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Different patterns of bacterial communities have been reported in the airways and gastrointestinal tract of asthmatics when compared to healthy controls. However, the blood microbiome of asthmatics is yet to be investigated. Therefore, we aimed to determine whether a distinct serum microbiome is observed in asthmatics by metagenomic analysis of serum extracellular vesicles (EVs). We obtained serum from 190 adults with asthma and 260 healthy controls, from which EVs were isolated and analyzed. The bacterial composition of asthmatics was significantly different from that of healthy controls. Chao 1 index was significantly higher in the asthma group, while Shannon and Simpson indices were higher in the control group. At the phylum level, Bacteroidetes was more abundant in asthmatics, while Actinobacter, Verrucomicrobia, and Cyanobacteria were more abundant in healthy controls. At the genus level, 24 bacterial genera showed differences in relative abundance between asthmatics and controls, with linear discriminant analysis scores greater than 3. Further, in a diagnostic model based on these differences, a high predictive value with a sensitivity of 0.92 and a specificity of 0.93 was observed. In conclusion, we demonstrated distinct blood microbiome in asthma indicating the role of microbiome as a potential diagnostic marker of asthma.

The root-knot nematode (Meloidogyne incognita) is an important pathogen in crop cultivation, however, few methods are available to control this parasitic roundworm. In this study, the nematicidal effects of approximately 30 Streptomyces strains isolated from soil samples of Mt. Naejang (Korea) were tested against Meloidogyne incognita, and the culture broth of the strains KRA-24 and KRA-28 exhibited approximately 75% and 85% insecticidal activity, respectively, in in vitro assays. In in vivo pot experiments, these strains reduced the number of nematodes in the soil and the number of egg masses in the roots of red peppers. The two strains also survived in the presence of insecticidal agents (0.1 to 3.0%) such as fosthiazate, ethoprophos and terbufos when they were used in parallel. The mixture of KRA-24 or KRA-28 culture broth and fosthiazate exhibited nematicidal effects that were similar to those observed when KRA-24 or KRA-28 were used alone. Our results clearly suggest that the Streptomyces strains KRA-24 and KRA-28 should be promoted as a biocontrol agent against Meloidogyne incognita.

We compared the cultivation performance and genetic characteristics of Korean (Sugwawon No. 301) and Chinese strains (Huangguan No. 1) of the brown alga Saccharina japonica at an aquaculture farm in Korea. The number of cortical layer cells and the area of the cortical cells were measured for different size groups (5, 15, 30, 50, 100, 150 cm) of the two strains in a Korean culture farm from January to June 2018. We found better performance of the Sugwawon No. 301 strain in Korean waters, possibly due to increased flexibility as a result of the different cell arrangement of the two strains. The differences in cell shape (spherical or rectangular) and arrangement (irregular or constant layered) of the two strains began to appear at lengths of 50 cm where the Sugwawon No. 301 strain had greater numbers of smaller cells than Huangguan No. 1. In addition we established six sequence simple repeats (SSR) genetic markers that clearly distinguish these two strains of S. japonica. Based on our observations, we speculate that differences in the cell arrangement between strains of S. japonica were determined by genetic variation and morphological properties of the two kelp strains.

A LC-MS-guided screening led to the isolation of two new streptimidone derivatives (2 and 3) containing a glutarimide ring and two glutarimide ring-opened compounds (4 and 5) along with a known glutarimide-containing polyketide, streptimidone (1) from Streptomyces sp. W3002 strain. Their structures were elucidated by MS and NMR spectroscopic analyses and by comparison with data from the literature. Compound 2 is a non-hydroxylated analog at the C-5 position of streptimidone. The structure of 3 was determined as a streptimidone derivative possessing the α, β-unsaturated ketone moiety at positions C-5 and C-6. Compound 4 had similar chemical shifts and splitting patterns with 3, but the glutarimide ring is opened. Compound 5 closely resembles that of 4 with the only difference being the existence of an additional methoxy group instead of an amide group. Streptimidone (1) and 3 showed weak cytotoxic activity against three human cancer cell lines, respectively.

Background Current guidelines for the treatment of asthma and chronic obstructive pulmonary disease overlap (ACO) recommend initial treatment using inhaled corticosteroids (ICSs) plus 1 or more bronchodilators. Objective To clarify which therapeutic effect is better between the ICS + long-acting β2 agonist (LABA) and ICS + LABA + long-acting muscarinic antagonist (LAMA) treatment in patients with ACO. Methods We conducted a multicenter, 48-week, randomized, noninferiority trial. Patients with ACO were enrolled if they were treated with a moderate to high dose of ICS + LABA. In total, 303 patients were involved in the present trial, with 149 receiving ICS + LABA + LAMA. The primary end point was the time to first exacerbation. Secondary outcomes included changes in FEV1, forced vital capacity, FEV1/forced vital capacity ratio, asthma control, blood eosinophil count, and fractional exhaled nitric oxide. Results In the ICS + LABA treatment group, 29 of 154 patients (18.83%) experienced exacerbation, whereas 28 of 149 patients (18.79%) experienced exacerbation in the ICS + LABA + LAMA treatment group. The results of this noninferiority study were ultimately inconclusive (hazard ratio, 1.1; 95% CI, 0.66-1.84). However, the patients treated with the addition of LAMA showed significant improvements in FEV1 and forced vital capacity (P Conclusions Although this study was unable to conclude that ICS + LABA treatment is not inferior to ICS + LABA + LAMA in terms of exacerbation, it is obvious that the ICS + LABA + LAMA treatment group had improved lung function in ACO.

Korean Saccharina japonica is highly valued, both for human consumption and abalone feed. For the stable production of abalone feed, fresh seaweed biomass is required throughout the year. However, currently, the production of farmed Saccharina is limited by environmental conditions such as temperature, irradiance, and nutrient availability between August and November. Due to shortages experienced in supply, the production of early-season biomass can be highly profitable and, therefore, some famers attempt to start their cultivation activities before prevailing, surface seawater temperatures (SST) are optimal. However, attempting to cultivate too early, can lead to total crop failure. Young kelp sporophytes are easily destroyed between 18 and 22 °C SST, which can occur during the early nursery period when the materials are confined to tanks. This study investigated the growth of S. japonica thalli and photosynthetic quantum yield (Fv/Fm) under five temperatures (i.e., 18–26 °C, at 2° increments) and five irradiances (i.e., 5, 10, 20, 40, and 80 μmol photons m−2 s−1). This was undertaken for four different size groups of sporophyte thalli (i.e., 0.25, 1, 5, 10 mm). There were different responses of the initial groups of S. japonica showing different tolerances to temperature and irradiance. In general, the smaller plants (1 mm) were more tolerant of sub-optimal conditions than their larger cohorts. These results indicated the optimum temperature and irradiance ranges for different size groups of S. japonica thalli which, if adopted in management protocols, could contribute to enhanced profitability and a more stable and evenly distributed production of Saccharina raw materials over an entire annual basis.

Cultivations of the brown seaweed, Saccharina japonica, was developed, promoted, and industrialized in the 1970s and is highly valued in Korea both for human consumption and as a feed for the rapidly developing abalone farming industry. Production has continually increased, and approximately 442,637 tonnes wet weight was harvested in Korea in 2015. Abalone is a highly sought-after delicacy in Korea, and the abalone farming industry has been developed based on a stable production of seaweed. Korean abalone farmers prefer to feed their stock on locally cultured S. japonica; however, between August and November, the supply of farmed S. japonica declines because higher seawater temperatures reduce S. japonica productivity. In an attempt to overcome this temperature-induced period of low production, cultivation trials with a strain of S. japonica selected to withstand higher cultivation temperatures were undertaken. Strain selection involved using individual parent thalli that were found to remain productive under high seawater temperature (26 °C) at Wando. Male and female gametophytes were isolated through 3 cycles of free-living gametophyte culture to produce the F3 strain used in the production trials. Production trials using the selected strain were initiated every month between December 2014 and March 2015. This delayed the initiation of culture beyond the latest initiation time currently used by farmers (December). Delaying initiation of cultivation resulted in delayed maximum growth compared to the control. Growth of the F3 strain continued for up to 3 months longer than normally achieved on farms for non-selected thalli. The mean length, growth rate, and biomass were also greater than those achieved by the control strain. The use of the F3 strain of S. japonica coupled with delayed initiation of culture can therefore be used to help to ensure a stable year round algal feed supply for abalone industry in Korea.

Aims The purpose of this study is to investigate IL-6/STAT3 inhibitory activity using lactic acid bacteria (LABs) isolated from Gajuknamu kimchi. Methods and Results Six lactic acid bacteria (LABs) were isolated from Gajuknamu kimchi and identified through 16S rRNA sequencing. Among them, the culture broth of Lactococcus lactis KR-050L inhibited IL-6-induced STAT3 luciferase activity. Fifteen compounds were isolated from the EtOAc extract of culture broth though column chromatography and preparative HPLC, and they were identified as 2,5-diketopipperazine structures by spectroscopic analyses (MS, 1H- and 13C-NMR). They also showed inhibitory activities on IL-6 induced STAT3 activation, and showed the different in activity according to the presence of a phenylalanine residue, hydroxyl groups and isometric structure. Conclusions The six new LABs isolated from Gajuknamu kimchi, and L. lactis KR-050L was selected as candidate IL-6/STAT3 inhibitors. The activity levels of 15 2,5-DKPs isolated from L. lactis KR-050L were verified. Significance and Impact of the Study This study constitutes the first attempt to isolate various LABs from Gajuknamu kimchi and to discover IL-6/STAT3 inhibitors in the EtOAc extract of L. lactis KR-050L culture broth. Moreover, our data provide useful biochemical information regarding the commercialization of L. lactis isolated from Gajuknamu kimchi as an approach to use functional foods for the treatment of various diseases via IL-6/STAT3 activation. This article is protected by copyright. All rights reserved.

The interleukin-6 (IL-6) family of cytokines plays a key role in the pathogenesis of rheumatoid arthritis and osteoporosis through the regulation of bone formation and resorption. In this study, it was observed that ethanol extract of Salvia plebeia R.Br. (S.P-EE) inhibited IL-6-induced signaling cascade including phosphorylation of JAK2/STAT3 and ERK. Subsequently, it was examined whether S.P-EE treatment could recover bone loss in ovariectomized (OVX) mice. Indeed, S.P-EE exhibited both preventive and therapeutic effect on OVX-induced bone loss in trabecular microarchitecture along with significant increase in bone mineral density and content. To understand the mechanism of action of S.P-EE in bone metabolism, the effect of S.P-EE on osteoclast differentiation and activity was investigated. S.P-EE significantly inhibited RANKL-induced osteoclast differentiation by suppressing phosphorylation of MAPK and Akt, and expression of NFATc1 and osteoclast marker genes. S.P-EE also inhibited bone-resorbing activity of osteoclasts. Furthermore, isolation and identification of the active compounds which are responsible for the inhibitory effect of S.P-EE on osteoclast differentiation was carried out. Six major flavonoids and plebeiolide A-C were isolated and examined their effects on osteoclast differentiation. Luteolin and hispidulin, and plebeiolide A and C, not B exhibited potent inhibitory activity on RANKL-induced osteoclast formation.

Lee, J.-S.; Son, D.-H.; Lee, S.-H.; Myeong, H.-H.; Cho, J.-S.; Lee, J.-C.; Lee, J.-Y.; Park, C.-S., and Kim, J.-W., 2020. Canonical correspondence analysis ordinations and competitor, stress tolerator, and ruderal strategies of coastal dune plants in South Korea. Journal of Coastal Research, 36(3), 528–535. Coconut Creek (Florida), ISSN 0749-0208.The distribution of plant communities in the sand dunes of the SW coasts of South Korea was studied, along with environmental factors and plant traits, by canonical correspondence analysis (CCA). The competitor, stress tolerator, and ruderal (CSR) ecological strategies were also evaluated. The coastal sand dune plants were classified into two plant trait groups in the CCA biplot diagram. First, vegetation was correlated with leaf dry weight, canopy height, specific leaf area, leaf dry matter content, and lateral spread. Second, it was correlated with the flowering period and flowering start. Coastal sand dune plants were classified into three soil factor groups in axes 1 and 2 of the CCA biplot diagram. First, the vegetation was correlated with total nitrogen (T-N), K+, Silt, Mg2+, Na+, and clay contents. Second, it was correlated with Ca2+. Third, it was correlated with the sand contents. To elucidate the relative significance of competition, stress, and disturbance in the distribution process of plant communities, the CSR distribution model was adopted. Many coastal plants (12 species) showed competitor-ruderal/competitor-stress-tolerant-ruderal strategies: Artemisia fukudo, Atriplex gmelinii, Carex kobomugi, Calystegia soldanella, Digitaria ciliaris, Linaria japonica, Messerschmidia sibirica, Oenothera bieenis, Salicornia europaea, Salsola komarovii, Suaeda glauca, and Suaeda maritima. The four species with stress-tolerant-competitor/competitor-stress-tolerant-ruderal strategies were Carex pumila, Imperata cylindrica var. koenigii, Limonium tetragonum, and Zoysia sinica. Conyza canadensis, Ischaemum anthephoroides, Phragmites communis, and Vitex rotundifolia displayed competitor/stress-tolerant-competitor, competitor-ruderal, stress-tolerant-competitor, and C/SC strategies, respectively. The differences in distribution and restoration patterns of the CCA diagrams and CSR triangles may be attributable to different adaptions of plant traits or soil factors.

Hand-foot-and-mouth disease, a highly contagious viral infection, occurs more common in children than in adults. However, there was a recent outbreak of Coxsackievirus A6-induced infection with an atypical presentation among the adult population. Stevens–Johnson syndrome is a severe mucocutaneous disease characterized by extensive necrosis and detachment of the epidermis, and this condition is commonly caused by medications. Herein, we describe a 30-year-old male patient taking allopurinol for the management of gout. The patient presented with numerous erythematous papules, vesicles, and patches with mucosal eruptions on the whole body, oral mucositis, and fever, and he was finally diagnosed with hand-foot-and-mouth disease.

Ampelopsis brevipedunculata(Maxim.) Trautv. (AB), a traditional East Asian medicine, exhibits protective effects against several inflammatory diseases. Our search for an inhibitor of IL-6-induced JAK2/STAT3 activation revealed that AB ethanolic extract (ABE) had a significant inhibitory effect on IL-6-induced STAT3 expression in Hep3B cells. The isolation and purification of an EtOAc-soluble fraction of ABE (ABEA) using reversed-phase high-performance liquid chromatography (RP-HPLC) afforded 17 compounds. The structures of these compounds (1-17) were elucidated based on1H and13C nuclear magnetic resonance (NMR) spectroscopy as well as electrospray-ionization mass spectrometry (ESI-MS) data. ABE and ABEA were screened by a luciferase assay using Hep3B cells transfected with the STAT3 reporter gene. ABEA exhibited potent inhibitory effects on IL-6-induced STAT3 expression; moreover, these effects arose from the inhibition of the phosphorylation of the STAT3, JAK2, and ERK proteins in U266 cells. In addition, the compounds isolated from ABEA were measured for their inhibitory effects on IL-6-stimulated STAT3 expression. Of the compounds isolated, betulin showed the greatest inhibitory effects on IL-6-induced STAT3 activation in the luciferase assay (IC50value: 3.12μM). Because of its potential for inhibiting STAT3 activation,A. brevipedunculatacould be considered a source of compounds of pharmaceutical interest.

The brown seaweeds, Sargassum fusiforme and Sargassum fulvellum are edible and highly valued in Korea. During the summer season, phytal animals graze heavily on young algal blades and holdfasts of these two species and substantially reduce harvestable biomass. Acid and salinity treatments are used to efficiently control infestations by phytal animals in other cultivated algae (e.g., Pyropia), but their effects on the removal of phytal animals in Sargassum cultivation are unknown. Here we investigated the effects of pH (range 2 ~ 13) and salinity (range 0 ~ 44 psu) on the removal of two major phytal animals, Caprella scaura and Gammaropsis utinomi, associated with S. fusiforme and S. fulvellum. We also examined the optimum quantum yield (F v/F m) of algae in the same experimental conditions to quantify the tolerance of algae to acid and salinity treatments. The two animal species experienced more than 80 % mortality at pH lower that pH 4 and the extreams of salinity (0 ~ 10 and 44 psu) after a 5-min of immersion. The quantum yield of S. fulvellum was not significantly different from controls (ambient pH and salinity) within the pH 4–10 range and within the 7–40 psu salinity range. Similarly, the quantum yield of S. fusiforme was not significantly different from controls within the pH 3–11 range and the 0–44 psu salinity range. Therefore, if pH and salinity conditions outside these ranges were used in commercial Sargassum culture, the removal of the two animal species would be higher, but quantum yield of algae would be substantially reduced. These results indicate that pH and salinity treatments could be used effectively to remove the phytal organisms without significantly affecting optimum quantum yield in S. fusiforme and S. fulvellum.

The kelp Undariopsis peterseniana (Kjellman) Miyabe et Okamura is warm water tolerant and consequently there is currently considerable interest in developing commercial cultivation techniques for this species in Korea. Undaria is an important species for both the abalone industry and for commercial seaweed mariculture. In an attempt to extend the culture period of Undaria we bred and cultured hybrid kelp that is a cross between free-living gametophytes of U. peterseniana and Undariopsis pinnatifida. The morphological characteristics of the F1 hybrid sporophytes were intermediate between those of the parent plants having shallow pinnated blades and forming both sorus and sporophyll. A F2 generation was produced by isolating zoospores from sorus and sporophyll separately from a F1 hybrid thallus. Using free-living gametophyte seeding and standard on-growing techniques, F0 (female U. pinnatifida and male U. peterseniana), F1, and F2 gametophytes were cultured from December 2011 to May 2013. The morphological differences between the F1 and F2 generations were assessed. The F2 hybrids were found to have longer pinnate blades and narrower midribs than the F1 hybrid and only formed sporophylls. Growth and morphology of F2 hybrids originating from the sporophyll or sorus of the F1 hybrids were not morphologically different from each other. Both of the F2 hybrids exhibited late maturation, with the early stages of sporophylls appearing in April. This late maturation of F2 hybrids is beneficial in the development of hybrid strains that extend the period of availability of Undaria for abalone feed and cultivation in Korea.

Ecklonia cava and Ecklonia stolonifera are perennial brown algae that form sea forests off the coast of Korea. Both species are cultured to supply a summer feed for the abalone industry. Recent expansion of the abalone industry in Korea has been bringing an increase in demand for fresh algal supply. Zoospores of the two algae were seeded in October 2006 on seed frames coiled with 100 m of seed fibers. After 2 months of indoor culture and 2 months of intermediate culture, growth and production of the two algae were compared during their main cultivation period from March 2007 to June 2008, in the culture ground in Wando, Korea (34°26′18.68″ N, 127°05′43.88″ E), in situ. Growth rate of E. cava and E. stolonifera was 1.058 and 3.089 mm day−1, respectively. The mean production of E. stolonifera obtained from the culture ropes was ca. 12 kg wet wt. m−1 of culture rope while production of E. cava was ca. 3 kg wet wt. m−1 of culture rope. The difference in production was attributed from the different growth strategies of the two algae, with only E. stolonifera being able to regenerate blades from the holdfast. The ability to regenerate blades from the holdfast therefore makes E. stolonifera the preferred species for biomass production for abalone feeds. In a 120-day feeding experiment, growth rate, weight gain, and survival rate of abalone showed that E. cava and E. stolonifera feeds could provide an alternative feed to Saccharina japonica during summer months.

Inhibiting interleukin-6 (IL-6) has been postulated as an effective therapy in the pathogenesis of several inflammatory diseases. In this study, seven flavonoids were isolated from the methanol extracts of Psoralea corylifolia by bioactivity-guided fractionation. The structures of bakuchiol (1), bavachinin (2), neobavaisoflavone (3), corylifol A (4), corylin (5), isobavachalcon (6), and bavachin (7) were determined by spectroscopic analysis (1H-, 13C- NMR and MS). We demonstrated that compounds 1-7 showed an inhibitory effect on IL-6-induced STAT3 promoter activity in Hep3B cells with IC50 values of 4.57 ± 0.45, 3.02 ± 0.53, 2.77 ± 0.02, 0.81 ± 0.15, 1.37 ± 0.45, 2.45 ± 0.13, and 4.89 ± 0.05 µΜ, respectively. These compounds also inhibited STAT3 phosphorylation induced by IL-6 in Hep3B cells. Overall, several flavonoids from P. corylifolia might be useful remedies for treating inflammatory diseases by inhibiting IL-6-induced STAT3 activation and phosphorylation.

The occurrence of encrusting colonies of the hydroid Obelia geniculata on farmed Saccharina japonica was examined between December 2007 and July 2008 at a Saccharina farm at Wando on the southwestern coast of Korea. The growth stages of S. japonica can be divided into two phases: an active growth phase from February to the end of May and a decay phase from June to July. There was a significant increase in the level of incrustation by colonies on fronds (measured as the percentage of fronds with encrusting colonies) between February and July (p

The kelp Undariopsis peterseniana is warm-water-tolerant, and consequently, there is currently considerable interest in developing commercial cultivation techniques for this species in Korea. U. peterseniana plants have been successfully transferred to the northern coast of Korea beyond their original habitat in Jeju Island (33°30′08.65″N, 126°55′39.02″E). In this study, we cultured a hybrid kelp consisting of a cross between free-living gametophytes of U. peterseniana and U. pinnatifida in an attempt to extend the culture period of Undaria which is an important species for both the abalone industry and for commercial seaweed mariculture for human food applications. Morphological characters and cultivation period were compared between the parent thalli and the hybrid. The cultivation experiment was conducted in Wando, on the southern coast of Korea (34°26′18.68″N, 127°05′43.88″E). The morphological characteristics of the hybrid thalli were intermediate between the two species having shallow pinnated blades and a reduced reproductive organ. Hybrid thalli showed faster growth rates, 1.5 times greater biomass, and a longer cultivation period than the parent thalli. The hybrid strain possessed characteristics that indicate it could be used as an alternative kelp source to supply the abalone feed industry.

The relationship between median grain size of sediments and the abundance in the wild of green algal mats (Ulva prolifera) on the intertidal flats of Muan, Korea, were investigated. The impact of substratum particle size on the growth and survival of germlings was examined in the laboratory. In the wild, the average annual density of algal mats was 7,950 ind m−2. The algal mats mainly occurred in sands and exhibited patchy distribution. Statistical analysis indicates significant spatial analysis differences and a significant relationship between density and the ratio of sands to silts, suggesting that the distribution and density of this species were related to particle size. In laboratory experiments, the survival rate of U. prolifera germlings was the lowest value (22%) on sediments with a median grain size of 63–125 μm. Laboratory experiments have generally shown a positive relationship between attachment or survival of the alga and substratum particles size. Our laboratory results indicate a clear link between germling settlement/survival and substratum particle size. These results explain the spatial differences in abundance observed in the field in relation to the distribution and ratio of sands to silt on the Muan flats.

곰피 (Ecklonia stolonifera Okamura)는 다시마목 미역과의 다년생 해조로서 점심대의 깊은 곳에서 자라며, 해중림을 구성하는 해조로 연안생태계에서 1차생산자로서의 중요한 기능을 담당하고 있다 (Kang and Yoo, 1993; Brown and Lamare, 1994). 곰피는 우리나라 동해안의 특산으로 부산 연안에서 포항인근까지 분포하였으나 서식지가 점점 확대되어 현재는 남해안 여수 지역까지 자생지가 확인되었다. 대형 갈조류인 곰피는 다시마와 형태적으로 매우 유사한 해조류로 알긴산뿐 아니라 순도가 높은 푸코이단을 다량 함유하고 있으며, 곰피의 푸코이단은 트롬빈 활성이 높아 헤파린의 약 1.4배에 달한다고 알려져 있다 (Lee et al., 1995). 또한 여름철 고수온기 동안 전복의 먹이원으로 이용하거나, 해중림 조성을 통한 연안어장 생산력 증대 및 해양 환경 정화 등에 활용이 가능하다. 대형 갈조류는 산업적 활용성 또는 생태계의 기여도 등에서 높은 잠재성을 가지고 있기 때문에 이들의 원활한 바이오매스 확보에 필수적인 효율적 증식방법 개발을 위한 다양한 시도들이 이루어지고 있다.이형세대교번을 하는 다시마과 해조류는 배우체 세대에서 암․수배우체를 각각 분리하여 무성적으로 증식시킬 수 있으며, 환경조절을 통하여 배우자형성을 유도할 수 있음이 알려져있다 (Luning, 1980). 또한 최근 칠레에서는 전복 먹이원으로써 다시마류의 대량양식에 실내에서 대량 증식시킨 유리 배우체를 이용하는 방안이 개발된 바 있으며 (Westermeier et al., 2006; Edding and Tala, 2003), 국내에서는 감태의 유리배우체 재생 및 성숙 유도 조건이 밝혀진 바 있다 (Wi et al., 2008).현재 우리나라에서 행해지고 있는 미역이나 다시마 등의 양식방법은 매년 성숙된 모조로부터 방출된 유주자를 기질에 채묘하여 양식하는 방법을 사용하고 있으나 이러한 종묘생산 방법을 대규모 곰피 양식에 적용할 경우 많은 양의 모조가 사용되어야 하므로 자연군락의 훼손을 야기할 가능성이 있다 (Wi et al., 2008). 그러나 유리배우체의 배양에 의한 방법은 소량의 성숙 엽체만으로도 많은 양의 배우체 확보가 가능하며, 클론 배우체의 유도를 통한 우량 품종의 교배와 선발육종의 기초를 다질 수 있는 매우 효율적인 방법이라 할 수 있다. 최근 Hwang et al. (2009)은 곰피 유주자 채묘에 의한 양식기법을 확립하여 가이식과 양성조건을 위한 적정조건을 보고한 바 있는데, 곰피의 유리배우체 배양에 의한 안정적인 종묘확보가 이루어질 수 있다면 해중림 조성용 또는 전복 먹이원으로 곰피의 안정적인 양식생산을 꾀할 수 있을 것이다. 따라서 본 연구에서는 곰피 암수 배우체의 분리 배양 및 생장과 성숙 조건을 파악하여 안정적인 유리배우체의 배양조건을 확립함으로써 효율적인 곰피 양식의 기초를 마련하고자 하였다.

Previously, three extracellular proteases, Vpr, PepT, and subtilisin were identified from Bacillus subtilis KCTC 3014. To confirm the activity of Vpr, two recombinant Vpr proteins, full Vpr with TTG (pGST-fTTG-Vpr) and full Vpr with ATG (pGST-fATG-Vpr) as an initiation codon were expressed using a pGEX-2T vector encoding glutathione S-transferase (GST) in Escherichia coli. Vpr was produced in two forms, occurring as four spots on a 2-DE gel, 68 and 75 kDa proteins with similar pI values (4.0 ∼ 4.5). Activity was detected in a fibrin zymography at the expected molecular size of 68 kDa (mature form) processed from full Vpr. However, the recombinant 75 kDa of GST-fVpr did not exhibit activity. Replacement of the TTG codon with ATG led to 1.9-fold increased enzyme activity in 68 kDa. Interestingly, the expression of GSTVpr resulted in the proteolytic degradation of the protein and no GST fusion Vpr protein was detected.

Physicochemical responses to acidification in Pythium porphyrae, the causative organism of red rot disease in Porphyra, were investigated. The acid tolerance of P. porphyrae mycelia under pH 4 (acidic condition) condition increased significantly compared with that of the mycelia under pH 8 (condition equivalent to seawater) condition. Free amino acid levels in the mycelia decreased 1.3–8.8-fold under pH 4 condition. However, some free amino acids such as the d-cysteinolic acid-like component, phosphoethanolamine, glutamic acid, aminoadipic acid and methionine increased 2.6–21.7-fold under the same condition. Proton flux on the mycelia exposed to pH 8 increased significantly compared with the mycelia exposed to pH 4. The patterns of proteins present (examined by two-dimensional polyacrylamide gel electrophoresis) differed among the pH conditions. These results suggest that P. porphyrae acquires acid tolerance and is able to adapt to the changing pH conditions. This has significant implications for the use of acidic fungicide treatment for prevention of red rot disease on Porphyra farms.

By disruption of the pullulan synthetase gene (pul) of Aureobasidium pullulans IMS822 KCTC11179BP, we constructed a mutant strain, A. pullulans NP1221, which produced a pure beta-glucan exopolysaccharide. The mutant NP1221 was white, whereas the wild-type strain produced a black dye. When we compared fermentation kinetics between wide-type and mutant strains, the mutant NP1221 did not produce pullulan. Substrate uptake rate and beta-glucan production were similar in both strains. However, the biomass yield of mutant NP1221 was 2.3-fold (9.2 g l(-1)) greater than that of wild-type.

The dietary fiber and crude fiber contents of different thallus regions (blade, stipe, and holdfast) in three brown algae (Laminaria japonica, Ecklonia stolonifera, and E. cava) were detennined at different ages, and then compared with one another. On a dry matter basis, the soluble dietary fiber (SDF) content was highest () in the holdfast of 2-year old L. japonica, and the insoluble dietary fiber (IDF), total dietary fiber (TDF), and crude fiber (CF) contents were highest in the holdfast of 2-year old E. cava at , , , respectively. The IDF, TDF, and CF contents of these three species were measured in the following order: holdfast > stipe > blade, and the SDF contents of L. japonica exhibited the reverse of this trend. The TDF/CF ratio of 1-year old L. japonica, E. stolonifera, and E. cava was greater than was observed in the corresponding 2-year old samples. This is, to the best of our knowledge, the first report demonstrating that the holdfasts of L. japonica, E. stolonifera, and E. cava are rich in dietary fiber contents, especially IDF, TDF, and CF.

The mass cultivation of Ecklonia stolonifera Okamura was studied as a possible summer feed for the abalone industry in Korea for the period between August and November when Undaria and Laminaria are not available. Experiments were conducted to investigate the optimal conditions for artificial seed production and mass cultivation of this species. Seedlings of E. stolonifera were reared in an indoor tank for 60 days until they were around 500 μm in length. Following indoor tank culture, the seedlings were transferred in situ to a nursery culture area for 2 months, before begin transferred to the main grow-out area. The maximum growth and development of young thalli in nursery culture area occurred at 2 m depth, whilst maximum growth of thalli in the main culture area occured at 1.5 m depth. Production of E. stolonifera was between 3 and 9 kg wet wt. m−1 in the first year of culture after seeding and 12 to 13 kg wet wt. m−1 in the second year of culture, after management (depth control and fouling organism removal, etc.) of the holdfast. The relationship between optimal water depth for culture and underwater irradiance during the E. stolonifera cultivation was defined as: y = −0.331x + 8.198 (r 2 = 0.9903). The growth rates achieved in this trial indicate that E. stolonifera cultures could produce sufficient biomass to supply summer feed for the Korean abalone industry.

Induction of regeneration and maturation for the free-living gametophytes of Ecklonia cava Kjellman was conducted under various temperature, irradiance and photoperiod conditions. Culture conditions for female or male gametophyte fragments were 4 temperature (5, 10, 15 and ), 4 irradiance (5, 10, 20 and ) and 3 photoperiod (14:10, 12:12 and 10:14 h L:D). Female and male gametophyte fragments were maintained in active regeneration without reaching sexual maturity under , , 12:12h or 10:14h (L:D) and , , 14:10h (L:D), respectively. Sexual maturation of female and male gametophytes facilitated under , , 14:10h (L:D) and , , 10:14h (L:D), respectively. These results provide basic informations to control the regeneration or maturation of the free-living gametophytes for artificial seed production of E. cava.

Infection rates of pinhole disease in farmed Undaria pinnatifida (Harvey) Suringar were examined between November 2002 and April 2003 at an Undaria farm at Wando on the southwestern coast of Korea. The growth stages of U. pinnatifida can be defined as: a growth phase from December to the end of February, and a decay phase from March to April. There was a significant increase in the level of infection with pinhole disease (measured as the number of pinholes per cm2) between December and April (P < 0.01). Infection progressed from the upper part of the frond in December to the basal part in April. The infection rate of pinhole disease in relation to growth phase on farms over time is limited by the harvest of the seaweed crop at the end of the cultivation period in April. The midrib, sporophyll and stipe showed no signs of infection at any time during the cultivation period. The reduced physiological activity and tissue aging that occurred over time in the sporophytic life phase is the probable cause of the increasing infection rate of the disease.

The summer algal flora and community of 15 uninhabited islands in Dochodo, southwestern coast of Korea, were investigated from 9 June to 16 September 2007. A total 53 species (10 green, 14 brown and 29 red algae) of marine algae were identified. Among 15 uninhabited islands, the number of species observed was the highest as 33 species at Jeongdo and Gyeongchido the least as 24 species at Hugdo and Mido. The dominant species were Enteromorpha compressa, Ishige okamurae, Gloiopeltis furcata, Ulva pertusa and Sargassum thunbergii. The algal zonation of intertidal zone was figured out by Gloiopeltis furcata, Caulacanthus okamurae – Ulva pertusa, Sargassum thungergii – Gelidium amansii, Sargassum horneri from upper to lower zone. The flora investigated could be classified into six functional groups such as coarsely branched form (39.6%), filamentous form (25.8%), sheet form (14.1%), thick leathery form (9.9%), jointed calcarious form (5.6%) and crustose form algae (4.9%).

Codium fragile (Suringar) Hariot is an edible green alga farmed in Korea using seed stock produced from regeneration of isolated utricles and medullary filaments. Experiments were conducted to reveal the optimal conditions for nursery culture and out-growing of C. fragile. Sampling and measurement of underwater irradiance were carried out at farms cultivating C. fragile at Wando, on the southwestern coast of Korea, from October 2004 to August 2005. Growth of erect thalli and underwater irradiance were measured over a range of depths for three culture stages. During the nursery cultivation stage (Stage I), growth rate was greatest at 0.5 m depth (0.055 ± 0.032 mm day−1), where the average midday irradiance over 60 days was 924 ± 32 μmol photons m−2 s−1. During the pre-main cultivation stage (Stage II), the greatest growth rate occurred at a depth of 2 m (0.113 ± 0.003 mm day−1) with an average irradiance of 248 ± 116 μmol photons m−2 s−1. For the main cultivation stage (Stage III) of the alga, thalli achieved the greatest increase in biomass at 1 m depth (7.2 ± 1.0 kg fresh wt m−1). These results suggest that optimal growth at each cultivation stages of C. fragile could be controlled by depth of cultivation rope.

Different sampling methods can often produce different results when cyanobacterial blooms are intense and there are surface scums. Accordingly, five commonly used sampling methods and four sampling times were compared for monitoring of Oscillatoria and Microcystis populations. The different methods and times led to significantly different results for cyanobacterial biomass and its variance. Sampling methods that tended to be more specific to surface scums resulted in much higher and more variable measurements. Although sampling later in the day greatly reduced variances, the measured biomass was much lower than at earlier times of day, which was attributed to cyanobacterial downward migration in the afternoon. These trends were common for Oscillatoria and Microcystis. In order to overcome such problems, the median and median absolute deviation (MAD) were proposed, instead of the arithmetic mean and standard deviation (SD), for presenting a central tendency of measured values. Although the mean ± SD has been widely used to express a central tendency, it is too sensitive to outlying values that are very common during cyanobacterial blooms. The large differences in the mean values between the sampling methods and times were reduced by using the median values. Furthermore, the median ± MAD revealed the real data distribution more effectively.

The nutritive value of Capsosiphon fulvescens (C. Agardh) Setchell et Gardner, a new developing species for cultivation of marine macroalgae in Korea, was assessed by comparison with common edible green seaweed Ulva prolifera (Oeder) J. Agardh collected from Korea and Japan, based on analysis of its chemical composition. The contents of protein and of total, essential, and free amino acids of C. fulvescens were significantly higher than those of U. prolifera, whereas those of lipid, carbohydrate, and total dietary fibres were significantly lower. There were no significant differences in the moisture content between Capsosiphon and Ulva species. The main minerals of Capsosiphon and Ulva are Na, Mg, K, Ca, and Al, making up approximately 97–99 % of the total minerals. The contents of Na and V of Capsosiphon were significantly greater than those of Ulva, while those of Mn and Sr were significantly lower in Capsosiphon. The contents of retinal (27.8 μg g−1 dry wt) and ascorbic acid (0.28 mg g−1 dry wt) of Capsosiphon were significantly higher than those of Ulva, but the contents of cobalamin were lower. These results suggest that C. fulvescens has greater potential to be used as human food and as an ingredient in formulated food.

The effects pH and salinity on the brown alga Sargassum fulvellum were investigated in the context of the removal of two major associated animals, Caprella scaura and Gammaropsis utinomi. Optimum quantum yield (Fv/Fm) of S. fulvellum was also examined in the same experimental conditions as an index of stress. Experiments on pH and salinity tolerances of the two animal species indicated that mortality was more than 80% at extremes of pH (2, 3, 4, 10, 11, 12) and salinity (0, 3.5, 7, 10, 44 psu) after a 5 min treatment. Lethal time (LT50) from pH 2 to pH 4 was less than 90 sec in C. scaura, and less than 70 sec in G. utinomi. From 0 to 10 psu, LT50 was less than 20 sec in C. scaura and less than 60 sec in G. utinomi. The quantum yield of S. fulvellum was not significantly different from controls within the pH range 4~10, and within the salinity range of 7-40 psu. When exposed to pH and salinity conditions outside these ranges, the effect of these factors on the removal of two animal species was higher, but quantum yield was highly reduced. These results indicate that the optimal conditions for removing the animal species without affecting optimum quantum yield were pH 4-10, and salinities 7-10 psu and 44 psu.

Sargassum fulvellum is a brown alga recently introduced to the seaweed cultivation industry in Korea. There is current interest in the commercial scale of aquaculture of this species. For the artificial seeding and cultivation of this alga, growth and maturation were investigated from September 2002 to August 2003. Indoor culture experiments for maturation induction were also conducted at temperatures of 5, 10, 15, 20 and 25 °C and irradiances of 20, 50, 80 and 100 µmol photons m−2 s−1 under 16:8 h (L:D) photoperiod. Within a given culture test range, higher temperature and irradiance levels favoured the maturation of receptacles in S. fulvellum. Using temperature and irradiance control for thalli, artificial seed production of this species could be done one month earlier than thalli matured in nature. Under natural condition, receptacle formation of the plants began in February, and the eggs were released from March to April. For mature thalli of 200 g wet wt., artificial seeding was complete enough for attachment on seed strings of 100 m. Mean production obtained from the artificial seeding technique in situ was 3.0 kg wet wt m−1 of culture rope during the cultivation period.

The fungal parasite Pythium porphyrae is the causative organism of red rot disease in Porphyra cultivation farms. The detection of P. porphyrae from dried Porphyra yezoensis sheets was achieved using the species-specific primers PP-1 (5′-TGTGTTCTGTGCT-CCTCTCG-3′) and PP-2 (5′-CCCAAATTGGTGTTGCCTCC-3′) with the polymerase chain reaction (PCR). The DNA sequence (707 bp) of PCR product was found to be identical to that amplified from ITS rDNA extracted from a type species of P. porphyrae (IFO 30800, The Institute of Fermentation, Osaka, Japan). Quantities of the product amplified varied with the time when samples were harvested after the occurrence of red rot disease in Porphyra farms. This simple, rapid, and inexpensive method should have great applications in furthering quality control and determination of quality ranking in the Porphyra processing industry.

Codium fragile is commercially farmed in Korea by natural blooming zygote attachment. Experiments found optimum conditions for artificial seed production and nursery culture of C. fragile by asexual reproduction. Isolated utricles and medullary filaments were regenerated to erect thalli using both indoor and outdoor culture experiments. Under the indoor culture conditions, irradiance was an important factor to control the development of erect thalli. Formation of erect thallus from the isolated medullary filaments in the indoor culture was induced after 30 days under and . The detachment of isolated utricles and medullary filaments from the substrates of seed strings was reduced by exposure to the air during 2 hrs before the indoor culture of seed strings. The maximum growth and development of erect thalli in the nursery culture was induced at a water depth of 0.5 m. Depending on the substrates of the seed strings the growth of erect thalli was not significantly different (p>0.05).

Codium fragile (Suringar) Hariot, an edible green alga is farmed in Korea by natural blooming zygotes attachment. Experiments were conducted to reveal the conditions for artificial seed production of C. fragile by sexual and asexual reproduction. Growth was compared between zygotes attachment (sexual reproduction) and isolated utricles with medullary filaments (asexual reproduction). Zygotes and isolated utricles with medullary filaments were cultured under different light conditions (10, 20, 40, 60 and ) and temperatures (5, 10, 15, 20 and ) under 16:8LD. Maximum growth of zygote was under and after 13 days culture. Maximum regeneration of isolated medullary filament was per one isolated utricle under and after 15 days culture. After intermediate culture during two months in the field, morphogenesis occurred in both sexual and asexual reproduction, and growth of young thalli was not significantly different (p>0.05) between the both reproduction methods. Even though seed production of C. fragile is possible in both sexual and asexual reproduction, the mass artificial seed production of asexual reproduction is much more effective than that of sexual reproduction that is too much affected by maturity.

Bacillus subtilis C9 effectively degrades aliphatic hydrocarbons up to a chain length of C19 and produces a lipopeptide-type biosurfactant, surfactin, yet it has no genetic competency. Therefore, to obtain a transformable surfactin producer, the sfp gene cloned from B. subtilis C9 was integrated into the chromosome of B. subtilis 168, a non-surfactin producer, by homologous recombination. The transformants reduced the surface tension of the culture broth from 70.0 mN/m to 28.0 mN/m, plus the surface-active compound produced by the transformants exhibited the same Rf value as that from B. subtilis C9 and authentic surfactin in a thin-layer chromatographic analysis. The integration of the sfp gene into the chromosome of B. subtilis 168 was confirmed by Southern hybridization. Like B. subtilis C9, the transformants readily degraded n-hexadecane, although the original strain did not. It was also statistically confirmed that the hydrocarbon degradation of the transformants was highly correlated to their surfactin production by the determination of the correlation coefficient (r2 = 0.997, P0.01). Therefore, these results indicate that the surfactin produced from B. subtilis enhances the bioavailability of hydrophobic liquids.

This study aimed to develop viable enzymes for bioconversion of resveratrol-glucoside into resveratrol. Out of 13 bacterial strains tested, Lactobacillus kimchi JB301 could completely convert polydatin into resveratrol. The purified enzyme had an optimum temperature of 30-40°C and optimum pH of pH 5.0 against polydatin. This enzyme showed high substrate specificities towards different substrates in the following order: isorhaponticin>>polydatin>>mulberroside A>oxyresveratrol-3-O-glucoside. Additionally, it rarely hydrolyzed astringin and desoxyrhaponticin. Based on these catalytic specificities, we suggest this enzyme be named stilbene glucoside-specific β-glucosidase. Furthermore, polydatin extracts from Polygonum cuspidatum were successfully converted to resveratrol with a high yield (of over 99%). Stilbene glucoside-specific β-glucosidase is the first enzyme isolated from lactic acid bacteria capable of bio-converting various stilbene glucosides into stilbene.

The detection and quantitative analysis of Pythium porphyraezoospores was performed by PCR using PP-1 and PP-2 primers specific tothe internal transcribed spacer region of P. porphyrae. To estimatethe amount of fungal zoospores of P. porphyrae, an internal standardplasmid (pPPISC) containing a modified DNA fragment was constructed. Both ends of this fragment were complementary to the PCR primers. Amplification using primers PP-1 and PP-2 produced DNA fragments ofapproximately 700 and 400 bp from the target DNA of P. porphyraezoospores and from the pPPISC, respectively. To perform quantitativePCR, known quantities of pPPISC were added to reaction mixturescontaining the experimental DNAs extracted from zoospores. After aco-amplification reaction, the two different sized PCR products wereseparated by agarose gel electrophoresis and visualized by ethidium bromidestaining. The number of zoospores was estimated by comparing thefluorescence intensities of the PCR products using a charge-coupled deviceimage analyzer. The results show that competitive PCR using P.porphyrae specific primers and competitor pPPISC are useful tools for thequantitative analysis of P. porphyrae zoospores in seawater from Porphyra cultivation farms.

A new zymography for detecting nisin-like acidic bacteriocins was developed using a tricine–sodium dodecyl sulfate (SDS) gel and an acidic gel matrix (pH 4.0). After electrophoresis, proteins in the tricine gel were electrotransferred to an optimal pH-conditioned gel matrix (OP–CGM). The OP–CGM was overlaid with indicator cells ( Bacillus cereus ) embedded in nutrient broth soft agar (0.8%, w/v). Antibacterial activity shown as a growth inhibition using B. cereus was detected at approximately 3.8 kDa. Because nisin is unstable in buffers at pH values over 6.0, the common electrophoretic systems, SDS–polyacrylamide gel electrophoresis and tricine gel, are not suitable for detection of nisin-like acidic bacteriocins.

SUMMARY:Pythium sp., the causative organism of red rot disease, isolated from Porphyra yezoensis from Wando in Korea was compared with those from Miyagi, Aichi, and Fukuoka Prefectures by means of electrophoretic patterns of soluble proteins and isozymes. Native and sodium dodecylsulfate–polyacrylamide gel electrophoretic patterns of soluble proteins revealed small but significant differences among the four isolates. Among the 14 detected enzymes, catalase (CAT), carboxylesterase (CE), glucose-6-phosphate dehydrogenase (G6PD), glucosephosphate isomerase (GPI), isocitrate dehydrogenase (IDH), malate dehydrogenase (MDH), superoxide dismutase (SOD), and xanthine dehydrogenase (XDH) possessed isozymes. The dissimilarity value of isozyme banding patterns for four isolates by unweighted pair-group method using arithmetic averages (UPGMA) cluster analysis was highest (0.51) between the isolate from Fukuoka and those from Miyagi and Aichi, and lowest (0.19) between the isolate from Miyagi and that from Aichi. Cluster analysis of data from isozyme banding patterns grouped the isolates from Miyagi and Aichi. These results indicate that the isolates from Miyagi and Aichi are similar, but each isolate from Wando and Fukuoka differed slightly from those of Miyagi and Aichi.