In this study, a repeated yeast integrative plasmid (R-YIp) harboring Cre/loxP system was constructed to integrate various gene expression cassettes into the yeast chromosome. The R-YIp system contains a reusable selective marker (CgTRP1), loxP sequence, and target sequence for integration. Therefore, many gene expression cassettes can be integrated into the same position of the same yeast chromosome. In the present study, several model enzymes involving xylan/xylose metabolism were examined, including endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) and xylitol dehydrogenase (XYL2). Efficient expression of these genes was obtained using two promoters (GAL10p and ADH1p) and various plasmids (pGMF-GENE and pAMF-GENE plasmids) were constructed. The XYLP, XYLB, GRE3, and XYL2 genes were efficiently expressed under the control of the GAL10 promoter. Subsequently, R-YIps containing the GAL10p-GENE-GAL7t cassette were constructed, resulting in pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids. These plasmids were sequentially integrated into chromosome VII of a Saccharomyces cerevisiae strain by repeated gene integration and selective marker rescue. These genes were integrated by the R-YIp system and were stably expressed in the yeast transformants to produce active recombinant enzymes. Therefore, we expect that the R-YIp system will be able to overcome current limitations of the host cells and allow selective marker selection for the integration of various genes into the yeast chromosome.
Objective: To evaluate the effects on microbial diversity and biochemical parameters of gradually increasing temperatures, from $5^{\circ}C$ to $25^{\circ}C$ on corn silage which was previously fermented at ambient or low temperature. Methods: Whole-plant corn silage was fermented in vacuum bag mini-silos at either $10^{\circ}C$ or $20^{\circ}C$ for two months and stored at $5^{\circ}C$ for two months. The mini-silos were then subjected to additional incubation from $5^{\circ}C$ to $25^{\circ}C$ in $5^{\circ}C$ increments. Bacterial and fungal diversity was assessed by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) profiling and biochemical analysis from mini-silos collected at each temperature. Results: A temperature of $10^{\circ}C$ during fermentation restricted silage fermentation compared to fermentation temperature of $20^{\circ}C$. As storage temperature increased from $5^{\circ}C$ to $25^{\circ}C$, little changes occurred in silages fermented at $20^{\circ}C$, in terms of most biochemical parameters as well as bacterial and fungal populations. However, a high number of enterobacteria and yeasts (4 to $5\;log_{10}$ colony forming unit/g fresh materials) were detected at $15^{\circ}C$ and above. PCR-DGGE profile showed that Candida humilis predominated the fungi flora. For silage fermented at $10^{\circ}C$, no significant changes were observed in most silage characteristics when temperature was increased from $5^{\circ}C$ to $20^{\circ}C$. However, above $20^{\circ}C$, silage fermentation resumed as observed from the significantly increased number of lactic acid bacteria colonies, acetic acid content, and the rapid decline in pH and water-soluble carbohydrates concentration. DGGE results showed that Lactobacillus buchneri started to dominate the bacterial flora as temperature increased from $20^{\circ}C$ to $25^{\circ}C$. Conclusion: Temperature during fermentation as well as temperature during storage modulates microorganism population development and fermentation patterns. Silage fermented at $20^{\circ}C$ indicated that these silages should have lower aerobic stability at opening because of better survival of yeasts and enterobacteria.
Objective: This study aimed to gain deeper insights into the dynamic changes in spoilage fungi populations during fermentation and the influence of traditional additives on silage quality. Methods: Elephant grass (Pennisetum purpureum) was prepared without any additive (control), and with the addition of 0.5% salt, and 0.5% salt-0.2% sugar mixture. The fungal community was then determined using a classic culturing method and high-throughput sequencing at 0, 5, 15, and 60 days after ensiling. Results: The results showed that the fungal community of elephant grass silage varied significantly between the natural fermentation without any additive and the two additive groups. The diversity and relative abundance of spoilage molds in the control group were much higher than those in the two treatment groups (p<0.05). Three species of yeasts (Candida sp., Pichia sp., Trichosporon sp.) and four spoilage molds (Fusarium sp., Aspergillus sp., Muco sp. and Penicillin sp.) were the predominant fungi in elephant grass during natural fermentation from 0 to 60 days, which were found to be significantly decreased in salt and sugar additive groups (p<0.05). Meanwhile, the diversity and relative abundance of undesirable molds in the 0.5%-salt additive group were the lowest among all groups. Conclusion: Adding salt and sugar, particularly 0.5% salt, is a promising effective approach to reduce the amount of undesirable fungi thus, improving the silage quality of elephant grass in northern Vietnam.
Background: The purpose of this study was to investigate the anti-oral microbial activity and anti-inflammatory effects of rosmarinic acid (RA) in lipopolysaccharide (LPS)-stimulated MC3T3-E1 osteoblastic cells on a titanium (Ti) surface during osseointegration, and to confirm the possibility of using RA as a safe natural substance for the control of peri-implantitis (PI) in Ti-based dental implants. Methods: A disk diffusion test was conducted to confirm the antimicrobial activity of RA against oral microorganisms. In order to confirm the anti-inflammatory effects of RA, inflammatory conditions were induced with 100 ng/ml of LPS in MC3T3-E1 osteoblastic cells on the Ti surface treated with or without 14 ㎍/ml of RA. The production of nitric oxide (NO) and prostaglandin E2 (PGE2) in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface was confirmed using an NO assay kit and PGE2 enzyme-linked immunosorbent assay kit. Reverse transcription polymerase chain reaction and western blot analysis were performed to confirm the expression of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in total RNA and protein. Results: RA showed weak antimicrobial effects against Streptococcus mutans and Escherichia coli, but no antimicrobial activity against the bacteria Aggregatibacter actinomycetemcomitans and the fungus Candida albicans. RA reduced the production of pro-inflammatory mediators, NO and PGE2, and proinflammatory cytokines, TNF-α and IL-1β, in LPS-stimulated MC3T3-E1 osteoblastic cells on the Ti surface at the protein and mRNA levels. Conclusion: RA not only has anti-oral microbial activity, but also anti-inflammatory effects in LPS-stimulated MC3T3-E1 osteoblasts on the Ti surface, therefore, it can be used as a safe functional substance derived from plants for the prevention and control of PI for successful Ti-based implants.
Niu, Jun Li;Wei, Lian Qing;Luo, Yuan Qing;Yang, Wen Ting;Lu, Qi Cheng;Zheng, Xin Xia;Niu, Yu Jie;Sheng, Wen;Cheng, Hong;Zhang, Wen Ju;Nie, Cun Xi
Animal Bioscience
/
v.34
no.4
/
pp.680-691
/
2021
Objective: This study was conducted to investigate the effect of fermented cottonseed meal (FCSM) on growth performance, carcass traits, and fat deposition in white-feather broiler chickens. Methods: A total of 480 male one-day-old white-feather broiler chickens were selected randomly and divided into four groups with six replicates of 20 chickens in each. The experimental chickens were fed diets including 3%, 6%, or 9% FCSM fermented by Candida tropicalis until 42 days old. In the experiment, the chickens of the control group were fed soybean meal. Results: FCSM supplementation linearly decreased the feed conversion ratio from d 15 to 21 and d 36 to 42, respectively (p<0.05). The percentage of carcass and semi-eviscerate increased in response to dietary FCSM supplementation at d 21 (p<0.05). The percentage of eviscerated and semi-eviscerate of 3FCSM was higher than that in other groups at d 35 (p<0.05). At the age of 42 d, the percentage of carcass increased in a quadratic way among increasing FCSM in diets (p<0.05). The subcutaneous fat thickness linearly decreased with the increasing levels of FCSM at d 21 (p<0.05). Gompertz and Logistic functions provided a better fit on abdominal fat and subcutaneous fat, respectively. The best fitted equation predicted that the maximum growth rate of abdominal fat weight and subcutaneous fat thickness occurred at d 28. FCSM had no significant effects on the shape of growth curve of abdominal fat weight and subcutaneous fat thickness, but reduced the height of the curve. Birds receiving the 6FCSM diet for 21 d had smaller adipocyte surface and lower serum glucose as well as triglyceride concentration. Conclusion: FCSM is beneficial for broiler chickens as it positively affects their growth and carcass in addition to altering their fat deposition.
Jeong-Su Moon;Min-Young An;Mal-Geum Park;Jong-Soo Lee
The Korean Journal of Mycology
/
v.51
no.1
/
pp.39-49
/
2023
This study aimed to screen and investigate the microbiological characteristics, enzyme activity, and physiological functionality of unrecorded wild yeasts from the Wolsung valley of South Deogyu mountain and Jinjamcheon in Daejeon city, Korea. Candida sorboxylosa WSC25-4 (NNIBRFG47336), Arxula adeninivorans WSC6-3 (NNIBRFG47335), Farysizyma taiwaniana JRC6-2 (NNIBRFG47339), Saturnispora dispora JTS19-1 (NNIBRFG47334), Vanderwaltozyma polyspora JJCH5-3 (NNIBRFG47337), and Vanrija fragicola SW-9 (NNIBRFG47338) have not been previously recorded. None of these formed spores or pseudo-mycelium, and four strains, including C. sorboxylosa WSC25-4, were global in shaped. Except for V. polyspora JJCH5-3, all strains grew well in the yeast extract-peptone-dextrose (YPD), potato-dextrose and yeast malt extract media. A. adeninivorans WSC6-3 and S. dispora JTS19-1 grew well in 40% glucose-containing YPD medium and V. fragicola SW-9 in 5% NaCl-containing YPD medium. The supernatant of A. adeninivorans WSC6-3, F. taiwaniana JRC6-2, and V. fragicola SW-9 exhibited 58.0, 59.0, and 59.0% of glucoamylase activity, respectively. C. sorboxylosa WSC25-4 also produced extracellular alkaline protease. Antioxidant activity of the supernatant from F. taiwanian JRC6-2 was high (79.8%), while the other five unrecorded yeasts demonstrated elevated antiaging superoxide dismutase (SOD)-like activity (99%). Anti-gout xanthin oxidase inhibitory activity of S. dispora JTS19-1 and V. fragicola SW-9 was also elevated at 79 and 80%, respectively.
Sun Woo Park;Ji Young Park;Hyoung Soo Choi;Hyunju Lee
Pediatric Infection and Vaccine
/
v.31
no.1
/
pp.46-54
/
2024
Purpose: This study aimed to identify the pathogens of bloodstream infection in children with underlying hemato-oncologic diseases, analyze susceptibility patterns, compare temporal trends with those of previous studies, and assess empirical antimicrobial therapy. Methods: Retrospective review study of children bacteremia in hemato-oncologic diseases was conducted at Seoul National University Bundang Hospital from January 2013 to July 2023. Results: Overall, 98 episodes of bacteremia were observed in 74 patients. Among pathogens isolated, 57.1% (n=56) were Gram-positive bacteria, 38.8% (n=38) were Gram-negative bacteria, and 4.1% (n=4) were Candida spp. The most common Gram-positive bacteria were coagulase-negative staphylococci (n=21, 21.4%) and Staphylococcus aureus, (n=14, 14.3%) whereas the most common Gram-negative bacteria were Klebsiella pneumoniae (n=16, 16.3%) and Escherichia coli (n=10, 10.2%). The susceptibility of Gram-positive bacteria to penicillin, oxacillin, and vancomycin was 11.5%, 32.7%, and 94.2%, respectively and the susceptibility of Gram-negative bacteria to cefotaxime, piperacillin/tazobactam, imipenem, gentamicin, and amikacin was 68.6%, 80%, 97.1%, 82.9%, and 91.4%, respectively. Methicillin-resistant S. aureus was detected in 1 strain and among Gram-negative strains, extended spectrum β-lactamase accounted for 28.9% (12/38). When analyzing the antibiotic susceptibility and empirical antibiotics, the mismatch rate was 25.5% (n=25). The mortality rate of children within 30 days of bacteremia was 7.1% (n=7). Conclusions: Empirical antibiotic therapy for bacteremia in children with hemato-oncologic diseases should be based on the local antibiogram in each institution and continuous monitoring is necessary.
In recent industrialization and development due to information and communication technology, modern women in modern society are exposed to physical and mental health due to numerous stresses. Popular inflammations are attributable to a decrease in lactic acid bacteria, frequent antibiotic use, and a decrease in immunity. It is necessary to develop products that are helpful and reflected. The inner care gel currently introduced on the market can increase beneficial bacteria and maintain a healthy y-zone. The inner gel contains a hydrogel component. 90% is made up of water, and other components act as support for supporting water and are formed through crosslinking between polymer chains. Hydroxyethyl cellulose (HEC) is a hydroxyethyl ethylenetel of cellulose. The purpose of use is to act as a binder, an emulsion stabilizer, a viscosity enhancer (water-soluble), and a film forming agent. CA (crosslinker) is a crosslinking agent and serves to bind. Hydrogel in the beauty field acts as a film forming agent that gently wraps around the skin by forming a thin film and serves as an emulsion stabilizer that helps to prevent separation of other raw materials. It also acts as a thickener by increasing viscosity in cosmetics. In addition, it is used for glucose monitoring, nursing care, cell transplantation, and wound treatment in the bio field. Currently, it is understood that no products using functional hydrogel have been released, so in this study, a Y zone care hydrogel solution was manufactured to find out the antibacterial properties of the functional hydrogel, and a new solution was developed. As a result, it was confirmed that the appropriate Ph was applied to the Y zone, and after culturing Candida albicans in PDB medium, all three products of the Y zone care hydrogel solution showed an antibacterial effect of 0.5-1.0mm
Qian Fan;Yini Xu;Yingping Xiao;Caimei Yang;Wentao Lyu;Hua Yang
Animal Bioscience
/
v.37
no.7
/
pp.1213-1224
/
2024
Objective: Enterotypes (ETs) are the clustering of gut microbial community structures, which could serve as indicators of growth performance and carcass traits. However, ETs have been sparsely investigated in waterfowl. The objective of this study was to identify the ileal ETs and explore the correlation of the ETs with growth performance and carcass traits in Muscovy ducks. Methods: A total of 200 Muscovy ducks were randomly selected from a population of 5,000 ducks at 70-day old, weighed and slaughtered. The growth performance and carcass traits, including body weight, dressed weight and evidenced weight, dressed percentage, percentage of apparent yield, breast muscle weight, leg muscle weight, percentage of leg muscle and percentage of breast muscle, were determined. The contents of ileum were collected for the isolation of DNA and 16S rRNA gene sequencing. The ETs were identified based on the 16S rRNA gene sequencing data and the correlation of the ETs with growth performance and carcass traits was performed by Spearman correlation analysis. Results: Three ETs (ET1, ET2, and ET3) were observed in the ileal microbiota of Muscovy ducks with significant differences in number of features and α-diversity among these ETs (p<0.05). Streptococcus, Candida Arthritis, and Bacteroidetes were the presentative genus in ET1 to ET3, respectively. Correlation analysis revealed that Lactococcus and Bradyrhizobium were significantly correlated with percentage of eviscerated yield and leg muscle weight (p<0.05) while ETs were found to have a close association with percentage of eviscerated yield, leg muscle weight, and percentage of leg muscle in Muscovy ducks. However, the growth performance of ducks with different ETs did not show significant difference (p>0.05). Lactococcus were found to be significantly correlated with leg muscle weight, dressed weight, and percentage of eviscerated yield. Conclusion: Our findings revealed a substantial variation in carcass traits associated with ETs in Muscovy ducks. It is implied that ETs might have the potential to serve as a valuable biomarker for assessing duck carcass traits. It would provide novel insights into the interaction of gut microbiota with growth performance and carcass traits of ducks.
Kim, Jong-Wan;Cho, Il-Sik;Lee, Shin-Jae;Kim, Tae-Woo;Chang, Young-Il
The korean journal of orthodontics
/
v.36
no.1
s.114
/
pp.55-62
/
2006
Orthodontic mini-implants, despite its usefulness as anchorage, have some limits such as loosening. Therefore, various shapes and lengths have been studied. The aim of this study is to analyze the shape and length of mini-implants mechanically. The shapes of mini-implants (1.6 mm, Dual Top, Jeil Medical Co., Seoul, Korea) were cylindrical and taper. The lengths of mini-implants were 6 mm and 8 mm. The tested groups were 5 groups (cylindrical 6 mm, cylindrical 8 mm, taper 6 mm, taper 8 mm and taper 8 mm modified whose thread is reduced from the middle to upper part). All were inserted and removed on the polyurethane foam with the torque measured. During insertion and removal, the taper shape needed higher torque than the cylindrical shape, and the 8 mm length than the 6 mm length (p<0.001). The taper 6mm group showed superior insertion torque (p<0.001) and similar removal torque to the cylindrical 8 mm group. The taper 8 mm modified group with gradually reduced threads, showed continuous high removal torque after the peak. The initial mechanical stability can be provided by the tapered shape and also, affected by length and thread design.
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