. All isolates were identified as Fusarium succisae based on their morphological characteristics on carnation leaf agar (CLA) medium and DNA sequences of the translation elongation factor 1-alpha gene (1). Macroconidia and sporodochia were sparsely produced on CLA medium. Microconidia were abundant, borne in false heads, oval or allantoid and sometimes pyriform, and measured 4.2 to 13 × 2.2 to 5.4 µm. Chlamydospores were absent. The EF-1α gene was amplified from three isolates by PCR assay and the amplification products were sequenced (2). The nucleotide sequences obtained were deposited in GenBank with accession numbers KF514658, KF514659, and KF514660. BLASTn analysis showed 99% homology with the EF-1α sequence of F. succisae NRRL13613 (GenBank Accession No. AF160291). Pathogenicity tests were conducted with inoculation of flowers on Coreopsis verticillata. Spore suspension was prepared by flooding 7-day-old cultures on potato dextrose agar with sterilized 2% (w/v) sugar solution. When the plants started to have buds, the isolates were inoculated by placing one drop (20 µl) of spore suspension (1 × 106 spores ml-1) into the buds. Fifteen buds of the plants were arranged into three replications. The control was treated with sterilized 2% sugar solution. Inoculated plants were kept in a greenhouse at 25/20°C (12 h/12 h). Three weeks after inoculation, the symptoms were observed on buds with mycelial production. Control plants had no mycelia on buds. F. succisae was re-isolated from the inoculated flowers. To our knowledge, this is the first report of flower rot of thread-leaf coreopsis caused by F. succisae. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (2) K. O'Donnell et al. Proc. Nat. Acad. Sci. 95:2044, 1998.

Effects of Korean ginseng berry extract (GB0710) on penile erection: evidence from in vitro and in vivo studies.

Several reports have promoted the root-derived Korean red ginseng (KRG; Panax ginseng) as alternative treatment for erectile dysfunction (ED), and ginsenosides are known to be the principal active ingredients of ginseng. Recent studies showed that ginseng berries produce more ginsenosides than KRG; thus, we investigated the ability of the Korean ginseng berry extract GB0710 to relax the penile corpus cavernosum smooth muscle (CCSM) in this study. As a comparative control, the results were compared to those obtained using KRG. In addition, possible mechanisms of action for GB0710 were investigated. While KRG and GB0710 both displayed dose-dependent relaxation effects on precontracted rabbit CCSM in vitro, GB0710 was shown to be more potent than KRG. The GB0710-induced relaxation could be partially reduced by removing the endothelium. In addition, pre-treatment with several nitric oxide (NO) inhibitors significantly inhibited the relaxation of muscle strips. Furthermore, administration of GB0710 increased intracavernosal pressure (ICP) in a rat in vivo model in both a dose- and duration-dependent manner. Intracellular NO production in human microvascular endothelial cells could be induced by GB0710 and inhibited by N(G)-monomethyl-L-arginine. In conclusion, GB0710 had a greater relaxation effect on rabbit CCSM than did KRG extract, and increased ICP in a rat model in both a dose- and a duration-dependent manner. This relaxing effect might be mediated by NO production.

Effects of Korean ginseng berry extract on sexual function in men with erectile dysfunction: a multicenter, placebo-controlled, double-blind clinical study.

Ginseng is beneficial for many aspects of human physiology, including sexual function. In this study, we have evaluated the efficacy and safety of an extract of ginseng berry, which has a ginsenoside profile distinct from other parts of the plant, on sexual function in men with erectile dysfunction. In all, 119 men with mild-to-moderate ED participated in a multicenter, randomized, double-blind, parallel, placebo-controlled clinical study. They were administered 4 tablets of either standardized Korean ginseng berry (SKGB, 350 mg ginseng berry extract per tablet), or placebo, daily, for 8 weeks. Efficacy was assessed with the International Index of Erectile Function (IIEF)-15 and premature ejaculation diagnostic tool (PEDT) at the end of the 4th and 8th week. We observed that the total and each of the individual domain scores of IIEF-15 increased from 40.95 ± 7.05 to 46.19 ± 12.69 significantly in the SKGB by the 8th week (P<0.05). The erectile function domain of IIEF changed slightly from 17.17 ± 2.57 to 18.59 ± 5.99 in the SKGB group by the 8th week (P<0.05). In addition, PEDT scores significantly improved from 9.14 ± 4.57 to 7.97 ± 4.4 and 7.53 ± 4.26 in the SKGB group after 4 and 8 weeks of treatment (P<0.05). Safety markers including hormone and lipid in the blood were assessed at the end of the 4th and 8th week and they remained unchanged. Oral administration of the SKGB extract improved all domains of sexual function. It can be used as an alternative medicine to improve sexual life in men with sexual dysfunction.

Basal STAT3 activities are negatively correlated with tumor size in papillary thyroid carcinomas.

BACKGROUNDS: Signal transducer and activators of transcription-3 (STAT3) plays a critical role in promoting survival and cell growth as well as facilitating angiogenesis and metastasis in several cancers. AIM: This investigation focused on evaluation of STAT3 activities in human papillary thyroid cancers (PTC). METHODS: STAT3 activities of nuclear extracts of tumor tissue were measured from 35 PTC patients using enzyme- linked immunosorbent assay-based kits. RESULTS: STAT3 activities of PTC tissues were significantly lower than those of surrounding normal thyroid tissues [0.36 (interquartile range 0.24-0.72) vs 0.50 (0.29-1.11) arbitrary units, p<0.01]. We further analyzed the association between STAT3 activity and clinicopathologic factors in PTC tissue. Tumors with size ≥2 cm displayed significantly lower STAT3 activities than those <2 cm [0.25 (0.21-0.37) vs 0.53 (0.37-0.61) arbitrary units, p<0.01]. Notably, tumor size was inversely correlated with STAT3 activities in T1799A BRAF mutation-positive cases (Rs=-0.58, p<0.05), but not mutation-negative cases. CONCLUSIONS: STAT3 activities of PTC measured via DNA binding are suppressed in contrast to other human cancers. Tumor size larger than 2 cm is the only clinicopathologic parameter associated with low STAT3 activity. Moreover, tumor size appears inversely correlated with STAT3 activity, specifically in T1799A BRAF mutation-positive cases.

First Report of Internal Fruit Rot of Sweet Pepper in Korea Caused by Fusarium lactis.

In June 2010, an internal fruit rot of sweet pepper (Capsicum annuum L.) fruit was found in a commercial greenhouse in Ilsan City, Korea. Disease incidence reached approximately 5% of 30 tons of harvested peppers. Affected fruits commonly did not show external symptoms. However, when the fruit was cut open, an internal rot and pinkish gray mycelium were observed on the seeds and the inner surface of fruit. Discolored soft patches, browning, and necrosis were observed on the outer surface of some fruits. Fragments (5 × 5 mm2) of the affected tissues were surface sterilized with 1% NaOCl for 30 s and then rinsed twice in sterile distilled water. The pieces were placed on water agar and incubated at 25°C for 4 to 6 days. Twenty-nine Fusarium isolates were obtained from 12 diseased fruits and maintained on synthetic low nutrient agar (SNA) at 10°C. The isolates were cultured on carnation leaf agar (CLA) and SNA at 23°C with 12 h of near-ultraviolet light per day for 14 days. Microconidia were abundant, borne in short, zig-zag chains or false heads, obovoid or clavate with a flattened base, and measured 4.3 to 7.1 × 2.2 to 3.3 µm. Macroconidia were sparse, thin walled, slender, straight to slightly curved, and measured 32 to 48 × 2.8 to 3.9 µm. Sporodochia were rare on CLA and chlamydospores were absent. The translation elongation factor 1-alpha (EF-1α) gene was amplified from four isolates (SPF01, SPF09, SPF16, and SPF22) by PCR assay using ef1 and ef2 primers (2), and the 700-bp amplification products were sequenced. The nucleotide sequences were deposited in GenBank (Accession Nos. JF411956 to JF411959). BLAST analysis showed 98% homology with the EF-1α sequence of Fusarium lactis NRRL25200 (GenBank Accession No. AF160272). All isolates were identified as F. lactis based on morphological and molecular characteristics (1). Pathogenicity tests of the four isolates were conducted by inoculating flowers on plants of the orange pepper cv. Orange Glory (3). A spore suspension was prepared by flooding 5-day-old cultures on potato dextrose agar with sterile distilled water. When the plants started to flower, each flower was inoculated by placing 20 µl of spore suspension (1 × 106 conidia/ml) on each flower. Four isolates of F. lactis were each inoculated onto three flowers on each of seven plants. Flowers from the same number of plants inoculated with sterile distilled water were used as the control treatment. Inoculated plants were kept in a greenhouse at 25°C by day and 20°C by night. Sixty days after inoculation, mature fruits were harvested and cut open to check for internal rot. Approximately 70% of inoculated fruits showed internal rot and pinkish gray mycelial growth on the inner surface of the fruits. No symptoms were observed on the control fruits. Fungal cultures resembling F. lactis were reisolated from inoculated fruits for all four isolates, fulfilling Koch's postulates. F. lactis has been reported on sweet pepper in the Netherlands and Canada (3). To our knowledge, this is the first report of internal fruit rot of sweet pepper caused by F. lactis in Korea. Although disease severity was low in this greenhouse, the economic impact on sweet pepper could be significant because the disease can reduce the quality, quantity, and market value of pepper fruits. References: (1) H. I. Nirenberg and K. O'Donnell. Mycologia 90:434, 1998. (2) K. O'Donnell et al. Proc. Natl. Acad. Sci. USA 95:2044, 1998. (3) J. Yang et al. Can. J. Plant Pathol. 31:47, 2009.

Effects of gamma irradiation and electron beam irradiation on quality, sensory, and bacterial populations in beef sausage patties.

This study compared effects of gamma ray (GR) and electron beam (EB) irradiation on quality (TBARS value, hardness, color), sensory characteristics, and total bacterial populations in beef sausage patties during accelerated storage at 30 degrees C for 10days. Beef sausage patties were vacuum-packaged and irradiated by GR and EB at 0, 5, 10, 15, and 20kGy at room temperature. The results of quality evaluation showed that the effects of GR irradiation were similar (p0.05) to EB irradiation on lipid oxidation, hardness, color and sensory scores of the beef sausage patties. However, GR-irradiated samples had lower (p<0.05) total bacterial counts than EB-irradiated samples after irradiation, and during storage regardless of irradiation dose. The results indicate that use of GR irradiation up to 10kGy on patties should be useful in reducing bacterial populations with no adverse effect on quality and most of sensory characteristics (color, chewiness, and taste).

A New Disease, Bacterial Leaf Spot of Rape, Caused by Atypical Pseudomonas viridiflava in South Korea.

In March 2007, a bacterial leaf spot of rape (Brassica napus var. oleifera) was observed in fields near Seogwipo City, Jeju Province, South Korea. Symptoms on leaves included white and corky-brown spots and sometimes water-soaked spots on the lower leaf surface. Seven bacterial isolates (BC2495-BC2497 and BC2506-BC2509) were recovered on trypticase soy agar (TSA) from leaf spot lesions surface sterilized in 70% ethyl alcohol for 1 min. Isolates were gram-negative, aerobic rods with one to three flagella. Pathogenicity was evaluated on 2-week-old rape plants by spot and spray inoculation. Bacteria were grown on TSA for 48 h at 25°C. Five microliters of bacterial suspension in sterile distilled water (1 × 105 CFU/ml) were spot inoculated on pinpricked positions of five detached leaves for each isolate. The detached leaves were incubated in a plastic box with high humidity at 20°C. Spot-inoculated surfaces turned white 48 h after inoculation followed by a brownish discoloration. A bacterial suspension in sterile distilled water (100 ml at 1 × 105 CFU/ml) was sprayed onto three plants for each isolate. Plants were maintained in a growth chamber at 20°C and 90% relative humidity. Isolates induced identical symptoms 2 weeks after spray inoculation as those originally observed on rape in the fields. Bacteria were reisolated 18 days after inoculation from diseased lesions surface sterilized in 70% ethyl alcohol for 1 min. Pathogenicity of the reisolated bacteria was confirmed by spot inoculation as described above. No symptoms were noted on detached leaves and intact plants inoculated with sterilized distilled water. Using the Biolog Microbial Identification System, Version 4.2 (Biolog Inc., Hayward, CA), the isolates were identified as Pseudomonas viridiflava with a Biolog similarity index range of 0.52 to 0.72 after 24 h. Results of LOPAT tests (2) of isolates were identical to that of atypical P. viridiflava reported by Gonzalez et al. (1). Levan production and pectolytic activity of the isolates were variable. All isolates were positive for tobacco hypersensitivity and negative for oxidase reaction and arginine dihydrolase production. The 16S rDNA region (1,442 bp) of the isolates (GenBank Accession Nos. HM190218-HM190224; P. viridiflava CFBP2107T = HM190229), amplified by using universal PCR primers, shared 100% sequence identity with atypical P. viridiflava (GenBank Accession No. AM182934) (1). The gyrB sequence (638 bp) from the isolates (GenBank Accession Nos. HM190232-HM190238; P. viridiflava CFBP2107T = HM190239), amplified by using previously reported PCR primers (3), had a distance index value range of 0.029 to 0.031 with that of the P. viridiflava CFBP2107T (=BC2597) as determined by Jukes-Cantor model using MEGA Version 4.1 (4). On the basis of phenotypic characteristics and the sequences, the seven isolates were identified as atypical P. viridiflava. The disease is named "bacterial leaf spot". To our knowledge, this is the first report of bacterial leaf spot of rape caused by atypical P. viridiflava. References: (1) A. J. Gonzalez et al. Appl. Environ. Microbiol. 69:2936, 2003. (2) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (3) H. Sawada et al. J. Mol. Evol. 49:627, 1999. (4) K. Tamura et al. Mol. Biol. Evol. 24:1596, 2007.

Effects of oral epigallocatechin gallate supplementation on the minimal erythema dose and UV-induced skin damage.

BACKGROUND/AIMS: Excessive exposure to UV radiation causes acute adverse effects like sunburn and photosensitivity reactions and is involved in the induction and development of skin cancer. It has been reported that antioxidants have photoprotective effects against solar UV radiation. We investigated the effect of oral epigallocatechin gallate (EGCG), a powerful antioxidant in green tea, on the minimal erythema dose (MED) and UV-induced skin damage. METHOD: Female HWY/Slc hairless rats were fed the normal diet supplemented with 1,500 ppm EGCG for 8 weeks; then, the MED was determined and visual scores and transepidermal water loss were assessed to evaluate the severity of UV-induced skin damage. RESULTS: At week 8 of the study, the use of dietary EGCG significantly increased MED. UV-radiation-induced sunburn severity and alterations in epidermal barrier function were also attenuated by the supplementation of EGCG. CONCLUSION: Regular intake of EGCG strengthens the skin's tolerance by increasing MED and thus prevents UV-induced perturbation of epidermal barrier function and skin damage. These results suggest that EGCG is a potent candidate for systemic photoprotection.

Wildfire of Soybean Caused by Pseudomonas syringae pv. tabaci, a New Disease in Korea.

In 2006 and 2007, a new bacterial disease was observed in field-cultivated soybeans in Boeun District and Munkyung City of Korea. The disease caused severe blighting of soybean (Glycine max) leaves. Soybean leaves in fields showed yellowish spots with brown centers. Brown and dead areas of variable size and shape were surrounded by wide, yellow haloes with distinct margins. Spots might coalesce and affected leaves fell readily. Seven bacterial strains were isolated from chlorotic areas of soybean leaves and all produced white colonies on trypticase soy agar. With the Biolog Microbial Identification System, version 4.2, (Biolog Inc., Hayward, CA) all strains and Pseudomonas syringae pv. tabaci CFBP2106T were identified as P. syringae pv. tabaci with a Biolog similarity index of 0.28 to 0.52 and 0.48 after 24 h. Pathogenicity of the strains (three plants per strain) on soybean leaves at the V5 stage (cv. Hwanggeum) was confirmed by rub inoculation with bacterial suspensions (1 × 108 CFU/ml) in sterile distilled water on the lesions cut 1 cm long on the upper side of the leaves with razor blades and by pinprick on 3-week-old leaves of tobacco (Nicotiana tabacum cv. Samsun) in the greenhouse. Wildfire symptoms on the soybean leaves and faint halos on tobacco leaves were observed 4 days after inoculation. The identification of reisolated bacterial strains was confirmed with the metabolic fingerprintings on Biolog. LOPAT tests (1) and phenotypic characteristics (3) of the strains were similar to those of the CFBP2106T. Colonies were levan positive, oxidase negative, potato soft rot negative, arginine dihydrase negative, and tobacco hypersensitivity negative. All strains were gram-negative, aerobic rods with a polar flagellum. Strains were negative for esculin hydrolysis, gelatin liquefaction, urea production, accumulation of poly-ß-hydroxy butyrate, starch hydrolysis, ornithine dihydrolase, lysine dihydrolase, growth at 37°C, utilization of geraniol, benzoate, cellobiose, sorbitol, trehalose, l-rhamnose, and adonitol. Positive reactions were catalase and arbutin hydrolysis, utilization of sorbitol, d-arabinose, and dl-serine. The strains were variable in utilization of mannitol, sucrose, and d-arabinose. The 1,472-bp PCR fragments of strains, BC2366 (GenBank Accession No. FJ755788) and BC2367 (No. FJ755789) was sequenced using 16S rDNA universal primers (2). The sequences shared 100% identity with the analogous sequences of P. syringae pv. glycenea (GenBank Accession No. AB001443) available in NCBI databases. Based on the phenotypic, genetic, and pathological characteristics, all strains were identified as P. syringae pv. tabaci. To our knowledge, this is the first report of P. syringae pv. tabaci causing wildfire on soybean in Korea. References: (1) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (2) I.-S. Myung et al. Plant Dis. 92:1472, 2008. (3) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001.

First Report of Bacterial Canker of Tomato Caused by Clavibacter michiganensis subsp. michiganensis in Korea.

In 2007, a new bacterial disease was observed in greenhouse-cultivated cherry tomatoes in Cheorwon and Iksan provinces, Korea. The disease caused severe wilt of tomatoes (Solanum lycopersicum cv. Koko). Infected young petioles were curled downward. Margins of the leaves rolled upward and whole leaves were distorted. Stem cankers had reddish or dark brown cavities. Vascular tissues in stems cut longitudinally were brown to deep brown, but no bird's eye lesions were observed. Eight bacterial strains recovered from the stems of wilted tomatoes produced yellow colonies on nutrient broth-yeast extract agar and pink colonies on triphenyl tetrazolium chloride. Pathogenicity of the strains (three plants per strain) on 18-day-old tomatoes (cv. Koko) was confirmed by clip inoculation of petioles of second leaves and spray inoculation with bacterial suspensions (1 × 108 CFU/ml) in sterile distilled water. Wilt and canker symptoms were observed 2 weeks after inoculation. Symptoms produced by both inoculation methods were systemic and localized. Clip inoculation of tomatoes resulted in wilt, defoliation, and open stem cankers, whereas small, white spots (2 to 3 mm in diameter) and sometimes water-soaked, dark brown-to-black lesions on the leaf margins were observed with spray inoculation. Bacteria were reisolated from stems and leaves of the inoculated plants and their identities confirmed by direct PCR using specific primer set CMM5/CMM6 (1). No symptoms were observed on negative control plants inoculated with sterile water. All strains were gram-positive aerobic rods with no polar flagella. Strains were positive for esculin hydrolysis, gelatin liquefaction, H2S production from peptone, utilization of citrate and succinate, and acid from d(+)mannose and negative for starch hydrolysis, casein hydrolysis, methyl red reaction, acid from inulin, mannitol, d(+)-melezitose and d(-)sobitol, and utilization of acetate, formate, lactate, propionate, and ribose. Identification as C. michiganensis subsp. michiganensis was confirmed using 16S rDNA universal primers fD1 and rP2 (4) and internal primers (3). The 1,439-bp PCR fragment of strain BC2643 was sequenced (GenBank Accession No. EU685335) and compared with reference C. michiganensis subspecies strains in GenBank: AM410696 (C. michiganensis subsp. michiganensis), AM410693 (C. michiganensis subsp. tessellarius), AM410697 (C. michiganensis subsp. nebraskensis), AM410694 (C. michiganensis subsp. sepedonicus), and AM410695 (C. michiganensis subsp. insidiosus). The sequence had a similarity index of 0.999 calculated by Juke-Cantor model (2) with the 16S rRNA sequence of C. michiganensis subsp. michiganensis (AM410696). The fragment size of eight strains amplified by PCR using CMM5/CMM6 (1) was identical to that of the C. michiganensis subsp. michiganensis reference strain KACC20122. On the basis of the physiological, genetic, and pathological characteristics, all strains were identified as C. michiganensis subsp. michiganenesis. To our knowledge, this is the first report of C. michiganensis subsp. michiganenesis causing bacterial canker on tomato in Korea. References: (1) J. A. Dreier et al. Phytopathology 85:464, 1995. (2) S. Kumar et al. Brief. Bioinform. 5:50, 2004. (3) S. W. Kwon et al. Int. J. Syst. Bacteriol. 47:1061, 1997. (4) W. G. Weinsburg et al. J. Bacteriol. 173, 697, 1991.

Histopathologic changes of acellularized xenogenic carotid vascular grafts implanted in a pig-to-goat model.

INTRODUCTION: The present study was designed to determine the in vivo patency and recellularization pattern of acellularized small-diameter xenogenic arterial grafts. We implanted acellularized porcine carotid arteries in bilateral carotid arteries of goats and microscopically analyzed the recellularization pattern of these grafts with the recipient's cells over time. MATERIAL AND METHODS: Carotid arteries of pigs weighing 30-40 kg were harvested and decellularized with hypertonic saline followed by sodium dodecyl sulfate. Acellularized porcine carotid vascular xenografts (0.4-0.5 cm in diameter) were prepared into 4 cm-long segments and implanted bilaterally in the carotid arteries of 10 black-haired goats. The in vivo patency of the implanted acellularized xenogenic grafts was evaluated at regular intervals by color Doppler ultrasonography. The goats were sacrificed at predetermined intervals (1 week, 1 month, 3 months, 6 months, 12 months after implantation), two animals at each interval. Upon retrieval, visual inspections and histopathologic examinations of the grafts were performed. To identify smooth muscle cells and functioning endothelial cells, immunohistochemical staining for alpha-smooth muscle actin and von Willebrand factor were also performed. RESULTS AND CONCLUSIONS: All experimental animals survived the observation period. Nineteen out of 20 implanted grafts showed patency with no thrombi. Microscopic analysis revealed that the grafts were completely covered with the hosts' endothelial cells, beginning from anastomotic sites. The grafts were gradually recellularized with recipients'cells including fibroblasts, myofibroblasts and smooth muscle cells. In conclusion, this study suggested that acellularized xenogenic vascular grafts can be a good alternative for the small-diameter vascular graft.

Comparative analysis of frozen and acellularized vascular xenografts.

BACKGROUND: We implanted frozen and acellularized porcine xenograft vessels as small-diameter arterial grafts in goats and comparatively analyzed the explanted grafts by gross observation and by light microscopy at predetermined periods. MATERIALS AND METHODS: Porcine carotid arteries were harvested and immediately stored within a tissue preservation solution at -70 degrees C in a freezer designated for frozen xenograft vessels. The acellularized xenograft vessels were prepared with NaCl-SDS solution and stored frozen until use. One pair of porcine xenograft vessels were used to compare the frozen and acellularized grafts in the bilateral carotid arteries in one goat. The grafts were implanted for one, 3, and 6 months in three animals. Periodic ultrasonographic examinations were performed during the observation period. Explanted grafts were analyzed by gross observation, and by light microscopy. RESULTS: All animals survived the experimental procedure without specific problems. Ultrasonographic examinations showed excellent patency in all grafts during the observation period. Gross observations revealed nonthrombotic patent smooth lumens. Microscopic examinations of the explanted grafts showed satisfactory cellular reconstruction to the 6-month stage. Although more inflammatory responses were observed in the early phase of implantation of frozen xenografts than of acellularized xenografts, there was no evidence of significant rejection of the frozen xenografts. CONCLUSION: These findings suggest that porcine vessel xenografts, regardless of them being acellularized or simply frozen xenografts, can be acceptably implanted in goats as a form of small-diameter vascular graft.

Biological control agent of common scab disease by antagonistic strain Bacillus sp. sunhua.

AIMS: To identify an antagonistic strain against Streptomyces scabiei and to characterize the antibiotic agent. The efficacy of the isolated strain in controlling common scab disease was also evaluated. METHODS AND RESULTS: A bacterial strain antagonistic against S. scabiei was isolated from the soil of a potato-cultivating area. This bacterium was identified as a Bacillus species by 16S rRNA gene sequence analysis and was designated Bacillus sp. sunhua. Antibiotics produced by this strain were proven to be stable within a broad pH range and at high temperatures. The culture broth was extracted with ethyl acetate, and then the crude extract was applied to HPLC. Two compounds were isolated and identified as iturin A and macrolactin A by 1H-NMR, 13C-NMR, HMBC, HMQC and mass spectrometer. The culture broth of Bacillus sp. sunhua had a suppressive effect on common scab disease in a pot assay, decreasing the infection rate from 75 to 35%. This strain also suppressed Fusarium oxysporum, the pathogen of potato dry rot disease. CONCLUSIONS: Bacillus sp. sunhua was shown to inhibit S. scabiei effectively. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report demonstrating that macrolactin A and iturin A inhibit S. scabiei. This study demonstrated the possibility of controlling potato scab disease using Bacillus sp. sunhua.

Terrein: a new melanogenesis inhibitor and its mechanism.

Terrein is a bioactive fungal metabolite whose effects are almost unknown. In this study, we found for the first time that terrein has a strong hypopigmentary effect in a spontaneously immortalized mouse melanocyte cell line, Mel-Ab. Treatment of Mel-Ab cells with terrein (10-100 microM) for 4 days significantly reduced melanin levels in a dose-dependent manner. In addition, terrein at the same concentration also reduced tyrosinase activity. We then investigated whether terrein influences the extracellular signal-regulated protein kinase (ERK) pathway and the expression of microphthalmia-associated transcription factor (MITF), which is required for tyrosinase expression. Terrein was found to induce sustained ERK activation and MITF down-regulation, and luciferase assays showed that terrein inhibits MITF promoter activity in a dose-dependent manner. To elucidate the correlation between ERK pathway activation and a decreased MITF transcriptional level, PD98059, a specific inhibitor of the ERK pathway, was applied before terrein treatment and found to abrogate the terrein-induced MITF attenuation. Terrein also reduced the tyrosinase protein level for at least 72 h. These results suggest that terrein reduces melanin synthesis by reducing tyrosinase production via ERK activation, and that this is followed by MITF down-regulation.

Home care artificial heart monitoring system via internet.

The availability of a remote management system, which provides both physiological-related information about the patient and device-related information about the implanted device, would be helpful during in vivo experiments or clinical trials involving artificial heart implantation. In order to be able to monitor the course of the in vivo experiment continuously regardless of the patient's location, an internet-based remote monitoring system was developed, which can monitor physiological-related information such as pressure (AoP, LAP, RAP, PAP) and flow data, as well as device-related information such as current, direction and pump operating conditions. The home care artificial heart monitoring system which we developed consists of four main components, which are the transcutaneous information transmission system (TITS), local monitoring station (LMS), data server station (DSS), and client monitoring station (CMS). The device-related information and physiological-related information can be transmitted in real time from a patient in a remote non-clinical environment to the specialist situated in a clinic depending on the current capabilities and availability of the internet. The local monitoring station situated at the remote site is composed of a data acquisition and preprocessing unit connected to a computer via its RS-232 port, and which communicate using a Java-based client-server architecture. The remote monitoring system so developed was used during an in vivo experiment of the artificial heart implantation for 2 months and performed successfully according to design specifications.

Tissue-engineered acellularized valve xenografts: a comparative animal study between plain acellularized xenografts and autologous endothelial cell seeded acellularized xenografts.

BACKGROUND: Acellularized valve xenografts are considered a promising way of overcoming the inherent limitations of current prosthetic valves. The aim of this study was to compare the biological responses of an autologous endothelial cell seeded acellularized xenograft (AAX) and a plain acellularized xenograft (PAX) implanted in the pulmonary valve leaflet in the same animal. METHODS: Endothelial cells were isolated and cultured from the jugular vein of goats. Porcine valve leaflets were acellularized with Nacl-SDS, and for AAX, leaflets were then seeded with autologous endothelial cells. A PAX and an AAX were implanted as double pulmonary valve leaflet replacement in the same animal in a goat model (n = 6). After sacrifice, the implanted valve leaflet tissues were retrieved and analyzed visually and under a light microscope. RESULTS AND CONCLUSIONS: Six animals were sacrificed as scheduled during the short-term (6 and 24 hours), mid-term (1 week and 1 month) and long-term (3 and 6 months). Gross and ultrasonographic examinations revealed good valve function with no thrombosis but with slight thickening. Microscopic analysis of the leaflets showed abundant cellular ingrowth into the acellularized leaflets over time. The role of endothelial cell seeding remains controversial. This animal experiment demonstrates the practical feasibility of using acellularized valve xenografts.

A chronic heart failure model by coronary artery ligation in the goat.

The availability of a reliable heart failure model in large animals is important. We report upon our efforts to develop a chronic heart failure model in seven goats using sequential ligation of the left anterior descending (LAD) coronary artery and its diagonal branch. After anesthesia and left thoracotomy, the LAD artery was ligated, and the diagonal vessel at the same level was ligated one hour later. Cardiac measurements were performed with a thermodilution catheter and by ultrasonography. Two months after the operation, the same measurements were made and animals were sacrificed for postmortem examinations of their hearts. Hemodynamic measurements, except cardiac output, showed no significant changes immediately after the coronary artery ligation. Echocardiographic measurements showed significant changes in the ejection fraction and fractional shortening without changes in left ventricular dimensions. Wall motion analyses demonstrated variable degrees of anteroseptal dyskinesia and akinesia in all animals immediately after coronary artery ligation. Five animals have undergone hemodynamic and ultrasonographic studies 2 months after coronary artery ligation. The results obtained from these animals showed significant increases in central venous pressure, right ventricular pressure, pulmonary artery pressure, and pulmonary artery capillary wedge pressure, and a significant decrease in cardiac output. Increases in left ventricular dimensions and decreases in ejection fraction with fractional shortening in ultrasonographic studies were also observed. Pathologically, well-demarcated thin-walled anteroseptal infarcts, with chamber enlargement, were clearly seen with dilatation of the heart chambers in all specimens. Based on this study, we conclude that goats, like sheep, can provide a reliable model of chronic heart failure by coronary artery ligation and in view of the many advantages offered by goats, we believe that this animal model will be useful for cardiac experimentation.

Autologous cardiomyocyte transplantation using a biodegradable polymer scaffold.

BACKGROUND: To achieve a more reliable way of transplanting cardiomyocytes, we conducted an autologous cardiomyocyte transplantation using a biodegradable scaffold, instead of a syringe injection, as a vehicle for transporting cells in an ovine myocardial infarction model. MATERIALS AND METHODS: A myocardial infarction was created in sheep using sequential ligation of the homonymous artery and its diagonal branch. Autologous cardiomyocytes from the right ventricular infundibulum were cultured and seeded onto a biodegradable polymer scaffold. Three months after creating myocardial infarction, the two animals were re-anesthetized and cardiomyocyte-seeded scaffolds were implanted in the infarcted area. The animals were kept alive for a further month, and then sacrificed for postmortem heart examinations. Light microscopic analysis and an immunohistochemical study for myoglobin were performed. RESULTS: On postmortem gross examinations, the polymer scaffolds were visible in the background of well-demarcated thin-walled anteroseptal myocardial infarcts. Microscopic analysis showed abundant myoglobin-stained cells between the fiber strands of the polymer scaffolds. However, there is a possibility that some of these cells might have been giant cells reacting to foreign material. CONCLUSION: The transplantation of cultured autologous cardiomyocytes into an infarct region using a biodegradable scaffold instead of syringe injection provides another promising option for cardiomyocyte transplantation, which warrants further study.