• 대한미생물학회지
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• 제11권1호
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• pp.1-11
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• 1976

Escherichia coli(ATCC 11115)에 실험실등에서 상용하는 여러가지 항균제를 시간별로 처리, 그 변화양상을 전자현미경으로 관찰한 바 그 결과를 요약하면 다음과 같다. 1) 대조군은 3층의 단일막으로 형성된 세포벽에 둘러쌓여 있으며 세포질은 전자밀도가 낮은 nucleoid와 ribosme들이 산재하여 있음을 관찰할 수 있었다. 2) 70% ethanol용액 처리군은 핵물질을 관찰할 수 없었고 세포질은 세포 중앙부로 응집되어 있었으며 세포벽의 외부에서는 bleb 들을 관찰 할 수 있었다. 3) 3% $H_2O_2$ 용액 처리군은 세포내용물의 변화는 70% ethanol 처리군과 대동소이(大同小異)하였으나 세포벽에서는 심한 굴곡현상이 관찰되었다. 4) 5% lysol 용액처리군은 세포질 및 핵물질 부위가 완전히 구분되어 나타났으며 세포질내의 ribosome과립들은 시간이 경과할수록 그 응집현상이 심하였고 세포 외부에는 ribosome 양 과립들이 부착하고 있음이 관찰되었다. 5) 1% DDEGH 용액 처리군은 세포질의 응집 및 세포막과 세포벽이 뚜렷이 관찰되지 않았으며 세포외부에 세포내용물과 동일한 물질로 생각되는 물질이 부착되어 있음을 관찰할 수 있었다. 6) 고압멸균 처리군은 세포막 및 세포벽의 파괴, 탈락 및 세포내용물의 유출현상이 관찰되었다.

• The Plant Pathology Journal
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• 제17권6호
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• pp.315-324
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• 2001

Tobacco (Nicotiana tabacum cvs.Xanthi-nc and NC 82) plants infected with Tobacco mosaic virus (TMV) were examined ultrastructurally. Local lesions produced by TMV were sunken and withered. The plants were subjected to temperature shift (TS), a method to produce programmed cell death (PCD), by placing the infected plants initially at high temperature (35$^{\circ}C$) for 2 days and then shifting them to greenhouse temperature (22-27$^{\circ}C$). As a result, expanded lesions around the original necrotic lesions were produced. The expanded area initially had no symptoms, but it withered and became necrotic 15 h after TS. No ultrastructural changes related to PCD were noted at 0 h after TS in Xanthi-nc tobacco tissues as well as in healthy and susceptible tobacco tissues infected with TMV, At 6 h after TS, chloroplasts were convoluted and cytoplasm began to be depleted; however no necrotic cells were found. At 17 h after TS, ground cytoplasm of affected cells was completely depleted and chloroplasts were stacked together with bent cell wall or dispersed in the intracellular space. Necrotic cells were also observed, containing virus particles in the necrotic cytoplasm. There were initially two types of symptoms in the expanded lesions: chlorosis and non-chlorosis (green). Abundant TMV particles and X-bodies were only found in the chlorotic tissue areas. These results suggest that PCD by TMV infection may start with the wilting of cells and tissues before necrotic lesion formation.

• The Plant Pathology Journal
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• 제15권1호
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• pp.1-7
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• 1999

The production of soybean cyst nematode (SCN), Heterodera glycines, including female formation and fecundity was much higher in SCN race 14 (R14) than in race 3 (R3) in susceptible soybean cultivars Bragg (intolerant), Lee74 (moderately tolerant), and PI 97100 (highly tolerant). The nematode body was also significantly larger in R14 than in R3 at 20 days after inoculation, but the further nematode growth appeared to be slower in R14 than in R3, resulting in no significant difference between the two races at 30 days after inoculation. Within each race, no significant difference was observed in the growth and reproduction among the soybean cultivars tested. Syncytial areas near the nematode lip regions (infection sites) were measured for each soybean cultivar-SCN race combination. R14 induced significantly larger syncytia than R3. Bragg had relatively larger syncytia than Lee74 and PI 97100, but the difference among the soybean cultivars was minimal or not significantly different. Syncytium occupation in the stelar region differed only between PI 97100 and the other two cultivars, which may be somewhat, but not exactly, related to tolerance levels. Syncytial cytomplasm was degenerated more with R14 and in Bragg than with R3 and in Lee74 and PI 97100, respectively. In light microscopy, degenerated syncytia were characterized by depleted and loose cytoplasm with less plastids than normal-looking (intact) syncytia which had dense syncytial cytoplasm. Electron microscopy revealed that degenerated syncytia contained highly vacuolated cytoplasm with degenerated plastids. The above results suggest that structural characteristics of syncytia may match the nematode growth and reproduction.

• 대한수의학회지
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• 제29권3호
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• pp.215-222
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• 1989

In order to investigate the morphological characteristics of epididymal duct of the squirrel, the histological and histochemical studies were carried out. The results obtained were summarized as follows: The epididymal duct can be divided into 9 segments by histological and histochemical features. Segments 1 to 5 were located in the head, segments 6 and 7 in the body, and segments 8 and 9 in the tail of the epididymis. The apical cells were numerous in the segment 1. Clear cells which has a compact, deeply staining nucleus and a characteristically clear cytoplasm were scattered in the epithelium throughout the duct. Interepithelial clear cells which had PAS-positive granules tended to increase in number caudally. Strong PAS-positive reaction was detected at the intralumen of the segments 3,8 and 9. Acid phosphatase activity was relatively high in the basal cytoplasm of the segment 7, and then in the supranuclear region of the segments 8 and 9. Alkaline phosphatase activity was weakly positive or negative except the segments 3 and 4. ATPase activity was strong in the free surface of the epithelium in the head and the entire cytoplasm in the body and tail, a,nd SDH activity was generally weak except for the body where it was more intense.

• Parasites, Hosts and Diseases
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• 제44권4호
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• pp.331-341
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• 2006

Actin binding proteins play key roles in cell structure and movement particularly as regulators of the assembly, stability and localization of actin filaments in the cytoplasm. In the present study, a cDNA clone encoding an actin bundling protein named as AhABP was isolated from Acanthamoeba healyi, a causative agent of granulomatous amebic encephalitis. This clone exhibited high similarity with genes of Physarum polycephalum and Dictyostelium discoideum, which encode actin bundling proteins. Domain search analysis revealed the presence of essential conserved regions, i.e., an active actin binding site and 2 putative calcium binding EF-hands. Transfected amoeba cells demonstrated that AhABP is primarily localized in phagocytic cups, peripheral edges, pseudopods, and in cortical cytoplasm where actins are most abundant. Moreover, AhABP after the deletion of essential regions formed ellipsoidal inclusions within transfected cells. High-speed co-sedimentation assays revealed that AhABP directly interacted with actin in the presence of up to $10{\mu}M$ of calcium. Under the electron microscope, thick parallel bundles were formed by full length AhABP, in contrast to the thin actin bundles formed by constructs with deletion sites. In the light of these results, we conclude that AhABP is a novel actin bundling protein that is importantly associated with actin filaments in the cytoplasm.

• Parasites, Hosts and Diseases
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• 제22권1호
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• pp.30-36
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• 1984

An ultrastructural study on the mature spermatoBoa oi Cloptorchis sineitsis was carried out. For this study, the liver cukes were collected from the livers of rabbits and rats artificially infected with the metacercariae obtained from the fresh water fish, Pseudorasbora parve. Six-month old worms were used. The collected liver fiukes were washed with 0.85% saline solution and then immediately moved to cold 2% glutaraldehyde buffered with 0.1M Millonig's phosphate buffier (pH 7.4) . The materials were dissected into appropriate pieces in the fixative about 30 minutes after beginning of the fixation. Two hours later the materials containing the seminal receptacle were rinsed several times with the buffier and were secondarily fixed with cold, bugeyed 1% osmium tetroxide(OsO4) for 2 hours. The fully mixed tissue blocks Ivere dehydrated in a series of graded concentrations of acetone and were embedded in Epon 812 mixture. Thin sections obtained from LKB-5 ultramicrotome were stained with uranyl acetate and Reynold's lead citrate. Observations of the sections were carried out with JEM-100CX II electron microscope, In general, the mature sperm was long thread-like form with a sickle-shaped head. According to the longitudinal sectioned view of the sperm tail, the nucleus seemed to be spirally coiled and run a little far along the tail. The acrosome was not observed. The cytoplasm of the tail was biflagellated as usual in trematodes. Unlike other platyhelminth spermatozoa, the sperm tail of Clenorchis sinensis showed the ${\ulcorner}9+2{\lrcorner}$ in the microtubular arrangement. The mitochondria with poorly developed cristae were observed throughout the middle piece. The middle piece of the tail showed dull ladder or triangular shapes with the two flagella at the bottom. But, the principal piece of the tail was slightly flattened cylindrical shape with two aagella within the cytoplasm. The end piece was uniflagellated. It was not clearly identised whether the end piece was subdivided into two by aagellum or the lengths of the two aagella were different. The glycogen granules were rich in the cytoplasm throughout the lenght of the spermatozoa. These granules might be the energy source for the movement of the spermatosoa.

• Applied Microscopy
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• 제31권4호
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• pp.325-331
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• 2001

돌가자미의 피부 상피층을 다층상피층으로 지지세포, 선세포 그리고 과립세포들로 구성된다. 상피층은 지지세포의 형태와 구조에 따라 표면층, 중간층 및 기저층으로 구분 할 수 있었다. 지지세포들의 세포질은 피질부와 수질부로 나누어지는데 피질부에는 미세섬유의 발달이 뚜렷하다. 점액세포들은 단세포선으로 상피의 표면층과 중간층에서 관찰된다. 점액세포의 점액물질은 중성이며, carboxylated mucosubstance의 당단백 질로 확인되었다. 곤봉상세포는 세포질에 잘 발달된 활면소포체와 골지체를 가진다. 과립세포는 주로 중간층과 기저층에 존재하고, 세포질은 막을 가진 전자밀도가 높은 과립들이 차지한다. 색소세포는 세포질에 존재하는 함유물의 전자밀도에 따라 세 종류로 구분할 수 있었으며, 색소세포 근처에서 신경종말을 관찰할 수 있었다.

• Asian Pacific Journal of Cancer Prevention
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• 제16권7호
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• pp.2777-2783
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• 2015

Background: Nestin is associated with neoplastic transformation. However, the mechanisms by which nestin contributes regarding invasion and malignancy of gastric adenocarcinoma (GAC) remain unknown. Recent studies have shown that the epithelial-mesenchymal transition (EMT) is important in invasion and migration of cancer cells. In the present study, we aimed to investigate the expression of nestin and its correlation with EMT-related proteins in GAC. Materials and Methods: The expression of nestin and EMT-related proteins was examined in GAC specimens and cell lines by immunohistochemistry and Western blotting. Clinicopathological features and survival outcomes were retrospectively analyzed. Results: Positive nestin immunostaining was most obviously detected in the cytoplasm, nucleus or both cytoplasm and nucleus of tumor cells in 19.2% (24/125) of GAC tissues, which was significantly higher than that in normal gastric mucosa tissues (1.7%, 1/60) (p=0.001). Nestin expression was closely related to several clinicopathological factors and EMT-related proteins (E-cadherin, vimentin and Snail) and displayed a poor prognosis. Interestingly, simultaneous cytoplasmic and nuclear nestin expression correlated with EMT-related proteins (E-cadherin, vimentin and Snail) (p<0.05) and lymph node metastasis (p=0.041) and a shorter survival time (p<0.05), but this was not the case with cytoplasmic or nuclear nestin expression. Conclusions: Nestin, particularly expression in both cytoplasm and nucleus, might be involved in regulating EMT and malignant progression in GAC, with potential as an unfavorable indicator in tumor diagnosis and a target for clinical therapy.

• 한국패류학회지
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• 제14권2호
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• pp.149-159
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• 1998

In order to observe the anticellulolytic localization in the epithelia of the digestive tract such as esophagus, crop, and intestine of a Korean land snail N. samarangae, a cytochemical method and a immunogold labelling method were applied. For the cytochemical study on the cellulase activity, Benedict reaction method applied. And for the immunocytochemical study, the rabbit serum immunoglobuins (IgG) was obtained from the rabbits injected with cellulase which was extracted from body fluid of the snail. The digestive tract tissues of N. samarangae were fixed with 4% paraformaldehyde and 2% OsO4 and embedded in Lowicryl K4M at -40$^{\circ}C$ under UV light (360 nm). The thin sections were loaded on the nickel grids and stained with the serum IgG and protein A-gold complex (particle size: 10 nm). Observations were undertaken with transmission electron microscope (Jeol, JEM-1010). The epithelium of the digestive tract was consisted of five types of cells. In the cytochemical study, the reaction products were found along the periphery of the vacuoles derived from the Bebedict reaction. In the immunocytochemical study, the protein-A gold particles were selectively labelled in Type 1, Type 3 and Type 4 cells in intestinal tissue. membranes of rER, in the surrounding cytoplasm of the rER and secretory granules, and in the apical cytoplasm of the cells. On the material being secreted from the apical cytoplasm was also labelled with the immunogold particles. The all results obtained throughtout present study suggest that the intestinal epithelium of the snail N. samarangae seretes cellulase as one of digestive enzymes.

• IMMUNE NETWORK
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• 제10권2호
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• pp.35-45
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• 2010

Background: Expression of recombinant antibodies and their derivatives fused with other functional molecules such as alkaline phosphatase in Escherichia coli is important in the development of molecular diagnostic reagents for biomedical research. Methods: We investigated the possibility of applying a well-known Fos-Jun zipper to dimerize $V_H$ and $V_L$ fragments originated from the Fab clone (SP 112) that recognizes pyruvate dehydrogenase complex-E2 (PDC-E2), and demonstrated that the functional zipFv-112 and its alkaline phosphatase fusion molecules (zipFv-AP) can be produced in the cytoplasm of Origami(DE3) trxB gor mutant E. coli strain. Results: The zipFv-AP fusion molecules exhibited higher antigen-binding signals than the zipFv up to a 10-fold under the same experimental conditions. However, conformation of the zipFv-AP seemed to be influenced by the location of an AP domain at the C-terminus of $V_H$ or $V_L$ domain [zipFv-112(H-AP) or zipFv-112(L-AP)], and inclusion of an AraC DNA binding domain at the C-terminus of VH of the zipFv-112(L-AP), termed zipFv-112(H-AD/L-AP), was also beneficial. Cytoplasmic co-expression of disulfide-binding isomerase C (DsbC) helped proper folding of the zipFv-112(H-AD/L-AP) but not significantly. Conclusion: We believe that our zipFv constructs may serve as an excellent antibody format bi-functional antibody fragments that can be produced stably in the cytoplasm of E. coli.