• The Korean Journal of Pain
• /
• v.9 no.2
• /
• pp.326-335
• /
• 1996

Background: Though a number of studies have described the distribution of substance P(SP)-like immunoreactivity in the spinal cord, they have been focused on lamina I and II of the dorsal horn and there are little morphological studies on the topographic distribution and ultrastructure of the SP immunoreactive neurons especially in the ventral horn of the spinal cord. this study was conducted to identify distribution pattern of SP immunoreactive neurons and to difine the synaptic organization of their processes in ventral horn of the thoracic cord of the cat by preembbeding immunocytochemical method using SP antiserum. Methods: Five adults cats of either sex were used and deeply anesthetized by intramuscular injection of ketamine. After removal of the spinal cord, samples of thoracic cord were taken and placed in fresh fixative at $4^{\circ}C$ for 2 hours. Transverse sections $50{\mu}m$ thick were processed using the preembbeding immunocytochemical method and incubated consecutively in the specific primary antibody and the 10% normal goat serum, the rabbit anti-substance P antiserum, the biotin-labelled goat anti-rabbit IgG and finally the avidin-biotin-peroxidase complex. The processed tissue sections were throughly washed and stained in the black with 1% uranyl acetate. Section were examined on a electron microscope. Results: 1) SP immunoreactive neurons were observed in the gray matter around central canal. 2) In lamina I and II SP immunoreactivity was observed in both myelinated and unmyelinated nerve fibers, but in ventral horn only in the unmyelinated nerve fibers. 3) SP immunoreactive axon terminals with small round and large dense core vesicles made chemical synapses onto the dendrites of motor neurons in the ventral horn. Conclusion: SP immunoreactive neurons might play an important role in modulation of motor neurons in the ventral horn of the thoracic cord of the cat.

• Journal of Environmental Science International
• /
• v.17 no.11
• /
• pp.1227-1233
• /
• 2008

The effect of freeze drying and fixatives in post-treating freeze drying on the morphological properties of the rose (Rosa hybrida L.) petal were investigated for the production of high quality of freeze dried rose. The morphology including form and color of the dried flowers of cut rose were depended on the drying methods. The drying time was extended due to their density and water content, and was shorter in the freeze drying than that in the natural and hot air drying. Freeze dried process for dried flowers took 2 days in a freeze dryer and did not cause shrinkage or toughening of rose petal being dried, preserving its natural shape and color. The diameter of freeze dried flowers showed little reduction compared to fresh flowers. In Hunter color values of petals of freeze dried flowers, L and a values were high and showed little variations in comparison to fresh petals. Freeze drying led to a noticeable increase in anthocyanin contents in petals, suggesting that anthocyanin contents play an important role in the acquisition of freezing tolerance. Exposure of flowers to freeze drying was accompanied by an increase in the carotenoid content. In the post-treating freeze drying, epoxy resin, a fixative, applied alone or in combination to petals of freeze dried flowers showed efficient coating for the protection from humidity and sunlight. Combined application of epoxy and acetone to freeze dried petals permitted maintenance of natural color and excellent tissue morphology, showing color stability and shiny texture in surface of petals. These findings suggest that application of fixatives to freeze dried rose petals improves the floral preservation and epoxy coating provides good quality in the freeze dried flower product.

• Parasites, Hosts and Diseases
• /
• v.22 no.1
• /
• pp.30-36
• /
• 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.

• The Korean Journal of Pain
• /
• v.29 no.3
• /
• pp.158-163
• /
• 2016

Background: Phenol and alcohol have been used to ablate nerves to treat pain but are not specific for nerves and can damage surrounding soft tissue. Lidocaine at concentrations > 8% injected intrathecal in the animal model has been shown to be neurotoxic. Tests the hypothesis that 10% lidocaine is neurolytic after a peri-neural blockade in an ex vivo experiment on the canine sciatic nerve. Methods: Under ultrasound, one canine sciatic nerve was injected peri-neurally with 10 cc saline and another with 10 cc of 10% lidocaine. After 20 minutes, the sciatic nerve was dissected with gross inspection. A 3 cm segment was excised and preserved in 10% buffered formalin fixative solution. Both samples underwent progressive dehydration and infusion of paraffin after which they were placed on paraffin blocks. The sections were cut at $4{\mu}m$ and stained with hemoxylin and eosin. Microscopic review was performed by a pathologist from Henry Ford Hospital who was blinded to which experimental group each sample was in. Results: The lidocaine injected nerve demonstrated loss of gross architecture on visual inspection while the saline injected nerve did not. No gross changes were seen in the surrounding soft tissue seen in either group. The lidocaine injected sample showed basophilic degeneration with marked cytoplasmic vacuolation in the nerve fibers with separation of individual fibers and endoneurial edema. The saline injected sample showed normal neural tissue. Conclusions: Ten percent lidocaine causes rapid neurolytic changes with ultrasound guided peri-neural injection. The study was limited by only a single nerve being tested with acute exposure.

• Asian Pacific Journal of Cancer Prevention
• /
• v.13 no.4
• /
• pp.1305-1310
• /
• 2012

Breast cancer cells undergo transformation when they spread into surrounding tissues. Studies have shown that cancer cells undergo surface alterations and interact with the surrounding microenvironment during the invasion process. The aim of the present study was to analyse these cancer cell surface alterations and interactions of cancer cells and stroma. Twenty 1-methyl-1-nitrosourea-induced breast cancer samples taken from five rats were fixed in McDowell-Trump fixative and then washed in 0.1 M phosphate buffer. The samples were then treated with osmium tetroxide before being washed in distilled water and subsequently dehydrated through graded ethanols. The dehydrated samples were immersed in hexamethyldisilazane (HMDS), then following removal of excess HMDS, the samples were air dried at room temperature in a dessicator. The dried samples were mounted onto specimen stubs and coated with gold coater before being viewed under a scanning electron microscope. We detected the presence of membrane ruffles on the surface of cancer cells and the formation of unique surface membrane protrusions to enhance movement and adhesion to the surrounding stroma during the process of invasion. Advancing cancer cells demonstrated formation of lamellipodia and invadopodia. The stroma at the advancing edge was desmoplastic with many collagen fibres laid down near the cancer cells. Our data suggest that all of these abnormalities could act as hallmarks of invasiveness for breast cancer.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.7 no.3
• /
• pp.115-120
• /
• 1974

Secondary amines are known as one of the precursors of nitrosamines which are potent carcinogenic compounds of man and animals. Nitrosamines are formed when both secondary amines and nitrite are present. The nitrites are occurred naturally in vegetables, fruits and many others, and frequently used in fish product as a color fixative or a preservative. In this paper, to know the formation of nitrosamines in fish meat, the changes of dimethylamine (DMA) content during the heat treatment such as drying, roasting and fish cake processing are discussed. The results showed that generally DMA increased considerably during heat processing. During drying, DMA in cuttle fish increased 16 times higher, while in Alaska pollack doubled than the fresh. In the roasted mackerel, the DMA content appeared 6 times higher than the fresh, while 9 times higher in canning. In fish cake from yellow corvenia, DMA increased 4 times higher than the fresh. On the contrary diethylamine and diphenylamine were not found in this experiment.

• Proceedings of the Korea Society of Environmental Toocicology Conference
• /
• 2003.10a
• /
• pp.174-174
• /
• 2003

There are many synthetic chemicals, such as di(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP), used in chemical reaction processes in industry. The establishment of toxicity and detection of synthetic chemicals that may pose a genetic hazard in our enviornment is subjects of great concern at present DEHP, a ubiquitous phthalate plasticizer, induces a wide range of developmental and reproductive toxicities in mammals. DEHP belongs to the large diverse class of peroxisome proliferator compounds, which include herbicides, hypolipidemic drugs. DBP is a plasticizer used to products containing nitrocellulose, polyvinyl acetate, and polyvinyl chloride such as food wraps and blood bags. DBP is also used in cosmetics as a solvent and fixative for perfumes, a suspension agent for solids, an antifoamer, a skin emollient, and hair spray The present study was performed to examine patterns of gene expression in MCF-7 cells following DEHP and DBP exposure. Changes in gene expression were determined by microarray analysis using KISTCHIP-400 including 401 endocrine related genes based on public database and research papers. Of the genes analysis, we determined that genes detected by array showed a 2-fold or greater change in their expression level(increase or decrease). The results of this study demonstrate that a number of genes were differentially expressed in MCF-7 cells but these changes were not significant. Therefore, we keep going this study using microarray analysis and future studies will examine changes of gene expression on time-course and does treatment in variable cell lines.

• The Korean Journal of Cytopathology
• /
• v.13 no.2
• /
• pp.66-69
• /
• 2002

In diagnosing a brain tumor, it is essential to obtain samples from many areas of the tumor. Although there are reports about the suitability of material obtained by cavitron ultrasonic surgical aspirator(CUSA), there is a paucity of reports regarding conventional intraoperative suction. This study was performed to evaluate the usefulness of the suction fluid and the effect of different hemolytic fixatives. Intraoperative suction fluid was obtained from 2 pituitary adenomas and 2 choroid plexus carcinomas. In two cases of mixed astro-oligodendroglioma, one of glioblastoma multiforme and 3 of meningioma, the fluid was collected by CUSA. Each sample was divided into four bottles for the different fixatives such as 0.1N HCI, 10% acetic acid, 95% alcohol, and no additive. All cases were evaluated by the both cytologic smear and cell block preparations, and were reviewed with concomitant histologic diagnosis. The result showed a good correlation between the cytologic study and the histologic diagnosis and 95% alcohol was found to be superior to other fixatives in ceil preservation.

• The Korean Journal of Cytopathology
• /
• v.18 no.1
• /
• pp.29-35
• /
• 2007

This study purposed to evaluate a $CellPrep^{(R)}$ (CP) of liquid-based cytology (LBC) to search for a less expensive and automated alternative cytologic preparation technique applicable to usually encountered cytologic specimens. Cervicovaginal direct-to-vial split samples from 457 gynecologic patients, 40 body fluid samples, and 34 urine samples were processed with the CP technique and the results were compared with those of currently used $ThinPrep^{(R)}$ (TP) method. Both CP and TP methods provide evenly distributed thin layers of cells with little cellular overlaps or significant obscuring elements in most of cases. Staining quality of both preparations showed a little difference due to the difference of fixative solutions without significant distractions in cytologic interpretation. On the supposition that TP was a gold standard, sensitivity, specificity, positive predictive value, and negative predictive value of CP cytology were 89%, 98%, 86%, and 99% in the cervicovaginal smear, 89%, 82%, 80%, and 90% in body fluid, and all of these values were 100% in urine samples. To testify the availability of immunohistochemistry on CP preparations, cytokeratin, vimentin, and Ki-67 were applied on body fluid specimens, and all of these antibodies were specifically stained on targeted cells. Conclusively, the CP method gave comparable results to those of TP in terms of smear quality and cytologic diagnostic evaluation, and was available on immunohistochemistry. The CP method could offer a cost-effective and automated alternative to the current expensive techniques of liquid-based cytology on popular cytologic materials including cervicovaginal, body fluid, and urine specimens.

• Applied Microscopy
• /
• v.22 no.2
• /
• pp.118-126
• /
• 1992

The pathway and time course of fucose-containing glycoprotein synthesis and intracellular translocation in osteoclasts of the mice maxillary alveolar bone were investigated by electron microscopic radioautography. Male Balb-C mice weighing 17gm were anesthetized with Nembutal and injected via the external jugular vein with 2.5 mCi of $L-[6-^{3}H]-fucose$ (specific activity 16.8 mCi/mmol) in 0.1 ml of sterile saline solution. At 5, 10, 20, 35 minutes and 8 hours after administration of the $^{3}H-fucose$, animals were killed by intracardiac perfusion of 30ml of 2% glutaraldehyde in a modified Tyroid solution, pH 7.4. The maxillae were then removed and further fixed in Karnovsky fixative for an additional 3-4 hours. After rinsing in 0.1M cacodylate buffer for 10 minutes, the maxillae were demineralized for 2 weeks at $4^{\circ}C$ in ethylene diamine tetra acetate containing 2% glutaraldehyde. The first interdental areas were mesiodistally sectioned into slices of 1mm thickness and postfixed in osmium tetroxide. Tissues were then dehydrated and embedded in Poly Bed. To prepare electron microscopic radioautography, the dipping method of Kopriwa (1973) was employed. Thin sections were coated with a crystalline monolayer of ILford $L_4$ photographic emulsion. After exposure for 4 months at $4^{\circ}C$, the sections were developed Kodak Microdol-X and Phenidon (for compact grains), fixed in 30% sodium thiosulfate, stained with uranyl acetate and lead citrate and examined in the electron microscope (JEOL 1200 EX). At 5, 10 and 20 minutes after injection, $^{3}H-fucose$ was concentrated in Golgi cisternae of the osteoblasts. By 35 minutes the labels were observed over small vesicles in the suprannclear area of osteoclasts. At 8 hours, numerous silver grains were located on the ruffled border and cell membrane of osteoclasts. These results indicate that fucose molecules are added in the Golgi apparatus and small vesicles appear to be responsible for translocation of the glycoproteins to the marginal portion of osteoblasts. The glycoproteins are distributed on the osteoclast cell surface and especially over the ruffled border.