• Journal of Microbiology and Biotechnology
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• v.26 no.1
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• pp.160-170
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• 2016

The increasing spread of antibiotic-resistant pathogens has raised the interest in alternative antimicrobial treatments. In our study, the functionally active gram-negative bacterium bacteriophage CP933 endolysin was produced in Nicotiana benthamiana plants by a combination of transient expression and vacuole targeting strategies, and its antimicrobial activity was investigated. Expression of the cp933 gene in E. coli led to growth inhibition and lysis of the host cells or production of trace amounts of CP933. Cytoplasmic expression of the cp933 gene in plants using Potato virus X-based transient expression vectors (pP2C2S and pGR107) resulted in death of the apical portion of experimental plants. To protect plants against the toxic effects of the CP933 protein, the cp933 coding region was fused at its Nterminus to an N-terminal signal peptide from the potato proteinase inhibitor I to direct CP933 to the delta-type vacuoles. Plants producing the CP933 fusion protein did not exhibit the severe toxic effects seen with the unfused protein and the level of expression was 0.16 mg/g of plant tissue. Antimicrobial assays revealed that, in contrast to gram-negative bacterium E. coli (BL21(DE3)), the gram-positive plant pathogenic bacterium Clavibacter michiganensis was more susceptible to the plant-produced CP933, showing 18% growth inhibition. The results of our experiments demonstrate that the combination of transient expression and protein targeting to the delta vacuoles is a promising approach to produce functionally active proteins that exhibit toxicity when expressed in plant cells.

• Journal of Plant Biology
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• v.36 no.4
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• pp.357-362
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• 1993

To elucidate the origin of secondary growth in the rhizome of B. ternaturn, the developmental changes of vascular cambium was observed in ultrastructural features. The vascular cambium was gradually differentiated from procambium as in seed plants, but the cambial activity did not persist very long so that the cambial cells became a dormant state like fossil cryptogams. Dense cytoplasm of procambial cells became progressively sparse during the growth, and the tiny vesicles were fused to form numerous small vacuoles and then a few large vacuoles. These gradual changes and the occurrence of storage materials which was associated with the developmental stages might support the progressive differentiation of the cambial cells. In addition, the cessation of cambial activity could be indicated by the facts that late vascular cambial cells accumulate large lipid bodies and show very small peripheral cytoplasm and unlikely thickened cell wall, compared to other meristematic cells. Therefore. the vascular cambium showed the characteristics of both seed plants and fossil cryptogams from the view point of cambial ontogeny and activity.tivity.

• Applied Microscopy
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• v.24 no.1
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• pp.86-101
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• 1994

The development of synovial membrane from knee joint was studied by electron microscope in human fetuses ranging from 20mm to 260mm crown rump length (40days to 30weeks of gestational age). At 40mm fetus, developing synovial tissue was observed in homogenous interzone as a vascular mesenchyme around the periphery. The primitive joint space was appeared after the intermediate layer of the interzone in direct contact with chondrogenic layer at 60mm fetus. Differentiation of the synovial membrane coincided with clarification of the joint cauity. When dilatation of the synovial cavity occurred, the two types of synovial cells were well endowed with rough endoplasmic reticulum. At 100mm fetus, type A cells with a markedly attenuated cytoplasm were found as well as those cells which contained pinocytotic vesicles and vacuoles. By 150-200mm fetuses a majority of the intimal cells were type B. These cells were characterized by abundant rough endoplasmic reticulum and well developed Golgi complex. In contrast, A-type cell had numerous filopodia, pinocytotic vesicles lysosomes, and large vacuoles containing amorphous material. At 260mm fetus, the intimal cells were well developed and plentiful. The most marked difference between the synovial membrane of full-term fetus and adult was the large amount of collagen in the latter. During fetal period, the B-cells were most numerous cell type in the intimal cells. The B-cells were clearly distinguishable from the A-cells by their content of extensive rough endoplasmic reticulum and well developed Golgi complex.

• Applied Microscopy
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• v.36 no.2
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• pp.109-118
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• 2006

The physicochemical properties of damaged hair by irradiation of ultraviolet-B (UV-B) have been investigated by using transmission electron microscope and scanning electron microscope. The range of irradiation of hair irradiated for expectative 6 hours, 12 hours, 24 hours and 48 hours with stimulated ultraviolet ray. The treated hairs showed characteristic morphological damage pattern in the cornified cell of matrix and the cuticle following time past. The various sized vacuoles in the endocuticle of the cuticular cells was formed. The statistically significant differences in diameter of cuticular cell were observed in terms of tranverse swelling by formation of vacuoles. The hair cortex and matrix undergo long term exposure to UV-B radiation. The macrofibrils of cortex appeared to be affected most by UV-B, although the morphology and volume of melanin granule was not changed. The physicochemical destruction of hair matrix and cuticular cells is largely accelerated by long term irradiation of UV-B.

• The Plant Pathology Journal
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• v.26 no.2
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• pp.139-148
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• 2010

A virus causing stunt, yellowing, severe mosaic, malformation symptoms on leaves and uneven development and malformation on fruits of zucchini was prevalent around Goseong, Gyeongsangnam-do, Korea. A survey conducted (2004) in the Goseong area revealed about 20% virus infection rate. The disease causative identified as Cucumber mosaic virus (CMV-Z1) was further characterized. The isolate induces mosaic symptoms on Cucumis sativus, while severe mosaic, stunt and malformation on C. pepo. Thin section analyses have shown that virus inclusions are formed in the cuticle layers as well as epidermal, parenchyma and collenchymas cells in virus-infected Nicotiana tabacum. CMV-Z1 isolate induced specific cytoplasmic inclusion bodies such as irregular clumps (IC), crystal (Cr) and irregular chloroplasts (ICh). IC was made up of virus particles interspersed with a darkly stained amorphous material and found both in the cytoplasm and vacuoles, whereas ICh and Cr were rarely found in the vacuoles. The genome of CMV-Z1 RNA-1 consists of 3359 nucleotide (nt) encoding 1a protein of 993 amino acids (aa). The CMV-Z1 RNA-2 was 3050 nt in length containing 2a (857 aa) and 2b (110 aa), while RNA-3 encoding 3a movement protein (279 aa) and coat protein (218 aa) was 2215 nt in length. Phylogenetic analyses of nucleotide sequences of CMV-Z1 isolate appeared it is more closely related to subgroup IA than to subgroup IB or II.

• Biomedical Science Letters
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• v.7 no.2
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• pp.53-58
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• 2001

We studied antitrichomoniasis with the extract of Gentiana scabra val buergeri, which may be effective in treating infectious diseases. The growth inhibition against T. vaginalis became optimal when tile extract concentration was 0.7 mg/ml and the cells were seeded at a density of 3$\times$10$^5$ per well. After incubation for 12, 24, 36, and 48 hrs, respectively, the number of cells were each 5$\times$10$^5$, 1$\times$10$^5$, 1$\times$10$^5$, and none, respectively. Under the electron microscope, the experimental group showed that the nucleus, karyosomes, and chromatin were weaker than those in the control group. After incubating for 3 hrs, the cells were destroyed completely, and only a remnant remained. The hydrogenosomes disappeared almost. The vacuoles and autophagic vacuoles increased. The cells became regressive form.

• Journal of Microbiology and Biotechnology
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• v.21 no.4
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• pp.412-420
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• 2011

Secretion of cellobiase occurred in a brefeldin A (BFA) uninhibited manner in the filamentous fungus Termitomyces clypeatus. Fluorescence confocal microscopy revealed that application of the drug at a concentration of 50 ${\mu}g$/ml caused arrest of Spitzenkorper assembly at the hyphal tip. This resulted in greater than 30% inhibition of total protein secretion in the culture medium. However, the cellobiase titer increased by 17%, and an additional 13% was localized in the vacuolar fraction en route secretion. The secretory vacuoles formed in the presence of the drug were also found to be bigger (68 nm) than those in the control cultures (40 nm). The enzyme secreted in the presence and absence of BFA revealed a single activity band in both cases in native PAGE and had similar molecular masses (approx. 120 kDa) in SDS-PAGE. The BFA enzyme retained 72% of native glycosylation. It also exhibited a higher stability and retained 98% activity at $50^{\circ}C$, 93.3% activity at pH 9, 63.64% activity in the presence of 1M guanidium hydrochloride, and 50% activity at a glucose concentration of 10 mg/ml in comparison to 68% activity, 75% activity, 36% activity, and 19% activity for the control enzyme, respectively. The observations collectively aimed at the operation of an alternative secretory pathway, distinct from the target of brefeldin A, which bypassed the Golgi apparatus, but still was able to deliver the cargo to the vacuoles for secretion. This can be utilized in selectively enhancing the yield and stability of glycosidases for a successful industrial recipe.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.18 no.1
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• pp.221-233
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• 2005

Background and Objectives : Despite of high prevalence, Tinnitus remains obscure because the cause and mechanism is poorly understood. In the absence of a suitable animal model, past investigations tool place in humans. Recently, the animal model with sodium salicylate ototoxicity is broadly used because of its reversibility. Balyangtongum-Tang had been used clinically to treat tinnitus and other hearing problems. We investigated the effects of Balyangtongum-Tang on cochlear morphologic change induced by sodium salicylate ototoxicity and were to find out its therapeutic effects on ototoxicity in rat model. Materials and Methods : Healthy ten Sprague-Dawley rats were divided into normal(2), control(4) and sample(4) groups. The sample group was given extract of Balyangtongum - Tang(1cc/100g) once a day for 4 days. After 3 hours when last medication were given, the sample and control groups were injected intraperitoneally with sodium salicylate(500mg/kg). We observed the cochlear morphologic changes of rats every 1, 2, 3 and 5 hours after injection. Results : The electron microscopic finding of outer hair cell shows some changes in the curticular plate and cytoplasm. Some vacuoles were found in the control and sample groups. Vacuolization in the curticular plate and cytoplasm of the sample group after 3, 5 hours were similar to the control group. But the curticular plate of the sample group after 1, 2 hours did not from vacuole. On the other hand the control group after 1, 2 hours formed vacuoles in the curticular plate. Light microscopic findings of cochlear duct in control and sample groups didn't find any difference. Conclusion : The results suggest that extract of Balyangtongum-Tang reduces the morphologic changes induced by sodium salicylate ototoxicity and the effects are remarkable in frist 2 hours. However the effective times are different with previous studies, it seems to due to the difference of tolerance and sensitivity of laboratory animals.

• Parasites, Hosts and Diseases
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• v.51 no.5
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• pp.537-544
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• 2013

The present study was performed to observe histopathological changes in tissues of Bithynia siamensis goniomphalos (Gastropoda, Bithyniidae) incubated in crude extract solutions of camellia (Camellia oleifera) seed and mangosteen (Garcinia mangostana) pericarp, and furthermore to estimate the molluscicidal effects of 2 plant substances. Substantial numbers of bithyniid snails were incubated in various concentrations of 2 plant solution for 24 hr. As the positive control, snails incubated in various concentrations of niclosamide, a chemical molluscicide, were used. The histopathological findings were observed in sectioned snail specimens of each experimental and control groups. The results showed that both camellia and mangosteen extracts had molluscicidal effects at 24 hr with 50% lethal concentration ($LC_{50}$) at concentrations of 0.003 and 0.002 g/ml, respectively, while niclosamide had $LC_{50}$ at concentrations 0.599 ppm. B. siamensis goniomphalos snail tissues (foot, gill, and digestive system) showed disruption of columnar muscle fibers of the foot, reduction of the length and number of gill cilia, numerous mucous vacuoles, and irregularly shaped of epithelial cells. Irregular apical and calciferous cells, dilatation of the digestive gland tubule, and large hemolymphatic spaces, and irregular apical surfaces, detachment of cilia, and enlargement of lysosomal vacuoles of epidermis were also shown in all groups. By the present study, it is confirmed that 2 plants, camellia and mangosteen, are keeping some substance having molluscicidal effects, and histopathological findings obtained in this study will provide some clues in further studies on their action mechanisms to use them as natural molluscicides.

• Clinical and Experimental Reproductive Medicine
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• v.42 no.4
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• pp.156-162
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• 2015

Objective: To investigate fertilization and embryo quality of dysmorphic mature oocytes with specific morphological abnormalities obtained from intracytoplasmic sperm injection (ICSI). Methods: The fertilization rate (FR) and embryo quality were compared among 58 dysmorphic and 42 normal form oocytes (control 1) obtained from 35 consecutive ICSI cycles, each of which yielded at least one dysmorphic mature oocyte, performed over a period of 5 years. The FR and embryo quality of 441 normal form oocytes from another 119 ICSI cycles that did not involve dysmorphic oocytes served as control 2. Dysmorphic oocytes were classified as having a dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body (PB). Results: The overall FR was significantly lower in dysmorphic oocytes than in normal form oocytes in both the control 1 and control 2 groups. However, embryo quality in the dysmorphic oocyte group and the normal form oocyte groups at day 3 was similar. The FR and embryo quality were similar in the oocyte groups with a single abnormality and multiple abnormalities. Specific abnormalities related with a higher percentage of top-quality embryos were dark cytoplasm (66.7%), abnormal PB (50%), and cytoplasmic vacuoles (25%). Conclusion: The fertilization potential of dysmorphic oocytes in our study was lower, but their subsequent embryonic development and embryo quality was relatively good. We were able to define several specific abnormalities related with good or poor embryo quality.