• Molecular & Cellular Toxicology
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• v.3 no.1
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• pp.23-30
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• 2007

The effects of high energy short wave solar radiation on human skin have received much publicity as the major cause of accelerated skin ageing and of skin cancers. To meet public demand, the cosmetic industry has developed sun protection factor products, which contain a variety of so-called "UV filters", among others benzophenone (BP) and its metabolites are the widely used UV filters. UV filters are also used to prevent UV light from damaging scents and colors in a variety of cosmetics products and to protect of plastic products against light-induced degradation. There are many variants of BP in use. In this respect, to regulate and to evaluate the hazardous effect of BP-type UV filters will be important to environment and human health. The genotoxicity of 7 BP-type UV filters was evaluated in L5178Y $(tk^{+/-})$ mouse lymphoma cells in vitro. BP, benzhydrol, 4-hydroxybenzophenone 2-hydroxy-4-methoxybenzophenone and 2, 4-dihydroxybenzophenone did not induce significant mutation frequencies both in the presence and absence of metabolic activation system. 2, 2'-Dihydroxy-4-methoxybenzophenone appeared the positive results at the highest dose, i.e. 120.4 ${\mu}g/mL$ only in the absence of metabolic activation system. And also, 2, 3, 4-trihydroxybenzophenone revealed a significant increase of mutation frequencies in the range of 138.1-207.2 ${\mu}g/mL$ in the absence of metabolic activation system and 118.3-354.8 ${\mu}g/mL$ in the presence of metabolic activation system. Through the results of MLA with 7 BP-type UV filters in L5178Y cells in vitro, we may provide the important clues on the genotoxic potentials of these BP-type UV filters.

• Journal of Life Science
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• v.24 no.10
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• pp.1144-1150
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• 2014

The eIF4E protein is the key regulator of translation initiation. The interaction of eIF4E with eIF4G triggers the translation of mRNA, and several proteins interrupt this association to modulate translation. Human 4E-T is one of the eIF4E-binding partners that represses the translation of bound mRNAs, and it is involved in the transport of eIF4E to processing bodies (P-bodies). Although Clast4, the mouse homolog of human 4E-T, might play critical roles in the regulation of translation, its properties are not well known. In this report, we deciphered the properties of Clast4 by determining its phosphorylation state, binding to eIF4E, and effects of overexpression on cell proliferation. Clast4 was phosphorylated by protein kinase A (PKA) in vivo on several residues of its amino terminus. Nevertheless, the PKA phosphorylation of Clast4 appeared to have no effect on either its eIF4E-binding ability or localization. Clast4 interacted with eIF4E1 and CPEB. The conserved eIF4E-binding sequence in Clast4, $YXXXXL_{\phi}$, was important for binding eIF4E1A but not eIF4E1B. Similar to that of another well-known eIF4E regulator, the eIF4E binding protein (4E-BP), the overexpression of Clast4 decreased cell proliferation. These results suggest that Clast4 acts as a global translation regulator in cells.

• Plant Breeding and Biotechnology
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• v.5 no.4
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• pp.334-343
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• 2017

Eclipta prostrata and E. alba are annual herbal medicinal plants and have been used as Chinese medicinal tonics. Both species are widely distributed in tropical and subtropical regions as well as in Korea. Both species have similar morphological features but E. alba has smoother leaf blade margins compared with E. prostrata. Although both species are utilized as oriental medicines, E. prostrata is more widely used than E. alba. Morphological semblances have confounded identification of either species. Here, we report the complete chloroplast genomes of both species to provide an authentication system between the two species and understand their diversity. Both chloroplast genomes were 151,733-151,757 bp long and composed of a large single copy (83,285-83,300 bp), a small single copy (18,283-18,346 bp), and a pair of inverted repeats (25,075-25,063 bp). Gene annotation revealed 80 protein coding genes, 30 tRNA genes and four rRNA genes. A phylogenetic analysis revealed that the genus Eclipta is grouped with Heliantheae tribe species in the Asteraceae family. A comparative analysis verified 29 InDels and 58 SNPs between chloroplast genomes of E. prostrata and E. alba. The low chloroplast genome sequence diversity indicates that both species are really close to each other and are not completely diverged yet. We developed six DNA markers that distinguish E. prostrata and E. alba based on the polymorphisms of chloroplast genomes between E. prostrata and E. alba. The chloroplast genome sequences and the molecular markers generated in this study will be useful for further research of Eclipta species and accurate classification of medicinal herbs.

• Development and Reproduction
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• v.4 no.2
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• pp.215-220
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• 2000

Most of endocrine disrupters (EDs) have been reported to exhibit estrogenic or anti-androgenic activity and thereby may disrupt reproductive development in human or wildlife. This study was performed to investigate the effects of estrogen (E$_2$), bisphenol (BP) and octylphenol (OP) on the mouse Leydig cell line (TM3). TM3 originated from testis of 11~13-daly-old BALB/c nu/＋ mice was cultured in DMEM supplemented with 10％ FBS alone or medium with estrogen (E$_2$), bisphenol (BP) and octylphenol (OP; 1 pM, 1 nM, 1 $\mu$M, 1 mM, respectively) for 48 hours. After culture, total cell number and viability were assessed by heamocyto-meter and trypan blue stain. Expression of cytochrome P450scc (CYPscc) mRNA whose product is involved in steroid hormone biosynthesis and estrogen receptor $\alpha$(ER $\alpha$) mRNA were detected by RT-PCR. As a result, treatment of TM3 with E$_2$, BP and OP(1 mM, respectively) significantly decreased the viability but not all of groups as high as 1 $\mu$M. Exposure of TM3 to OP significantly reduced the total cell number but not E$_2$ or BP. The expression of CYPscc mRNA was slightly reduced in BP (1 nM, 1 $\mu$M) and significantly decreased in OP (1 nM, 1 $\mu$M) treated TM3, except E$_2$ group. But the expression of ER $\alpha$ mRNA was sightly increased in all treated groups. In conclusion, BP and OP (high concentration) might inhibit steroidogenesis by decreasing the CYPscc mRNA expression in the mouse testis. These results suggest that BP and OP might impair spermatogenesis and subsequently disturb testicular function.

• Animal cells and systems
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• v.15 no.1
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• pp.29-36
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• 2011

Insect lysozymes are basic, cationic proteins synthesized in fat body and hemocytes in response to bacterial infections and depolymerize the bacterial cell wall. The c-type lysozyme of the insect Spodoptera litura (SLLyz) is a single polypeptide chain of 121 residues with four disulfide bridges and 17 rare codons and is approximately 15 kDa. The full-length SLLyz cDNA is 1039 bp long with a poly(A) tail, and contains an open reading frame of 426 bp long (including the termination codon), flanked by a 54 bp long 5' UTR and a 559 bp long 3' UTR. As a host for the production of high-level recombinant proteins, E. coli is used most commonly because of its low cost and short generation time. However, the soluble expression of heterologous proteins in E. coli is not trivial, especially for disulfide-bonded proteins. In order to prevent inclusion body formation, GST was selected as a fusion partner to enhance the solubility of recombinant protein, and fused to the amplified products encoding mature SLLyz. The expression vector pGEX-4T-1/rSLLyz was then transformed into E. coli BL21(DE3)pLysS for soluble expression of rSLLyz, and the soluble fusion protein was purified successfully. Inhibition zone assay demonstrated that rSLLyz showed antibacterial activity against B. megaterium. These results demonstrate that the GST fusion expression system in E. coli described in this study is efficient and inexpensive in producing a disulfide-bonded rSLLyz in soluble, active form, and suggest that the insect lysozyme is an interesting system for future structural and functional studies.

• Microbiology and Biotechnology Letters
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• v.31 no.2
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• pp.111-116
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• 2003

The xylA gene of Bacillus stearothermophilus No. 236 encoding $\beta$-xylosidase was cloned and sequenced previously. The transcriptional start site of the xylA gene cloned in E. coli was identified to be the guanine (G) by primer extension analysis. This supports that the expression of xylA gene is also directed in the E. coli cells by the previously determined transcription initiation signals, -10 sequence (CATAAT) and -35 sequence (TTGTTA) separated by 12 bp. To increase the expression of $\beta$-xylosidase, firstly the spacer region of xylA promoter was extended from 12 to 17 bp, and then the -10 and -35 elements were converted into their respective consensus sequences. The mutant promoters thus obtained were tested for their activities in both the E. coli and B. subtilis host cells. The change of the length of the spacer region from 12 to 17 bp resulted in a 1.6- and 2.5-fold increase in promoter strength in comparison with the wild type promoter in E. coli and B. subtilis cells, respectively. Also, strength of the promoter with the fourth T to A transversion on its -35 element increased in the transcription level by about 35 times compared with that of wild-type promoter. However, surprisingly the 5' end C-to-T transition of the -10 hexamer showed a 5- to 15-fold reduction in $\beta$-xylosidase activity in both E. coli and B. subtilis. Together, the present data demonstrated that the 5' end nucleotide C of the -10 sequence CATAAT and the fourth nucleotide A of the -35 hexamer are two most critical nucleotides for the promoter activity in the context of the xylA promoter.

• Animal Bioscience
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• v.35 no.11
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• pp.1752-1759
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• 2022

Objective: Effects of direct-fed Enterococcus faecium plus bacteriophages (EF-BP) were investigated as potential substitutes for pharmacological ZnO for weanling pigs. Methods: Dietary treatments were supplementations to a basal diet with none (NC), 3,000-ppm ZnO (PC), 1×10¹⁰ colony-forming units of E. faecium plus 1×10⁸ plaque-forming units (PFU) of anti-Salmonella typhimurium bacteriophages (ST) or 1×10⁶ PFU of each of anti-enterotoxigenic Escherichia coli K88 (F4)-, K99 (F5)-, and F18-type bacteriophages (EC) per kg diet. In Exp 1, twenty-eight 21-day-old crossbred weanling pigs were individually fed one of the experimental diets for 14 days and euthanized for histological examination on intestinal mucosal morphology. In Exp 2, 128 crossbred weanling pigs aged 24 days were group-fed the same experimental diets in 16 pens of 8 piglets on a farm with a high incidence of post-weaning diarrhea. Results: None of the diarrheal score or fecal consistency score (FCS), average daily gain (ADG), gain: feed ratio, structural variables of the intestinal villus, and goblet cell density, differed between the EF-BP (ST+EC) and NC groups, between EF-BP and PC, or between ST and EC, with the exception of greater gain: feed for EF-BP than for PC (p<0.05) during days 7 to 14 (Exp 1). In Exp 2, ADG was less for EF-BP vs PC during days 0 to 7 and greater for EF-BP vs NC during days 7 to 14. FCS peaked on day 7 and declined by day 14. Moreover, FCS was less for EF-BP vs NC, did not differ between EF-BP and PC, and tended to be greater for ST vs EC (p = 0.099). Collectively, EF-BP was comparable to or slightly less effective than PC in alleviating diarrhea and growth check of the weanling pigs, with ST almost as effective as PC, when they were group-fed. Conclusion: The E. faecium-bacteriophage recipe, especially E. faecium-anti-S. typhimurium, is promising as a potential substitute for pharmacological ZnO.

• Korean Journal of Veterinary Service
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• v.28 no.1
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• pp.71-79
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• 2005

In this study, a total of 2,119 samples was taken from bovine feces and carcass from March 2002 to December 2003. And those were examined for the presence of enterohemorrhagic E coli O157:H7. The properties of the isolates were characterized for biochemical features, serotypes, virulence genes and antimicrobial susceptibility. Forty five strains($3.7\%$) of E coli O157:H7 were isolated from 1,208 fecal samples and were not detected in carcass using immunomagnetic separation technique and selective media. In multiplex PCR using stx1, stx2, eaeA and hlyA primers, the amplified bands at 180 bp, 255bp, 384bp and 534bp were observed, respectively. In antimicrobial susceptibility test, all isolates were susceptible to amoxicillin/clavulanic acid and cefazolin. The isolates were most resistant to sulfisoxazole($24.4\%$), followed by streptomycin($22.2\%$), tetracycline($20.0\%$). Eight strains($17.8\%$) of 45 isolates showed the multi-resistant patterns with over 3 drugs.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.357-363
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• 2013

The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.

• Journal of Veterinary Science
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• v.19 no.6
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• pp.817-826
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• 2018

The bursa of Fabricius (BF) is a central humoral immune organ unique to birds. Four bursal peptides (BP-I, BP-II, BP-III, and BP-IV) have been isolated and identified from the BF. In this study, the immunoadjuvant activities of BPs I to IV were examined in mice immunized with H9N2 avian influenza virus (AIV) vaccine. The results suggested that BP-I effectively enhanced cell-mediated immune responses, increased the secretion of Th1 (interferon gamma)- and Th2 (interleukin-4)-type cytokines, and induced an improved cytotoxic T-lymphocyte (CTL) response to the H9N2 virus. BP-II mainly elevated specific antibody production, especially neutralizing antibodies, and increased Th1- and Th2-type cytokine secretion. BP-III had no significant effect on antibody production or cell-mediated immune responses compared to those in the control group. A strong immune response at both the humoral and cellular levels was induced by BP-IV. Furthermore, a virus challenge experiment followed by H&E staining revealed that BP-I and BP-II promoted removal of the virus and conferred protection in mouse lungs. BP-IV significantly reduced viral titers and histopathological changes and contributed to protection against H9N2 AIV challenge in mouse lungs. This study further elucidated the immunoadjuvant activities of BPs I to IV, providing a novel insight into immunoadjuvants for use in vaccine design.