• Title/Summary/Keyword: Histological techniques

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Micropropagation of a rare plant species, Astragalus membranaceus Bunge var. alpinus N. (희귀식물 제주황기의 미세번식)

  • Han, Mu Seok;Noh, Seol Ah;Kwak, Myung Cheol;Moon, Heung Kyu
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.100-106
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    • 2014
  • In order to develop an efficient in vitro micropropagation technique for a rare plant species, Astragalus membranaceus Bunge var. alpinus N., shoot proliferation and in vitro or in vivo rootings were conducted and hyperhydrated leaf generated from cultures was histologically observed. During shoot induction, no distinct effect on multiple shoot induction was found between BA and kinetin treatment. BA enhanced the number of internodes, whereas kinetin stimulated shoot elongation. Hyperhydrated leaf composed of bigger cells and retarded palisade parenchyma and showed irregular cell arrangement compared to normal leaf. Especially starch content in hyperhydrated leaf was significantly reduced. The best rooting rate was achieved by B5 medium among three different medium (B5, MS and WPM) and 0.1mg/L IBA treatment induced the highest rooting ratio (80%). No statistical difference was induced by explant types (apical bud or axillary bud) in terms of rooting ratio. In vivo cutting induced rooting rate up to 65% by 0.5% IBA/Talc powder treatment. Although in vivo rooting rate was less efficient compared to in vitro rooting, better survival rate was observed after soil acclimatization. Present study suggested that above micropropagation techniques can be used for rapid multiplication as well as in vitro or in vivo conservation of the species.

AN EXPERIMENTAL STUDY OF ELECTROPHYSIOLOGICAL AND HISTOLOGICAL ASSESSMENT ON THE INJURY TYPES IN RABBIT INFERIOR ALVEOLAR NERVE (가토의 하치조 신경 손상 형태에 따른 전기생리학적 및 조직학적 변화에 관한 실험적 연구)

  • Lee, Jae-Eun;Lee, Dong-Keun
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.18 no.4
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    • pp.679-700
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    • 1996
  • Inferior alveolar nerve dysfunction may be the result of trauma, disease, or iatrogenic injury. Inferior alveolar nerve injury is inherent risk in endodontic therapy, orthognathic surgery of the mandible, and extraction of mandibular teeth, particularly the third molars. The sensory disturbances of inferior alveolar nerve associated with such injury have been well documented clinical problem that is commonly evaluated by several clinical sensory test including Tinels sign, Von Frey test(static light touch detection), directional discrimination, two-point discrimination, pin pressure nociceptive discrimination, and thermal test. These methods used to detect and assess inferior alveolar nerve injury have been subjective in nature, relying on the cooperation of the patients. In addition, many of these techniques are sensitive to differences in the examiners experience and skill with the particular technique. Data obtained at different times or by different examiners are therefore difficult to compare. Prior experimental studies have used electro diagnostic methods(sensory evoked potential) to objectively evaluate inferior alveolar nerve after nerve injury. This study was designed with inferior alveolar nerve of rabbit. Several types of injury including mind, moderate, severe compression and perforation with 19 gauze, 21 gauze needle and 6mm, 10mm traction were applied for taking the sesory evoked ppterntial. Latency and amplitude of injury rabbit inferior alveolar nerve were investigated with sensory evoked potential using unpaired t-test. The results were as follows : 1. Intensity of threshold (T1) was $128{\pm}16{\mu}A$ : latency, $0.87{\pm}0.07$ microsecond : amplitude, $0.4{\pm}0.1{\mu}V$ : conduction velocity, 23.3 m/s in sensory evoked potential of uninjured rabbit inferior alveolar nerve. 2. Rabbit inferior alveolar nerve consists of type II and III sensory nerve fiber. 3. Latency was increased and amplitude was decreased in compression injury. The more injured, the more changed in latency and amplitude. 4. Findings in perforation injury was similar to compression injury. Waveform for sensory evoked potential improved by increasing postinjured time. 5. Increasing latency was prominent in traction injury rabbit inferior alveolar nerve. 6. In microscopic histopathological findings, significant degeneration and disorganization of the internal architecture were seen in nerve facicle of severe compression and 10mm traction group. From the above findings, electrophysiological assessment(sensory evoked potential) of rabbit injured inferior alveolar nerve is reliable technique in diagnosis and prognosis of nerve injury.

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A study of Expression of TGF-β1, c-Myc, Erb-B2 and Thymosin-β4 Gene in Alcoholic Liver Damage Tissue. (알코올성 간 손상 조직에서 TGF-β1와 c-Myc, Erb-B2, Thymosin-β4 유전자 발현 융합 연구)

  • Kim, Jean-Soo;Choi, Sang-Ki
    • Journal of the Korea Convergence Society
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    • v.9 no.5
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    • pp.91-97
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    • 2018
  • This study has been conducted to see the expression of $TGF-{\beta}_1$, c-Myc, Erb-B2 and $Thymosin-{\beta}_4$ genes in ethanol - damaged liver tissues. Experimental groups were divided into 2 groups, one where damaged liver was caused by 25% ethanol and normal group administered with purified water. Results of test showed the expression of $TGF-{\beta}_1$, c-Myc, and $Thymosin-{\beta}_4$ genes was higher in the experimental group treated with 25% ethanol than in the normal group. Erb-B2 gene was not expressed clearly. Thus, it is considered that we can expect to utilize $TGF-{\beta}_1$, c-Myc 및 $Thymosin-{\beta}_4$ as auxiliary data and find clinical meanings of diagnosis on hepatic diseases, In addition to serologic and histological examination by convergence examining the gene expression status by molecular diagnostic techniques in liver-related disease prevention and diagnosis through results of this study.

Effects of SIS/PLGA Porous Scaffolds and Muscle-Derived Stem Cell on the Formation of Tissue Engineered Bone (SIS/PLGA 담체와 근육유래 줄기세포를 이용한 생체조직공학적 골재생)

  • Kim Soon Hee;Yun Sun Jung;Jang Ji Wook;Kim Moon Suk;Khang Gilson;Lee Hai Bang
    • Polymer(Korea)
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    • v.30 no.1
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    • pp.14-21
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    • 2006
  • Tissue engineering techniques require the use of a porous biodegradable/bioresorbable scaffold, which server as a three-dimensional template for initial cell attachment and subsequent tissue formation in both in vitro and in vivo. Small intestinal submucosa (SIS) has been investigated as a source of collagenous tissue with the potential to be used as biomaterials because of its inherent strength and biocompatibility. SIS-loaded poly(L-lactide-co-glicolide)(PLGA) scaffolds were prepared by solvent casting/particle leaching. Characterizations of SIS/PLGA scaffold were carried out by SEM, mercury porosimeter, and so on. Muscle-derived stem cells can be differentiated in culture into osteoblasts, chondrocytes, and even myoblasts by the controlling the culture environment. Cellular viability and proliferation were assayed by 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyltetrazolium-bromide(MTT) test. Osteogenic differential cells were analyzed by alkaline phosphatase(ALP) activity. SIS/PLGA scaffolds were implanted into the back of athymic nude mouse to observe the effect of SIS on the osteoinduction compared with controlled PLGA scaffolds. Thin sections were cut from paraffin embedded tissues and histological sections were conducted hematoxylin and eosin (H&E), Trichrome, and von Kossa. We observed that bone formatioin of SIS/PLGA hybrid scaffold as natural/synthetic scaffold was better thean that of only PLGA scaffold. It canb be explained that SIS contains various kinds of bioactive molecules for osteoinduction.

The Effect of Hair Growth and Distribution by Sophorae Radix, Panax ginseng, Salvia miltiorrhiza BUNGE Water Extracts (고삼, 인삼 및 단삼 혼합물에 의한 모발의 성장과 분포에 미치는 영향)

  • Hwang, Cho-Won;Hwang, Jae-Wan;Kim, Sang-Tae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.36 no.3
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    • pp.215-219
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    • 2010
  • In this study, we investigated effects of Monegy (mixture of Sophorae Radix, Panax ginseng, Salvia miltiorrhiza BUNGE) on epilate-induced hair-loss in dorsal region of C57/BL6 mice and external structure of human hair. For morphological and histological analysis in scalp of epilate-induced hair-loss animal model, we utilized several microscopic techniques, such as confocal laser scanning microscopy (CLSM) and LAS 4000. Confocal analysis showed the distribution of FITC-conjugated Monegy and penetration depth compared with normal and control group. Furthermore, when Monegy was topically administrated onto a C57BL6 mouse, it penetrated very well. The fluorescence intensity was increased upto 205 and 113 folds compared to normal and control group, respectively. Also, area of fluorescence was increased to upto 255 to 127 folds compared to normal and control group. Broad scale area of fluorescence in dermis region was observed in the Monegy-treated mice. Furthermore, Monegy induced upto 75% hair repair against depilation. It might be promoted via the induction of growth factors in hair follicle.

Gross, organoleptic and histologic assessment of cadaveric equine heads preserved using chemical methods for veterinary surgical teaching

  • Rodrigo Romero Correa;Rubens Peres Mendes;Diego Darley Velasquez Pineros;Aymara Eduarda De Lima;Andre Luis do Valle De Zoppa;Luis Claudio Lopes Correia da Silva;Ricardo de Francisco Strefezzi;Silvio Henrique de Freitas
    • Journal of Veterinary Science
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    • v.25 no.2
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    • pp.29.1-29.11
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    • 2024
  • Background: Preservation of biological tissues has been used since ancient times. Regardless of the method employed, tissue preservation is thought to be a vital step in veterinary surgery teaching and learning. Objectives: This study was designed to determine the usability of chemically preserved cadaveric equine heads for surgical teaching in veterinary medicine. Methods: Six cadaveric equine heads were collected immediately after death or euthanasia and frozen until fixation. Fixation was achieved by using a hypertonic solution consisting of sodium chloride, sodium nitrite and sodium nitrate, and an alcoholic solution containing ethanol and glycerin. Chemically preserved specimens were stored at low temperatures (2℃ to 6℃) in a conventional refrigerator. The specimens were submitted to gross and organoleptic assessment right after fixative solution injection (D0) and within 10, 20, and 30 days of fixation (D10, D20, and D30, respectively). Samples of tissue from skin, tongue, oral vestibule, and masseter muscle were collected for histological evaluation at the same time points. Results: Physical and organoleptic assessments revealed excellent specimen quality (mean scores higher than 4 on a 5-point scale) in most cases. In some specimens, lower scores (3) were assigned to the range of mouth opening, particularly on D0 and D10. A reduced the range of mouth opening may be a limiting factor in teaching activities involving structures located in the oral cavity. Conclusions: The excellent physical, histologic, and organoleptic characteristics of the specimens in this sample support their usability in teaching within the time frame considered. Appropriate physical and organoleptic characteristics (color, texture, odor, and flexibility) of the specimens in this study support the use of the method described for preparation of reusable anatomical specimens.

Autometallography for Zinc Detection in the Central Nervous System (중추신경계통내 분포하는 Zinc의 조직화학적 동정)

  • Jo, Seung-Mook;Gorm, Danscher;Kim, Sung-Jun;Park, Seung-Kook;Kang, Tae-Cheon;Won, Moo-Ho
    • Applied Microscopy
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    • v.30 no.4
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    • pp.347-355
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    • 2000
  • Zinc is one of the most abundant oligoelements in the living cell. It appears tightly bound to some metalloproteins and nucleic acids, loosely bound to some metallothioneins or even as free ion. Small amounts of zinc ions (in the nanomolar range) regulate a plentitude of enzymatic proteins, receptors and transcription factors, thus rolls need accurate homeostasis of zinc ions. Zinc is an essential catalytic or structural element of many proteins, and a signaling messenger that is released by neural activity at many central excitatory synapses. Growing evidences suggest that zinc may also be a key mediator and modulator of the neuronal death associated with transient global ischemia and sustained seizures, as well as perhaps other neurological disease stoles. Some neurons have developed mechanisms to accumulate zinc in specific membrane compartment ('vesicular zinc') which can be evidenced using histochemical techniques. Substances giving a bright colour or emitting fluorescence when in contact with divalent metal ions are currently used to detect them inside cells; their use leads to the so called 'direct' methods. The fixation and precipitation of metal ions as insoluble salt precipitates, their maintenance along the histological process and, finally, their demonstration after autometallographic development are essential steps for other methods, the so called 'indirect methods'. This study is a short report on the autometallograhical approaches for zinc detection in the central nervous system (CNS) by means of a modified selenium method.

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Comparative Histological Study on the Parafollicular Cells of Mammals (각종 포유동물 갑상선내의 소포방세포에 관한 비교조직학적 연구)

  • 고정식;박상윤
    • The Korean Journal of Zoology
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    • v.23 no.2
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    • pp.89-108
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    • 1980
  • Comparative studies on the parafollicular cells of the some mammalian species from five different orders were carried out; i.e., man from Primates, cattle, pig, and black goat from Artiodactyla, dog from Carnivora, rabbit from Lagomorpha, rat, mouse, and squirrel from Rodentia. For this study, various special techniques for the parafollicular cells, including Grimelius' silver impregnation method (Sawicki and Bajko, 1974), Singh's argentaffin method (Singh, 1964), HCl-toluidine blue stain (Sawicki, 1971), and HCl-lead hematoxylin stain (Solcia et al., 1969), were applied. Authors obtained the following results: 1. Number of parafollicular cells in the same area of thyroid tissues are significantly different from species to species. Number of cells were largest in dog and less cells were found in the following orders; rat, squirrel, mouse, rabbit, cattle, pig, black goat and finally the smallest number in man. 2. Distribution of parafollicular cells within thyroid gland are significantly different from portion to portion in case of cattle, rabbit, squirrel and mouse, but it is not significant in dog, man, pig, black goat and rat (see Table 1-1 and 1-2). 3. In dog, clustered parafollicular cells are located usually in the interfollicular space, and groups of parafollicular cells are located in the para-and/or inter-follicular positions in rabbit. But in the other animals parafollicular cells are found solitarily in the intra-and/or para-follicular positions. 4. The shape of parafollicular cells shows oval to round contour in dog, but it is polymorphic, for example, spindle, conical, oval, round or elongated with cytoplasmic processes, in the other animals. 5. Size of parafollicular cells is larger in cattle, dog and pig, smaller in rat, mouse and squirrel, and medium-size in rabbit, man and black goat. 6. Parafollicular cells of pig, cattle, dog and squirrel are observed to contain densely packed granules, whereas those of mouse, rat and man contain relatively scanty granules. 7. Parafollicular cells of all the mammals show more or less positive reaction to Grimelius' argyrophile silver impregnation method, HCl-toluidine blue stain and HCl-lead hematoxylin stain, whereas they show negative reaction to argentaffin method (see Table 2). 8. Considering the above finding, it is concluded that there are species differences in the distribution, location and shape of parafollicular cells, and infer that preferable staining method should be selected for reliable detection of parafollicular cells, beacuse staining methods applied on the cells in this study show variable reactions according to species.

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Histological studies on in vitro Propagation of Pulsatilla koreana Nakai (할미꽃 기내증식(器內增殖)에 관(關)한 조직학적(組織學的) 연구(硏究))

  • Lee, Man-Sang;Oh, Ki-Hong
    • Korean Journal of Medicinal Crop Science
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    • v.1 no.2
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    • pp.137-157
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    • 1993
  • This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

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