• Title/Summary/Keyword: DFI

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TLR-1, TLR-2, and TLR-6 MYD88-dependent signaling pathway: A potential factor in the interaction of high-DNA fragmentation human sperm with fallopian tube epithelial cells

  • Zahra Zandieh;Azam Govahi;Azin Aghamajidi;Ehsan Raoufi;Fatemehsadat Amjadi;Samaneh Aghajanpour;Masoomeh Golestan;Reza Aflatoonian
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.1
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    • pp.44-52
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    • 2023
  • Objective: The DNA integrity of spermatozoa that attach to fallopian tube (FT) cells is higher than spermatozoa that do not attach. FT epithelial cells can distinguish normal and abnormal sperm chromatin. This study investigated the effects of sperm with a high-DNA fragmentation index (DFI) from men with unexplained repeated implantation failure (RIF) on the Toll-like receptor (TLR) signaling pathway in human FT cells in vitro. Methods: Ten men with a RIF history and high-DFI and 10 healthy donors with low-DFI comprised the high-DFI (>30%) and control (<30%) groups, respectively. After fresh semen preparation, sperm were co-cultured with a human FT epithelial cell line (OE-E6/E7) for 24 hours. RNA was extracted from the cell line and the human innate and adaptive immune responses were tested using an RT2 profiler polymerase chain reaction (PCR) array. Results: The PCR array data showed significantly higher TLR-1, TLR-2, TLR-3, TLR-6, interleukin 1α (IL-1α), IL-1β, IL-6, IL-12, interferon α (IFN-α), IFN-β, tumor necrosis factor α (TNF-α), CXCL8, GM-CSF, G-CSF, CD14, ELK1, IRAK1, IRAK2, IRAK4, IRF1, IRF3, LY96, MAP2K3, MAP2K4, MAP3K7, MAP4K4, MAPK8, MAPK8IP3, MYD88, NFKB1, NFKB2, REL, TIRAP, and TRAF6 expression in the high-DFI group than in the control group. These factors are all involved in the TLR-MyD88 signaling pathway. Conclusion: The MyD88-dependent pathway through TLR-1, TLR-2, and TLR-6 activation may be one of the main inflammatory pathways activated by high-DFI sperm from men with RIF. Following activation of this pathway, epithelial cells produce inflammatory cytokines, resulting in neutrophil infiltration, activation, phagocytosis, neutrophil extracellular trap formation, and apoptosis.

Influences of Temperature and Density on the Feeding Growth, and Blind-side Malpigmentation of Fry Starry Flounder Platichthys stellatus (강도다리(Platichthys stellatus) 치어의 섭식, 성장 및 흑화 발현에 있어 온도와 밀도의 영향)

  • Kang, Duk-Young;Kim, Won-Jin;Kim, Hyo-Chan;Chang, Young Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.47 no.5
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    • pp.588-596
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    • 2014
  • To examine the influences of water temperature and stocking density on feeding, growth and blind-side hypermelanosis of the starry flounder Platichthys stellatus, we performed an experiment with fry at two densities: 2 and 8 fish/L. The fry had a total length of $3.2{\pm}0.1cm$, body weight $0.6{\pm}0.1g$, and pigmented patches (pigmented ratio $2.6{\pm}0.4%$) on the blind side. Duplicate experiments were conducted in 93.7 L glass tanks for 120 days, from July to November. We determined daily food intake (DFI), food efficiency (FE), growth, survival rate, ratio of hypermelanic fish, and pigmented area rate on the blind side at 60-day intervals. The DFI was less than 50 mg/fish/day, and growth was delayed from July to September (water temperature [WT]> $20^{\circ}C$). After October, when WT < $20^{\circ}C$, the DFI increased significantly and the growth was accelerated, suggesting that the rearing temperature of starry flounder in artificial facilities should be < $20^{\circ}C$. While the FE, and survival did no differ between the two density groups, DFI and growth were significantly higher at 2 fish/L than at 8 fish/L. There was no difference in the pigmented area ratio between the two density groups. Although the hypermelanosis was not correlated with body size, the malpigmentation increased with growth. These results suggest that a high stocking density is not the main cause of blind-side hypermelanosis, although it can accelerate hypermelanosis in the starry flounder.

Influence of Water Temperature, Background Color, and Light Intensity in Feeding, Growth and Blind-Side Hypermelanosis of Starry Flounder, Platichthys stellatus (강도다리, Platichthys stellatus의 먹이섭식, 성장 및 무안측 체색발현에 있어 수온, 수조색상 및 조도의 영향)

  • Kang, Duk-Young;Kim, Won-Jin;Kim, Hyo-Chan;Chang, Young Jin
    • Korean Journal of Ichthyology
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    • v.26 no.3
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    • pp.185-193
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    • 2014
  • To find the influence of water temperature, tank color and illumination in feeding, growth and blind-side hypermelanosis of starry flounder, Platichthys stellatus, in the present study, we performed a series of temperature, background color and illumination intensity test for 180 days (From June to December). The test was done in duplicate at 100 fish/tank (430% of initial covering area [PCA]) with the selected ordinary juvenile flounder (TL $17.3{\pm}0.5cm$, BW $82.5{\pm}0.2g$). The rearing was performed in darkgreen FRP aquarium tanks ($H100cm{\times}L100cm{\times}W100cm$; bottom area $1m^2$) lighten with average 1,000 lux and 230 lux, and in white FRP aquarium tank ($H100cm{\times}L100cm{\times}W100cm$; bottom area $1m^2$) lighten with average 230 lux of light intensity. We investigated correlation of daily food intake (DFI) with water temperature and salinity, and compared the influences of background colors and light intensity in DFI, food efficiency (FE), growth, survival rate, and ratio of malpigmented blind-side area and ambicolored fish ratio. In DFI, although it was not related with salinity, the amount was significantly decreased under 0.5 g/fish/day in summer and winter season, but was significantly increased over 1.5 g/fish/day in autumn season showing from $10^{\circ}C$ to $20^{\circ}C$ in water temperature. In background and illumination test, DFI, FE and survival rate showed no difference among three groups. The ratios of malpigmented blind-side area and ambicolored fish were also not significantly different among three groups, indicating that the blind-side hypermelanosis of starry may be governed not by background color (or light intensity) but by a genetics external trait inherited from parents.

The Secondary Island Flap Using Omental Vascular Carrier in Rats (백서의 대망을 혈관경으로 이식하여 생성시킨 이차적 도서형 피판)

  • Lew, Dae-Hyun;Tark, Kwan-Chul;Lew, Jae-Duk
    • Archives of Reconstructive Microsurgery
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    • v.5 no.1
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    • pp.16-23
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    • 1996
  • The omental pedicle based on right gastroepiploic vessels is designed new experimental model for prefabrication(revasculirization) of skin flaps in rats. A $2.5{\times}4cm$ pack of omentum with right gastroepiploic vessels was transferred under a bipediceld panniculocutaneous flap which is $2.5{\times}8cm$ size. At day 7, all four margin was divided and the flap was rasied as an secondary island flap connected only by its vascular pedicle, then the composite flap sutured back in place. The flap perfusion was examined by dermofluorometry and flap survival area was measured at day 12. The Secondary island flap demonstrated a dye fluorescence index(DFI%) of $31.38{\pm}12.33$ and survival rate $80.47{\pm}9.61$ The survival rate was increased when DFI% and contact surface between vascular carrier and skin flap was increased. An india ink injection and histologic examination provided visual evidence of revasculization. The omental pedicle is a promising and safe model for revasculirization of other tissues.

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Efficient isolation of sperm with high DNA integrity and stable chromatin packaging by a combination of density-gradient centrifugation and magnetic-activated cell sorting

  • Chi, Hee-Jun;Kwak, Su-Jin;Kim, Seok-Gi;Kim, Youn-Young;Park, Ji-Young;Yoo, Chang-Seok;Park, Il-Hae;Sun, Hong-Gil;Kim, Jae-Won;Lee, Kyeong-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.43 no.4
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    • pp.199-206
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    • 2016
  • Objective: This study was carried out to investigate the correlations of the sperm DNA fragmentation index (DFI) with semen parameters and apoptosis, and to investigate the effects of density-gradient centrifugation (DGC) and magnetic-activated cell sorting (MACS) on reducing the proportion of sperm with DNA fragmentation and protamine deficiency. Methods: Semen analysis and a sperm DNA fragmentation assay were performed to assess the correlations between semen parameters and the DFI in 458 semen samples. Sperm with progressive motility or non-apoptosis were isolated by DGC or MACS, respectively, in 29 normozoospermic semen samples. The effects of DGC or MACS alone and of DGC and MACS combined on reducing the amount of sperm in the sample with DNA fragmentation and protamine deficiency were investigated. Results: The sperm DFI showed a significant correlation (r=-0.347, p< 0.001) with sperm motility and morphology (r=-0.114, p< 0.05) but not with other semen parameters. The DFI ($11.5%{\pm}2.0%$) of semen samples was significantly reduced by DGC ($8.1%{\pm}4.1%$) or MACS alone ($7.4%{\pm}3.9%$) (p< 0.05). The DFI was significantly further reduced by a combination of DGC and MACS ($4.1%{\pm}1.3%$, p< 0.05). Moreover, the combination of DGC and MACS ($1.6%{\pm}1.1%$, p< 0.05) significantly reduced the protamine deficiency rate of semen samples compared to DGC ($4.4%{\pm}3.2%$) or MACS alone ($3.4%{\pm}2.2%$). Conclusion: The combination of DGC and MACS may be an effective method to isolate high-quality sperm with progressive motility, non-apoptosis, high DNA integrity, and low protamine deficiency in clinical use.

Comparative Evaluation of Real-Time PCR and Conventional Culture Method Using Two Selective Agars for the Detection of Cronobacter spp. in Powdered Infant Formula and Dried Pumpkin (조제분유와 건조호박에서 Cronobacter spp. 검출을 위한 두 가지 선택배지와 Real-time PCR의 비교검증)

  • Kim, Hong-Seok;Shin, Minjung;Chon, Jung-Whan;Lim, Jong-Soo;Kim, Young-Ji;Kim, Dong-Hyeon;Chang, Ho-Seok;Kim, Hyunsook;Om, Ae-Son;Oh, Deog-Hwan;Song, Kwang-Young;Seo, Kun-Ho
    • Journal of Food Hygiene and Safety
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    • v.31 no.6
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    • pp.439-444
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    • 2016
  • In the present study, the performance of culture methods using two selective agars and real-time PCR were compared for selective isolation of Cronobacter in powdered infant formula and dried pumpkin. Two food samples were spiked with the pathogen and then preenriched in distilled water. A small portion of preenrichment (10 mL) was incubated in Enterobacteriaceae enrichment both, followed by inoculation onto Druggan-Forsythe-Iversen agar (DFI agar) and Cronobacter sakazakii chromogenic plating agar (R&F agar). The preenrichment and enrichment (1 mL each) was used in real-time PCR assay. In powdered infant formula (PIF), no statistical difference was observed between both culture methods and real-time PCR with preenrichemt (p > 0.05). However, the number of positives obtained by R&F agar and real-time PCR was much higher than that of culture method using DFI agar in dried pumpkin (p < 0.05). In particular, R&F agar yielded a significantly greater selectivity than DFI agar in dried pumpkin (p < 0.05). Real-time PCR and R&F agar, which are currently recommended by US FDA, could be used as an alternative detection tools for the isolation of Cronobacter in PIF and ingredient of child foods such as dried pumpkin that has high number of competing natural microflora.

On the new mold structure with multi-point gate for filling-balance mold (다점 핀포인트 금형에서 균형충전이 가능한 사출금형 구조)

  • Kwon, Youn-Suk;Jeong, Yeong-Deug
    • Design & Manufacturing
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    • v.2 no.5
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    • pp.25-29
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    • 2008
  • Almost all injection molds have multi-cavity runner system for mass production, which are designed with geometrically balanced runner system in order to accomplish filling balance between cavity to cavity during processing. However, even though geometrically balanced runner is used, filling imbalance have been observed. Filling imbalance could be decreased by modifying processing conditions such as injections rate, mold temperature, injection pressure, melt temperature that are related to shear, viscosity. In this study, a series of experiment was conducted to investigate filling imbalance variation when modifying runner layout and ploymer and to determine which processing condition influences as the primary cause of filling imbalance in geometrically balanced runner system. The filling imbalance was desreased up to result range of $3{\leq}DFI{\leq}8(%)$ by using a new runner system for balanced filling.

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Optimization of Processing on Filling Balance of the HR3P Mold Structure (균형충전을 위한 HR3P 금형 구조에서의 공정의 최적화)

  • Kwon, Youn-Suk;Jeong, Yeong-Deug
    • Journal of the Korean Society for Precision Engineering
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    • v.26 no.3
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    • pp.98-102
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    • 2009
  • Almost all injection molds have multi-cavity runner system for mass production, which are designed with geometrically balanced runner system in order to accomplish filling balance between cavity to cavity during processing. However, even though geometrically balanced runner is used, filling imbalance has been observed. Filling imbalance could be decreased by modifying processing conditions such as injections rate, mold temperature, injection pressure, melt temperature that are related to shear, viscosity. In this study, a series of experiment was conducted to investigate filling imbalance variation when modifying runner layout and polymer and to determine which processing condition influences as the primary cause of filling imbalance in geometrically balanced runner system. The filling imbalance was decreased up to result range of $3{\leq}DFI{\leq}8(%)$ by using a new runner system for balanced filling.

Influence of Background Color and Substratum on the Blind-side Hypermelanosis in Starry Flounder Plathchthys stellatus (강도다리(Platichthys stellatus) 흑화 발현에 미치는 수조색깔 및 자갈기질의 영향)

  • KIM, Won-Jin
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.3
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    • pp.841-847
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    • 2016
  • To study the influence of background color and substratum on hypermelanosis of starry flounder, we compared the daily food intake (DFI), the feed efficiency (FE), the survival, the growth, the ratio of pigmented area on the blind side and the ratio of hypermelanic fish duplicately reared for 180 days in dark-green FRP aquarium (control), white FRP aquarium together with dark-green substratum. The ratio of pigmented area on the blind side was significantly higher at the dark-green group than at the white group. DFI, FE and growth were higher in the dark-green substratum. Pigmented area rate and ratio of hypermelanic fish were significantly higher at the dark green group than at the high dark-green substratum. The results suggest that bright tank color and substratum bottom could inhibit the hypermelanosis.

Isolation and Genotyping of Enterobacter sakazakii from Powdered Infant Formula Manufactured in Korea

  • Yoo, Mi-Kyung;Kim, Suk-Shin;Oh, Sang-Suk
    • Food Science and Biotechnology
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    • v.14 no.6
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    • pp.875-877
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    • 2005
  • Presence of Enterobacter sakazakii, occasional pathogen of powdered infant formula causing rare, but life-threatening diseases such as neonatal meningitis, bacteremia, necrotizing enterocolitis, and necrotizing meningoencephalitis after ingestion was examined in 45 powdered infant formula products manufactured in Korea using chromogenic Druggan-Forsythe-Iversen (DFI) medium, and isolates were identified with API 20E. Ent. sakazakii was isolated from three products. Ent. sakazakii isolates were genotyped by RAPD-PCR using two random primers, and their banding patterns were compared.