• 제목/요약/키워드: immunohistochemical methods

검색결과 765건 처리시간 0.035초

p53 Protein Expression Area as a Molecular Penumbra of Focal Cerebral Infarction in Rats

  • Hong, Hyun-Jong;Park, Seung-Won;Kim, Young-Baeg;Min, Byung-Kook;Hwang, Sung-Nam;Suk, Jong-Sik
    • Journal of Korean Neurosurgical Society
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    • 제38권4호
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    • pp.293-298
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    • 2005
  • Objective : The authors investigate the spatial characteristics of apoptotic genes expressed around the focal cerebral infarction, and attempted to explain the penumbra with them. Methods : A delayed focal cerebral infarction was created in twelve adult Sprague-Dawley rats. We performed the immunohistochemical staining for the apoptosis, bcl-2 and p53 proteins and measured the local cerebral blood flow [CBF] at the infarction core area and peri-infarct area pre- and intra-operatively. The peri-infarct area was divided into six sectors by distance from the infarction border. Results : The size [$mm^2$] of apoptosis, bcl-2, and p53 areas were $3.1{\pm}1.2$, $4.7{\pm}2.1$, and $6.8{\pm}2.4$, respectively. Apoptosis, bcl-2 or p53 positive cells were concentrated at the peri-infarct area adjacent to the infarction core. Their numbers reduced peripherally, which was inversely proportional to the local CBF. The p53 area seems to overlap with and larger than the ischemic penumbra. Conclusion : The p53 positive area provides a substitutive method defining the penumbra under the molecular base of knowledge.

MSG와 phenylalanine 처리가 흰쥐 시상하부와 대뇌피질에 미치는 영향 (The Effect of Treatment with MSG(monosodium L-glutamate) and/or Phenylalanine on the Hypothalamus and Cerebral Cortex of the Rats)

  • 김명순;이창현;김용준
    • 한국임상수의학회지
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    • 제16권2호
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    • pp.265-271
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    • 1999
  • These experiments were performed to investigate the effect of saline, monosodium L-glutamate(MSG), MSG-phenylalanine, and/or phenylalanine on TH-immunoreactivity in arcuate nucleus of hypothalamus and hind limb area of cerebral cortex in rats using the immunohistochemical methods. The result were as follows: 1. TH-immunoreactive neurons in hind limb area of cerebral cortex and arcuate nucleus of hypothalamus were decreased in MSG treated group compared to the saline treated group and also lesions in arcuate nucleus and median eminence of hypothalamus were increased with MSG treatment 2. TH-immunoreactive neurons in median eminence and arcuate nucleus were increased in phenylalanine treated group compared to the saline treated group and also neurons were more increased in lamina V of hind limb area of cerebral cortex with phenylalanine treatment. 3. TH-immunoreactive neurons in median eminence and arcuate nucleus were decreased in MSG-phenylalanine treated group compared to the phenylalanine treated group and increased compared to the MSG treated group. In lamina V of hind limb area of cerebral cortex, TH-immunoreactive neurons were more decreased in MSG-phenylalanine treated group than that of the phenylalanine treated group, and more increased than that of MSG treated group. These experiments indicated that TH-immunoreactive neurons in hypothalamus and cerebral cortex were increased due to the activation of phenylalanine and decreased by suppressing activation of phenylalanine through MSG treatment.

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Response of masticatory muscles to passive stretch stimulus - from perspectives of functional appliances

  • Pae, Eung-Kwon
    • 대한치과교정학회지
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    • 제42권2호
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    • pp.64-72
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    • 2012
  • Objective: The aims of this study were to examine whether a passive stretch stimulus by means of a functional appliance induces changes in the fiber composition of masticatory muscles and whether these changes are similar to the changes in stretched limb muscle fibers by using RT-PCR, western blot, and immunohistochemical assays. Methods: Five male New Zealand White rabbits were fitted with a prefabricated inclined plane on the maxillary central incisors to force the mandible forward (- 2 mm) and downward (- 4 mm). Further, 1 hind limb was extended and constrained with a cast so that the extensor digitorum longus (EDL) was stretched when the animal used the limb. The animals were sacrificed aft er 1 week and the masseter, lateral pterygoid, and EDL were processed and compared with those from control animals (n = 3). Results: The stretched EDL had a significantly higher percentage of slow fibers, whereas the stretched masticatory muscles did not show changes in the composition of the major contractile proteins aft er 7 days. Conclusions: The transition of fiber phenotypes in response to a stretch stimulus may take longer in the masticatory muscles than in the limb muscles.

사람의 타액선에서 proline-rich protein의 발현양상과 기능적 단백 구조에 대한 분자생물학적 연구 (A MOLECULAR BIOLOGICAL STUDY ON THE EXPRESSION PATTERN AND FUNCTIONAL PROTEIN STRUCTURES OF PROLINE-RICH PROTEINS IN HUMAN SALIVARY GLANDS)

  • 주재용;이석근;박영욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제28권1호
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    • pp.31-41
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    • 2002
  • Proline-rich proteins (PRPs) are major components of human saliva. In order to know the biological roles of PRPs, we explored the expression pattern and functional protein structures of PRPs by the immunohistochemical and various molecular biological methods. Polyclonal antibody against human gPRP was generated from rabbit by the injection of oral exfoliated cells specially treated by urea and SDS buffer. The PRPs began to be expressed both in the acinar cells and ductal cells from the EIDS (Early Intermediate Developmental Stage) of fetal salivary glands and became intense in the salivary epithelium in the LDS (Late Developmental Stage) and adult salivary glands. The polyclonal antibody against the gPRP showed the cross-reactivity with aPRP and bPRP, these results were relevant to the high homology among subtypes of PRP. However, the simulated protein structures of PRPs showed the characteristic repetitive whorling domains except the N-terminal signal peptide. The whorling domains were also contained the multiple amino acids of glutamine and glycine, which may provide the receptor binding or cross-linking sites of PRPs.

체외 배양된 SNU-1 세포주에서 transglutaminase C antisense inhibition이 일으키는 세포핵질 변화 (NUCLEAR MATRIX CHANGES BY THE ANTISENSE INHIBITION OF TRANSGLUTAMINASE C IN IN VITRO CULTURE OF SNU-1 CELLS)

  • 장재현;이석근;박영욱
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제29권2호
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    • pp.86-94
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    • 2003
  • It has been known that transglutaminase C (TGase C, TGase II) is directly participated in the DNA organization of chromosome, and affects the cellular processes such as proliferation, differentiation, and apoptosis of cells, but still not known what mechanism is working on. In this study, the cytogenetic and the immunohistochemical methods were used to observe the TGase C expression in the nuclear chromosome of the proliferating cells, especially in mitotic stage. The human gastric adenocarcinoma (SNU-1) cell line was used for immunohistochemistry and antisense inhibition study in vitro. The present study was also aimed to disclose the efficiency of antisense inhibition by using antisense oligonucleotide DNA labeled with fluorescence, and found that anti-TGase C probe was diffusely infiltrated into the cytoplasm and the nucleus of the cell. By the antisense inhibition the nuclei of SNU-1 cells became rough nuclear shape, as they were greatly reduced in TGase C immunoreactivity both for the normal and apoptotic SNU-1 cells. However, it is clearly presumed that the TGase C directly interacts with the chromosome of SNU-1 cells and it may play an important role in the division and organization of the chromosome during the mitotic stage.

Wound Healing Effect of 525 nm Green LED Irradiation on Skin Wounds of Male Sprague Dawley Rats

  • Cheon, Min-Woo;Park, Yong-Pil
    • Transactions on Electrical and Electronic Materials
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    • 제11권5호
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    • pp.226-229
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    • 2010
  • Many methods exist that promote wound healing, including light therapy, which consists of light beams that assist the human body in treating and sterilizing wounds, as well as regenerating cells. Irradiation with specific wavelengths of either laser or LED light has been shown to induce beneficial proliferation of fibroblasts that, depending on the size of the wound, can be effective in promoting wound healing. The experiments in this study utilized 8 week old 250~300 g Male Sprague Dawley Rats (ILAR Code: NTacSam:SD) and included a non-irradiation group and a 525 nm green LED irradiation group (n of each group = 7). In experiments animals were allowed to rest for 24 hours after wounds had been excised, which was followed by non- irradiation or 525 nm green LED irradiation therapy one hour per day for 9 days. Immunohistochemical staining was conducted for cytokeratin in order to precisely measure the defect size. In addition, Masson's trichrome staining was utilized in order to compare levels of collagen between the 525 nm green LED irradiation group and the non-irradiation group. Animals exposed to 525 nm green LED irradiation (p<0.05) healed at a faster rate and had increased collagenosis compared with the non-irradiated control group. Thus, treatment with 525 nm green LED irradiation had a beneficial effect on wound healing and should be considered as a possible alternative to low power laser treatment.

천마가 중대뇌동맥 폐쇄 흰쥐의 신경세포 자연사에 미치는 영향 (Effect of Gastrodiae Rhizoma on Apoptosis in Cerebral Infarction Induced by Middle Cerebral Artery Occlusion in Rats)

  • 윤유석;이종수
    • 한방재활의학과학회지
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    • 제19권3호
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    • pp.1-13
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    • 2009
  • Objectives : This study evaluates neuroprotective effect of Gastrodiae Rhizoma on apoptosis in the cerebral infarct. Methods : Cerebral infarct was induced by the middle cerebral artery occlusion (MCAO) for 2 hours with intraluminal thread method in Sprague-Dawley rats. Then ethanol extract of Gastrodiae Rhizoma was administered orally for 3 days. Infarct area and volume were evaluated with TTC staining. Neuronal apoptosis was identified with TUNEL labeling. Apoptosis modulatory effect was observed with immunohistochemical Bax, Bcl-2, iNOS, and MMP-9 expressions in penumbra. Results : 1. Ethanol extract of Gastrodiae Rhizoma reduced infarct size partly and volume significantly of in the MCAO rat brain. 2. Ethanol extract of Gastrodiae Rhizoma reduced TUNEL positive cell ratio in the penumbra of MCAO rat brain significantly. 3. Ethanol extract of Gastrodiae Rhizoma suppressed Bax, iNOS and MMP-9 expression in the penumbra of MCAO rat brain significantly. 4. Ethanol extract of Gastrodiae Rhizoma did not change Bcl-2 expression in the penumbra of MCAO rat brain. But expression ratio of Bcl-2 against Bax was increased in the Gastrodiae Rhizoma group. Conclusions : These results suggest that Gastrodiae Rhizoma plays an anti-apoptotic neuroprotective effect through suppression of Bax, iNOS, and MMP-9 expressions and relative up-regulation of Bcl-2 in the ischemic brain tissue.

치자(梔子)가 강제수영부하시험에서 Corticotropin-Releasing Factor, c-fos와 Tyrosine Hydroxylase에 미치는 영향 (Effects of Gardeniae Fructus on Corticotropin-Releasing Factor, c-fos and Tyrosine Hydroxylase in Forced Swimming Test)

  • 박찬혁;이태희
    • 대한한의학방제학회지
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    • 제17권1호
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    • pp.163-173
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    • 2009
  • Objectives : The goal of this study was to investigative the effect of Gardeniae Fructus (GF)as antidepressant in the forced swimming test(FST) model rats. Methods : The expressions of corticotropin-releasing factor(CRF), tyrosine hydroxylase (TH) and c-fos were measured by immunohistochemical method at paraventricular nucleus(PVN), locus coeruleus (LC) and ventral tegmental area(VTA). Results : The duration of immobility in FST was significantly decreased in the GF 100mg/kg groups (p<0.05). CRF expression was significantly decreased at PVN in the GF 100 mg/kg and 400mg/kg treated group in comparison with the control group, respectively (p<0.01). c-fos expression was decreased at PVN in the GF 100 mg/kg treated group with no significance. TH expression was significantly decreased in the GF 100 mg/kg and 400mg/kg treated group in comparison with the control group, at LC and VTA respectively (p<0.001). Conclusion : According to the results, it can be considered that Gardeniae Fructus has antidepressant effect by showing the reduction of immobility in FST through the reduction of CRF, TH expression.

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목단피(牧丹皮)가 강제수영부하시험에서 Corticotropin-Releasing Factor, c-Fos 와 Tyrosine Hydroxylase에 미치는 영향 (Effects of Moutan Cortex on Corticotropin-Releasing Factor, c-Fos and Tyrosine Hydroxylase in Forced Swimming Test)

  • 성영석;이태희
    • 대한한의학방제학회지
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    • 제17권1호
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    • pp.129-140
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    • 2009
  • Objectives : The goal of this study was to investigate the effect of Moutan Cortex as antidepressant in forced swimming test(FST) model. Methods : The expressions of corticotropin-releasing Factor(CRF), c-Fos and tyrosine hydroxylase(TH) were measured with immunohistochemical method at paraventricular nucleus (PVN), locus coeruleus(LC) and ventral tegmental area(VTA). Results : The duration of immobility in the forced swimming test was significantly decreased in the Moutan Cortex 100 mg/kg treated group in comparison with the control group(p<0.01). CRF and c-Fos expressions at PVN were decreased in the Moutan Cortex 100mg/kg treated group in comparison with the control group. But only the expression of c-Fos was shown the significance(p<0.05). TH expressions at LC and VTA were significantly decreased in the Moutan Cortex 100mg/kg and 400mg/kg treated group in comparison with the control group(p<0.001). Conclusion : According to the results, Moutan Cortex has the antidepressant effect by showing the reduced immobility through the reduction of c-Fos expression at PVN and the reduction of TH expression at LC and VTA.

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Carcinogen-DNA and Protein Adducts-Markers of Exposure and Risk

  • Sanetella, Regina M.
    • 한국독성학회:학술대회논문집
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    • 한국독성학회 2002년도 국제심포지움
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    • pp.1-19
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    • 2002
  • It is well established that the initiating event in chemical carcinogenesis is the binding of reactive carcinogens to DNA. Thus, a number of analytic methods have been developed for determining levels of carcinogen-DNA adducts in humans as a marker of individual exposure and, potentially, of risk for cancer development. In addition, reactive carcinogens also bind to protein suggesting protein adducts can be used as a surrogate for DNA adducts in some situations. We have developed monoclonal and polyclonal antibodies to carcinogen-DNA and protein adductis and highly sensitive ELISA and immunohistochemical assays for determining levels of adducts in human tissues. These studies have demonstrated higher levels of adducts in those with higher exposure as a result of workplace, dietary, chemotherapy, environmental of lifestyle (smoking) exposures. Elevated levels of adducts have been found in lung and liver cancer cases compared to controls. We have also used DNA adducts to determine efficacy of an antiosidant vitamin intervention. DNA adduct studies have demonstrated very different levels of damage in those with similar exposure levels. These interindividual differences are likely the result genetic differences in capacity to activate carcinogens, detoxify reactive intermediates and repair DNA adducts once formed. We are currently investigating the relationship between polymorphisms in a number of these genes to determine their relationship to adduct levels as well as their ability to confer increased risk for cancer development. The ability to identify high risk individuals will allow the targeting of screening and preventive strategies to those most likely to benfit.

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