Pathological Properties of Cryptococcus pseudolongus on the Mycelia and Fruit Body of Lentinula edodes

Recently, Cryptococcus pseudolongus has been reported as a new pathogen of shiitake (Lentinula edodes). However, its pathological properties are not much known. To further understand its impact on the mushroom, we investigated the pathogen’s interactions with the mycelium of shiitake, histopathological properties, host range, and sensitivity to diverse antifungal agents. The strain C. pseudolongus DUCC 4014 inhibited the mycelial growth of L. edodes strain (cultivar Sanjo 701ho) and caused browning in the mycelia confronted with the yeast on PDA. Spray inoculation of the yeast caused an abnormal browning symptom on the cap and/or gills of three shiitake cultivars grown on sawdust media in vinyl bags.Scanning electron microscopic images of the abnormally browned parts of shiitake fruit body illustrated that mushroom tissues were loosed and dispersed in the middle and edge of the cap and the arrangement of basidiospores borne on basidia in the gills was disturbed compared to those of normal shiitake fruit body. Spray inoculation also led to developing abnormal browning on the harvested fruit body, indicating C. pseudolongus could be a problem during mushroom storage. But the yeast was not able to induce abnormal browning on mushrooms of Pleurotus ferulae, Pleurotus fostreatus, and Agaricus bisporus. But it induced browning only on button mushroom (A. bisporus) when they were inoculated after wounding. Tests with 16 kinds of fungicides revealed that the cell growth of C. pseudolongus could be inhibited by benzalkonium chloride at MIC 7 lg/ml and benomyl at MIC 3 lg/ml.

Pathological Properties of Cryptococcus pseudolongus on the Mycelia and Fruit Body of Lentinula edodes

Recently, Cryptococcus pseudolongus has been reported as a new pathogen of shiitake (Lentinula edodes). However, its pathological properties are not much known. To further understand its impact on the mushroom, we investigated the pathogen’s interactions with the mycelium of shiitake, histopathological properties, host range, and sensitivity to diverse antifungal agents. The strain C. pseudolongus DUCC 4014 inhibited the mycelial growth of L. edodes strain (cultivar Sanjo 701ho) and caused browning in the mycelia confronted with the yeast on PDA. Spray inoculation of the yeast caused an abnormal browning symptom on the cap and/or gills of three shiitake cultivars grown on sawdust media in vinyl bags.Scanning electron microscopic images of the abnormally browned parts of shiitake fruit body illustrated that mushroom tissues were loosed and dispersed in the middle and edge of the cap and the arrangement of basidiospores borne on basidia in the gills was disturbed compared to those of normal shiitake fruit body. Spray inoculation also led to developing abnormal browning on the harvested fruit body, indicating C. pseudolongus could be a problem during mushroom storage. But the yeast was not able to induce abnormal browning on mushrooms of Pleurotus ferulae, Pleurotus fostreatus, and Agaricus bisporus. But it induced browning only on button mushroom (A. bisporus) when they were inoculated after wounding. Tests with 16 kinds of fungicides revealed that the cell growth of C. pseudolongus could be inhibited by benzalkonium chloride at MIC 7 lg/ml and benomyl at MIC 3 lg/ml.

Antiplatelet Effect of Cudraxanthone L Isolated from Cudrania tricuspidata via Inhibition of Phosphoproteins

Cudrania tricuspidata (C. tricuspidata) is a deciduous tree found in Japan, China and Korea. The root, stems, bark and fruit of C. tricuspidata has been used as traditional herbal remedies such as eczema, mumps, acute arthritis and tuberculosis. In this study, we investigated the potential efficacies of this natural compound by focusing on the inhibitory effect of cudraxanthone L (CXL) isolated from the roots of C.tricuspidata on human platelet aggregation. Our study focused on the action of CXL on collagen-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding, intracellular calcium mobilization, fibronectin adhesion, dense granule secretion, and thromboxane A 2 secretion. In addition, we investigated the inhibitory effect of CXL on thrombin-induced clot retraction. Our results showed that CXL inhibited collagen-induced human platelet aggregation, intracellular calcium mobilization, fibrinogen binding, fibronectin adhesion and clot retraction without cytotoxicity. Therefore, we confirmed that CXL has inhibitory effects on human platelet activities and has potential value as a natural substance for preventing thrombosis.

Antiplatelet Effect of Cudraxanthone L Isolated from Cudrania tricuspidata via Inhibition of Phosphoproteins

Cudrania tricuspidata (C. tricuspidata) is a deciduous tree found in Japan, China and Korea. The root, stems, bark and fruit of C. tricuspidata has been used as traditional herbal remedies such as eczema, mumps, acute arthritis and tuberculosis. In this study, we investigated the potential efficacies of this natural compound by focusing on the inhibitory effect of cudraxanthone L (CXL) isolated from the roots of C.tricuspidata on human platelet aggregation. Our study focused on the action of CXL on collagen-stimulated human platelet aggregation, inhibition of platelet signaling molecules such as fibrinogen binding, intracellular calcium mobilization, fibronectin adhesion, dense granule secretion, and thromboxane A 2 secretion. In addition, we investigated the inhibitory effect of CXL on thrombin-induced clot retraction. Our results showed that CXL inhibited collagen-induced human platelet aggregation, intracellular calcium mobilization, fibrinogen binding, fibronectin adhesion and clot retraction without cytotoxicity. Therefore, we confirmed that CXL has inhibitory effects on human platelet activities and has potential value as a natural substance for preventing thrombosis.

Solanum lycopersicum (tomato) ethanol extract elicits anti-inflammatory effects via the nuclear factor kappa B pathway and rescues mice from septic shock

Solanum lycopersicum, commonly known as tomato, is widely used in raw, cooked, or liquid forms because it contains nutritional compounds that are beneficial for human health, including carotenoids, lycopene, ascorbic acid, vitamins, and minerals. The tomato is perhaps the most widely studied fruit, especially with respect to its cardioprotective effects. In this study, we aimed to identify the anti-inflammatory mechanisms by which the tomato elicits its anti-inflammatory properties. We treated murine macrophage RAW 264.7 cells with a tomato ethanol extract and performed various biochemical assays including nitric oxide inhibition, cell viability, RNA extraction, expression of pro-inflammatory mediators and cytokines, and immunoblotting, as well we assessed cell survival rates. Our results have shown for the first time that a tomato ethanol extract treatment can suppress nitric oxide production in a dose-dependent manner without cytotoxicity. Moreover, it inhibits the expression of pro-inflammatory mediators and cytokines and elicits its anti-inflammatory effects via the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and mitogen-activated protein kinase (MAPK) pathways. In addition, administration of tomato syrup potently rescued mice from septic shock induced by lipopolysaccharide injection. Collectively, our results elucidate details regarding the anti-inflammatory mechanisms of tomato.

Ribosomal Intergenic Spacer 1 Based Characterization of Button Mushroom (Agaricus bisporus) Strains

Breeding the button mushroom requires genetic information about its strains. This study was undertaken to genetically characterize four domestically bred button mushroom strains (Saea, Saejung, Saedo, Saeyeon cultivars) and to assess the possibility of using the intergenic spacer 1 (IGS1) region of rDNA as a genetically variable region in the genetic characterization. For the experiment, 34 strains of Agaricus bisporus, two strains of A. bitorquis, and one strain of A. silvaticus, from 17 countries were used. Nucleotide sequence analysis of IGS1 rDNA in these 37 Agaricus strains confirmed that genetic variations exist, not only among the four domestic strains, but also between the four domestic strains and foreign strains. Crossing two different haploid strains of A. bisporus seems to generate genetic variation in the IGS1 region in their off-spring haploid strains. Phylogenetic analysis based on the IGS1 sequence revealed all A. bisporus strains could be differentiated from A. silvaticus and A. bitorquis strains. Five genetic groups were resolved among A. bisporus strains. Saejung and Saeyeon cultivars formed a separate genetic group. Our results suggest that IGS1 could be complementarily applied in the polymorphism analysis of button mushroom.

Genetic and Biochemical Characterization of Monokaryotic Progeny Strains of Button Mushroom (Agaricus bisporus)

To promote the selection of promising monokaryotic strains of button mushroom (Agaricus bisporus) during breeding, 61 progeny strains derived from basidiospores of two different lines of dikaryotic parental strains, ASI1038 and ASI1346, were analyzed by nucleotide sequencing of the intergenic spacer I (IGS I) region in their rDNA and by extracellular enzyme assays. Nineteen different sizes of IGS I, which ranged from 1,301 to 1,348 bp, were present among twenty ASI1346-derived progeny strains, while 15 different sizes of IGS I, which ranged from 700 to 1,347 bp, were present among twenty ASI1038-derived progeny strains. Phylogenetic analysis of the IGS sequences revealed that different clades were present in both the ASI10388- and ASI1346-derived progeny strains. Plating assays of seven kinds of extracellular enzymes (beta-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease) also revealed apparent variation in the ability to produce extracellular enzymes among the 40 tested progeny strains from both parental A. bisporus strains. Overall, this study demonstrates that characterization of IGS I regions and extracellular enzymes is useful for the assessment of the substrate-degrading ability and heterogenicity of A. bisporus monokaryotic strains.

First Report of Leaf Rust Caused by Puccinia caricis in Farfugium japonicum in Korea

Farfugium japonicum is used in traditional medicine and as an edible herb in China and Korea. In July 2013, leaf spots were observed in F. japonicum seedlings at Ulleung Island, Gyeongsangbuk Province, Korea. Early symptoms on the leaf adaxial surface included roughly circular yellow spots that later developed brown, necrotic centers. The aecia were hypophyllous, cupulate, yellowish, 180~430 microm in diameter, clustered, and erumpent with a peridium with a recurved margin. The aeciospores were globoid, 14~17 x 13~16 microm, light yellow or colorless, and densely verrucose. The 28S rDNA sequence of the isolate was identical to each other and shared 99% identity with Puccinia caricis. This is the first report of rust caused by P. caricis in F. japonicum in Korea or elsewhere in the world.

Characterization of Myrothecium roridum Isolated from Imported Anthurium Plant Culture Medium

During an investigation of microorganisms and pests in plant culture media from imported anthurium pots, a fungal isolate (DUCC4002) was detected. Based on its morphological characters including colony shape on potato dextrose agar, the microstructures of spores observed by light and scanning electron microscopy and the results of phylogenetic analysis using an internal transcribed spacer rDNA sequence, the fungal isolate was identified as Myrothecium roridum. Pathogenicity testing on anthurium leaves revealed that the fungus could colonize and produce sporodochia on the inoculated leaves. This is the first report of M. roridum detected in imported plant culture medium in Korea.

Extracellular Enzyme Activities of the Monokaryotic Strains Generated from Basidiospores of Shiitake Mushroom

To obtain basic information on the biochemical property of basidiospores of shiitake mushroom (Lentinula edodes), the ability of producing extracellular enzyme was assessed using a chromogenic plate-based assay. For the aim, amylase, avicelase, beta-glucosidase, CM-cellulase, pectinase, proteinase, and xylanase were tested against monokaryotic strains generated from forty basidiospores of two different parental dikaryotic strains of shiitake mushroom, Sanjo-101Ho and Sanjo-108Ho. These two parental strains showed different degree of extracellular enzyme activity. No identical patterns of the degree of enzyme activity were observed between monokaryotic strains and parental strains of the two shiitake cultivars. The degree of extracellular enzyme activity also varied among monokaryotic strains of the two shiitake cultivars. Our results showed that dikaryotic parental strains of shiitake mushroom produce monokaryotic basidiospores having very diverse biochemical properties.

Detection of Extracellular enzymes Activities in Various Fusarium spp

Thirty seven species of Fusarium were evaluated for their ability of producing extracellular enzymes using chromogenic medium containing substrates such as starch, cellobiose, CM-cellulose, xylan, and pectin. Among the tested species Fusarium mesoamericanum, F. graminearum, F. asiaticum, and F. acuminatum showed high beta-glucosidase acitivity. Xylanase activity was strongly detected in F. proliferatum and F. oxysporum. Strong pectinase activity was also found in F. oxysporum and F. proliferatum. Amylase activity was apparent in F. oxysporum. No clear activity in cellulase was found from all the Fusarium species tested.

Characterization of Sclerotinia sclerotiorum Isolated from Paprika

A fungal isolate collected from infected paprika (Capsicum annuum var. grossum) was characterized as Sclerotinia sclerotiorum based on its ability of sclerotium formation, physiological and molecular properties. When the isolate was grown on potato dextrose agar, oatmeal agar, and malt extract agar, it grew most well on PDA. Optimal temperature and pH for its growth were 25degrees C and pH 7, respectively. The fungal isolate produced sclerotia on PDA within 10 days, and the color and shape of the sclerotia were similar to those of S. sclerotiorum . The ITS rDNA regions including ITS1 and ITS2 and 5.8S sequences were amplified using ITS1F and ITS4 primers from the genomic DNAs of the paprika isolate and other known pathogenic S. sclerotiorum isolated from different crops in Korea, and their nucleotide sequences were determined. Sequence comparison analysis showed the ITS rDNA of the paprika isolate shares 100% sequence identity with those of S. sclerotiorum isolated from red pepper, lettuce and a S. sclerotiorum isolate registered in GenBank DNA database. Neighbor joining analysis based on the ITS rDNA sequence revealed the paprika isolate has very close phylogenetic relationships with known Sclerotinia sclerotiorum isolates. This is the first report that S. sclerotiorum has been found associated with paprika rot in paprika growing countries.

Classification and Clinical Evaluation of Gastrointestinal Stromal Tumor: Immunohistochemical Expression of CD117, CD34, alpha- Smooth Muscle Actin, S-100

PURPOSE: Gastrointestinal stromal tumors (GISTs) represent a distinct and the most important subset of, mesenchymal tumors of the GI tract. Stromal tumors of the gastrointestinal tract have long been a source of confusion and controversy, with regard to their classification, differentiation, criteria of malignancy and prognostic features. METHODS: The 26 case studies of patients treated for a Gastrointestinal mesenchymal tumor, including leiomyomas, leiomyosarcomas and GISTs, between 1994 and 2002 at Keimyung University Hospital, were evaluated retrospectively. The cases were confirmed as leiomyomas, schwannomas, or GISTs by pathological re-examination. 20 of the cases were diagnosed as GISTs, from the pathological examination, and were chosen for the evaluation of their clinicopathological and immunohistochemical characteristics, using CD34, CD117, alpha-SMA and S-100 done. RESULTS: The new diagnoses of the mesenchymal tumors were a leiomyoma in 3 cases, a schwannoma in 3 and gastric stromal tumors in all 20. The immunohistochemical studies were positive for CD117 and CD34 in 95 and 75% of the gastric stromal tumors, respectively. The histopathological findings showed 5 benign tumors, 3 borderline tumors, and 12 malignant tumors in the 20 patients. CONCLUSION: The immunohistochemical marker (CD117) for KIT is a specific marker for GISTs among the tumors occurring in the stomach, and can be used to distinguish GISTs from true leiomyomas and gastric schwannomas. We also found that severe cellularity, atypism, intratumoral hemorrhage and necrosis, large size and a high mitotic count correlate with malignant behaviour and a poor prognosis.

Imaging with Magnetization Transfer Technique on the Intracranial Tumors

PURPOSE: To measure the magnetization transfer ratio(MTR) of intracranial tumors and to evaluate the difference of the character and the border of tumors on magnetization transfer image. MATERIALS AND METHODS: We prospectively reviewed magnetization transfer imaging(MTI) findings of intracranial tumors of 13 patients. 1.0T MRI machine was used. T1 weighted image(TRITE=700/14) and T1 weighted magnetization transfer image were obtained. The offset and bandwidth of magnetization transfer pulse were 1000 Hz and 250Hz, respectively. Postcontrast images were also obtained. MTR was measured at lesion area. RESULTS: The mean of normal MTR of white matter and gray matter in 7 normal volunteers were 37% and 29%, respectively. The MTR of each tumor was as follows;metastatic tumors(mean 16.3%), meningioma(mean 27.3%), oligodendroglioma(mean 20.2%), glomus jugulare tumor(mean 17.3%), gliomatosis cerebri(mean 27. 0%). The contrast of lesions were better at magnetization transfer image than at T1 weighted image. CONCLUSION: The MTR of the intracranial tumors is lower than normal tissue. Measurement of MTR at muirpie areas within the tumor can be useful for evaluation of indistinct border of the tumor.

Magnetization Transfer Ratio of Brain Tissue: Normal Value and Effect of TR/TE

PURPOSE: Magnetization transfer imaging(MTI) is a new imaging contrast technique. Our MT pulse sequence is designed as fixed time interval between echo and MT pulse. This study was peformed to evaluate the influence of variations in TR/TE on MTR in T1 weighted image of normal brain tissue on this kind of MT pulse sequence. MATERIALS AND METHODS: Seven healthy volunteers in twenties of age as the objectives, MRI was taken under various TR/TE(TR/TE ;700/14, 650/14, 750/14, 700/20 and 1500/20 msec). MTR was calculated from signal intensities measured at the same point in both pre and post MT images and statisticslly analyzed. The MR imager used in this study was 1.0T Magnetom 42SP(Siemens, Erlangen, Germany) and the parameters of additional MT pulse sequence were offset 1000Hz and bandwidth 250Hz, and posteriorly located to echo with 7.7 msec fixed interval. Offset of this MT pulse was variable. RESULT: In white matter of brain tissue from a normal person, MTR was 34-39%(average 37%) for TR and TE of 700/14 in T1WI and 33-36%(average 35%) for TR/TE of 650/14, and 34-38%(average 35%) for TR/TE of 750/14 which showed no statistical difference. However, in case of 1500/20 of TR/TE, MTR was 26-28%(average 26%) which is statistically significant. With TR/TE of 700/14 as the standrd value, the MTR of gray and white matter were 37% and 29% respectively, showing a definite difference of statistical means. Signal from CSF in ventricles is rarely influenced by MT pulse. CONCLUSION: Conclusively, a subtle variation in TR/TE in T1WI has little influence on MTR but wide range of variation in TR/TE as in proton density image induces significant difference in MTR on this kind of MT pulse sequence. Therefore, the exchangeable usage of MTR data would be possible in narrow range of TR/TE change but difficult in wide range of variation.

MRI Study about the Early Changes of Lumbar Disk Degeneration using Magnetization Transfer Contrast (MTC)

PURPOSE: To obtain magnetization transfer ratio(MTR) of the annulus fibrosus and nucleus pulposus and to assess the feasibility of utilizing the changes of these MTRs as an early indicator of disk degeneration. MATERIALS AND METHODS: MR images of lumbar spine with magnetization transfer(MT) technique in 42 patients were obtained. spin echo techniques (600/14) with same TR/TE with 1KHz off-resonance saturation were employed in 1.0T MR system. MTRs were calculated in two regions, anterior annulus fibrosus and nucleus pulposus, and the results were compared between the normal and degenerative disks, from grade I to IV, on T2-weighted images. RESULTS: MTRs of the nucleus pulposus were 17.6% in the normal disks, and 26.7%, 28.4%, 29.1%, 29.7% in degenerative disks, from grade I to IV, respectively, with a significant difference(P<0.05). On the other hand, MTRs in the annulus fibrosus were 30.2% in the normal disks and 31.5%, 33.2%, 32.1% and 35.6% in degenerative disks, from grade I to IV, respectively, without significant difference. CONCLUSION: Since MTRs are significantly higher in degenerative nucleus pulposus than those of the normal disks, increased MTRs in the nucleus pulposus can be used as an early sign of the degeneration of the nucleus pulposus.