• Title/Summary/Keyword: Cell membrane integrity

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Antimicrobial activity of the hexane extract of Stachys sieboldii MIQ leaf (초석잠의 잎 추출물의 항균 활성)

  • 류병호;박법규
    • Journal of Life Science
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    • v.12 no.6
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    • pp.803-811
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    • 2002
  • The present study was carried out for research and development of natural antimicrobial from extract of Stachys sieboldii MIQ against food borne bacteria. The hexane extract of Stachys sieboldii MIQ at 250$\mu\textrm{g}$/$m\ell$ per disc showed 15 ~ 20 mm inhibition zone against gram positive and gram negative barteria. Minimum inhibitory concentration (MIC) of hexane extract was 250${\mu}g$/$m\ell$against Bacillus cereus, 250~500${\mu}g$/$m\ell$against Listeria monocytogenes, 500${\mu}g$/$m\ell$ against Staphylococcus aureus, Psedomonas aeruginosa. Observation by transmission electron microscope, showed that disruption of the cell wall assumed to be due to the bactericidal activity. In addition, the membrane integrity of the sensitive cells was disrupted by exposure to Stachys sieboldii MIQ extract on the D-$\beta$-galatosidase activity as substrate of O-nitrophenol-$\beta$-D-galacto-pyranoside. The hexane extract of Stachys sieboldii MIQ was very stable on the pH and thermal stability.

Analysis of Membrane Integrity, DNA Fragmentation and Mitochondrial Function in Pig Spermatozoa Sorted by Flowcytometer

  • Kim, In-Cheul;Han, Deug-Woo;Lee, Sung-Won;Ryu, Jae-Weon;Choi, Eun-Ji;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.32 no.2
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    • pp.123-126
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    • 2008
  • The objective of this study was to determine the potential hazardous effects of sorting process by flowcytometry on the quality of boar spermatozoa by flowcytometer. Freshly collected boar semen was diluted and divided into two groups; control none sorted and sorted. Sperms in sorted group were processed with flowcytometer for cell sorting with $100\;{\mu}M$ nozzle under the 20 psi pressure. Measurements on each parameter were made at two time points, 0hr (right after sorting) and 24 hr post sorting. Although there was a tendency of lower viability in sorted group than none sorted control group, the percentage of live cells in control ($75.83{\pm}6.92\;&\;59.53{\pm}10.34$) was not significantly different from sorted ($59.70{\pm}7.37\;&\;43.97{\pm}3.76$) at both 0 and 24 hr post sorting. However, sorted sperm showed significantly lower mitochondrial function compared to the control at both 0 h ($79.37{\pm}3.22\;vs.\;63.50{\pm}10.05$) and 24 hr ($67.27{\pm}3.22$ vs. $46.97{\pm}5.37$) time points (p<0.007). Sperm DNA fragmentation rate was significantly lower in control ($22.0{\pm}7.04$) than that of sorted ($32.27{\pm}7.49$) at 24 hr time point (p<0.0002). Taken together, these data suggested thatsorting process by flowcytometer may have influenced sperm motility rather than viability. Also high speed sperm sorting by flowcytometer has significant effects on DNA fragmentation on elapsed time after sorting.

Effect of supplementation of feed with Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls

  • Shah, Syed Mazhar Hussain;Ali, Shujait;Zubair, Muhammad;Jamil, Huma;Ahmad, Nazir
    • Journal of Animal Science and Technology
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    • v.58 no.7
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    • pp.25.1-25.6
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    • 2016
  • Background: The current study was designed to investigate the effect of supplementation of Flaxseed (Linumusitatisimum) oil on libido and semen quality of Nilli-Ravi buffalo bulls. Methods: In this study, 12 adult healthy bulls kept at the Semen Production Unit, Qadirabad district Sahiwal, were used. These bulls were divided into three equal groups, A, B and C. Group A was kept as control, while in groups B and C supplementation of feed was provided by using flaxseed oil @125 ml/day and 250 ml/day,respectively for 12 weeks. Two ejaculates per animal were collected at 0 day then 5th, 6th, 7th, 8th, 9th, 10th, 11th and 12th week of treatment. In this way a total 216 samples were taken, and each semen sample was evaluated for color, volume, mass activity, percent motility, sperm cell concentration per ml, percentage of live sperm, and plasma membrane integrity. Libido of bulls was also evaluated before every collection. Results: Analysis of data revealed that these parameters were significantly (P < 0.01) increased in flax oil treated animals as compared to control. Conclusion: It was concluded from the present study that flax seed oil has beneficial effects on reproductive health of buffalo bull.

Relief of the negative effects of heat stress on semen quality, reproductive efficiency and oxidative capacity of rabbit bucks using different natural antioxidants

  • El-Ratel, Ibrahim Talat;Attia, Kandil Abdel Hai;El-Raghi, Ali Ali;Fouda, Sara Fikry
    • Animal Bioscience
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    • v.34 no.5
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    • pp.844-854
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    • 2021
  • Objective: The potential of extra virgin olive oil (EVOO), betaine (BET), and ginger (GIN), as natural antioxidants, in reducing negative effects of heat stress on physiological responses, antioxidant capacity, semen quality and fertility of bucks under heat stress were investigated. Methods: Forty adult Animal Production Research Institute line rabbit bucks were distributed randomly into four experimental treatments of ten rabbits each. The first treatment was fed the commercial pellet diet (CPD) without supplementation and served as a control. The other three treatments were fed CPD supplemented with EVOO (300 mg), BET (1,000 mg), and GIN (200 mg) per kg diet for 3 consecutive months during the summer season. Results: Supplementation of EVOO, BET, or GIN improved (p<0.05) the sexual desire, progressive motility, vitality, intact acrosome and membrane integrity, sperm cell concentration, sperm outputs and fertility. Seminal plasma total proteins, globulin, total antioxidant capacity, glutathione and glutathione S-transferase, and initial fructose increased (p<0.05), while total lipids, aspartate and alanine aminotransferases and malondialdehyde decreased (p<0.05) compared with the control. In comparing the natural antioxidants treatments, GIN evoked the largest improvement. Conclusion: The inclusion of GIN (200 mg/kg diet) appeared to improve the sexual desire, semen quality and oxidative stress of bucks. This may be a beneficial supplement for the management of rabbit bucks used in natural mating or artificial insemination.

Effects of Antioxidants Treatment on the Cryopreservation of Human Hematopoietic Stem Cells (인간 조혈모 줄기세포의 냉동보존에 미치는 항산화제의 영향)

  • Kim, Eung-Bae;Hong, Soon-Gab;Do, Byung-Rok;Kim, Kyung-Suk;Lee, Joon-Yeong
    • Development and Reproduction
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    • v.12 no.1
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    • pp.67-76
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    • 2008
  • Oxidative damage resulted from reactive oxygen species (ROS) is one of the main causes for the decrease of the viability during in vitro culture and cryopreservation process. This experiment was performed to determine the effects of antioxidants on the human hematopoietic stem cell (HSC) during cryopreservation procedure. HSCs cultured in vitro with or without antioxidants were frozen and then examined for stem cell potential after thawing. The cell viability of thawed HSC was increased in $\alpha$-tocopherol and ascorbic acid treatment group compared to control group ($62.7{\pm}8.0%$) and it was higher in 150 uM $\alpha$-tocopherol treatment group ($70.5{\pm}7.0%$). No significant difference was observed in the membrane integrity in all groups. In auto-differentiation rate, no significant difference was appeared in all groups, but was lower in 150 uM $\alpha$-tocopherol ($7.3{\pm}2.6%$) compared to control group ($10.1{\pm}1.6%$). These results demonstrate that treatment of antioxidants improves the efficiency of cryopreservation for HSC and $\alpha$-tocopherol may be considered effective antioxidant for the protective effect on HSC.

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Antifungal Activity of Silver Salts of Keggin-Type Heteropolyacids Against Sporothrix spp.

  • Mathias, Luciana Da Silva;Almeida, Joao Carlos De Aquino;Passoni, Luis Cesar;Gossani, Cristiani Miranda David;Taveira, Gabriel Bonan;Gomes, Valdirene Moreira;Vieira-Da-Motta, Olney
    • Journal of Microbiology and Biotechnology
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    • v.30 no.4
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    • pp.540-551
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    • 2020
  • Sporotrichosis is a chronic and subacute mycosis causing epidemiological outbreaks involving sick cats and humans in southeastern Brazil. The systemic disease prevails in cats and in humans, with the symptoms restricted to the skin of immunocompetent individuals. Under these conditions, the prolonged treatment of animals and cases of recurrence justify the discovery of new treatments for sporotrichosis. This work addresses the antifungal activity of silver salts of Keggin-type heteropolyacid salts (Ag-HPA salts) such as Ag3[PW12O40], Ag6[SiW10V2O40], Ag4[SiW12O40] and Ag3[PMo12O40] and interactions with the antifungal drugs itraconazole (ITC), terbinafine (TBF) and amphotericin B (AMB) on the yeast and mycelia forms of Sporothrix spp. Sporothrix spp. yeast cells were susceptible to Ag-HPA salts at minimum inhibitory concentration (MIC) values ranging from 8 to 128 ㎍/ml. Interactions between Ag3[PW12O40] and Ag3[PMo12O40] with itraconazole and amphotericin B resulted in higher antifungal activity with a reduction in growth and melanization. Treated cells showed changes in cell membrane integrity, vacuolization, cytoplasm disorder, and membrane detachment. Promising antifungal activity for treating sporotrichosis was observed for the Ag-HPA salts Ag3[PMo12O40] and Ag3[PW12O40], which have a low cost, high yield and activity at low concentrations. However, further evaluation of in vivo tests is still required.

Follow-up of Exogenous DNA by Sperm-mediated Gene Transfer via Liposome

  • Cho, Hwang-Yun;Chung, Ki-Hwa;Kim, Jin-Hoi
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.10
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    • pp.1412-1421
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    • 2002
  • To examine the feasibility of using a sperm vector system for gene transfer, we have investigated the binding and the uptaking of foreign DNA into the sperm nucleus by PCR, in situ hybridization and LSC. We have also examined the transportation of exogenous DNA into oocytes by immunofluorescene via PCR. Sperm cells were incubated with DNA/liposome complexes (1:4 ratio) in fertilization medium with BSA or without BSA. In situ hybridization demonstrated that the transfection rate of sperm cells with and without BSA was 41 and 68% respectively, when the cells were treated with liposome/DNA complexes and 13% for DNA alone. LSC analysis showed that the binding of exogenous DNA was greatly reduced by DNase I treatment which digests DNA bound onto spermatozoa, suggesting that some of the DNA was internalized into the sperm membrane. To find out whether transfected DNA was internalized into sperm intracytomembrane, sperm DNA was amplified by inverse PCR. No PCR products were detected from sperm cells, indicating that the foreign DNA was simply bound onto the sperm membrane. To investigate transfer rates of exogenous DNA into oocytes via sperm cells, we used immunofluorescene method to follow the distribution of foreign DNA via spermatozoa: a few exogenous DNA was located in the cytoplasm of early embryos (13/60, 21.7% for DNA+/liposome+/BSA) and was not located in the pronucleus and/or nucleus. These results suggest that most of the transfected sperm cells could carry the foreign DNA into the egg by in vitro fertilization, but that the transferred DNA is degraded in the developing embryos without stable integration into the zygote genome. Therefore, we have directly injected with transfected sperm cell into oocyte cytoplasm and observed that some of the exogenous DNA was detected in preimplantation embryonic cytoplasm and expressed at preimplantation stages, suggesting that exogenous DNA in early zygote has their integrity. In this study, we have not identified a noble mechanism that interfering transportation of foreign DNA into zygote genome via spermatozoa. Our data, however, demonstrated that inverse PCR and immunofluorescene methods would be used as a new tool for follow-up of gene distribution in oocyte via sperm cells.

Changes in the $Ca^{2+}\;and\;Mg^{2+}$ - dependent Adenosine Triphosphatase Activity and Ultrastructure of Marine Fishes by Partial Freezing III. Changes in the Ultrastructure of Muscle Tissues of Yellowtail during Low-temperature Preservation (a해산어의 부분동결에 의한 $Ca^{2+}\;및\;Mg^{2+}$ -dependent Adenosin Triphosphatase 활성 및 근섬유의 미세구조 변화 III. 저온저장 과정중 방어 근육조직의 미세구조의 변화)

  • 최경호;박찬성
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.6
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    • pp.629-636
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    • 1991
  • Yellowtail fishes(Seriola quinqueeradiata) were submitted to the storages using ice-cooling($0^{\circ}C$), partial freezing($-3^{\circ}C$) and freezing $-20^{\circ}C$) method. Changes in the structures of muscle during storage at different temperatures were investigated. The ice-cooling and partial freezing storage caused early decomposition of glycogen granules and mitochondrial inner membrane, but it was accorded to much slower manner comparing with that of ice-cooling storage. The scars of ice crystals were appeared after three days of storage. The number and size of the crystal increased as progressing of the storage. They were circular and mostly located between fibers. When using the freezing storage, glycogen granules were mostly found from the muscle cell even after fourteen days of storage. Mitochonidral inner membrane maintained their integrity. The scars of ice crystals were also found, however, different from those of partial freezing storage. Their existing sites were random and their shapes were irregular. In many cases, they located in the fiber and had keen edges. Fibers were broken mostly at the Z-lines on fourteen days of storage. From these results, it was concluded that partial freezing storage can repress autolytic enzymic action and can reduce the physical damage from ice crystals which is caused by freezing.

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Foliage Contact Herbicidal Activity of Dehydrocostus lactone Derived from Saussurea lappa (목향(Saussurea lappa) 유래 Dehydrocostus lactone의 경엽 접촉 살초 활성)

  • Cho, Kwang-Min;An, Xue-Hua;Chon, Jae-Kwan;Kim, Hyo-Sun;Chun, Jae-Chul
    • Korean Journal of Weed Science
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    • v.30 no.4
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    • pp.421-428
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    • 2010
  • A foliage contact herbicidal substance was separated from ethyl ether fraction in n-hexane extract of Saussurea lappa roots and identified as dehydrocostus lactone [(3aS,6aR,9aR,9bS)-3,6,9-trimethylidene-3a,4,5,6a,7,8,9a,9b-octahydroazuleno[5,4-d]furan-2-one](DHCL). When DHCL at 4,000 ppm was foliage-applied to two grasses and two broadleaf plants, greater than 85% necrotic injury was obtained from large crabgrass, maize and soybean, whereas only about 40% necrotic injury appeared in black nightshade, indicating that DHCL has no gross morphological selectivity, but shows difference in contact response among the plant species tested. Conductivity in incubation medium of the leaf disks treated with DHCL increased as the incubation time continued. Relatively low contact injury in black nightshade as compared with the other three plant species tested was attributed to decrease in absorption of DHCL due to relatively high amount of cuticle. DHCL did not require light in the herbicidal action and there were no inhibitory effects on seed germination and cell elongation. Acetyl-CoA carboxylase activity was inhibited by 30% and 58% at $100\;{\mu}M$ and $1000\;{\mu}M$ DHCL, respectively. These results suggested that the herbicidal action of DHCL was related with inhibition of fatty acid synthesis which in turn caused to weaken cell membrane integrity.

Comparison of AndroMed and Triladyl Extender for Freezing of Jeju Native Black Bull Semen (제주흑우 정액 동결을 위한 AndreoMed와 Triladyl 희석제의 동결성 비교)

  • Cho, Sang-Rae;Cho, In-Chel;Han, Sang-Hyun;Kang, Tae-Young;Cho, Won-Mo;Chae, Hyun-Seok;Kim, Nam-Young;Park, Yong-Sang;Kang, Yong-Jun;Kim, Young-Hoon;Ko, Yeoung-Gyu;Kim, Hyun-Jong;Ko, Moon-Suck
    • Reproductive and Developmental Biology
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    • v.37 no.3
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    • pp.155-159
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    • 2013
  • This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of $2{\times}10^8/ml$ by doubling in every 30 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm ($93.27{\pm}1.62%$) than frozen-thawed sperm ($73.34{\pm}3.27%$). However, there were no significant differences between fresh and frozen-thawed dead cell rate ($7.35{\pm}2.63$ vs, $13.71{\pm}2.85$). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different ($72.86{\pm}2.83$ vs, $81.47{\pm}2.48$), similarly, the dead cell rates was similar ($18.41{\pm}3.42%$ and $17.26{\pm}4.25$). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.