• Title/Summary/Keyword: Morphological Recovery

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Cryopreservation of in Vitro Grown Axillary Shoot-tip Meristems of Lycium chinense by Vitrification

  • Yang, Byeong-Hun;Song, Min-Jung;Ahn, In-Suk;Kim, Hyun-Tae;Park, Young-Goo
    • Journal of Korean Society of Forest Science
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    • v.95 no.5
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    • pp.585-590
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    • 2006
  • In vitro-grown axillary shot-tip meristems of Lycium chinense Mill. from cold-acclimated plant were successfully cryopreserved using a vitrification technique. After loading for 15 minutes with a mixture of 2.0 M glycerol and 0.4 M sucrose ($20^{\circ}C$), small segments (1-2 mm, 3-4 mm, and 5-6 mm) were cut from axilary buds and exposed to the cryoprotectant solution containing 30% glycerol, 15% ethylen glycol,15% dimethyl sulfoxide (DMSO), and 0.4 M sucrose at $0^{\circ}C$ for 30-120 minutes prior to direct plunge into liquid nitrogen (LN). After rapid thawing ($40^{\circ}C$), the segments were washed with MS medium containing 1.2 M sucrose for 0-35 minutes, and then transferred onto recovery-growth medium. The highest survival rate (about 90%) was obtained with cold-hardening treatment, and cryopreserved explants were successfully recovered to plantlets. No abnormal morphological changes were observed with the recovered plants after cryopreservation.

Unraveling the Role of Cytochrome P450 as a Key Regulator Lantipeptide Production in Streptomyces globisporus

  • Da-Ran Kim;Su In Lee;Youn-Sig Kwak
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.566-574
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    • 2023
  • The aim of this study was to investigate the regulation of lantipeptide production in Streptomyces globisporus SP6C4, which produces the novel antifungal lantipeptides conprimycin and grisin, and to identify the role of cytochrome P450 (P450) in tis regulation. To investigate the regulation of lantipeptide production, we created gene deletion mutants, including ΔP450, ΔtsrD, ΔlanM, ΔP450ΔtsrD, and ΔP450ΔlanM. These mutants were characterized in terms of their morphology, sporulation, attachment, and antifungal activity against Fusarium oxysporum. The gene deletion mutants showed distinct characteristics compared to the wild-type strain. Among them, the ΔP450ΔlanM double mutant exhibited a recovery of antifungal activity against F. oxysporum, indicating that P450 plays a significant role in regulating lantipeptide production in S. globisporus SP6C4. Our findings highlight the significant role of P450 in the regulation of lantipeptide production and morphological processes in S. globisporus. The results suggest a potential link between P450-mediated metabolic pathways and the regulation of growth and secondary metabolism in SP6C4, thereby highlighting P450 as a putative target for the development of new antifungal agents.

The Effect of Multi-lamellar Emulsion (MLE) on Skin Barrier Function: Can an Improve Permeability Barrier Provide a Solution for Itching due to Skin Barrier Malfunction\ulcorner

  • Youm, Jong-kyung;Kim, Yang-hee;Park, Byeong-deog;Jeong, Se-kyoo;Park, Eung-ho;Ahn, Sung-ku;Lee, Seung-hun
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.765-779
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    • 2003
  • Physiological lipid mixtures comprised of cholesterol, ceramide and free fatty acid better maintain epidermal homeostasis and have been recently used for dermatoses induced by skin barrier damage, for example for atopic dermatitis and xerotic skin. Itching and dry atopic dermatitis of the skin may be related to altered skin barrier function. In a previous study, the use of multi-lamellar emulsion (MLE), which is a lipid mixtures containing cholesterol, pseudoceramide and free fatty acid, has been shown to accelerate the recovery of the epidermal permeability barrier. In this study, we assessed the efficacy of MLE compared with a currently used anti-itch moisturizer (AIM), the active ingredients of which are menthol and camphor, on barrier recovery after barrier disruption. To clarify the effect of MLE and AIM after acute barrier perturbation, we measured the relation between transepidermal water loss (TEWL) and the barrier recovery rate at 3, 6, 24, and 48 hours after tape stripping hairless mice and then observed changes in the stratum corneum (SC), including the intercellular lipid structure and secretion of lamellar bodies, by electron microscopy. MLE treated skin recover skin barrier function more rapidly, and AIM treated skin delayed barrier repair. Morphological changes in the epidermis, of MLE treated skin revealed well-conserved lipid multi-lamellar structures at 24 h after tape stripping, whereas AIM treated skin showed altered lamellar bilayers within the SC interstices at 48 h. In addition, MLE treated skin showed an increase in the number of LBs and in their secretions and a decrease in the number of SC layers versus AIM treated skin. These results suggest that MLE may accelerate the production of an epidermal permeability barrier in hairless mice by increasing the number and secretion of LB and improve the dryness and itch associated with an altered epidermal permeability barrier.

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A Mutant Arthrospira platensis M20CJK3 Showing Enhanced Growth Rate and Floatation Activity (생장 및 부상성이 향상된 남세균 돌연변이 균주 Arthrospira platensis M20CJK3)

  • Yoo, Chan;Kim, Choong-Jae;Choi, Gang-Guk;Ahn, Chi-Yong;Choi, Jong-Soon;Oh, Hee-Mock
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.268-274
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    • 2009
  • A photosynthetic cyanobacterium Arthrospira platensis, well known for health food supplement, was studied as a target species for atmospheric $CO_2$ removal as well as biomass production. Although the biomass of A. platensis was massively produced in many countries, the recovery cost of its biomass is still high. The purpose of this study was to develop the A. platensis mutant strains which have enhanced growth rate and floatation activity to reduce the recovery cost. A. platensis KCTC AG20590 was treated with 0.24% ethyl methanesulfonate (EMS) for 20 min at room temperature. The mutant strain A. platensis M20CJK3 was finally selected by its morphological and physiological features. The morphology of the mutant A. platensis M20CJK3 was changed from loose-coiled form to tight-coiled form showing short pitch. The growth and $CO_2$ uptake rate of A. platensis M20CJK3 were improved about 15% and 17% compared with A. platensis KCTC AG20590, respectively. The floatation activity of A. platensis M20CJK3 was enhanced in 2-fold compared with that of A. platensis KCTC AG20590. Soluble proteins extracted from two strains were analyzed by two dimensional electrophoresis (2-DE) and MALDI-TOF MS/MS. Among 15 protein spots induced in 2-DE analysis, two spots were the proteins related to photosynthesis and electron transfer system of the other cyanobacteria. As a consequence, it seems that the tight-coiled mutant A. platensis M20CJK3 has an advantage of high growth rate and floatation activity which are beneficial for the mass cultivation and recovery.

Observation of External Injury and Morphological Movement for Analysis of Recovery Possibility after Storage of the Juvenile Sea Cucumber, Apostichopus japonicus (돌기해삼(Apostichopus japonicus)의 보관 후 회복 가능성 파악을 위한 외상 및 형태학적 변화의 관찰)

  • KIM, Tae-Ik;SON, Maeng-Hyun;CHO, Jae-Kwon;JIN, Young-Guk
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.4
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    • pp.1006-1013
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    • 2016
  • The observed external injury, movement and survival rate according to storage and recovery of the juvenile sea cucumber, Apostichopus japonicus(wet weight $1.0{\pm}0.2g$). The investigated application possibility of external injury(grade I~IV) and movement variation(buccal tentacle, motion, obversion, movement) for estimate of survival rate. The survival rate was observed through the recovery after storage of the sea cucumber in various water temperature(10, 15, 20, $25^{\circ}C$) and hours(3, 6, 12, 24 hour). Grade of external injury lower in the 24 hour experiment group of $20^{\circ}C$ water temperature and 12 hour experiment group of $25^{\circ}C$ water temperature. Buccal tentacle and movement strong related survival rate and external injury. In the case of $20^{\circ}C$ water temperature, survival rate was observed decrease 24 hours experiment. $25^{\circ}C$ water temperature was high to 6 hours but 12 hours experiment group observed decrease of the survival rate. The lethal time(LT10) of the juvenile sea cucumber was 15.73 hours($20^{\circ}C$) and 5.57 hours($25^{\circ}C$). The results of this study provided various measurement method of survival rate according to transportation of the juvenile sea cucumber for release.

Enhanced antidiabetic efficacy and safety of compound K/β-cyclodextrin inclusion complex in zebrafish

  • Nam, Youn Hee;Le, Hoa Thi;Rodriguez, Isabel;Kim, Eun Young;Kim, Keonwoo;Jeong, Seo Yule;Woo, Sang Ho;Lee, Yeong Ro;Castaneda, Rodrigo;Hong, Jineui;Ji, Min Gun;Kim, Ung-Jin;Hong, Bin Na;Kim, Tae Woo;Kang, Tong Ho
    • Journal of Ginseng Research
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    • v.41 no.1
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    • pp.103-112
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    • 2017
  • Background: 20(S)-Protopanaxadiol 20-O-D-glucopyranoside, also called compound K (CK), exerts antidiabetic effects that are mediated by insulin secretion through adenosine triphosphate (ATP)-sensitive potassium ($K_{ATP}$) channels in pancreatic ${\beta}$-cells. However, the antidiabetic effects of CK may be limited because of its low bioavailability. Methods: In this study, we aimed to enhance the antidiabetic activity and lower the toxicity of CK by including it with ${\beta}$-cyclodextrin (CD) (CD-CK), and to determine whether the CD-CK compound enhanced pancreatic islet recovery, compared to CK alone, in an alloxan-induced diabetic zebrafish model. Furthermore, we confirmed the toxicity of CD-CK relative to CK alone by morphological changes, mitochondrial damage, and TdT-UTP nick end labeling (TUNEL) assays, and determined the ratio between the toxic and therapeutic dose for both compounds to verify the relative safety of CK and CD-CK. Results: The CD-CK conjugate ($EC_{50}=2.158{\mu}M$) enhanced the recovery of pancreatic islets, compared to CK alone ($EC_{50}=7.221{\mu}M$), as assessed in alloxan-induced diabetic zebrafish larvae. In addition, CD-CK ($LC_{50} =20.68{\mu}M$) was less toxic than CK alone ($LC_{50}=14.24{\mu}M$). The therapeutic index of CK and CD-CK was 1.98 and 9.58, respectively. Conclusion: The CD-CK inclusion complex enhanced the recovery of damaged pancreatic islets in diabetic zebrafish. The CD-CK inclusion complex has potential as an effective antidiabetic efficacy with lower toxicity.

Panax ginseng (Korea Red Ginseng) repairs diabetic sensorineural damage through promotion of the nerve growth factor pathway in diabetic zebrafish

  • Nam, Youn Hee;Moon, Hyo Won;Lee, Yeong Ro;Kim, Eun Young;Rodriguez, Isabel;Jeong, Seo Yule;Castaneda, Rodrigo;Park, Ji-Ho;Choung, Se-Young;Hong, Bin Na;Kang, Tong Ho
    • Journal of Ginseng Research
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    • v.43 no.2
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    • pp.272-281
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    • 2019
  • Background: Diabetic sensorineural damage is a complication of the sensory neural system, resulting from long-term hyperglycemia. Red ginseng (RG) has shown efficacy for treatment of various diseases, including diabetes mellitus; however, there is little research about its benefit for treating sensorineural damage. Therefore, we aim to evaluate RG efficacy in alloxan-induced diabetic neuromast (AIDN) zebrafish. Methods: In this study, we developed and validated an AIDN zebrafish model. To assess RG effectiveness, we observed morphological changes in live neuromast zebrafish. Also, zebrafish has been observed to have an ultrastructure of hair-cell cilia under scanning electron microscopy. Thus, we recorded these physiological traits to assess hair cell function. Finally, we confirmed that RG promoted neuromast recovery via nerve growth factor signaling pathway markers. Results: First, we established an AIDN zebrafish model. Using this model, we showed via live neuromast imaging that RG fostered recovery of sensorineural damage. Damaged hair cell cilia were recovered in AIDN zebrafish. Furthermore, RG rescued damaged hair cell function through cell membrane ion balance. Conclusion: Our data suggest that RG potentially facilitates recovery in AIDN zebrafish, and its mechanism seems to be promotion of the nerve growth factor pathway through increased expression of topomyosin receptor kinase A, transient receptor potential channel vanilloid subfamily type 1, and mitogen-activated protein kinase phosphorylation.

The Effects of Herbal Acupuncture with Chijadaehwangtang Applied to the Gansu($BL_{18}$) on D-galactosamine-induced Liver Injury in Rats (농도별(濃度別) 치자대황탕(梔子大黃湯) 간유혈(肝兪穴)($BL_{18}$)약침(藥鍼)이 D-galactosamine에 의해 유발된 간손상(肝損傷) 백서(白鼠)에 미치는 영향(影響))

  • Lee, Hyuk-Jae;Ryu, Chung-Ryul;Cho, Myeong-Rae
    • Journal of Acupuncture Research
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    • v.25 no.6
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    • pp.95-107
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    • 2008
  • Objectives : The main purpose of this research is to observe the effects of herbal acupuncture(HA; herbal acupuncture) with Chijadaehwangtang applied to the Gansu($BL_{18}$) on D-galactosamine-induced liver injury in rats. Methods : According to HA concentration, the experimental rats were divided 5 groups(control group, saline group, CP-1, CP-2, CP-3 group). In the control group, we first injected D-galactosamine and then didn't treated. In the saline group, we first injected D-galactosamine and then injected saline. In the CP-1, CP-2, CP-3 group, we first injected D-galactosamine and then injected HA with Chijadaehwangtang applied to the Gansu($BL_{18}$), each 25.3mg/kg, 12.7mg/kg, 5.1mg/kg. HA with Chijadaehwangtang was treated at $20{\mu}{\ell}$ every second day, total 10 times in 20 days. We observed the changes of $\gamma$-GTP, GOT, GPT, total bilirubin, LDH, ALP, total cholesterol, triglyceride, SOD, Catalase and hepatic tissues. Results : 1. In the changes of $\gamma$-GTP, GOT contents, as compared with control group, CP-2 group was significantly decreased. 2. In the changes of GPT content, CP-1, CP-2, CP-3 groups as compared with control group were significantly decreased. 3. In the changes of LDH content, CP-3 group as compared with control group was significantly decreased. 4. In the changes of ALP content, all groups as compared with control group were significantly decreased. 5. In the changes of SOD, Catalase contents, CP-2 group as compared with control group was significantly increased. 6. In the morphological and histopathological changes of hepatic tissues, CP-2, CP-3 groups as compared with control group were improved. Conclusions : we observed HA with Chijadaehwangtang applied to the Gansu($BL_{18}$) can be effective treatment on hematological recovery and regenerative process in morphological liver change. Further studies about their underlying mechanism and concentrations may be needed to use HA with Chijadaehwangtang for liver injury clinically.

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Evaluation of Extended Canine Semen after Different Filtration Treatment (개 희석 정액의 다양한 filtration 처리 후 정자평가)

  • Kim, Yong-Jun;Kim, Jin-Young;Kim, Sue-Hee;Lee, Young-Jun
    • Journal of Veterinary Clinics
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    • v.24 no.4
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    • pp.577-583
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    • 2007
  • It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

The Effect of Ultrasound Treatment on Regeneration Nerve and Muscle on Injured Peripheral Nerve of Rats Model (말초신경 손상 백서모델에서 초음파 치료의 신경 및 근육 재생효과)

  • Cheong, Mee-Sun;Kim, Kyoung-Yoon
    • The Journal of Korean Physical Therapy
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    • v.17 no.4
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    • pp.589-600
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    • 2005
  • The purposes of this study is to discuss and analyze the effect on the recovery from cut in sciatic nerve. This study used 9 weeks male rats of Sprague-Dawley family. Rat in groups 4 were treated with pulsed therapeutic ultrasound for 3 minutes. 3 times weekly at 3MHz respectively (intensity; $0.2W/cm^2,\;0.5W/Cm^2,\;10W/cm^2$); rat in group 1 received placebo ultrasound. In addition, changes of serum aspartate amino-transferase(AST) and creatine phosphokinase(CPK) levels were also demonstrated with diameter of individual muscle fasciculate and number of muscle fiber in each of three types of muscles located in gastrocnemius, soles. The results of comparing the changes in groups are as follows; 1. We found out that hypertrophic epineurium was present in sciatic nerve injured ultrasound treatment of groups. 2. In the gastrocnemius morphological investigation of the group I (control group), severe muscle atrophy were observed at the 7th days of the sciatic nerve injury. however, muscle atrophy of the group IV ($1.0W/cm^2$) were slightly recovered at the 14th days after treatment ultrasound. At the 28th days, muscular fibers were formed in polygon and were significantly recovered. 3. C-fos immunoreactive of the group II ($0.2W/cm^2$), III ($0.5W/cm^2$) were remarkably increased at the 1th day after treatment of ultrasound. Group IV were markedly deceased. 4. Brain-Derived Neurotrophic Factor(BDNF) immunoreactive of the group II, III were increased after 7 days of the sciatic nerve injury. Group IV were markedly increased from 14th days to 28th days after treatment of ultrasound. 5. A significant increase of serum AST levels were demonstrated in control group. However, serum AST levels of massage groups were significantly decreased compared to that of control group in followed order ; ($0.2W/cm^2<0.5W/cm^2<1.0W/cm^2$). 6. A significant increase of serum CK levels were demonstrated in control of group. However, serum CK levels of massage groups were significantly decreased compared to that of control group in followed order ; ($0.2W/cm^2<0.5W/cm^2<1.0W/cm^2$). The above results suggest that ultrasound treatment after peripheral nerve injury might reduce noxious stimuli, facilitate nerve recovery and effective in the functional improvement delaying muscle atrophy or degeneration.

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