Treatment of rainbow trout macrophages with glycyrrhizin (GL), an aqueous extract of licorice (Glycyrrhiza glabra), enhanced their respiratory burst activity. Maximal effects were seen using concentrations of 10-100 ug/ml. GL also modulated trout lymphocytes, increasing proliferation responses to the mitogen phytohemagglutinin two-fold over a range of GL concentrations. In addition, GL elicited the release of a macrophage activating factor (MAF) kom head kidney leukocytes, as assessed by the ability of generated supernatants to increase respiratory burst activity of target macrophages. MAF activity was most apparent using 100 ug/ml GL to induce MAF release and a 48 h incubation period with the target macrophages. Finally, GL was shown to enhance the release oF MAF in response to the mitogen concanavalin A. The results suggest that GL might modulate the innate defences in fish.
Concanavalin A ; Glycyrrhiza ; Glycyrrhizic Acid* ; Head Kidney ; Leukocytes* ; Lymphocytes ; Macrophages ; Oncorhynchus mykiss* ; Oncorhynchus* ; Respiratory Burst ; Trout

Animals ; Erythrocytes ; physiology ; Oncorhynchus mykiss ; physiology ; Oxygen ; Water ; chemistry

BACKGROUND: Polydeoxyribonucleotide (PDRN) influencing cellular growth and differentiation is recognized to promote wound healing by stimulating tissue repair. Although PDRN can be extracted from human placentas, PDRN medications have recently been extracted from the semen of trout (Oncorhynchus mykiss) and salmon (Oncorhynchus keta). The present study was designed to evaluate the wound healing effects of O. keta-derived PDRN for injection (Rejuvenex) and PDRN cream (Rejuvenex Cream) in comparison with those of O. mykiss-derived PDRN injection (Placentex). METHODS: Full-thickness skin defects were made on the back of mice (n=60). The mice were divided into the following four groups according to the dressing used for the wounds: O. mykiss-derived PDRN injection group, O. keta-derived PDRN injection group, O. keta-derived PDRN cream group, and normal saline soaked dressing group (control group). We analyzed the gross findings, wound sizes, histological findings, immunohistochemistry and enzyme-linked immunosorbent assays for the groups immediately after the treatment, and again after 4, 7, and 10 days of treatment. RESULTS: The wound healing effects were the greatest in the O. keta-derived PDRN injection and O. mykiss-derived PDRN injection groups, which showed similar scores, followed by the O. keta-derived cream and normal saline soaked dressing groups. CONCLUSION: The injection of PDRN extracted from O. keta was found to be as effective at healing full-thickness skin defects as the O. mykiss-derived PDRN injection, which is currently used in the clinic. Moreover, the O. keta-derived PDRN injection was also found to reduce the time required for wound healing.
Animals ; Bandages ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunohistochemistry ; Mice ; Oncorhynchus* ; Placenta ; Salmon ; Semen ; Skin ; Trout ; Wound Healing* ; Wounds and Injuries*

BACKGROUND: The effects of the dietary administration of two heat-inactivated whole bacteria from the Vibrionaceae family, singly or combined, on innate immune response of the rainbow trout were studied. The two bacteria (Pdp11 and 51M6), which were obtained from the skin of rainbow trout, showed in vitro characteristics that suggested they could be considered as potential fish probiotics. METHODS: The fish were fed four different diets: control (non-supplemented), or diets supplemented with heat-inactivated bacteria at 10(8) cfu/g Pdp11, 10(8) cfu/g 51M6 or with 0.5x10(8) cfu/g Pdp11 plus 0.5x10(8) cfu/g 51M6 for 4 weeks. Six fish were sampled at weeks 1, 2, 3 and 4, and then the main humoral (natural haemolytic complement activity and serum peroxidase content) and cellular innate immune responses (leucocyte peroxidase content, phagocytosis, respiratory burst and cytotoxicity) were evaluated. RESULTS: The serum peroxidase content and the natural haemolytic complement activity increased with time, reaching the highest values in the third and fourth weeks of feeding, respectively. The phagocytic ability of specimens fed the mixture of the two inactivated bacteria was significantly higher than in the controls after 2 and 3 weeks of treatment. The same activity increased significantly in rainbow trout fed the Pdp11 diet for 2 weeks or the 51M6 diet for 3 weeks. Respiratory burst activity was unaffected by all the experimental diets at all times assayed. Cytotoxic activity had significantly increased after 3 weeks in fish fed the 51M6 diet. CONCLUSION: Our results demonstrated the usefulness of incorporating inactivated probiotic bacteria into fish diets.
Bacteria ; Complement System Proteins ; Diet ; Flow Cytometry ; Humans ; Immunity, Innate ; Oncorhynchus mykiss ; Peroxidase ; Phagocytosis ; Probiotics ; Respiratory Burst ; Skin ; Vibrionaceae

PURPOSE: To evaluate the extent of tissue coagulation during interstitial laser photocoagulation (ILP) innormal bovine liver, using a diode laser unit and various parameters, and to determine whether the procedure isapplicable to clinical practice.. MATERIALS AND METHODS: Using an 18-gauge needle, experimental interstitiallaser photocoagulation (ILP) was carried out in normal bovine liver. On the basis of differing parameters, threegroups were established. For group I, a single photofiber with laser power of 1, 3 and 5 watts and an exposuretime of 60, 180, 300, 420 or 600 seconds was used. For groups II and III, four needles were fixed at a distance of1cm and 1.5cm ; in each case a needle fixation device was used, as well as a laser distributor for simultaneouslaser exposure of photofibers. As a control, four photofibers were placed as for groupIII, but to compare groups IIand III, each photofiber was exposed to a laser of 3 watts 300 seconds, without using a laser distributor. Toevaluate the range of tissue coagulation, specimens were analyzed both with regard to cross-sectional grossfindings and histopathologically . RESULTS: The largest diameter of thermal coagulation necrosis in Group I was15x15mm, and this was ball-shaped. Coalescence of coagulation between each photofiber was observed in Group II,and this was up to 25 mm in diameter. In Group III and controls, coalescence was not found, though the extent oftissue coagulation increased with increasing wattage and exposure time. The extent of charring at the center ofcoagulation also increased with increasing wattage. Smoke bubbles emanating from the coagulation area wereobserved, and during ILP involving a single photofiber, increased from 3 watts, applied for 300 seconds.CONCLUSION: Using an 8-gauge needle and a diode laser ILP, we have shown that a range of tissue coagulationacutely ablates normal bovine liver. In selective cases, the procedure could be applied to clinical trials.
Animals ; Lasers, Semiconductor* ; Light Coagulation* ; Liver* ; Necrosis ; Needles ; Smoke ; Trout

The purpose of this study was to evaluate the structural change of root surface and the occlusion of dentinal tubule following CO2 laser treatment. Seven extracted healthy human premolar werw curetted, sectioned, and four specimens were randomly assigned to each of 6 different treatment groups : 1) untreated EDTA etched control; 2) root plande only; 3) CO2 laser treated with 2W mode 6(10msec/pulse, 20pps) for 1 minute; 4) CO2 laser treated with 2W mode 6(10msec/pulse, 20pps) for 2 minutes; 5) CO2 laser treated with 2W mode 7(20msec/pulse, 20pps) for 1 minute; 6) CO2 laser treated with 2W mode 7(20msec/pulse, 20pps) for 2 minutes. Following the prescribed treatment, the specimens were prepared for SEM evaluation. Results showed that CO2 laser may be effective to occlude dentinal tubules tor dentin sensitivity treatment. The effect of dentinal tubule occlusion was enhanced with increasing the total energy level lased to specimen regardless of lasing mode. The structural changes of root surfaces were restricted to superficies, and these changes included fissuring, charring, crater formation over the smooth lava like texture. The charring and crater formation implying root damage was observed in the case of the longer duration of a pulse. The results of the present study suggests that the pulsed CO2 laser with shorter pulse duration and longer exposure time can be used effectively in order to obtain the optimal dentinal tubule occlusion with minimal root damage.
Bicuspid ; Dentin ; Dentin Sensitivity ; Edetic Acid ; Humans ; Lasers, Gas* ; Trout

The aim of this study is to investigate the status of the websites of plastic surgery clinics in Seoul, and to elucidate the factors that affect the page views. Directors of the clinics were asked by telephone as to whether their clinics have an website, and the rank of their sites were measured using the Navvy (ve r 2.75) program. The gene ral characteristics of each clinic and directors were surveyed using the list of members for the Korean association of clinical plastic surgeons. The characteristics of the websites were also surveyed and categorized into their accessibility, introduction, contents, interactions, and etc. The relationship between each variable, the possession of a homepage and the rank were analyzed by univariate analysis and the independent effects of these variables were examined by multiple logistic regression analysis. The results are as follows: 196 out of 252 clinics(77.8%) had a website and 77 of these sites were ranked below the 150,000th place. In multiple logistic regression analysis, younger directors had a higher likelihood of possessing a website, and the location of the clinic also significantly affected whether or not there was a website. The websites of the clinics located in Kangnam/Seocho-gu and run by more than 2 plastic surgeons had a higher tendency to rank be low the 150,000th place. However, the char acteristic s of the website did not aff ect the r ank distribution. Considering the above results, the characteristics of the clinics and surgeons may contribute greatly for the clinics to have a website and to have a high rank. In addition, the characteristics of the site may not be a major influencing fac tor.
Internet ; Logistic Models ; Seoul* ; Surgery, Plastic* ; Telephone ; Trout

In order to detect Infectious hematopoietic necrosis virus with immunological methods, the surface glycoprotein of a recent IHNV-Sn isolated from farmed rainbow trout ( Oncorhynchus mykiss ) in China was amplified and cloned into pET27b(+) vector (designated as pET27b-G ). The expression of recombinant plasmid pET27b-G in E. coli BL21(DE3) was induced and determined by SDS-PAGE analysis. The predicted molecular weight of glycoprotein protein was approximately 55 kD and was confirmed in this study. The inclusion body of glycoprotein was treated with urea at different urea concentrations, and dialyzed into PBS buffer. Purified glycoprotein with high concentration was obtained after dialyzed in the PBS buffer. Antisera against glycoprotein were produced from immunized rabbits. The prepared antisera could react specifically with both the recombinant glycoprotein and natural glycoprotein of the IHNV-Sn isolated in the test of indirect ELISA, and the titer against the recombinant glycoprotein was 1:20,000. IFA showed that the antisera can recognize the glycoprotein located on the surface of IHNV-Sn and IHNV reference strain. These results indicated that the expressed glycoprotein was immunogenical and antigenical and could be functional as the natural IHNV glycoprotein. These results established a foundation for further study on vaccine and rapid diagnosis of IHNV.
Animals ; Antibodies, Viral ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Fish Diseases ; immunology ; virology ; Gene Expression ; Glycoproteins ; genetics ; immunology ; Infectious hematopoietic necrosis virus ; genetics ; immunology ; Neutralization Tests ; Oncorhynchus mykiss ; Rabbits ; Rhabdoviridae Infections ; immunology ; veterinary ; virology ; Viral Proteins ; genetics ; immunology

Saprolegniasis is one of the most devastating oomycete diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated oomycete belongs to the member of S. parasitica, supported by its typical features including cotton-like mycelium, zoospores and phylogenetic analysis with internal transcribed spacer region. Pathogenicity of isolated S. parasitica was developed in embryo, juvenile, and adult zebrafish as a disease model. Host-pathogen interaction in adult zebrafish was investigated at transcriptional level. Upon infection with S. parasitica, pathogen/antigen recognition and signaling (TLR2, TLR4b, TLR5b, NOD1, and major histocompatibility complex class I), pro/anti-inflammatory cytokines (interleukin [IL]-1β, tumor necrosis factor α, IL-6, IL-8, interferon γ, IL-12, and IL-10), matrix metalloproteinase (MMP9 and MMP13), cell surface molecules (CD8⁺ and CD4⁺) and antioxidant enzymes (superoxide dismutase, catalase) related genes were differentially modulated at 3- and 12-hr post infection. As an anti-Saprolegnia agent, plant based lawsone was applied to investigate on the susceptibility of S. parasitica showing the minimum inhibitory concentration and percentage inhibition of radial growth as 200 μg/mL and 31.8%, respectively. Moreover, natural lawsone changed the membrane permeability of S. parasitica mycelium and caused irreversible damage and disintegration to the cellular membranes of S. parasitica. Transcriptional responses of the genes of S. parasitica mycelium exposed to lawsone were altered, indicating that lawsone could be a potential anti-S. parasitica agent for controlling S. parasitica infection.
Adult ; Cytokines ; Embryonic Structures ; Fresh Water ; Host-Pathogen Interactions ; Humans ; Interferons ; Interleukin-12 ; Interleukin-6 ; Interleukin-8 ; Korea* ; Major Histocompatibility Complex ; Membranes ; Microbial Sensitivity Tests ; Mycelium ; Oncorhynchus mykiss* ; Oomycetes ; Permeability ; Plants ; Saprolegnia* ; Tumor Necrosis Factor-alpha ; Virulence ; Zebrafish*

Germ cells make two major decisions when they move from an indeterminate state to their final stage of gamete production. One decision is sexual commitment for sperm or egg production, and the other is to maintain mitotic division or entry into meiosis. It is unclear whether the two decisions are made as a single event or separate events, because there has been no evidence for the presence of germ cell sex prior to meiosis. Here we report direct evidence in the fish rainbow trout that gonia have distinct sexuality. We show that dazl expression occurs in both male and female gonia but exhibits differential intracellular distribution. More strikingly, we show that boule is highly expressed in male gonia but absent in female gonia. Therefore, mitotic gonia possess sex, sperm/egg decision and mitosis/meiosis decision are two independent events, and sperm/egg decision precedes mitosis/meiosis decision in rainbow trout, making this organism a unique vertebrate model for mechanistic understanding of germ cell sex differentiation and relationship between the two decisions.
Animals ; Female ; Fish Proteins ; genetics ; Gene Expression Regulation ; Male ; Meiosis ; Oncorhynchus mykiss ; genetics ; physiology ; Ovary ; cytology ; metabolism ; Ovum ; cytology ; metabolism ; RNA, Messenger ; genetics ; metabolism ; RNA-Binding Proteins ; genetics ; Sex Determination Processes ; Spermatozoa ; cytology ; metabolism ; Testis ; cytology ; metabolism