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Metagenomic Analysis of Novel Lignocellulose-Degrading Enzymes from Higher Termite Guts Inhabiting Microbes

  • Nimchua, Thidarat;Thongaram, Taksawan;Uengwetwanit, Tanaporn;Pongpattanakitshote, Somchai;Eurwilaichitr, Lily
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.462-469
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    • 2012
  • A metagenomic fosmid library was constructed from genomic DNA isolated from the microbial community residing in hindguts of a wood-feeding higher termite (Microcerotermes sp.) collected in Thailand. The library was screened for clones expressing lignocellulolytic activities. Fourteen independent active clones (2 cellulases and 12 xylanases) were obtained by functional screening at pH 10.0. Analysis of shotgun-cloning and pyrosequencing data revealed six ORFs, which shared less than 59% identity and 73% similarity of their amino acid sequences with known cellulases and xylanases. Conserved domain analysis of these ORFs revealed a cellulase belonging to the glycoside hydrolase family 5, whereas the other five xylanases showed significant identity to diverse families including families 8, 10, and 11. Interestingly, one fosmid clone was isolated carrying three contiguous xylanase genes that may comprise a xylanosome operon. The enzymes with the highest activities at alkaline pH from the initial activity screening were characterized biochemically. These enzymes showed a broad range of enzyme activities from pH 5.0 to 10.0, with pH optimal of 8.0 retaining more than 70% of their respective activities at pH 9.0. The optimal temperatures of these enzymes ranged from $50^{\circ}C$ to $55^{\circ}C$. This study provides evidence for the diversity and function of lignocellulose-degrading enzymes in the termite gut microbial community, which could be of potential use for industrial processes such as pulp biobleaching and denim biostoning.

Involvement of PI3K/AKT and MAPK Pathways for TNF-α Production in SiHa Cervical Mucosal Epithelial Cells Infected with Trichomonas vaginalis

  • Yang, Jung-Bo;Quan, Juan-Hua;Kim, Ye-Eun;Rhee, Yun-Ee;Kang, Byung-Hyun;Choi, In-Wook;Cha, Guang-Ho;Yuk, Jae-Min;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.53 no.4
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    • pp.371-377
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    • 2015
  • Trichomonas vaginalis induces proinflammation in cervicovaginal mucosal epithelium. To investigate the signaling pathways in $TNF-{\alpha}$ production in cervical mucosal epithelium after T. vaginalis infection, the phosphorylation of PI3K/AKT and MAPK pathways were evaluated in T. vaginalis-infected SiHa cells in the presence and absence of specific inhibitors. T. vaginalis increased $TNF-{\alpha}$ production in SiHa cells, in a parasite burden-dependent and incubation time-dependent manner. In T. vaginalis-infected SiHa cells, AKT, ERK1/2, p38 MAPK, and JNK were phosphorylated from 1 hr after infection; however, the phosphorylation patterns were different from each other. After pretreatment with inhibitors of the PI3K/AKT and MAPK pathways, $TNF-{\alpha}$ production was significantly decreased compared to the control; however, $TNF-{\alpha}$ reduction patterns were different depending on the type of PI3K/MAPK inhibitors. $TNF-{\alpha}$ production was reduced in a dose-dependent manner by treatment with wortmannin and PD98059, whereas it was increased by SP600125. These data suggested that PI3K/AKT and MAPK signaling pathways are important in regulation of $TNF-{\alpha}$ production in cervical mucosal epithelial SiHa cells. However, activation patterns of each pathway were different from the types of PI3K/MAPK pathways.

New Azafluorenone Derivative and Antibacterial Activities of Alphonsea cylindrica Barks

  • Talip, Munirah Abdul;Azziz, Saripah Salbiah Syed Abdul;Wong, Chee Fah;Awang, Khalijah;Naz, Humera;Bakri, Yuhanis Mhd;Ahmad, Mohamad Syahrizal;Litaudon, Marc
    • Natural Product Sciences
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    • v.23 no.3
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    • pp.151-156
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    • 2017
  • A phytochemical study of Alphonsea cylindrica King (unreported) has led to the isolation of six alkaloids. The compounds were identified as kinabaline (1; azafluorenone alkaloid), muniranine (2), O-methylmoschatoline (3; oxoaporphine alkaloid), lysicamine (4), atherospermidine (5) and N-methylouregidione (6; 4, 5-dioxoaporphine alkaloid). The structures of the isolated compounds were determined based on the spectroscopic techniques and by comparison with data reported in the literature. Alkaloid 2 was isolated as a new derivative of azafluorenone while alkaloids 1, 3 - 6 were isolated for the first time from Alphonsea species. In addition, alkaloid 3 and 4 showed inhibition zone against Staphylococcus aureus, Pseudomonas aeruginosa and Bacillus cereus in disc diffusion test. The minimum inhibition concentration (MIC) values of lysicamine (4) against S. aureus, B. cereus and P. aeruginosa were found to be smaller than O-methylmoschatoline (3). Therefore, the reported antibacterial activity showed the potential of this plant as natural antibacterial agent and supported the documented traditional use of Alphonsea sp. in the treatment of diarrhea and fever.

Cultural characteristics of fungal species associated with deterioration or foxing of paper and chemical removal (종이변색균류의 배양적 특성 및 화학적 방법에 의한 변색제거)

  • Jo, Seong-Eun;Kim, Yong-Tae;Jeong, So-Yeong;Jo, Byeong-Muk;Lee, Jong-Gyu
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2009.04a
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    • pp.295-303
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    • 2009
  • The annals of Joseon dynasty, especially the volumes of King SeJong(1418-1450 A.D.), were heavily deteriorated by fungi. Investigations on the deteriorating and foxing fungi were carried out. Fungal structures on the beeswax, which were coated on the both side of Han-Ji, were suspected to be involved in the deterioration, and were observed by SEM. Isolation and culturing of these fungi were tried by scrubing swab samples and placing on the artificial media. Culture-independent approaches were used to identify the fungal strains associated with damages of beeswax and foxing of the paper by the analyses based on DNA sequences data from the specific ITS region of rDNA regions. In addition, well-known paper staining fungi(PSF), i.e., Aspergillus terreus var. terreus, Fusarium oxysporum, Chaetomium globosum, Cladosporium cladosporioides, and Alternaria solani, were compared in the mycelial growth and stain on beeswax and papers under different environmental conditions (temperature, light, moisture, etc). Fungal strains isolated from the air samples in the storage room and shelves were identified as Irpex sp., Arthrinium sacchari, Cladosporium tenuissimum, Aspergillus sclerotiorum, Sistotrema brinkmannii, and Hypoxylon bovei var. microsporum The isolated strains were compared in growth and stain patterns on beeswax and papers(Han-Ji, Hwa-Ji, and Yang-Ji) whether these can cause damage or foxing on the annals or not.

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Artificial induction and isolation of cadmium-tolerant soil bacteria

  • Lee, Sangman
    • Journal of Applied Biological Chemistry
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    • v.63 no.2
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    • pp.125-129
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    • 2020
  • Environmental pollution caused by various heavy metals is a serious global problem. To solve this problem, microbial bioremediation of contaminated metals has developed rapidly as an effective strategy when physical and chemical techniques are not suitable. In this study, cadmium (Cd)-tolerant soil bacteria were isolated via artificial induction in laboratory conditions instead of screening bacteria naturally adapted to metal-contaminated soils. Wild-type (WT) bacteria grown in uncontaminated soils were artificially and sequentially adapted to gradually increasing Cd concentrations of up to 15 mM. The resultant cells, named Soil-CdR15, survived at a Cd concentration of 10 mM, whereas WT cells failed to survive with 4 mM Cd on solid media for 2 d. In liquid media containing Cd, the SoilCdR15 cells grew with 15 mM Cd for 7 d, whereas the WT cells could not grow with 5 mM Cd. Both Soil-CdR15 and WT cells removed approximately 35% of Cd at the same capacity from liquid media containing either 0.5 or 1.0 mM Cd over 2 d. In addition to Cd, the Soil-CdR15 cells showed increased resistance to nickel, zinc, and arsenic compared to WT cells. The Soil-CdR cells were identified as Burkholderia sp. by partial sequencing of 16S rRNA. The data presented in this study demonstrate that isolation of heavy metal-tolerant microorganisms via artificial induction in laboratory conditions is possible and may be useful for the application of the microorganisms for the bioremediation of heavy metals.

Chiisanoside, A Lupane Triterpenoid from Acanthopanax Leaves, Stimulates Proliferation and Differentiation of Osteoblastic MC3T3-E1 Cells

  • Choi, Eun-Mi;Ding, Yan;Nguyen, Huu Tung;Park, Sang-Hyuk;Nguyen, Xuan Nhiem;Liang, Chun;Lee, Jung-Joon;Kim, Young-Ho
    • Natural Product Sciences
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    • v.14 no.1
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    • pp.21-26
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    • 2008
  • The leaves of Acanthopanax species have traditionally been used as a tonic and a sedative as well as in the treatment of rheumatism and diabetes. Chiisanoside is the major active lupane triterpenoid of Acanthopanax leaves. To investigate the bioactivities of chiisanoside, which act on bone metabolism, the effects of chiisanoside on the function of osteoblastic MC3T3-E1 cells were studied. Chiisanoside $(0.02{\sim}20\;{\mu}M)$ significantly increased the growth of MC3T3-E1 cells and caused a significant elevation of alkaline phosphatase (ALP) activity, collagen content, and nodules mineralization in the cells (P < 0.05). The effect of chiisanoside (2 ${\mu}M$) in increasing ALP activity was completely prevented by the presence of tamoxifen, suggesting that the effect of chiisanoside might be partly estrogen receptor mediated. Moreover, cotreatment of p38 inhibitor SB203580 or JNK inhibitor SP600125 inhibited chiisanoside-mediated ALP upregulation, suggesting that the induction of differentiation by chiisanoside is associated with increased activation of p38 and JNK mitogen-activated protein kinases. Our data indicate that the enhancement of osteoblast function by chiisanoside may result in the prevention for osteoporosis.

Relationship between the N-acetyl-β-D-glucosaminidase levels and the presence of mastitis pathogens in bovine mastitis milk samples (유우(乳牛) 유방염(乳房炎) 진단(診斷)에 있어서 N-acetyl-β-D-glucosaminidase 치(値)와 분리(分離) 원인균(原因菌)과의 관계(關係))

  • Kang, Byong-kyu;Nam, Hyang-mi;Son, Chang-ho
    • Korean Journal of Veterinary Research
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    • v.33 no.3
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    • pp.531-537
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    • 1993
  • A study was carried out to define the relationship between the N-acetyl-$\beta$-D-glucosaminidase(NAGase) levels and isolated pathogenic bacteria in 379 quarter fore milk of mastitis suspected samples collected in this clinics. All samples were tested the NAGase, California mastitis test(CMT), Somatic cell count(SCC) and bacterial culture. Except 111 from 379 samples, 268 bacteria-positive quarter fore milk samples were classified into the latent and mastitis infection group by SCC($500,000cells\;per\;m{\ell}$), and the mean NAGase levels($nmol/min/m{\ell}$) of each isolated pathogen in mastitis infection group were Staphylococcus aureus 3.067, Coagulase-negative staphylococci 4.083, Staphylococcus aureus 3.594, Str. uberis 3.513, Str. dysgalactiae 1.640, E coli 4.441 and gram negative rods 4.560, respectively. Most of the relationship between mean SCC and NAGase in each pathogen group were highly significant using a student t test(p<0.05). When the mastitis pathogens were classified into minor(Coagulase-negative staphylococci, Corynebacterium sp.) and major pathogen group(Staphylococcus aureus, Streptococcus agalactiae, Str. uberis, Str. dysgalactiae, gram negative rods), the NAGase levels were higher at major than minor pathogen group. On the other hand, when the mastitis milk samples were classified by SCC($500,000cells\;per\;m{\ell}$) and by the presence of pathogen(IDF scheme), the NAGase levels were also higher at the mastitis than latent infection. The possibility of combining SCC and NAGase data in order to give the more difinitive diagnosis is discussed.

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Complete Mitochondrial Genome of a Troglobite Millipede Antrokoreana gracilipes (Diplopoda, Juliformia, Julida), and Juliformian Phylogeny

  • Woo, Hyung-Jik;Lee, Yong-Seok;Park, Shin-Ju;Lim, Jong-Tae;Jang, Kuem-Hee;Choi, Eun-Hwa;Choi, Yong-Gun;Hwang, Ui Wook
    • Molecules and Cells
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    • v.23 no.2
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    • pp.182-191
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    • 2007
  • The complete mitochondrial genome of a troglobite millipede Antrokoreana gracilipes (Verhoeff, 1938) (Dipolopoda, Juliformia, Julida) was sequenced and characterized. The genome (14,747 bp) contains 37 genes (2 ribosomal RNA genes, 22 transfer RNA genes and 13 protein-encoding genes) and two large non-coding regions (225 bp and 31 bp), as previously reported for two diplopods, Narceus annularus (order Spirobolida) and Thyropygus sp. (order Spirostreptida). The A + T content of the genome is 62.1%, and four tRNAs ($tRNA^{Ser(AGN)}$, $tRNA^{Cys}$, $tRNA^{Ile}$ and $tRNA^{Met}$) have unusual and unstable secondary structures. Whereas Narceus and Thyropygus have identical gene arrangements, the $tRNA^{Thr}$ and $tRNA^{Trp}$ of Antrokoreana differ from them in their orientations and/or positions. This suggests that the Spirobolida and Spirostreptida are more closely related to each other than to the Dipolopoda. Three scenarios are proposed to account for the unique gene arrangement of Antrokoreana. The data also imply that the Duplication and Nonrandom Loss (DNL) model is applicable to the order Julida. Bayesian inference (BI) and maximum likelihood (ML) analyses using amino acid sequences deduced from the 12 mitochondrial protein-encoding genes (excluding ATP8) support the view that the three juliformian members are monophyletic (BI 100%; ML 100%), that Thyropygus (Spirostreptida) and Narceus (Spirobolida) are clustered together (BI 100%; ML 83%), and that Antrokoreana (Julida) is a sister of the two. However, due to conflict with previous reports using cladistic approaches based on morphological characteristics, further studies are needed to confirm the close relationship between Spirostreptida and Spirobolida.

Seasonal variation of fish assemblage in Sacheon marine ranching, the southern coast of Korea (사천바다목장해역 어류군집의 변동특성)

  • Kim, Young-Seup;Choi, Jung-Hwa;Kim, Jung-Nyun;Oh, Taeg-Yun;Choi, Kwang-Ho;Lee, Dong-Woo;Cha, Hyung-Kee
    • Journal of the Korean Society of Fisheries and Ocean Technology
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    • v.46 no.4
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    • pp.335-345
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    • 2010
  • Species composition and seasonal variation of fish assemblage in Sacheon marine ranching, Korea were examined. Sampling was seasonally made by a shrimp trawl from February 2009 to November 2009. A total of 55 species were collected. Of these, five species was dominant species comprising 51.1% of the total number of individuals. Nine species represented 57.3% of the total biomass. While total abundance (number of individuals) was high in spring, total biomass and species richness (number of species) and diversity were high in autumn. Cluster analysis, based on seasonally abundance data of the 21 most common species, showed that the species were separated into 4 different groups. Group A composed of Pholis nebulosa, Pleuronectes yokohamae, Hypodytes rubripinnis, Hexagrammos otakii, Tridentiger trigonocephalus and Paracercis sexfasciata, which were year round residents, group B Congr myriaster, Leiognathus nuchalis, and Platycephalus indicus, which were abundant in autumn, group C Sillago japonica, Pseudaesopia japonica, Zoarces gilli, Pholis fangi, and Pleuronichthys cornutus which were abundant in spring, and group D Sillago sihama, Cociella sp., Johnius grypotus, Cynoglossus joyneri, Cynoglossus robustus, and Pennahia argentata, which were abundant in summer. Principal component analysis revealed that seasonal variation in the fish assemblage was attributed to the abundance of temporal species such as conger and sand smelt fishes, which were abundant in summer, due to seasonal variation of water temperature.

A Review of the Clinical Study on Korean Medicine Treatment for Women with Chronic Pelvic Pain (만성 골반통 여성의 한의학적 치료에 관한 국내 임상 연구 고찰)

  • Moon, Hye-Yeon;Lee, Yoon-Jae;So, Min-Ji;Yu, Ok-Cheol;Choi, Sung-Ryul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.31 no.3
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    • pp.50-60
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    • 2018
  • Objectives: The purpose of this study was to investigate the treatment methods of chronic pelvic pain, which have no definite treatment methods, and to provide basic data on clinical trials and clinical practical guideline in Korean Medicine related to chronic pelvic pain. Methods: The key words such as 'pelvic pain', 'pelycalgia' are used for the research through 'OASIS, KTKP, RISS, DBpia, KISS'. Total 6 studies were analyzed. Results: 6 studies were chronic pelvic pain case reports. Case reports 6 studies covered a total of 11 cases. Of these, 4 cases related to adenomyosis, 3 cases of endometriosis, and 2 cases of ovarian cyst. However, there were many cases in which the causative disease was not clear. In addition, the majority of the studies reported on the progress of hospitalization, and all studies used herbal medicine, acupuncture and moxibustion. The duration of treatment ranged from 5 to 30 days for hospitalization and 5 months for outpatient treatment. The most commonly used meridian is Conception Vessel (任脈), and SP6 (三陰交) was the most frequently used acupuncture treatment. The herbal medicine Banchong-san (蟠蔥散) was generally prescribed. Conclusions: The effect on treatment of Korean Medicine for chronic pelvic pain had mostly positive results. However, the number of searched studies, which are insufficient for design or analysis of study, is very limited. We hope that this study will be useful for further clinical studies of chronic pelvic pain.