• Title/Summary/Keyword: ELK

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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.

Identification of Deer Antler Species Using Sequence Analysis and PCR-RFLP of Mitochondrial DNA (사슴 미토콘드리아 DNA의 염기서열 및 PCR-RFLP분석에 의한 녹용의 종 감별)

  • Shin, Ki-Hyun;Shin, Sung-Chul;Chung, Ku-Young;Chung, Eui-Ryong
    • Food Science of Animal Resources
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    • v.28 no.3
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    • pp.276-282
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    • 2008
  • It is estimated that over 80% of deer antlers produced in the world are consumed in Korea. However, mislabeling or fraudulent replacement of costly antlers with cheaper ones is one of the most common problems in the domestic antler market. Therefore, there is a great need for the development of technology to identify species of antlers. This study was carried out to develop an accurate and reliable method for the identification and authentication of species or subspecies of antlers using DNA sequence analysis and comparison of mitochondrial cytochrome band D-loop region genes among antlers of five deer species, Cervus elaphus sibericus, Cervus elaphus canadensis, Cervus nippon, Cervus elaphus bactrianus and Rangifer tarandus. A variable region of cytochrome band D-loop genes was amplified using PCR with specifically designed primers and sequenced directly. The cytochrome band D-loop region genes showed different DNA sequences between the species of antlers and thus it is possible to differentiate between species on the basis of sequence variation. To distinguish between reindeer (Rangifer tarandus) antlers and other deer antlers, PCR amplicons of the cytochrome b gene were digested with the restriction enzymes NlaIV and TaqI, respectively, which generates a species-specific DNA profile of the reindeer. In addition, samples of 32 sliced antlers labeled Cervus elaphus sibericus from commercial markets were collected randomly and the mt DNA D-loop region of these antler samples was sequenced. Among the antler samples investigated, only 62.5% were from Cervus elaphus sibericus, and others were from Cervus elaphus bactrianus (25.0%), elk (Cervus elaphus canadensis) and reindeer (Rangifer tarandus). Our results suggest that DNA sequencing of mt DNA and PCR-RFLP methods using NlaIV and TaqI enzymes are useful for the identification and discrimination of deer antler species by routine analysis.

Changes in Physio-Chemical Properties of Deer Meat Cold Storage at 4°C and -2°C (냉장저장중 사슴고기의 이화화적 성질의 변화)

  • Shin Teak-soon;Kang Han-seok;Kim Seon-ku;Lee Kil-wang
    • Journal of Life Science
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    • v.15 no.4 s.71
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    • pp.522-527
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    • 2005
  • A total of 5 female elk dEER $(220kg\pm10kg)$ were included in a study on the changes in physicochemical properties of deer meat during storage at $4^{\circ}C\;and\;-2^{\circ}C$. The deer was exposed to normal pre-slaughter handling and put under anesthesia before slaughtered. The loin and leg cuts were deboned from the carcass after 24hrs slaughter. The samples weighing approximately 300g were packaged using wrap packaging and stored for 3, 7, 11 and 15 days at $4^{\circ}C\;and\;-2^{\circ}C$. Water-holding capacity was decreased with increasing storage days at $4^{\circ}C\;or\;-2^{\circ}C$, respectively The deer meats kept at $-2^{\circ}C$ showed lower TBARS value than the meats kept at $4^{\circ}C$, and it was possible to extend the storage period of the meats. VBN values of the meats kept at $4^{\circ}C\;and\;-2^{\circ}C$ showed as edible values after storage for 15 days, although there were no significant differences among the storage temperature. pH values of loin and leg tended to be increased with the passage of storage time, and the values of the meats kept at $-2^{\circ}C$ was lower than that at $4^{\circ}C$. The change of meat softness was remarkable at $4^{\circ}C$, and the change at $-2^{\circ}C$ was slow. Therefore, it was effective to extend the storage period when the meats were kept at $-2^{\circ}C$. Color of the meats kept at $-2^{\circ}C$ was darker than that at $4^{\circ}C$, the index of red color was higher for the meats kept at $-2^{\circ}C$, and yellow color of meats kept at $-2^{\circ}C$ was more rapidly changed with the passage of storage time.