• Applied Microscopy
• /
• v.37 no.1
• /
• pp.43-52
• /
• 2007

In order to observe the localization of excretory, purified and infected antigenic protein in the tissue of Trichinella spiralis larvae, immunogoldlabeling methodology using IgG and protein A-gold complex was implemented. T. spiralis larvae obtained from rat muscle were initially cultured in medium, and secreted excretory antigen was collected for 1 or 3 days. Purified antigenic protein was obtained from homogenized T. spiralis larvae. Rabbits were then immunized with 1 or 3 days secreted excretory protein and purified 45 kDa protein, and IgG was purified from collected serum. Serum, against infected antigen, collected from rat on 1 and 4 weeks after infection with T. spiralis larvae, and IgG was purified from collected serum. T. spiralis larvae were embedded in Lowicryl HM20 medium. Then they were finally treated with immunized IgG and protein A-gold complex (particle size; 15 nm) and observed under electron microscope. In T. spiralis larvae tissue, the tissue antigen reacted with rabbit IgC antigen Day 1 secreted excretory protein, infected antigenic protein and purified 45 kDa protein. But different distribution pattern of labeled gold particles were observed. When Day 1 secreted excretoy protein was used, gold particle labeling was observed specifically on the cuticle, basal layer, esophagus interstitial matrix (EIM) and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. In a separate group of tissue, the antigen reacted with rabbit IgG against Day 3 secreted excretory protein. Labeled gold particles were specifically distributed on the surface layer of cuticle, EIM and ${\alpha}_0$ granules of stichocyte of the worm. In case of using infected antigenic protein, gold particle labeling was specifically distributed on the cuticle and EIM of the worm. When purifed 45 kDa protein was used gold particle labeling was specifically distributed on the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte of the worm. Therefore, excretory antigens appeared to originate from the cuticle and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte for the first day but the cuticle layer associated with globular proteins and ${\alpha}_0$ granules of stichocyte after 3 days and infected antigens appeared to originate from the cuticle for 1 and 4 weeks after infection. These results suggest that excretory and infection specific antigens are secreted into the cuticle, basal layer, EIM and ${\alpha}_0,\;{\alpha}_1$ granules of stichocyte and 45 kDa protein may be contained these specific antigens.

• Parasites, Hosts and Diseases
• /
• v.30 no.1
• /
• pp.25-32
• /
• 1992

Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as pri-mary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody(MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.

• Parasites, Hosts and Diseases
• /
• v.30 no.3
• /
• pp.227-230
• /
• 1992

Spirometra mansoni plerocercoid (sparganum) was incubated in saline at $4^{\circ}C{\;}or{\;}37^{\circ}C$ up to 100 hours. Protein contests in the excretory$.$secretory product (ESP) were rather constant (mean 7.7 mg of protein/gram of sparganum) in the preparations. Reducing SDS-PAGE of ESP showed similar protein subunit compositions with those in crude extract. Antigenic 36 and 31 kDa Proteins were major bands in ESP. ESP exhibited specific activities of protease(2.9~5.3 units/mg) at pH 6.0 and pH 7.5. Presence of protease activity in ESP may be a supporting evidence that hitherto known cysteiRe protease of sparganum is possibly secreted.

• Applied Microscopy
• /
• v.37 no.2
• /
• pp.83-91
• /
• 2007

The present study examined the epidermal changes of adhesive disks which occur during attachment in Parthenocissus tricuspidata using scanning and transmission electron microscopy. Several adhesive disks, each covered with a bract, develop from the shoot apical meristem during early development. In the initial stage, the adhesive disks are club-shaped and their upper and lower epidermis are indistinguishable. However, in the actively growing stage, they become spherical and both epidermis are clearly differentiated into the adventitious roots. Prior to wall attachment, the adhesive disks exhibit adaxial convex and abaxial concave shapes, and electron-dense substances are abundant in the vacuoles of epidermal cells. The peripheral area of the adhesive disk is adhered first to the wall surface, while the central area is drawn inward in a vacuum-like state during attachment. As the attachment progresses and the electron-dense substances continue to discharge, the upper and lower epidermis rapidly undergo deterioration and the disks shrink considerably. At this stage, structural changes of the lower epidermis occur much faster than in the upper one. The discharged substance is accumulated on the wall surface, and this aids the attachment of adhesive disks on the wall for long periods. In this manner, the shape and structure of the adhesive disk epidermis change drastically from initial growth to the mature stage. Further, the role of electron-dense substance and shrinkage of the disk during attachment has been discussed in Parthenocissus tricuspidata.

• Applied Microscopy
• /
• v.30 no.1
• /
• pp.101-111
• /
• 2000

In this study, the distribution of lectin receptors in Paragonimus westermani tissue was explored using colloidal gold label complexed with lectin WGA purified from wheat germ (Triticum vulgare). The lectin WGA gold complex, shown to recognize GlcNAc (N-acetylgalactosamine) and NeuNAc (N-acetylneuraminic acid) regions, was applied to detect binding sites in Lowicryl HM 20 sections viewed under electron microscope. Labeled sections of the metacercaria revealed gold particles specifically distributed on the tegumental syncytium and lamella of the excretory canal. Labeling of young adult tissue was then quantified and compared to that of adult worm tissue. Adult worm tissue sections resulted in specific gold particle distribution on the lamella of caecal epithelium and excretory canal. These results indicate that lectin WGA receptors are located in the tegumental syncytium and lamella of the excretory canal of the metacercariae, and in the lamella of the caecum and excretory canal of the young adult and adult. Therefore, the GlcNAc and NeuNAc regions in the tegumental syncytium appear to be functionally associated with cell-recognition and protection from the immune system of the host, and linked with membrane transport and absorption of nutrients in the lamella of the excretaory canal and caecal epithelia.

• Korean Journal of Environment and Ecology
• /
• v.26 no.2
• /
• pp.192-199
• /
• 2012

This study was to evaluate the application of microhistological analysis to collect information on food materials of korean water deer from september to October 2006. For microhistological analysis we extracted type-materials of 4 plant species ($Ipomoea$ $batatas$, $Rubus$ $coreanus$, $Glycine$ $max$, and $Pueraria$ $thunbergiana$) from fecal pellets of korean water deer ($Hydropotes$ $inermis$) and then made preparations of the fecal materials for microscopic analysis. The fecal preparations were compared with reference preparation under the microscope. In microscopic analysis of fecal preparations, we could identify the four plant species by the followings: tip shape and morphology of trichomes, epidermal cell, morphological composition and shape of stomatal apparatus, arrangement of long-cell, and distribution of crystals in cells. According to the comparisons of the preparations, the morphology of trichome, epidermal cell and stomata are similar between both preparations of the fecal pellets and the references, respectively. This methodology will contribute to understanding feeding behavior and behavioural ecology by climate change of other hervibores as well as korean water deers.

• Journal of the Korean earth science society
• /
• v.28 no.2
• /
• pp.169-178
• /
• 2007

In this research we diagnosed the actual status of the 7th National science elective curriculum and suggested a way to select and organize the content of the new science elective curriculum. The first science education reform was grounded in the structuralism where the structure of discipline was valued above everything else. On the other hand, the second science education reform suggested alternative interpretations of students' opportunity to learn, putting a brake on the structuralist thinking. According to the survey result, the majority of the science elective courses are in need for revision because the contents are overcrowded, too difficult in light of students' learning readiness, failed to draw students' interest in science, and are overlapped and repeated among the 10th grade science, high school science I and II. In particular, Earth Science II and physics II are the most unfavorable courses among students. Thus, we recommended a fundamental change be made in the new curriculum in addition to the optimization of the content. In this paper, we suggested 'topic-centered content organization' for the science elective course I, i.e., Physics I, Chemistry I, Biology I and Earth Science I that is designed for both science track and non-science track students. Since curriculum provides students with an 'opportunity to learn', a curriculum study should focus on what the 'opportunity to learn' is that students ought to be offered. Based on the result of this study, we recommended one way to select and organize the content of high school elective curriculum.

• Applied Microscopy
• /
• v.37 no.2
• /
• pp.93-102
• /
• 2007

Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

• Parasites, Hosts and Diseases
• /
• v.29 no.3
• /
• pp.245-258
• /
• 1991

In order to determine the antigenic localization in the tissues of the adult Metagonimus yokegawai, immunogoldlabeling method was applied using serum immunoglobulins (IgG) of cats which were infected with isolated metacercariae from Plecoglossus altivelis. The sectioned worm tissue was embedded in Lowicryl HM 20 medium and stained with infected serum IgG and protein A gold complex (particle size: 12 nm) , It was observed by electron microscopy at each tissue of the worm. The gold particles were observed on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum. The gold particles were not observed on the basal lamina of the tegument, interstitial matrix of the parenchyma, the muscle tissue and mitochondria of the tegument. The gold particles were specifically labeled in the secretory granules in the vitelline cells. They were also labeled on the lumen of bladder and egg shell. The above findings showed that antigenic materials in the tissue of adult worms were specifically concentrated on the tegumental syncytium as well as cytoplasm of tegumental cells and epithelial lamella of the caecum.

• Parasites, Hosts and Diseases
• /
• v.29 no.1
• /
• pp.21-30
• /
• 1991

An immunohistochemical study was performed to demonstrate comparative antigenicity of each body structure of the liver fluke, Clonorchis sinensis, such as the digestive tract, reproductive organs, excretory system, tegument and suckers. Indirect immunoperoxidase technique was applied, rising formalin-fixed and paraffin-embedded sections of C. sinensis as the antigen. Pooled cat sera obtained 10 weeks after an experimental infection with C. sinensis and peroxidase-conjugated goat anti-cat IgG were used as the primary and secondary antibodies, respectively. The intensity of immunohistochemical stain was very sensitive upon the titers of the primary and secondary antibodies, and their optimum dilutions were found to be 1:1,000∼1:2,000 and 1:1,000, respectively. The intestinal epithelial cells, intestinal content and excretory bladder showed strong positive coloring reactions even at lower titer (1 : 2,000) of the primary antibody, whereas the uterine wall and eggs, vitelline glands, and male reproductive organs showed only weak positive reactions despite an increase in the antibody titer (1:1,000). On the other hand, the suckers, tegument, subtegumental cells and other parenchyme portions did not reveal any positive immunoperoxidase reaction at the same antibody titers. From the above results, it is highly suggested that the most potent antigenicity of C. sinensis occur from their excretory-secretory substances originated from the digestive and excretory organs.