PURPOSE: This study investigated the effect of COX-2 inhibitor on the expression of MMP-13 in the healing process of fracture. MATERIAL AND METHODS: Adult Sprague-Dawley rats were divided into two groups of twenty five rats each. Unilateral femoral shaft fractures were created artificially under displacement in all two groups. COX-2 inhibitor was only given to the experimental group from the postoperative day 1. At 2 weeks after fracture the rats were sacrificed and the callus from each group was used for histologic examination and real time RT-PCR for MMP-13 expression. RESULTS: Histologically, proliferation of osteoblasts and formation of osteoid was less abundant in the experimental group. In real time RT-PCR, the mean expression of MMP-13 is 2.84+/-2.50 in the control group compared with 1.16+/-1.05 in the experimental group. CONCLUSION: In the early stage of fracture healing, COX-2 inhibitor suppress the expression of MMP-13.
Adult ; Animals ; Bony Callus ; Displacement (Psychology) ; Fracture Healing ; Humans ; Osteoblasts ; Rats ; Rats, Sprague-Dawley

In Pleurotus sp., green mold, which is considered a major epidemic, is caused by several Trichoderma species. To develop a rapid molecular marker specific for Trichoderma spp. that potentially cause green mold, eleven Trichoderma species were collected from mushroom farms and the Korean Agricultural Culture Collection (KACC). A dominant fungal isolate from a green mold-infected substrate was identified as Trichoderma pleuroticola based on the sequences of its internal transcribed spacer (ITS) and translation elongation factor 1-a (tef1) genes. In artificial inoculation tests, all Trichoderma spp., including T. atroviride, T. cf. virens, T.citrinoviride, T. harzianum, T. koningii, T. longibrachiatum, T. pleurotum, and T. pleuroticola, showed pathogenicity to some extent, and the observed symptoms were soaked mycelia with a red-brown pigment and retarded mycelium regeneration. A molecular marker was developed for the rapid detection of wide range ofTrichoderma spp. based on the DNA sequence alignment of the ITS1 and ITS2 regions ofTrichoderma spp. The developed primer set detected only Trichoderma spp., and no cross reactivity with edible mushrooms was observed. The detection limits for the PCR assay of T. harzianum (KACC40558), T. pleurotum (KACC44537), and T. pleuroticola (CAF-TP3) were found to be 500, 50, and 5 fg, respectively, and the detection limit for the pathogen-to-host ratio was approximately 1:10,000 (wt/wt).

Soft rot on banana fruit caused by Rhizopus oryzae was identified for the first time in Korea. Colonies were white to light brown and formed numerous sporangiospores. Optimum temperature for mycelial growth was 30degrees C. Sporangia were globose and 30~200 microm. Sporangiophores were usually straight, 8~20 microm, and rhizoids usually in groups of 3~5. Columella were globose to sub-globose and 90~110 microm. Sporangiospores were sub-globose or oval and 4~10 microm. Based on its mycological characteristics, molecular analysis, and pathogenicity to host plants, this fungus was identified as Rhizopus oryzae Went & Prisen Geerligs. This is the first report of soft rot on banana caused by Rhizopus oryzae in Korea.
Fruit ; Fungi ; Humans ; Korea ; Light ; Musa ; Oryza ; Rhizopus ; Sporangia

PURPOSE: Methemoglobinemia is a disorder caused by over accumulation of methemoglobin in the red blood cells of circulating blood, prohibiting adequate supply of oxygen to organs. The seriousness of its clinical symptoms and its treatment methods are determined by the blood methemoglobin level. Therefore, we revealed the clinical relation between the blood methemoglobin level and oxygen saturation on a pulse oximeter. Then we tried to indirectly measure the blood methemoglobin level by using their relation, instead of checking its level through blood sampling. METHODS: The medical records of 39 patients who were admitted to the Chonnam University Hospital Emergency Medical Center due to acute methemoglobinemia between January 1, 2001, and June 30, 2005, underwent a prospective analysis. RESULTS: Among the total of 39 cases, there were 25 males (64.1%) and 14 females (35.9%). There were 15 cases (38.5%) of dapsone overdosage, 18 cases (46.1%) of aniline- type pesticide intoxication, and 6 cases (15.4%) of aniline gas inhalation. As for the main symptoms for admission to the emergency center, there were 8 cases involving on altered mental state, 7 involving dizziness, 3 involving cyanosis, 7 involving dyspnea; and 14 cases were nonsymptomatic. Relational analyses of arterial blood gas analysis results, pulse oximetry saturation levels, and blood methemoglobin levels of the admitted patients revealed that only the pulse oximetry saturation level was related to the blood methemoglobin level (p<0.001). CONCLUSION: When in doubt about the possibility of acute methemoglobinemia, differences in the oxygen saturation level on the pulse oximeter level can be used instead of repeated co-oximetry examinations, can be used to judge treatment responses.
Blood Gas Analysis ; Cyanosis ; Dapsone ; Dizziness ; Dyspnea ; Emergencies ; Erythrocytes ; Female ; Humans ; Inhalation ; Jeollanam-do ; Male ; Medical Records ; Methemoglobin* ; Methemoglobinemia* ; Oximetry ; Oxygen* ; Prospective Studies

A degenerated strain of Pleurotus eryngii KNR2312 was isolated from a commercial farm. Random amplified polymorphic DNA analysis performed on the genomic DNA of the normal and degenerated strains of this species revealed differences in the DNA banding pattern. A unique DNA fragment (1.7 kbp), which appeared only in the degenerated strain, was isolated and sequenced. Comparing this sequence with the KNR2312 genomic sequence showed that the sequence of the degenerated strain comprised three DNA regions that originated from nine distinct scaffolds of the genomic sequence, suggesting that chromosome-level changes had occurred in the degenerated strain. Using the unique sequence, three sets of PCR primers were designed that targeted the full length, the 5' half, and the 3' half of the DNA. The primer sets P2-1 and P2-2 yielded 1.76 and 0.97 kbp PCR products, respectively, only in the case of the degenerated strain, whereas P2-3 generated a 0.8 kbp product in both the normal and the degenerated strains because its target region was intact in the normal strain as well. In the case of the P2-1 and P2-2 sets, the priming regions of the forward and reverse primers were located at distinct genomic scaffolds in the normal strain. These two primer sets specifically detected the degenerate strain of KNR2312 isolated from various mushrooms including 10 different strains of P. eryngii, four strains of P. ostreatus, and 11 other wild mushrooms.
Agaricales ; Chromosomal Instability ; DNA* ; Genetic Markers* ; Pleurotus* ; Polymerase Chain Reaction

During reperfusion of skeletal muscle after ischemia, lipid mediators, mainly eicosanoids, are released and may have a role in the pathogenesis of reperfusion injury. To validate the role of eicosanoids in the ischemia-reperfusion induced functional deficits in skeletal muscle, we compared muscle edema and the changes of eicosanoid concentration in the rat hind limb after ischemia-reperfusion injury by application of tourniquet. After 4 hours of ischemia, reperfusion was established for 4 hours by releasing tourniquet. To assess tissue damage, edema, and wet/dry weight ratios were determined and the eicosanoid concnentrations were measured by the HPLC. The muscle edema and the release of cyclooxygenase metabolites were not induced by the ischemia itself rather they were significantly increased by reperfusion. Indomethacin treatment ameliorated limb edema and decreased the release of 6-keto-PGF1alpha, thromboxane B2, and PGE2 induced by reperfusion. But the inhibitory effect of indomethacin on edema (35%) was relatively low than the inhibitory effect on release of cyclooxygenase metabolites (up to 69%) by reperfusion. These results support the view that cyclooxygenase products may play a significant role in the formation of muscle injury by ischemia-reperfusion and suggest that nonsteroidal antiinflammatory agents might be partially beneficial to the management of acute limb ischemia-reperfusion injury.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; Chromatography, High Pressure Liquid ; Dinoprostone ; Edema* ; Eicosanoids* ; Extremities ; Indomethacin* ; Ischemia ; Muscle, Skeletal* ; Prostaglandin-Endoperoxide Synthases ; Rats ; Reperfusion ; Reperfusion Injury* ; Thromboxane B2 ; Tourniquets

Allelic differences in A and B mating-type loci are a prerequisite for the progression of mating in the genus Pleurotus eryngii; thus, the crossing is hampered by this biological barrier in inbreeding. Molecular markers linked to mating types of P. eryngii KNR2312 were investigated with randomly amplified polymorphic DNA to enhance crossing efficiency. An A4-linked sequence was identified and used to find the adjacent genomic region with the entire motif of the A locus from a contig sequenced by PacBio. The sequence-characterized amplified region marker 7-2299 distinguished A4 mating-type monokaryons from KNR2312 and other strains. A BLAST search of flanked sequences revealed that the A4 locus had a general feature consisting of the putative HD1 and HD2 genes. Both putative HD transcription factors contain a homeodomain sequence and a nuclear localization sequence; however, valid dimerization motifs were found only in the HD1 protein. The ACAAT motif, which was reported to have relevance to sex determination, was found in the intergenic region. The SCAR marker could be applicable in the classification of mating types in the P. eryngii breeding program, and the A4 locus could be the basis for a multi-allele detection marker.
Breeding ; Cicatrix ; Classification ; Dimerization ; DNA ; DNA, Intergenic ; Inbreeding ; Pleurotus ; Transcription Factors

Eight distinct bacteria were isolated form diseased mycelia of the edible mushroom, Pleurotus eryngii. 16S rDNA sequence analysis showed that the isolates belonged to a variety of bacterial genera including Bacillus (LBS5), Enterobacter (LBS1), Sphingomonas (LBS8 and LBS10), Staphylococcus (LBS3, LBS4 and LBS9) and Moraxella (LBS6). Among them, 4 bacterial isolates including LBS1, LBS4, LBS5, and LBS9 evidenced growth inhibitory activity on the mushroom mycelia. The inhibitory activity on the growth of the mushroom fruiting bodies was evaluated by the treatment of the bacterial culture broth or the heat-treated cell-free supernatant of the broth. The treatment of the culture broths or the cell-free supernatants of LBS4 or LBS9 completely inhibited the formation of the fruiting body, thereby suggesting that the inhibitory agent is a heat-stable compound. In the case of LBS5, only the bacterial cell-containing culture broth was capable of inhibiting the formation of the fruiting body, whereas the cell-free supernatant did not, which suggests that an inhibitory agent generated by LBS5 is a protein or a heat-labile chemical compound, potentially a fungal cell wall-degrading enzyme. The culture broth of LBS1 was not inhibitory. However, its cell-free supernatant was capable of inhibiting the formation of fruiting bodies. This indicates that LBS1 may produce an inhibitory heat-stable chemical compound which is readily degraded by its own secreted enzyme.
Agaricales ; Bacillus ; Bacteria ; DNA, Ribosomal ; Enterobacter ; Fruit ; Moraxella ; Ostreidae ; Pleurotus ; Sequence Analysis ; Sphingomonas ; Staphylococcus

This study is to compare the effect of wound healing using three different types of chitin, which include the shapes of sponge, velvet, thick non-woven fabrics, and thin non-woven fabrics. The sponge type had more capacity to absorb the first discharge of a wound than the velvet type and the two non-woven fabrics types. Instead of absorbing the discharge effectively, the velvet type showed a difficulty to take off the dressing stuff from a wound since it was solidly stuck to the wound. The sponge type showed less infiltration of inflammatory cells, producing angiogenesis and fibroblast faster than any other types. Next, the thick non-woven fabrics type was a little more effective than the thin non-woven fabrics type: However, there was no difference between two types. The velvet type sustained the infiltration of inflammatory cells for the longest duration, producing slower angiogenesis and fibroblast. In wound contraction and wound healing, the sponge type was most effective with statistical significance than any other types(p<0.05). The two types of non-woven fabrics did not present any statistical significance in wound contraction, wound epithelization and wound healing(p>0.05). In conclusion, the sponge type showed the best effectiveness to absorb the early discharge, facilitating the progress of inflammatory phase to increase the healing rate. It induced an early healing of wound caused by wound contraction rather than by wound epithelization.
Bandages ; Chitin ; Chitosan* ; Fibroblasts ; Porifera ; Wound Healing* ; Wounds and Injuries*