• Title/Summary/Keyword: Neutral-formalin

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Effects of Continuous Therapeutic Ultrasound on Subcutaneous Murine Melanoma (초음파치료가 종양세포에 미치는 효과)

  • Kim, Ji-Won;Lee, Kuk-Il;Han, Jae-Mun
    • Physical Therapy Korea
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    • v.4 no.1
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    • pp.11-19
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    • 1997
  • The use of therapeutic ultrasound(US) in humans with malignant neoplasms has been contraindicated in physical therapy practice. Some studies have shown that results after application of US differ according to tumor type and penetration depth. The purposes of this study were to determine the effects of US on melanoma in mice and to determine treatment dosage. Twenty-four female C57BL/6 mice, age 8 weeks. The right flank of all mice was shaved, and a 0.1 ml suspension of cells was injected subcutaneously into the animals' right flank. In this study, 24 subjects were randomly divided into three groups: experimental group 1(n=8), experimental group 2(n=8), control group(n=8). In the experimental group 1, animals received continuous 3 MHZ US treatment, administered at $2.0W/cm^2$ for five minutes. In experimental group 2, animals received continuous 3 MHz US treatment, administered at $1.0W/cm^2$ for 5 minutes. The control group received the same handling as other experimental groups, including rodent chow, water, US gel application but US head pressure without the power turned on. After 10 days treatment, all mice were killed with a potassium solution. Tumors were excised and weighed on an electrical balance and fixed in a 10% neutral buffered formalin solution. Tumor weights were smaller in experimental group 2(0.3838 g) than in the control group(0.6275 g). Tumor weights of the experimental group 1(0.015 g) were smaller than those of experimental group 2. Continuous therapeutic US decreased the weight of subcutaneous melanoma tumors in mice. The treatment dosage($2.0W/cm^2$) we suggest was more effective than earlier studies on decreasing tumor size with ultrasound.

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Light and Scanning Electron Microscopic Study on the Pelvic Epithelium of the Sheep Kidney (양(羊)의 신우상피에 대한 광학 및 주사전자현미경적 연구)

  • Kim, J.;Oh, S.J.;Chung, J.W.
    • Applied Microscopy
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    • v.15 no.2
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    • pp.98-110
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    • 1985
  • This study was performed to clarify the morphological structures of the epithelia of the renal papilla, renal pelvis and ureter of the sheep (Ovis aries L.) through the light and scanning electron microscopes, Tissue specimens were taken from the renal papilla (common renal papilla and peripelvic column) and the renal pelvis (pelvis proper and pelvic pouch) of the kidney and the ureter. For the light microscopy, tissue blocks were fixed in 10 % neutral buffered formalin and embedded in paraffin wax, serially sectioned at a thickness of $6{\mu}m$. These sections were stained with hematoxylin-eosin and periodic acid-Schiff reaction. For the scanning electron microscopy, tissue blocks were prefixed in 1% glutaral-dehyde-1.5% paraformaldehyde solution and postfixed in 1% osmium tetroxide solution, dehydrated in graded alcohol, transferred to isoamyl acetate, and then dried by the critical point dryer (Polaron E 3000). These dried tissues were coated with gold and observed with a scanning electron microscope (JSM-35C), The results were as follows: The apex of the common renal papilla was lined with simple columnar epithelium having many microvilli on its luminal surface. Lateral portion of the papilla was lined with stratified epithelium $2{\sim}3$ layers thick, and its superficial cells were microvillar cells having many microvilli. The epithelium lining the peripelvic column was $1{\sim}2$ layers thick. The superficial layer was made of the microvillar cells, but a few microplica cells were appeared in the region near the pelvic pouch. The epithelium of the pelvic pouch was $1{\sim}2$ layered transitional type, and its superficial cells were microplica cells. The epithelia of the pelvis proper and ureter were $4{\sim}6$ layered transitional type, and their superficial cells were typical facet cells existing many round depressions and ridges of cell membranes of the luminal side.

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Pathologic studies for porcine pleuropneumonia and serotypes of the agents in Jeju (제주지역 돼지 흉막폐렴에 대한 병리학적 연구 및 원인체의 혈청형)

  • Kim, Ki-Seung;Yang, Hyoung-Seok;Kim, Jae-Hoon
    • Korean Journal of Veterinary Service
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    • v.44 no.2
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    • pp.103-111
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    • 2021
  • Actinobacillus (A.) pleuropneumoniae is the etiological agent of a porcine pleuropneumonia and have great economic importance to the global swine industry. For recent 5 years, a total of 50 pleuropneumonia cases of 24 pig farms were selected from pig lungs submitted to the College of Veterinary Medicine, Jeju National University using polymerase chain reaction (PCR) analysis. Collected lungs were fixed in 10% neutral phosphate-buffered formalin and processed for histological examination. Serotypes of A. pleuropneumoniae in pneumonic lesions were analyzed by PCR methods. And the antimicrobial susceptibility of A. pleuropneumoniae isolates was determined by a disc diffusion test. Grossly, unilateral distribution of hemorrhagic or necrotic pneumonic lesions was more common than bilateral distribution in lungs. In peracute or acute cases, histopathologic changes were characterized by necrosis, hemorrhage, neutrophils infiltration, vascular thrombosis, widespread edema and fibrinous exudates. Following the acute response, macrophage infiltration, marked fibrosis around zonal necrotic areas, and marked fibrous pleuritis were characteristic in chronic cases. A total of 50 pleuropneumonia were associated with A. pleuropneumoniae serotype 5 in 46 cases (92%), serotype 2 in 3 cases (6%), and both 2 and 5 in 1 case (2%). More than 90% of collected isolates showed high sensitivity to ceftiofur, amoxicillin, and colistin. However, ampicillin, penicillin, and tylosin showed low susceptibility. The results of this study demonstrated that A. pleuropneumoniae serotype 5 was predominant at porcine pleuropneumonia cases in Jeju.

Alaria alata (Digenea: Diplostomidae) from Korean Raccoon Dog, Nyctereutes procyonoides koreensis, in Korea

  • Lee, Heon Woo;Hong, Eui Ju;Kim, Hyeon Cheol;Ryu, Si Yun;Park, Bae Keun
    • Parasites, Hosts and Diseases
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    • v.60 no.5
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    • pp.361-365
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    • 2022
  • We report a species of diplostomid fluke recovered from 3 carcasses of wild Korean raccoon dog, Nyctereutes procyonoides koreensis, in Korea. A total of 107 diplostomid flukes were recovered from the small intestines of Korean raccoon dogs, which were obtained from the Gangwon Wildlife Medical Rescue Center. Worms fixed with 10% neutral formalin were subjected to microscopic observation and those fixed in 70% ethanol were used for molecular genomic analysis. The worm was divided into 2 separate parts, forebody and hindbody, with a total length of 3,020-4,090 (3,855) ㎛ and a width of 1,210-1,770 (1,562) ㎛. The boat-shaped forebody has a pair of characteristic tentacular appendage, 2 suckers, holdfast organ, and vitelline follicles. The oval to cylindrical hindbody has reproductive organs. The ovary was round or elliptical and located in the anterior of the testes. Two large testes were slightly segmented and tandemly arranged, occupying almost half of hindbody. The short uterus contained a relatively small number of unembryonated eggs sized 130-140×85-96 ㎛. The partial sequence of 18S rRNA of this fluke was consistent with Alaria alata. Based on the morphological and molecular characteristics, the diplostomid flukes recovered from the small intestine of Korean raccoon dogs were identified as A. alata (Digenea: Diplostomidae).

Effects of BCG on the DNA Synthesis and Ultrastructure of Mouse Gastric Mucosal Epithelial Cells Inoculated with Ehrlich Carcinoma Cells (BCG가 Ehrlich 암세포를 이식한 생쥐의 위점막 상피세포의 DNA합성 및 미세구조에 미치는 영향)

  • Ko, Jeong-Sik;Ryoo, In-Sang;Park, Kyung-Ho;Park, Dae-Kyoon
    • Applied Microscopy
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    • v.39 no.3
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    • pp.205-218
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    • 2009
  • This experiment was performed to evaluate the morphological responses of the gastric epithelial cells of the mouse, inoculated with Ehrlich carcinoma cells in the inguinal area, following administration of BCG. Healthy adult ICR mice weighing 25 gm each were divided into normal and experimental groups (tumor control group and BCG-treated group). In the experimental groups, each mouse was inoculated with $1{\times}10^7$ Ehrlich carcinoma cells subcutaneously in the inguinal area. From next day after inoculations, 0.2 mL of saline or BCG (0.5 mL/25 g B.W.: $0.03{\times}10^8{\sim}0.32{\times}10^8$ CFU) were injected subcutaneously to the animals every other day, respectively. The day following the 7th injection of saline or BCG, each mouse was injected with a single dose of 0.7 ${\mu}Ci/g$ of methyl-$^3H$-thymidine (25 Ci/mmol, Amersham Lab., England) through tail vein. Seventy minutes after the thymidine injection, animals were sacrificed, and gastric tissues were taken and fixed in 10% neutral formalin. Deparaffinized sections were coated with autoradiographic emulsion EM-1 (Amersham Lab., England) in a dark room. The number of labeled epithelial cells in the gastric mucosae (mean number of labeled epithelial cells per 3.5 mm length of mucosa) were observed and calculated. And for electron microscopic observation, gastric tissues were prefixed with 2.5% glutaraldehyde-1.5% paraformaldehyde solution, followed by post-fixation with 1% osmium tetroxide solution. On the light microscopic study, gastric mucosae had no morphological changes following the injection of BCG. On the electron microscopic study, in the BCG-treated mice, myelin figures and multivesicular bodies within the gastric epithelial cells were observed more frequently than in those of the normal control ones. On the autoradiographic study, number of the labeled cells of normal control, tumor control and BCG-treated mice were 380.2 (${\pm}31.35$), 426.1 (${\pm}28.43$) and 301.8 (${\pm}34.63$), respectively. In the BCG-treated mice, poorly-labeled cells containing only a few silver grains of 3H-thymidine were observed more frequently as compared in those of the normal control and tumor control ones. From the above results, BCG may suppress the DNA synthesis of the gastric epithelial cells, but does not results severe fine structural defect on the gastric epithelial cells. These results suggest that BCG is expected as one of the effective supplemental anticancer drugs.

Recovery rate, growth and development of Heterophyopsis continua in experimental chicks (병아리에서 Heterophuopsis continua의 충체 회수율 및 성장 발육)

  • 홍성종;이순형
    • Parasites, Hosts and Diseases
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    • v.28 no.1
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    • pp.53-62
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    • 1990
  • The growth and developmental pattern of H. continua was observed after experimental infection of their metacercariae to chicks. The recovery rate of worms from the chicks at 1 to 28 days post-infection(PI) was 12.8% in average. The rate remained fairly high for early 4 days of infection but decreased thereafter rapidly till 28 days PI. Most of the nukes, 91.9%, were recovered from the ileum of the chicks. In metacercariae, genital organs such as the ovary, testes, seminal vesicle, seminal receptacle and genital sucker were recognizable. At one day PI Mehlis'gland appeared, and at 2 days follicular vitellaria were observed. At 3 days PI, eggs were formed in the uterine tubule and increased in number as the worm grew old. The worms reached $2,990{\;}{\mu\textrm{m}}$ in length and $525{\;}{\mu\textrm{m}}$ in width at 28 days PI. Genital organs developed rapidly in early stages of infection but slowly thereafter to 28 days Pl, whereas non-genital organs developed steadily through the infection period. It was proved by this experiment that chicks should be a moderately suitable final host of H. continua.

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THE EFFECTS OF IRRADIATION ON THE MANDIBULAR BONE OF RATS ON THE LOW CALCIUM DIET (방사선조사가 저칼슘식이 백서의 하악골에 미치는 영향에 관한 실험적 연구)

  • Hwang Eui-Hwan;Lee Sang-Rae
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.22 no.2
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    • pp.241-252
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    • 1992
  • The purpose of this study was to investigate the changes of morphology and structure of bone tissue in the irradiated mandibular bone in rats which were fed a low calcium diet. In order to carry out this experimant, 64 seven-week old Sprague-Dawley strain rats weighing about 150gms were selected and equally divided into one experimental group of 32 rats and one control group with the remainder. The experimental group and the control group were then subdivided into two groups when the rats reached ten-week old, 16 were assigned rats for each subdivided group, exposed to irradiation. The two irradiation groups received a single dose of 20 Gy in the jaws area only and irradiated with a cobalt-60 teletherapy unit. The rats in the control and experimental groups were serially terminated by fours on the 3rd, the 7th, the 14th, and the 21st day after irradiation. Following termination, both sides of the dead rats mandibular bodies were removed and fixed with 10% neutral formalin. One side of the mandibular body was radiographed with a soft X-ray apparatus. Thereafter, the obtained microradiographs were observed by a light microscope. The remaining side of the mandibular bone was further decalcified and embedded in paraffin as using the general method. The specimen sectioned and stained with hematoxylin and eosin, and Rabit Anti-Human Tumor Necrosis Factor-a, observed by a light microscope. The obtained results were as follows: 1. Microradiogram revealed that thinning of the cortex and a decrease in the trabecula of the interradicular bone and mandibular body were observed and noted from the start to finish throughout the experiment in the non-irradiated rats on the low calcium diet rather than in the non-irradiated rats on the normal diet. In microscopic observations, there were marked osteolytic changes in the center of the bone marrow. 2. Microradiogram revealed that thinning of the cortex and a decrease in the trabecula of interradicular bone and mandibular body were more marked after 7 days in the irradiated rats on the low calcium diet rather than in the non-irradiated rats on the low clacium diet. In microscopic observations, osteoblasts were decreased markedly after 7 days, and a few osteoclasts were observed. 3. Microradiogram revealed that thinning of the cortex and a decrease in trabecula of interradicular bone and the mandibular body were more marked from the strart to finish troughout the experiment in the irradiated rats on the low calcium diet rather than in the irradiated rats on the normal diet. In microscopic observations, there were no osteolytic changes noted in the irradiated rats on the normal diet. 4. In immunocytochemical findings, a few bone marrow cells including Tumor Necrosis Factor were observed in the irradiated rats on the normal diet, but was not observed in the rats on the low calcium diet.

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THE EFFECTS OF IRRADIATION ON THE PERIODONTAL TISSUES OF RATS WITH THE LOW CALCIUM DIET (방사선조사가 저칼슘식이 백서 치주조직에 미치는 영향에 관한 연구)

  • Choi Mun-Cheol;Lee Sang-Rae
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.22 no.2
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    • pp.223-234
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    • 1992
  • The purpose of this study was to investigate the changes of periodontal tissues in the irradiated mandibular bone in rats which were fed normal diet and low calcium diet In order to carry out this experiment, 64 seven-week old Sprague-Dawley strain rats weighing about 150gms were selected and equally divided into one experimental group of 32 rats and one control group with the remainder. The experimental group and the control group were then subdivided into two groups when the rats reached the age of 10 weeks, 16 rats were allotted for each subdivided group was composed of 16 rats and exposed to irradiation. The two groups were irradiated a single dose of 20Gy on the only jaw area and irradiated with a cobalt-60 teletherapy unit The rats in the control and experimental groups were serially dissected by fours on the 3rd, the 7th, the 14th, and the 21st day after irradiation. After each dissection, both sides of the dead rat mandibular bodies were removed and fixed with 10% neutral formalin. The specimens sectioned and observed in histopathological. histochemical. and immunocellular chemical methods. The obtained results were as follows: 1. In the mandibles of rats with low calcium diet the increased number of fibroblasts of periodontal ligaments. many small capillaries and irregular arrangement of loose collagen fibers were detected and the partial resorption of dentin and cementum could be found by the microscopic studies. 2. In the group of irradiated rats, degenerated periodontal tissues led to the condition of irregular arrangement of collagen fibers and the decreased number of fibroblasts. But this condition was somewhat restored after 21 days of experiment. 3. Periodontal tissues of the irradiated rat group with low calcium diet were destroyed earlier than those of the irradiated rat group with normal diet. Soon this condition was restored and then high cellularity and dense collagen fibers were observed. 4. Many periodontal cells bearing tumor necrosis factor could be clearly observed in the nonirradiated group of rats with normal diet, whereas could not be observed on the 7th day and reappeared on 14th day in the irradiated group of rats with normal diet. A few of them could be observed in the group of rats with low calcium diet, but they could be clearly observed in the both groups after 21 days of experiment.

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Detection of Pneumocvstis carinii by in situ hybridization in the lungs of immunosuppressed rats (면역억제 흰쥐에서 조직내교잡법을 이용한 페포자충의 검출)

  • Jin KIM;Jae-Ran YU;Sung-Tae HONG;Chang-Soo PARK
    • Parasites, Hosts and Diseases
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    • v.34 no.3
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    • pp.177-184
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    • 1996
  • In situ hybridization was performed to detect rat heumocwstis ca4nii in the lung sections. Rats were immunosuppressed by weekly subcutaneous injection of 10 mg/kg methylprednisolone. On the 6th, 8th and 9th week of immunosuppression, the lungs were removed and fled in 10% neutral formalin. A 22 base oligonucleotide probe complementary to p. carinii 5S ribosomal RMh was commercially synhesized and its 3' terminal was labeled wiH biotin. In situ hybridization was performed utilizing manual capillary action technolog)r on the Microprobe system. p. cnrinii were detected along the luminal surface of alveolar pneumocytes, in exudate of alveolar cavities, and also in secretory material of bronchioles. In the 6th week group, positive reaction was observed focally in the peripheral region of the lung sections, but the reaction was observed diffusely in the 8th or 9th week groups. In comparison with Grocott's methenamine silver stain, in situ hybridization technique can detect the organism rapidly, and can detect trophic forms very well. Furthermore, no nonspecific reaction with other pathogenic fungi and protozoa was recognized. Therefore, in situ hybridization can be a good technique to detect p. carinii in the lungs of infected rats.

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Dystrophin Degradation in Skeletal Muscles with Lipid Enrichment in Cattle (지방 침착률이 높은 식용소에서 나타난 골격근의 디스트로핀 소실)

  • Jeon, Sung-Hwan;Kim, Ah-Young;Lee, Eun-Mi;Lee, Eun-Joo;Hong, Il-Hwa;Hwang, Ok-Kyung;Jeong, Kyu-Shik
    • Journal of Life Science
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    • v.26 no.5
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    • pp.592-602
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    • 2016
  • This study investigated the muscular dystrophin levels in freely moving Australian cattle mainly fed grass, freely moving Korean cattle fed mainly a grain diet, and Korean cattle fed a grain diet but housed in a relatively limited space of a cow house. The total skeletal muscle specimens of 244 cattle were collected and immediately fixed in 10% neutral formalin. The same area was biopsied from the cattle in both countries. The findings showed that fatty infiltration is highly correlated with membrane-associated protein degradation in skeletal muscle, and that among several membrane-associated proteins, dystrophin showed the most significant reduction in expression in the cattle with fatty infiltration. Similarly, CD36 was more highly expressed in the cattle with fatty infiltration of skeletal muscle. Various breeding factors, such as oxidative stress; the presence of oxidized lipids in the diet; and environmental factors such as exercise, temperature and amount of time spent, may have critical effects on the degradation of normal cytoskeleton proteins, which are required for maintaining normal skeletal muscle architecture. Among the sarcolemma membrane-associated proteins, dystrophin is the most sensitive membrane protein that is involved muscular dystrophy and muscular degeneration. Thus, the present findings may be useful for studies on muscular dystrophy in humans or the pathogenesis of muscular diseases in animal models.