Molecular features, antioxidant potential, and immunological expression assessment of thioredoxin-like protein 1 (TXNL1) in yellowtail clownfish (Amphiprion clarkii).

Thioredoxin-like protein 1 (TXNL1) is a redox-active protein belonging to the thioredoxin family, which mainly controls the redox status of cells. The TXNL1 gene from Amphiprion clarkii (AcTXNL1) was obtained from a pre-established transcriptome database. The AcTXNL1 is encoded with 289 amino acids and is predominantly localized in the cytoplasm and nucleus. The TXN domain of AcTXNL1 comprises a34CGPC37 motif with redox-reactive thiol (SH-) groups. The spatial distribution pattern of AcTXNL1 mRNA was examined in different tissues, and the muscle was identified as the highest expressed tissue. AcTXNL1 mRNA levels in the blood and gills were significantly increased in response to different immunostimulants. In vitro antioxidant capacity of the recombinant AcTXNL1 protein (rACTXNL1) was evaluated using the ABTS free radical-scavenging activity assay, cupric ion reducing antioxidant capacity assay, turbidimetric disulfide reduction assay, and DNA nicking protection assay. The potent antioxidant activity of rAcTXNL1 exhibited a concentration-dependent manner in all assays. Furthermore, in the cellular environment, overexpression of AcTXNL1 increased cell viability under H2O2 stress and reduced nitric oxide (NO) production induced by lipopolysaccharides (LPS). Collectively, the experimental results revealed that AcTXNL1 is an antioxidant and immunologically important gene in A. clarkii.

Molecular cloning, expression analysis of interleukin 17D (cysteine knot cytokine) from Amphiprion clarkii and their functional characterization and NFκB pathway activation using FHM cells.

Interleukin 17D (IL-17D), a pro-inflammatory cytokine, is a signature cytokine of T helper 17 (Th17) cells. However, studies characterizing the functions of IL-17D in teleost are scarce. Therefore, we aimed to characterize the properties of IL-17D in Amphiprion clarkii. We performed spatial and temporal expression, AcIL-17D-mediated antibacterial and inflammatory gene expression, NFκB pathway-related gene expression analyses, and bacterial colony counting and cell protection assays. We found that AcIL-17D contains a 630 bp coding sequence and encodes 210 amino acids. The spatial expression analysis of AcIL-17D in 12 tissues showed ubiquitous expression, with the highest expression in the brain, followed by blood and skin. Temporal expression analysis of AcIL-17D in blood showed upregulated expression at 6 and 24 h (polyinosinic: polycytidylic acid and lipopolysaccharide), 12 h (all stimulants), and 48 h (polyinosinic: polycytidylic acid and Vibrio harveyi). AcIL-17D expression in the blood gradually decreased at later hours in response to all the stimulants. After treatment of fathead minnow (FHM) cells with different recombinant AcIL-17D concentrations, the downstream gene expression analysis showed increased expression of antimicrobial genes in the FHM cells, namely [NK-Lysin (NKL), Hepcidin antimicrobial peptide-1 (HAMP-1), Defensin-ß (DEFB1)] and some inflammatory genes such as IL-1ß, TNF-α, IL-11, and STAT3. Further nuclear factor κB (NFκB) subunits (NFκB1, NFκB2, RelA, and Rel-B) showed upregulated gene expression at 12 and 24 h. The bacterial colony counting assay using FHM cells showed lower bacterial colony counts in rAcIL-17D-treated cells than in control. Furthermore, the Water-Soluble Tetrazolium Salt (WST -1) assay confirmed the ability of rAcIL-17D in the protection of FHM cells from bacterial infection and conducted the Hoechst 33342 staining upon treatment with rAcIL-17D and rMBP. Therefore, our findings provide important insights into the activation of IL-17D pathway genes in FHM cells, the protective role of AcIL-17D against bacterial infection, and host defense mechanisms in teleost.

Effect of Zr Content on Phase Stability, Deformation Behavior, and Young's Modulus in Ti-Nb-Zr Alloys.

Ti alloys have attracted continuing research attention as promising biomaterials due to their superior corrosion resistance and biocompatibility and excellent mechanical properties. Metastable ß-type Ti alloys also provide several unique properties such as low Young's modulus, shape memory effect, and superelasticity. Such unique properties are predominantly attributed to the phase stability and reversible martensitic transformation. In this study, the effects of the Nb and Zr contents on phase constitution, transformation temperature, deformation behavior, and Young's modulus were investigated. Ti-Nb and Ti-Nb-Zr alloys over a wide composition range, i.e., Ti-(18-40)Nb, Ti-(15-40)Nb-4Zr, Ti-(16-40)Nb-8Zr, Ti-(15-40)Nb-12Zr, Ti-(12-17)Nb-18Zr, were fabricated and their properties were characterized. The phase boundary between the ß phase and the α'' martensite phase was clarified. The lower limit content of Nb to suppress the martensitic transformation and to obtain a single ß phase at room temperature decreased with increasing Zr content. The Ti-25Nb, Ti-22Nb-4Zr, Ti-19Nb-8Zr, Ti-17Nb-12Zr and Ti-14Nb-18Zr alloys exhibit the lowest Young's modulus among Ti-Nb-Zr alloys with Zr content of 0, 4, 8, 12, and 18 at.%, respectively. Particularly, the Ti-14Nb-18Zr alloy exhibits a very low Young's modulus less than 40 GPa. Correlation among alloy composition, phase stability, and Young's modulus was discussed.

Survival Rate and Hematological Responses with Temperature Changes of Red Spotted Grouper, Epinephelus akaara in South Korea.

The effect of sudden changes of water temperature (WT) on the survival rate and physiological responses of the red spotted grouper (Epinephelus akaara) were examined by manipulating WT control system for 9 days. Experimental condition was divided in two different regimes at low (from 10°C to 4°C, decreased 1℃/d) and high (from 28°C to 34°C, increased 1°C/d) WT. Survival rate of experimental fishes were observed, and determined the changes of hematological characteristics by analyzing plasma levels of cortisol, glucose, total protein, and electrolytes (Na(+), Cl-, K(+)). No mortality was observed until low WT 6°C (144 h) and high WT 32°C (96 h), and 100% mortality was observed at low WT 4°C (216 h) and high WT 35°C (171 h). Plasma levels of cortisol and glucose increased rapidly as decreasing WT, and the loss of swimming ability and respiration response was observed at low WT 7°C and high WT 34°C conditions.

Artificial Spawning Behavior and Development of Eggs, Larvae and Juveniles of the Red Spotted Grouper, Epinephelus akaara in Korea.

This study was conducted in order to examine the egg development in red spotted grouper, Epinephelus akaaraand the morphological development of its larvae and juveniles, and to obtain data for taxonomic research. This study was conducted in June 2013, and 50 male and female fish were used for the study. One hundred µg/kg of LHRHa was injected into the body of the fish for inducing spawning, and the fish were kept in a small-sized fish holder (2×2×2 m). Eggs were colorless transparent free pelagic eggs, 0.71-0.77 mm large (mean 0.74±0.02 mm, n=30), and had an oil globule. Hatching started within 27 h after fertilization. Pre-larvae that emerged just after hatching were 2.02-2.17 mm in total length (mean 2.10±0.11 mm), their mouth and anus were not opened yet, and the whole body was covered with a membrane fin. Post-larvae that emerged 15 days post hatching were 3.88-4.07 mm in total length (mean 3.98±0.13 mm), and had a ventral fin with two rays and a caudal fin with eight rays. Juveniles that were formed at 55 d post hatching, were 31.9-35.2 mm in total length (mean 33.6±2.33 mm), with red color deposited over the entire body, and black chromophores deposited in a spotted pattern. The number of fin rays, body color, and shape were the same as that in the adult fish.

Complete Genome Sequence of Nervous Necrosis Virus Isolated from Sevenband Grouper (Epinephelus septemfasciatus) in South Korea.

The draft genome sequence of the nervous necrosis virus (NNV) SGYeosu08, isolated from sevenband grouper (Epinephelus septemfasciatus) in Yeosu, South Korea, was cloned and analyzed. The full-length RNA1 was a 3,103-nucleotide-encoding region of RNA-dependent RNA polymerase, and the RNA2 encoding a coat protein was 1,433 nucleotides in length. This genome sequence might be useful in the development of an accurate diagnostic tool.

Effects of microstructure and deformation conditions on the hot formability of Ni-Ti-Hf shape memory alloys.

Ingots of Ni-Ti-Hf shape memory alloys were prepared by vacuum arc re-melting. Isothermal hot compression tests were conducted at temperatures ranging from 700 to 1000 degrees C and at strain rates from 10(-2) s(-1) to 1.0 s(-1). A decrease in the Ni content below 50.2 at.% significantly deteriorated the hot workability due to the formation of a brittle second phase. Also, the low Ni content alloy showed poor workability when the temperature exceeded 900 degrees C. Additional compression tests were conducted under various conditions to clarify the effects of the chemical composition, solidification anisotropy, and the strain rate.

Early Life History of the Sevenband Grouper, Epinephelus septemfasciatus from Korea.

This study is conducted to monitor the morphological developmental features of the egg development, larvae and juvenile of Epinephelus septemfasciatus, the fertilized eggs were gotton using artificial insemination. Matured parents are collected from marine caged fish farms in Geomun-ri, Samsan-myeon, Yeosu-si, Jeollanamdo Korea in June 2012. The fertilized eggs were pelagic eggs containing one oil globule, and measured 0.81~0.89 mm (0.85±0.04 mm, n=50) in diameter. In regard to rearing environment, the water temperature is 21.0~23.0°C and the salinity is 32.0~33.2‰. Hatching was observed from 48 hours after fertilization, the mouth and anus of prelarvae was not opened but had egg yolk at newly hatched. 4 days after hatching, the mouth and anus of postlarvae was opened and began to eat Rotifer and was measured 2.40~2.49 mm (2.45±0.03 mm n=10) in total length. 12 days after hatching, postlarvae was measured 3.77~4.67 mm (4.27±0.33 mm) in total length, its the second pole tide of dorsal fin and the first pole tide of pelvic fin was extended longitudinally. 71 days after hatching, juvenile was measured 40.5~45.4 mm (42.6±2.04 mm) in total length. Seven bands were observed in body, and pole tides of dorsal and pelvic fins were shortened.

Additional duplicated Hox genes in the earthworm: Perionyx excavatus Hox genes consist of eleven paralog groups.

Annelida is a lophotrochozoan phylum whose members have a high degree of diversity in body plan morphology, reproductive strategies and ecological niches among others. Of the two traditional classes pertaining to the phylum Annelida (Polychaete and Clitellata), the structure and function of the Hox genes has not been clearly defined within the Oligochaeta class. Using a PCR-based survey, we were able to identify five new Hox genes from the earthworm Perionyx excavatus: a Hox3 gene (Pex-Hox3b), two Dfd genes (Pex-Lox6 and Pex-Lox18), and two posterior genes (Pex-post1 and -post2a). Our result suggests that the eleven earthworm Hox genes contain at least four paralog groups (PG) that have duplicated. We found the clitellates-diagnostic signature residues and annelid signature motif. Also, we show by semi-quantitative RT-PCR that duplicated Hox gene orthologs are differentially expressed in six different anterior-posterior body regions. These results provide essential data for comparative evolution of the Hox cluster within the Annelida.

Analysis of reports on orphan lung diseases in Korean children.

PURPOSE: Orphan lung diseases are defined as lung diseases with a prevalence of 1 or less in 2,000 individuals. Despite an increase in the numbers of patients with such diseases, few studies on Korean children have appeared. To obtain epidemiologic and demographic data on these diseases, we systematically reviewed reports on pediatric orphan lung diseases in Korea over the last 50 years. METHODS: We reviewed 223 articles that have appeared since 1958 on orphan lung diseases in Korean children. These articles described a total of 519 patients aged between 0 and 18 years. We classified patients by year of publication, diagnosis, geographic region, and journal. RESULTS: Of 519 patients, 401 had congenital cystic lung diseases and 66 had bronchiolitis obliterans. About 80% of patients were described in reports published in three journals, Pediatric Allergy and Respiratory Disease (Korea), the Korean Journal of Pediatrics, and the Korean Journal of Thoracic and Cardiovascular Surgery, in which papers on 157 (30.2%), 138 (26.6%), and 111 (21.4%) patients appeared, respectively. The frequency of publication of case reports has increased since 1990. Of the 519 patients, 401 (77.3%) were from Seoul/Gyeonggi-do and 72 (13.9%) from Busan/Gyeongsangnam-do. CONCLUSION: The prevalence of pediatric orphan lung disease has increased since 1990, and some provinces of Korea have a higher incidence of these diseases than do others. Studies exploring the incidence of pediatric orphan lung diseases in Korea are needed for effective disease management.

Status of intestinal parasitic infections in a remote island, Yondo, Jeonranam-do: province.

This study was performed to observe the present status of intestinal parasitic infections in a remote island, Yondo, located in southern part of Jeonranam-do(province), Korea. In February and May 1988, total 1,011 individual stool samples were collected and examined for intestinal helminths and protozoa using formalin-ether centrifugal sedimentation technique. The results are summarized as follows: Of 1,011 ingabitants examined, 398(39.4%) were positive for intestinal parasites. Helminth positives were 372(36.8%), and protozoan cyst positives were 56(5.5%). Ten species of parasites were found. Trichuris trichiura revealed the highest infection rate of 27.5%, Ascaris lumbricoides 17.4%, Taenia sp. 5.8%, Entamoeba coli 3.3%, Giardia lamblia 1.5%, Endolimax nana 0.8%, Hymenolepis nana 0.4%, hookworm 0.2%, Trichostrongylus orientalis 0.2%, and Entamoeba histolytica 0.2%, respectively. The female group showed higher positive rate (44.0%) than males (34.7%). Also, higher positive rates were observed among adults as compared with the group younger than 10 years old. Average value of E.P.G. was 1,876(range 200-17,800) in A. lumbricoides positives, and 327(range 200-1,600) in T. trichiura positive cases. In helminth egg positive cases, single infection was 63.4%, double infection 34.7%, and triple infection 1.9%, respectively. Among protozoan cyst positives, single infection was 94.6%, and double infection was 5.4%. The present study revealed that the prevalence of intestinal parasites among inhabitants in Yondo island is still so high that special control measures should be performed.

[Ultrastructural studies on the surface of Paragonimus westermani metacercaria]

The present study was undertaken to demonstrate the surface structure of Paragonimus westermani metacercaria in Korea with special reference to the distribution of sensory papillae. Metacercariae were isolated from crayfish, one of the second intermediate host of P. westermani in Bogil island, Chollanam-do (Province), Korea, where has been known as an endemic area of human paragonimiasis. Isolated metacercariae were excysted and examined with light, scanning and transmission electron microscopes for morphological features. On the surface of metacercariae, three types of sensory papillae were identified. Large domed papillae (3-5 micrometer), which were covered with wrinkled plasma membrane of the worm, were distributed on the oral and ventral suckers only. On the oral sucker, these large domed papillae were 12-13 in number. On the other hand large domed papillae on the ventral sucker were constantly 6 in number and hexagonal in distribution. Small domed papillae (2-3 micrometer), of which surface was more smooth than those of large ones, were distributed symmetrically on the ventral (30-32 pairs) and dorsal surfaces (40-42 pairs). Ciliated papillae (0.8-1.5 micrometer) were observed about 5-6 in number around the oral sucker and 3-5 pairs each on the ventral and dorsal surface of the body. Single pointed spines covered the entire surface of the body except around the excretory pore. Spines on the anterior part of the body were 0.9-2.0 micrometer in length and 45-55/100 square micrometer in number, and were gradually reduced in length (0.4-1.4 micrometer) and in number (12-27/100 square micrometer) toward the posterior part. The body wall of P. westermani metacercariae was consisted with anucleated syncytium layer, fibrous interstitial layer and musclar layer. In the anucleated syncytium, biconcave (0.15-0.55 micrometer) and spherical (0.08-0.16 micrometer) secretory granules, which were transferred from epidermal cells via protoplasmic tubules, mitochondria and ribosomes, were observed. Spines originated around the basement membrane protruded externally. Epidermal cells were consisted with a nucleus and a cytoplasm, and connected to syncytium with protoplasmic tubules. In the cytoplasm many secretory granules, mitochondria, Golgi complex, endoplasmic reticula, ribosomes and lipid droplets were observed.

[Detection of IgG antibodies with immunofluorescent antibody technique in human trichomoniasis]

The indirect fluorescent antibody(IFA) test was used to detect serum IgG and IgM antibodies to Trichomonas vaginalis in 31 vaginal trichomoniasis, 7 candidiasis and in 20 non-infected healthy wonem with antigen prepared from axenic culture of Trichomonas vaginalis isolated from vulvovaginitis patient. The results were as follows: In 31 vaginal trichomoniasis the positive reactions of IgG antibody were 27 in the 1/8 dilution or higher and 4 in the 1/4 dilution whereas in healthy women the reaction showed signigicantly low as in the 1/4 dilution of below. The sensitivity and specificity of IFA test for IgG antibody to trichomonad antigen in this study were 87.1% and 100%, respectively. No significant difference of IgM antibody levels between vaginal trichomoniasis and healthy women was observed. No relation between the levels of IgG and IgM antibodies to trichomonad antigen by IFA test was observed. No relation between the time lapse and the level of serum IgG antibodies in IFA test of vaginal trichomoniasis was regarded. In conclusion the present study suggests that IFA test in trichomoniasis could be a useful tool for detection of anti-trichomonad IgG antibodies and applicable as an immundiagnostic method.

[Effect of flubendazole against Ascaris lumbricoides, Trichocephalus trichiurus and Enterobius vermicularis in infected children]

Effectiveness of flubendazole, a newly introduced as an anthelmintic in Korea, was evaluated in 28 ascariasis, 28 whipworm infections and 17 pinworm infections in primary school children. All patients were orally given by flubendazole of 500 mg once. Stools for roundworm and whipworm and anal swabs for pinworm infections were examined both before and 25 days after treatment. After flubendazole treatment, all ascariasis and pinworm infected cases were cured. On the other hand, in whipworm infections, the cure rate was 82.1 percent (23 cured of 28 infected), and the egg reduction rate was 67.1 percent, respectively. Headache (one case), dizziness (2 cases) and abdominal pain (3 cases) were noticed as untoward effects, but they were mild and disappeared within 24 hours. These results demonstrate the excellent effectiveness of flubendazole in treating these helminth infections.