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.

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.

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.

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.

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.

Post-transcriptional inhibition of lipopolysaccharide-induced expression of inducible nitric oxide synthase by Gö6976 in murine microglia.

Glia in the brain respond to various toxins with an increased expression of inducible nitric oxide synthase (iNOS) and an increased production of nitric oxide (NO). Here, we report that lipopolysaccharide (LPS)-induced expression of iNOS was down-regulated post-transcriptionally through the destabilization of iNOS mRNA by the indolocarbazole compound, Gö6976, in murine microglia. This Gö6976 effect is specific for iNOS since tumor necrosis factor alpha was unaffected by the compound. Interestingly, the post-transcriptional effects ascribed to Gö6976 were not observed with other inhibitors of protein kinase A, C (PKC), G, or protein tyrosine kinases. Instead, these kinases appear to affect the iNOS/NO system at the transcriptional level. In the past, Gö6976 has been reported to be a rather specific inhibitor of PKC in vitro. Results from our experiments, through prolonged treatment with phorbol esters and with the various PKC inhibitors including phorbol ester-insensitive PKC isotype inhibitor, suggest that the Gö6976-mediated post-transcriptional regulation of iNOS gene expression and NO production in microglia is not mediated through its reputed effects on PKC activity. Since the effects of various neurotoxins and certain neurodegenerative diseases may be manifested through alterations in the iNOS/NO system, post-transcriptional control of this system may represent a novel strategy for therapeutic intervention.

Time dependency of the action of nitric oxide in lipopolysaccharide-interferon-gamma-induced neuronal cell death in murine primary neuron-glia co-cultures.

We investigated the time-dependency of the action of nitric oxide (NO) on glia-mediated neuronal cell death. Cortical neuron-glia co-cultures were treated with lipopolysaccharide and interferon gamma (LPS/IFNgamma). The production of NO was first detectable 9 h after the exposure to LPS/IFNgamma and increased for up to 48 h. A significant neuronal cell death was observed 36-48 h after treatment with LPS/IFNgamma. The NO generated at the initial stage of NO synthesis (about 12 h) following exposure to LPS/IFNgamma was found to be critical for LPS/IFNgamma-induced neurotoxicity. Furthermore, the rate of NO production at the initial stage of NO synthesis was correlated linearly with the extent of neuronal cell death. These findings suggest that the maximal rate of NO synthesis, instead of the accumulated NO(2)(-) level, is a sensitive index for predicting endotoxin-induced cytotoxicity.

Sesquiterpene O-naphthoquinones from the root bark of Ulmus davidiana.

Three new sesquiterpene ortho-naphthoquinones, davidianones A, B and C, together with four known compounds, mansonones E, F, H and I, were isolated from the root bark of Ulmus davidiana. On the basis of spectral data including pulse field gradient two-dimensional NMR spectroscopy, the structures of new compounds were established as 3-hydroxymethyl-6,9-dimethylnaphtho(1,8-b,c)pyran-7,8-dione, 6-methoxycarbonyl-3,9-dimethylnaphtho(1,8-b,c)pyran-7,8-dione, 6-dimethoxymethyl-3,9-dimethylnaphtho(1.8-b,c)pyran-7,8-d ion e, respectively. Their antioxidative activities were evaluated by a thiobarbituric acid method using rat liver microsomes, with mansonone F showing the greatest activity.

Benzastatins A, B, C, and D: new free radical scavengers from Streptomyces nitrosporeus 30643. II. Structural determination.

The structures of benzastatins A, B, C, and D, new free radical scavengers, were determined by spectroscopic studies. Benzastatins A and B incorporate the para-aminobenzamide unit which is rare in fungal metabolites. Benzastatins C and D are unique alkaloids related to virantmycin; they contain the tetrahydroquinoline unit in the molecules.

Benzastatins A, B, C, and D: new free radical scavengers from Streptomyces nitrosporeus 30643. I. Taxonomy, fermentation, isolation, physico-chemical properties and biological activities.

In the course of screening for free radical scavengers, rare metabolites, benzastatins A and B having aminobenzamide skeleton and benzastatins C and D having tetrahydroquinoline skeleton, were isolated from the culture broth of streptomyces nitrosporeus 30643. They showed inhibitory activity against lipid peroxidation in rat liver microsomes. In the cell assay, benzastatins C and D inhibited glutamate toxicity in N18-RE-105 cells with EC50 values of 2.0 and 5.4 microM, respectively.

New diphenazines with neuronal cell protecting activity, phenazostatins A and B, produced by Streptomyces sp.

Phenazostatins A and B, new diphenazine compounds, were isolated from the culture broth of Streptomyces sp. 833 as new neuronal cell protecting substances which also showed free radical scavenging activity. In the cell assay, phenazostatins A and B inhibited glutamate toxicity in N18-RE-105 cells with EC50 values of 0.34 and 0.33 microM, respectively.

Nitrofen-induced congenital malformations of the heart and great vessels in rats: an animal model.

BACKGROUND: Nitrofen (2,4-dichloro-4'-nitrodiphenyl ether), a diapheny ether herbicide, is known to induce in rat fetuses a variety of congenital cardiovascular anomalies, together with diaphragmatic hernia and hydronephrosis. The purpose of the current study was to produce congenital cardiovascular anomalies in rat fetuses by oral nitrofen administration at the indicated doses and days of gestation, and to determine the characteristics of resulting nitrofen-induced cardiovascular anomalies. METHODS: All the observed fetuses were removed from pregnant Sprague-Dawley rats killed on the 21st day of gestation. They were preserved in 10% formalin, and dissection for examination was carried out under a dissecting microscope. RESULTS: The following results were based on dissecting microscopic findings of 482 offspring: (1) The 11th day of gestation was the most sensitive for nitrofen induction of congenital cardiovascular anomalies. The incidence of these was dose related. (2) Ventricular septal defect was the most common single cardiovascular anomaly, representing more than half of all such irregularities. The next most common were aortic arch anomalies and tetralogy of Fallot. (3) Cardiac anomalies derived from infundibular maldevelopment such as tetralogy of Fallot and pulmonary atresia with ventricular septal defect were observed only in the group treated with nitrofen on the 11th gestational day. (4) Aortic arch anomalies were very frequent; the great majority were anomalous right subclavian artery with left aortic arch. CONCLUSION: This animal model is suitable for further embryological investigation of the development of congenital cardiovascular anomalies.

Chronic heart failure model with sequential ligation of the homonymous artery and its diagonal branch in the sheep.

We report a reliable chronic heart failure model in sheep using sequential ligation of the homonymous artery and its diagonal branch. After a left anterior thoracotomy in Corridale sheep, the homonymous artery was ligated at a point approximately 40% of the distance from the apex to the base of the heart, and after 1 hour, the diagonal vessel was ligated at a point at the same level. Hemodynamic measurements were done preligation, 30 minutes after the homonymous artery ligation, and 1 hour after diagonal branch ligation. The electrocardiograms were obtained as needed, and cardiac function was also evaluated with ultrasonography. After a predetermined interval (2 months for five animals and 3 months for two animals), the animals were reevaluated in the same way as before, and were killed for postmortem examination of their hearts. All seven animals survived the experimental procedures. Statistically significant decreases in systemic arterial blood pressure and cardiac output and increases in pulmonary artery capillary wedge pressure were observed 1 hour after sequential ligation of the homonymous artery and its diagonal branch. Untrasonographic analyses demonstrated variable degrees of anteroseptal dyskinesia and akinesia in all animals. The data from animals at 2 months after coronary artery ligation showed significant increases in central venous pressure, pulmonary artery pressure, and pulmonary artery capillary wedge pressure. Left ventricular enddiastolic dimension and left ventricular end-systolic dimension on ultrasonographic studies were also increased. Electrocardiography showed severe ST elevation immediately after the ligation and pathologic Q waves were found at 2 months after ligation. The thin walled infarcted areas with chamber enlargement were clearly seen in the hearts removed at 2 and 3 months after ligation. In conclusion, we could achieve a reliable ovine model of chronic heart failure using a simple concept of sequential ligation of the homonymous and diagonal arteries. This animal model was comparable to the clinical situation.

A solar cell system for extension of battery run time in a moving actuator total artificial heart.

An implantable total artificial heart (TAH) system has strong dependence upon the external battery performance for operation. Even under sophisticated battery management control, the usable external battery performance continues to decrease, which limits TAH performance. One of the ways to overcome this energy problem is to use a solar system (SS). An SS can provide electrical power for the partial TAH drive or battery recharge. This article presents a new concept for use of the solar cell for obtaining double external battery performance. To achieve it, numeric simulations were carried out to obtain the proper magnitude of solar parameters. In the TAH used, the battery power for a cardiac output of 4-6 L/min is approximately 17 W/20 min. From simulated results, the optimal power and voltage of the SS were found to be 7 W, Voc = 27 V in the case of the 24 V motor. Each solar cell includes Voc = 0.5 V, Isc = 37 mA/cm2, FF (fill factor) = 0.77, and efficiency = 10%. Based on the simulation, the effect of solar power capacity on battery run time was studied. With use of 6.5 W SS (W 304 x H 245 x D 16 mm, 1.1 kg), battery performance decreased in vitro from 100% (fully charged) to > 55% vs 0% in the conventional battery system after 20 min operation. However, it dropped to below 20% when using W SS (W 192 x H 192 x D 16 cm, 0.6 kg). The results showed doubled battery run time could be obtained compared with a system without the SS. It was concluded that the proposed SS can be put to practical use as a future energy source for a TAH.