• Asian Pacific Journal of Cancer Prevention
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• v.15 no.6
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• pp.2571-2576
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• 2014

Background: Published studies on the association between the exonuclease 1 (EXO1) Glu589Lys polymorphism and cancer susceptibility have yielded conflicting results. Thus, a meta-analysis of published studies was performed to assess the possible association. Materials and Methods: All eligible case-control studies published up to January 2013 on the association between the EXO1 Glu589Lys polymorphism and cancer susceptibility were identified by searching PubMed, Web of Science, Science Direct and hand search. Either fixed-effect or random-effect models were used to calculate pooled odds ratios (ORs) with 95% confidence intervals (CIs) using the Comprehensive Meta-Analysis software version 2.2. Results: A total of 4,391 cancer cases and 4,339 controls from 10 studies were included. Overall, no significant association between the EXO1 Glu589Lys polymorphism and cancer susceptibility was observed in either genetic model. However; in subgroup analyses by cancer type, a significant association between EXO1 Glu589Lys and lung cancer risk was found (Lys vs Glu: OR=1.23, 95%CI=1.07-1.41, $p_{heterogeneity}$=0.05). Further, subgroup analysis by ethnicity indicated that there was a statistically increased cancer risk in Asians (Lys vs Glu: OR=1.42, 95%CI=1.30-1.55, $p_{heterogeneity}$=0.07; Lys/Lys vs Glu/Glu: OR=1.93, 95%CI=1.20-3.12, $p_{heterogeneity}$=0.01; Lys/Lys+Glu/Lys vs Glu/Glu: OR=1.52, 95%CI=1.37-1.68, $p_{heterogeneity}$=0.42; Lys/Lys vs Glu/Lys+Glu/Glu: OR=1.68, 95%CI=1.07-2.65, $p_{heterogeneity}$=0.02). However, significant association was absent in Caucasians. Conclusions: This meta-analysis suggests, for the first time, that the EXO1 Glu589Lys polymorphism is not associated with overall cancer susceptibility, although marginal associations were found for lung cancer and Asian subgroups. Additional well-designed studies with larger sample size focusing on different ethnicities and cancer types are needed to confirm these findings.

• Journal of the Korea Organic Resources Recycling Association
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• v.20 no.2
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• pp.66-75
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• 2012

The objective of this study was to assess the effects of methanogenic bacteria-activated leachate recirculation method for enhancing waste stabilization and landfill gas production from a solid waste landfill. To simulate a conventional landfill (Lys-A), a landfill recirculated only fresh leachate (Lys-B), and two landfills recirculated leachate after pretreating with ASBR (Lys-C and Lys-D), four lysimeters were operated over a period of 4 years. Lys-D was recirculated two times of pretreated leachate volume than that of Lys-C. In the case of the landfill recirculated only fresh leachate and the landfill recirculated leachate after pretreating with ASBR, methane productions were increased until about 600 days, but there were not effect of leachate recirculation for enhancing methane production after about 600 days. It was assumed that leachate recirculation into fewer biodegradable organic wastes had not effect to enhance landfill gas production. Lys-C and Lys-D showed the highest performance for enhancing cumulative methane yield as well as acceleration waste stabilization. In cumulative methane yield, Lys-C (35.51 mL $CH_4/g$ VS) and Lys-D (36.12 mL $CH_4/g$ VS) were much higher than Lys-A (28.37 mL $CH_4/g$ VS) and Lys-B (30.07 mL $CH_4/g$ VS). In case of between Lys-B and Lys-C with the same recirculation rate, COD concentration in Lys-C was more rapidly decreased compared with that in Lys-B. This was attributed to the presence of methanogenic bacteria as well as dilution of inhibitory substances by the methanogenic bacteria-activated leachate recirculation. Therefore, the landfill recirculated leachate after pretreating with ASBR was found to be the most appropriate operating techniques for enhancing waste stabilization and landfill gas production.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.3
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• pp.401-407
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• 2013

Two experiments were conducted to determine an optimum Lys:calorie ratio (g of total dietary Lys/Mcal of DE) for growing barrows and gilts in cool and hot seasons in a commercial environment. In Exp. 1, 96 barrows and 96 gilts were randomly allocated in 1 of 4 dietary treatments (2.7, 3.0, 3.3, 3.6 g of Lys/Mcal of DE). Each treatment had 12 replicate pens with 4 pigs per pen. The experiment lasted for 34 d in the cool season (March 12th to April 15th). Diets were based on corn-wheat-soybean meal. Lys:calorie ratio were attained by adjusting the amount of corn and soybean and supplementation of crystalline Lys. Total Lys intake and available Lys intake were increased (p<.05) as dietary Lys:calorie ratio increased. The BUN concentration on d 34 for barrows, and BUN change for barrows and gilts linearly increased (p<0.05) in response to increasing dietary Lys:calorie ratio. For gilts, back fat was decreased and then increased (Quadratically, p<0.05) as increasing dietary lys:calorie ratio. Exp. 2 had a similar design as Exp. 1 with the exception that Exp. 2 was conducted in hot season (June 30th to September 11th) for 42 d. Diet of Exp. 2 was the same as Exp. 1. Total Lys intake and available Lys intake increased (p<0.05) as dietary Lys:calorie increased. On d 42, the BUN concentration increased (p<0.05) in response to the increasing dietary Lys:calorie ratio. In conclusion, dietary Lys:calorie ratio of 2.7 g of Lys/Mcal of DE could satisfy the requirement of 25 to 50 kg growing pigs. Increasing dietary Lys:calorie ratio could increase BUN concentration in growing pigs.

• Animal cells and systems
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• v.4 no.2
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• pp.165-171
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• 2000

Ubiquitinated proteins in lysosomes were characterized by using two monoclonal antibodies (mAbs): LYS8-1, a mAb to lysosomal proteins, and NYA124, a mAb to ubiquitin. LYS8-1 stained lysosome-like vesicles in immunofluorescence microscopy of Amoeba proteus and Acanthamoeba castellanii. In immunoblotting, LYS8-1's antigens (LYS proteins) were detected as 68-kDa and 77-kDa proteins in A. proteus, and as 30-kDa and 39-kDa proteins in A. castellanii. In immunoprecipitation of A. castellanii, at least four distinct LYS proteins, LVS35p, LyS39p, LyS42p, and LYS46p, were detected and accumulated upon inhibition of lysosome functions but not upon that of 26S proteasome functions. They were all found to be ubiquitinated, and were recovered in the lysosome fractions in subcellular fractionation experiments. In chemical fractionation analyses, LYS35p and LYS39p were demonstrated to be peripherally associated with lysosome membrane, while LYS42p and LYS46p tightly bound to the membrane. These results suggest that the LYS proteins become associated to lysosomal membrane upon ubiquitination.

• The Plant Pathology Journal
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• v.36 no.4
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• pp.323-334
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• 2020

Lysin motif (LysM) proteins are reported to be necessary for the virulence and immune response suppression in many herbaceous plant pathogens, while far less is documented in woody plant pathogens. In this study, we preliminarily characterized the molecular function of a LysM protein LtLysM1 in woody plant pathogen Lasiodiplodia theobromae. Transcriptional profiles revealed that LtLysM1 is highly expressed at infectious stages, especially at 36 and 48 hours post inoculation. Amino acid sequence analyses revealed that LtLysM1 was a putative glycoprotein with 10 predicted N-glycosylation sites and one LysM domain. Pathogenicity tests showed that overexpressed transformants of LtLysM1 displayed increased virulence on grapevine shoots in comparison with that of wild type CSS-01s, and RNAi transformants of LtLysM1 exhibited significantly decreased lesion length when compared with that of wild type CSS-01s. Moreover, LtLysM1 was confirmed to be a secreted protein by a yeast signal peptide trap assay. Transient expression in Nicotiana benthamiana together with protein immunoblotting confirmed that LtLysM1 was an N-glycosylated protein. In contrast to previously reported LysM protein Slp1 and OsCEBiP, LtLysM1 molecule did not interact with itself based on yeast two hybrid and co-immunoprecipitation assays. These results indicate that LtLysM1 is a secreted protein and functions as a critical virulence factor during the disease symptom development in woody plants.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.11
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• pp.1623-1628
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• 2011

The effects of dietary lysine (Lys) at a fixed ratio to crude protein (CP) and essential amino acids (EAA) on early growth response of broilers were studied. Four diets were formulated to contain similar metabolizable energy (ME, 2,950 kcal/kg) but contained graded levels of incremental Lys (1.1, 1.2, 1.3 and 1.4%) while also increasing the dietary CP and EAA (methionine, methionine+ cystine, threonine and tryptophan) to maintain a constant ratio with Lys. Each diet was fed at random to 10 replicates of 6 chicks each throughout the experimental period (1-21 d). At the lowest concentration of Lys of 1.1% (19.04% CP), body weight gain (BWG) was lowest and feed conversion ratio (FCR) was poorest. The BWG increased and FCR decreased linearly as dietary Lys increased upto 1.3% (22.5% CP). Lowest feed consumption was observed in the dietary group that contained 1.1% Lys (19.04% CP) in the diet. Increasing the concentration of Lys to 1.2% (20.77% CP), significantly increased the feed consumption. The concentrations of protein, calcium, phosphorus and cholesterol in serum were not influenced by the variation in Lys contents in the diet. The humoral immune response as measured by antibody titre in response to SRBC inoculation was significantly lower in the diets containing 1.1% Lys compared to 1.4%. It is concluded that the Lys requirement of broilers is 1.3% (22.5% CP) during 0 to 21 days of age for eliciting optimum performance when a fixed ratio of Lys to CP (1:17.31) and essential AA is maintained (1:0.47 Met; 1:0.56 Thr; 1:0.17 Try).

• Journal of the Korean Magnetic Resonance Society
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• v.20 no.4
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• pp.102-108
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• 2016

With the rapid increase of data on protein-protein interactions, the need for delineating the 3D structures of huge protein complexes has increased. The protocols for determining nuclear magnetic resonance (NMR) structure can be applied to modeling complex structures coupled with sparse experimental restraints. In this report, I suggest the use of multiple rigid bodies for improving the efficiency of NMR-assisted structure modeling of huge complexes using CYANA. By preparing a region of known structure as a new type of residue that has no torsion angle, one can facilitate the search of the conformational spaces. This method has a distinct advantage over the rigidification of a region with synthetic distance restraints, particularly for the calculation of huge molecules. I have demonstrated the idea with calculations of decaubiquitins that are linked via Lys6, Lys11, Lys27, Lys29, Lys33, Lys48, or Lys63, or head to tail. Here, the ubiquitin region consisting of residues 1-70 was treated as a rigid body with a new residue. The efficiency of the calculation was further demonstrated in Lys48-linked decaubiquitin with ambiguous distance restraints. The approach can be readily extended to either protein-protein complexes or large proteins consisting of several domains.

• Journal of Microbiology and Biotechnology
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• v.10 no.3
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• pp.399-404
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• 2000

The adjuvant effect of a muramyl dipeptide (MDP) derivative, MDP-Lys(L18), on enhancing of antitumor immunity induced by X-irradiated tumor cells against highly metastatic B16-BL6 melanoma cells was examined in mice. Mice immunized intradermally (i.d.) with a mixture of X-irradiated B16-BL6 cells and MDP-Lys (L18) [Vac+MDP-Lys (L18)] followed by an intravenous (i.v.)inoculation of $10^4$ viable tumor cells 7 days after immunization, showed a significant inhibition of experimental lung metastasis of B16-BL6 melanoma cells. The most effective immunization for the prophylactic inhibition of tumor metastasis was obtained from the mixture of $100{\;}\mu\textrm{g}$ of MDP-Lys (L18) and $10^4$ X-irradiatied tumor vaccine. Furthermore, immunization of mice with Vac+MDP-Lys(L18), 3 days after tumor challenge, resulted in a significant inhibition of lung metastasis of B16-BL6 melanoma cells in an experimental lung metastasis model. Similarly, the administration of Vac+MDP-Lys(L18), 1 or 7 days after tumor removal, markedly inhibited tumor metastasis of B16-BL6 in a spontaneous lung metastasis model. When Vac+MDP-Lys (L18) was i.d. administered 3 days after subcutaneous (s.c.) inoculation of tumor cells ($5{\times}10^5/site$) on the back, mice treated with Vac+MDP-Lys(L18) showed inhibition of significantly tumor growth on day 20. These results suggest that MDP-Lys (L18) is able to enhance antitumor activity induced by X-irradiated tumor vaccine to reduce lung metastasis of tumor cells, and is a potent immunomodulating agent which may be applied prophylactically as well as therapeutically to treatment of cancer metastasis.

• Animal Bioscience
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• v.36 no.8
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

• Molecules and Cells
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• v.42 no.1
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• pp.79-86
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• 2019

Endolysins are bacteriophage-derived enzymes that hydrolyze the peptidoglycan of host bacteria. Endolysins are considered to be promising tools for the control of pathogenic bacteria. LysB4 is an endolysin produced by Bacillus cereus-infecting bacteriophage B4, and consists of an N-terminal enzymatic active domain (EAD) and a C-terminal cell wall binding domain (CBD). LysB4 was discovered for the first time as an L-alanoyl-D-glutamate endopeptidase with the ability to breakdown the peptidoglycan among B. cereus-infecting phages. To understand the activity of LysB4 at the molecular level, this study determined the X-ray crystal structure of the LysB4 EAD, using the full-length LysB4 endolysin. The LysB4 EAD has an active site that is typical of LAS-type enzymes, where $Zn^{2+}$ is tetrahedrally coordinated by three amino acid residues and one water molecule. Mutational studies identified essential residues that are involved in lytic activity. Based on the structural and biochemical information about LysB4, we suggest a ligand-docking model and a putative endopeptidase mechanism for the LysB4 EAD. These suggestions add insight into the molecular mechanism of the endolysin LysB4 in B. cereus-infecting phages.