• 대한약침학회지
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• 제19권4호
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• pp.319-328
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• 2016

Objectives: Advancements in nanotechnology have led to nanoparticle (NP) use in various fields of medicine. Although the potential of NPs is promising, the lack of documented evidence on the toxicological effects of NPs is concerning. A few studies have documented that homeopathy uses NPs. Unfortunately, very few sound scientific studies have explored the toxic effects of homeopathic drugs. Citing this lack of high-quality scientific evidence, regulatory agencies have been reluctant to endorse homeopathic treatment as an alternative or adjunct treatment. This study aimed to enhance our insight into the impact of commercially-available homeopathic drugs, to study the presence of NPs in those drugs and any deleterious effects they might have, and to determine the distribution pattern of NPs in zebrafish embryos (Danio rerio). Methods: Homeopathic dilutions were studied using high-resolution transmission electron microscopy with selected area electron diffraction (SAED). For the toxicity assessment on Zebrafish, embryos were exposed to a test solution from 4 - 6 hours post-fertilization, and embryos/larvae were assessed up to 5 days post-fertilization (dpf ) for viability and morphology. Toxicity was recorded in terms of mortality, hatching delay, phenotypic defects and metal accumulation. Around 5 dpf was found to be the optimum developmental stage for evaluation. Results: The present study aimed to conclusively prove the presence of NPs in all high dilutions of homeopathic drugs. Embryonic zebrafish were exposed to three homeopathic drugs with two potencies (30CH, 200CH) during early embryogenesis. The resulting morphological and cellular responses were observed. Exposure to these potencies produced no visibly significant malformations, pericardial edema, and mortality and no necrotic and apoptotic cellular death. Conclusion: Our findings clearly demonstrate that no toxic effects were observed for these three homeopathic drugs at the potencies and exposure times used in this study. The embryonic zebrafish model is recommended as a well-established method for rapidly assessing the toxicity of homeopathic drugs.

• 한국잡초학회지
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• 제17권3호
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• pp.281-287
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• 1997

다양한 재배조건에서 벼와 피의 파종 및 이앙후 7 일째에 oxyfluorfen 유제 0.08kg ai/ha을 경엽처리한 후 그들의 생장 및 해부학적 반응을 조사하고자 온실내 pot시험이 수행되었다. Oxyfluorfen은 피의 지상부, 지상부 신장 및 지상부 생체중을 각 재배조건에서 100% 억제시켰으나 직파벼는 0~64%, 이앙벼는 0~12% 각각 억제시켰다. 초장과 지상부생체중에 있어서 oxyfluorfen은 건답조건(16~33%)보다는 담수조건(53~63%) 에서, 토중파종벼(16~63%)보다는 표면파종벼(34~64%)를 더 억제시켰다. 검경에 의한 해부학적 반응에 있어서 벼의 엽육세포의 부분적 파괴 및 엽신의 두께 감소가 관찰되었고 표피 및 유관속초세포는 큰 변화를 보이지 않았다. 한편 피는 전재배조건에서 표피세포, 엽육세포 및 유관속초세포의 파괴 및 위축 그리고 엽신두께의 유의적인 감소가 있었다. 이러한 해부학적 반응은 담수조건에서 보다 건답조건에서 더 크게 나타났다. oxyfluorfen은 피에 대한 방제는 각 조건에서 완벽하게 방제하였으나 벼에 대한 약해는 피에 비해 상대적으로 작았고 건답조건과 토중파종경우에서 상대적인 안전성을 보였으나 근본적인 약해를 피하기는 어려울 것으로 판단되었다.

• 대한한의학방제학회지
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• 제25권4호
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• pp.483-497
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• 2017

Background : Behavioral stress has been suggested as one of the significant factors that is able to disrupt physiological systems and cause depression as well as changes in various body systems. The stressful events can alter cognition, learning, memory and emotional responses, resulting in mental disorders such as depression and anxiety. Results : We used a restraint stress model to evaluate the alteration of behavior and stress-related blood parameter. The animals were randomly divided into two groups of five animals each group. Furthermore, we assessed the change of body weight to evaluate the locomotor activity as well as status of emotional and anxiety in mice. After 7 days of restraint stress, the body weight had significantly decreased in the restraint stress group compared with the control group. We also observed stress-associated behavioral alterations, as there was a significant decrease in open field and forced swim test, whereas the immobilization time was significantly increased in the stress group compared to the control group. We observed the morphological changes of neuronal death and microglia by immunohistochemistry and western blot. In our study restraint stress did not cause change in neuronal cell density in the frontal cortex and CA1 hippocampus region, but there was a trend for an increased COX-2 and iNOS protein expression and microglia (CD11b) in brain, which is restraint stress. Conclusion : Our study, there were significant alterations observed in the behavioral studies. We found that mice undergoing restraint stress changed behavior, confirming the increased expression of inflammatory factors in the brain.

• 대한의용생체공학회:의공학회지
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• 제28권4호
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• pp.512-519
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• 2007

The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

• 한국잡초학회지
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• 제16권2호
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• pp.160-170
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• 1996

비광요구형 디페닐에테르계 화합물인 TOPE의 제초작용 생리를 알기 위하여 온실에서의 제초특성을 비롯한 식물색소대사, 전자전달계, 단백질 및 핵산대사 등에 미치는 TOPE.의 영향을 조사하였다. 작용발현 속도가 광요구형 디페닐에테르계보다는 늦지만 엽신부의 고사와 동시에 생육저해 및 생장점에서의 분열이상이 관찰되었으며 절취된 오이자엽에 처리될 경우 전해질 누출과 더불어 엽주변부의 심한 분열이상이 초래되었다. 그러나 2,4-D와 같은 오옥신 활성은 아니었다. 저광도에서의 엽록소 형성 저해정도는 관찰되지 않았으며 oxyfluorfen 과는 달리 protoporphyrin IX의 축적도 유기되지 않았으나, 카로티노이드 성분은 모두 비정상적으로 증가되었다. 탈수소효소의 활성 또는 미토콘드리아에서의 호흡활성은 일정농도에 서 증가되는 경향을 보이나 호흡저해제들은 TOPE에 의한 전해질 누출을 경감시키지 못하였다. 한편 광합성 전자전달 저해는 고농도에서만 관찰되었다. 오이의 전해질 누출과 분열이상을 지표로 한 TOPE와의 상호작용 실험에서 전해질 누출에 대하여 cycloheximide는 상승적으로, chloramphenicol, actinomycin-D, hydroxyurea는 길항적으로 작용하였고, cycloheximide, actinomycin-D, hydroxyurea 등은 자엽의 이상분열을 경감시켰다. 이는 TOPE가 핵산대사에 관여됨을 시사해 주는 것으로써 오이자엽 및 벼 근에 TOPE를 처리했을 때 RNA 및 단백질보다 DNA의 증가가 보다 빠른 시기에 일어났다. 따라서 TOPE가 처리되면 비정상적인 핵산대사 촉진으로 인해 제반 생리작용이 교란되고 이어 엽신부의 고사와 분열이상 등의 제초활성이 발현되는 것으로 생각된다.

• 한국환경과학회지
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• 제5권6호
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• pp.813-826
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• 1996

발아후 7일된 팥유식물에서 alumlnlum(Al)이 생장, 엽록소 함량, ALAD활성 및 뿌리와 경엽부의 형태에 미치는 영향을 조사하였다. 저농도(50, 100 $\muM)의$ Al처리에 의해 뿌리와 경엽부의 신장이 매우 감소되었으며 농도가 증가함에 따라 생장이 더욱 억제되었다. 따라서 생장억제는 농도의존적이었다. 뿌리신장은 Al 처리 24시간에서 감소되었으며 7일간의 저농도처리에 의해 억제효과가 회복되는 경향을 보여주었다. Al의 독성증상과 생장반응은 경엽부에 비해 뿌리에서 더 크게 나타났다. Hematoxylin 염색법에 의해 Al 분포를 조사한 결과 Al은 근단을 통해 표피와 피층세포에 축적되어 있음을 알 수 있었다. 한편 Al처리는 엽록소함량을 감소시켰으며, ALAD활성 또한 억제시켰다. 엽록소 함량과 ALAD 활성 간에는 양의 상관관계가 나타났다. Al 처리에 의한 뿌리의 형태변화를 보면 표피세포 및 피층세포의 변형 또는 파괴가 관찰되었으며, 뿌리직경과 피층의 체적도 매우 감소되었다. 경엽부에서도 Al처리는 직경과 세포크기의 감소를 보여주었다. 그러나 잎에서의 형태적 변화는 엽록체수와 크기변화 이외에는 거의 관찰되지 않았다. 이와 같은 결과에서 Al의 독성효과는 1차적으로 뿌리에서 나타나며, 뿌리형태의 변화는 뿌리의 생장패턴과 관련이 있음을 알 수 있었다. 따라서 Al은 팥유식물에서 특히 뿌리의 형태와 기능적 손상을 일으키는데 큰 영향을 미치는 것으로 생각된다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권17호
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• pp.7229-7234
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• 2014

Trichoderma spp. are known as a rich source of secondary metabolites with biological activity belonging to a variety of classes of chemical compounds. These fungi also are well known for their ability to produce a wide range of antibiotic substances and to parasitize other fungi. In search for new substances, which might act as anticancer agents, the overall objective of this study was to investigate the cytotoxic effects of Trichoderma harzianum and Trichoderma asperellum cultural filtrates against human cervical and breast cancer cell lines (HeLa and MCF-7 cells respectively). To achieve this objective, cells were exposed to 20, 40, 60, 80 and 100 mg/ml of both T. harzianum cultural filtrate (ThCF) and T. asperellum cultural filtrate (TaCF) for 24h, then the cell viability and the cytotoxic responses were assessed by using trypan blue and 3-(4,5-dimethylthiazol-2yl)-2,5-biphenyl tetrazolium bromide (MTT) assays. Morphological changes in cells were investigated by phase contrast inverted microscopy. The results showed that ThCF and TaCF significantly reduce the cell viability, have cytotoxic effects and alter the cellular morphology of HeLa and MCF-7 cells in a concentration dependent manner. A concentration of 80 and 100mg/ml of ThCF resulted in a sharp decline in the cell viability percent of HeLa and MCF-7 respectively (25.2%, 26.5%) which was recorded by trypan blue assay. The half-maximal inhibitory concentrations ($IC_{50}$) of ThCF and TaCF in HeLa and MCF-7 were recorded as 16.6, 12.0, 19.6 and 0.70mg/ml respectively by MTT assay. These results revealed that ThCF and TaCF have a substantial ability to reduce the viability and proliferation of human cervical and breast cancer cells.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2007년도 International Meeting of the Microbiological Society of Korea
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• pp.120-122
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• 2007

All living organisms use numerous signal-transduction pathways to sense and respond to their environments and thereby survive and proliferate in a range of biological niches. Molecular dissection of these signalling networks has increased our understanding of these communication processes and provides a platform for therapeutic intervention when these pathways malfunction in disease states, including infection. Owing to the expanding availability of sequenced genomes, a wealth of genetic and molecular tools and the conservation of signalling networks, members of the fungal kingdom serve as excellent model systems for more complex, multicellular organisms. Here, we employed Cryptococcus neoformans as a model system to understand how fungal-signalling circuits operate at the molecular level to sense and respond to a plethora of environmental stresses, including osmoticshock, UV, high temperature, oxidative stress and toxic drugs/metabolites. The stress-activated p38/Hog1 MAPK pathway is structurally conserved in many organisms as diverse as yeast and mammals, but its regulation is uniquely specialized in a majority of clinical Cryptococcus neoformans serotype A and D strains to control differentiation and virulence factor regulation. C. neoformans Hog1 MAPK is controlled by Pbs2 MAPK kinase (MAPKK). The Pbs2-Hog1 MAPK cascade is controlled by the fungal "two-component" system that is composed of a response regulator, Ssk1, and multiple sensor kinases, including two-component.like (Tco) 1 and Tco2. Tco1 and Tco2 play shared and distinct roles in stress responses and drug sensitivity through the Hog1 MAPK system. Furthermore, each sensor kinase mediates unique cellular functions for virulence and morphological differentiation. We also identified and characterized the Ssk2 MAPKKK upstream of the MAPKK Pbs2 and the MAPK Hog1 in C. neoformans. The SSK2 gene was identified as a potential component responsible for differential Hog1 regulation between the serotype D sibling f1 strains B3501 and B3502 through comparative analysis of their meiotic map with the meiotic segregation of Hog1-dependent sensitivity to the fungicide fludioxonil. Ssk2 is the only polymorphic component in the Hog1 MAPK module, including two coding sequence changes between the SSK2 alleles in B3501 and B3502 strains. To further support this finding, the SSK2 allele exchange completely swapped Hog1-related phenotypes between B3501 and B3502 strains. In the serotype A strain H99, disruption of the SSK2 gene dramatically enhanced capsule biosynthesis and mating efficiency, similar to pbs2 and hog1 mutations. Furthermore, ssk2, pbs2, and hog1 mutants are all hypersensitive to a variety of stresses and completely resistant to fludioxonil. Taken together, these findings indicate that Ssk2 is the critical interface protein connecting the two-component system and the Pbs2-Hog1 pathway in C. neoformans.

• 음성과학
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• 제10권3호
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• pp.7-27
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• 2003

Esophageal Speech uses the esophageal air during phonation. Fluent esophageal speakers frequently intake air in oral communication, but unskilled esophageal speakers are difficult with swallowing lots of air. The purpose of this study was to investigate the difference of acoustic characteristics of vowel and consonants production according to the speech proficiency level in esophageal speech. 13 normal male speakers and 13 male esophageal speakers (5 unskilled esophageal speakers, 8 skilled esophageal speakers) with age ranging from 50 to 70 years old. The stimuli were sustained /a/ vowel and 36 meaningless two syllable words. Used vowel is /a/ and consonants were 18 : /k, n, t, m, p, s, c, $C^{h},\;k^{h},\;t^{h},\;p^{h}$, h, I, k', t', p', s', c'/. Fundermental frequency (Fx), Jitter, shimmer, HNR, MPT were measured with by electroglottography using Lx speech studio (Laryngograph Ltd, London, UK). 36 meaningless words produced by esophageal speakers were presented to 3 speech-language pathologists who phonetically transcribed their responses. Fx, Jitter, HNR parameters is significant different between skilled esophageal speakers and unskilled esophageal speakers (P<.05). Considering manner of articulation, ANOVA showed that differences in two esophageal speech groups on speech proficiency were significant; Glide had the highest number of confusion with the other phoneme class, affricates are the most intelligible in the unskilled esophageal speech group, whereas in the skilled esophageal speech group fricatives resulted highest number of confusions, nasals are the most intelligible. In the place of articulation, glottal /h/ is the highest confusion consonant in both groups. Bilabials are the most intelligible in the skilled esophageal speech, velars are the most intelligible in the unskilled esophageal speech. In the structure of syllable, 'CV+V' is more confusion in the skilled esophageal group, unskilled esophageal speech group has similar confusion in both structures. In unskilled esophageal speech, significantly different Fx, Jitter, HNR acoustic parameters of vowel and the highest confusions of Liquid, Nasals consonants could be attributed to unstable, improper contact of neoglottis as vibratory source and insufficiency in the phonatory air supply, and higher motoric demand of remaining articulation due to morphological characteristics of vocal tract after laryngectomy.

• Applied Microscopy
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• 제27권1호
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• pp.13-30
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• 1997

The experiment was performed to study the morphological responses of the thymus of the mice, to antitumour agents (5-Fluorouracil or mitomycin C). Healthy adult mice weighing 25 gm each were divided into normal and experimental groups. 5-Fluorouracil (60 mg/kg) or Mitomycin-C $(400{\mu}g/kg)$ were injected subcutaneously to the animals every other day. Animals were sacrificed at 4 days and 7 days following the first injection. Pieces of the tissue taken from the thymus were prefixed with 2.5% paraformaldehyde-l.5% glutaraldehyde, followed by post-fixation with 1% osmium tetroxide. Ultrathin sections stained with uranyl acetate and lead citrate were observed with a JEM 100 CX-II electron microscope. The observed results were as follow: 1. Apoptoses of T-lymphocytes were observed more frequently in the thymus of the experimental groups than in those of a normal group. 2. In the experimental group, the plasma cells with distended cisternae of the granular endoplasmic reticulum and the eosinophile leukocytes were observed frequently. 3. In the experimental group, newly forming Hassall's corpurscles were observed frequently. 4. In the mitomycin-treated group, the epithelial reticular cells containing distended perinuclear cisternae, distended the granular endoplasmic reticula and pyknotic nuclei were observed in the cortico-medullary junctional area. 5. In the mitomycin-treated group, nuclear bodies with medium electron dense materials were often observed in the T lymphocyte. 6. In the 5-fluorouracil-treated groups, fused and dissolved tonofilament bundles and apoptotic bodies were observed in the some epithelial reticular cells in the medullary area. 7. In the 5-fluorouracil-treated groups, some elongated and bar-shaped lysosomes with electron lucent gap were often observed in the macrophages. 8. In the 5-fluorouracil-treated group, membrane complex of the smooth endoplasmic reticulum were ofen observed in the macrophage. From the above results, it was suggested that 5-fluorouracil or mitomycin could induce rapid involution of the thymus, and disturb maturation and differentiation of T lymphocytes, and, in turn, supress immunity.