• Applied Microscopy
• /
• v.31 no.2
• /
• pp.157-165
• /
• 2001

Sertoli cells in the normal adult testis are nondividing cells, which are relatively inconspicuous on cross section of the seminiferous tubule and comprise about 10% to 15% of the tubular cellular elements. Ultrastructurally, Sertoli cells have characteristic nucleoli, plasma membrane, and cytoplasmic components. The plasma membrane has two types of intercellular junctions which are developed at puberty: junctions between adjacent Sertoli cells and Sertoli cell-germ ceil junction. However, the ultrastructural findings of Sertoli cells in human fetus is not fully elucidate yet. In the present study, human fetal testes ($14\sim27$ weeks) obtained from artificially induced abortions legally without gross malformation were studied using transmission electron microscopy to make clear the differentiation process of Sertoli cells in human. In human fetal testes from 14 weeks to 27 weeks, the cell junctions of Sertoli-germ cells and Sertoli-Sertoli cells are desmosome like structure and not tight junction or desmosome. The Overall intracytoplasmic organelles of Sertoli cells are relatively sparse. The mitochondrias are relatively abundant but no developed cristae. And the rough endoplasmic reticuli are abundant and smooth endoplasmic reticuli are sparse. The amount of lipid droplets are regularly observed in human fetal Sertoli cells. No microfilaments or Charcot-Bottcher's crystalloids are present. From the results, Sertoli cells in human fetal testes are somewhat different ultrastructural findings with puberty or adult. However, to make clear the differentiation process of Sertoli cells in human, further study for 28 weeks to puberty is required.

• Applied Microscopy
• /
• v.24 no.2
• /
• pp.48-62
• /
• 1994

The acute irradiation effect on rat Purkinje cell was carried out. Anesthetized rats, weighing 200-250g each, were exposed their heads to the linear accelerator (ML-4MV) with the doses of 3,000 rads or 6,000 rads respectively. Irradiated rats were sacrificed by perfusion fixation under anesthesia, six hours, two days and six days following the irradiations. Rats were perfused with the fixative of 1% glutaraldehyde-1% paraformaldehyde solution (pH 7.4). Small pieces of cerebellar cortices were taken out. Tissue blocks were washed out, and were refixed in the 2% osmium tetroxide solution. After dehydration, tissues were embedded in the araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution, were examined with an electron microscope. The results observed were as follow; 1. Many dark Purkinje cells exhibited most severe cellular alterations on 6 hours. But after the 2 or 6 days, the cells exhibited only some alterations of cytoplasmic organelles. 2. Many granular and agranular endoplasmic reticula exhibited the fusion of cisterns. These reticular alterations were most severe on 6 hours following irradiation. But the alterations were hardly found on 6 days. 3. In the Golgi region, alterations including the adhesion of lamelliform cisterns, enlarged saccules, and increased number of vesicles, etc, were seen on 6 hours. But the Golgi complexes were almost recovered on 6 days. 4. Lysosomes were abundant on 6 hours or 2 days, but some residual bodies were found on 6 days. 5. Mitochondrial changes were also most severe at on hours, and they were recovered thereafter. From the results, it was concluded that the cerebellar Purkinje cells reacted to the high doses of irradiation by hyperactive protein synthesis, autolytic activities and energy metabolism. The reaction was most active in the early stage. It implies that motor-control function of Purkinje cells are severely disturbed in the early stage of irradiation.

• Journal of Veterinary Clinics
• /
• v.27 no.3
• /
• pp.232-239
• /
• 2010

Two cell lines derived from spontaneous canine mammary gland tumors were established and characterized. Mammary gland tumors from 9 years old pug and 9 years old toy-poodle dogs were collected by aseptic surgical resection and primary culture was performed. The histopathologic examination of tumors revealed adenocarcinoma and complex carcinoma and two dogs died from metastasis of the tumors. The tumor cells were subcultured over 60 times for more than 1 year and morphological consistency maintained. Light microscopic examination, growth curve, doubling time calculation, xenotransplantation to female nude mice, immunohistochemistry for wide spectrum keratin, vimentin, $\alpha$-smooth muscle actin and cytokeratin 8 was performed for characterization. The cell lines exhibited polygonal, elongated cell shape and cytoplasmic bridge and doubling time of 47.1 hrs and 18.6 hrs, respectively. Subcutaneous xenotransplantation to nude mice of the cells produced localized palpable mass within 4 weeks in 4 of 5 and 5 of 5 nude mice, respectively. In immunohistochemical examination one cell line showed strong positive against wide spectrum keratin and cytokeratin 8 and the other cell line showed strong positive against smooth muscle actin and cytokeratin 8. Additional characterization would be possible by investigator's needs and the cell lines may be useful for in vivo and in vitro studies of canine mammary tumor and adjuvant therapies.

• Childhood Kidney Diseases
• /
• v.12 no.1
• /
• pp.99-104
• /
• 2008

Microscopic polyangiitis(MPA) is a systemic necrotizing vasculitis that involves many organ systems including the skin, joint, kidneys, and lungs. In spite of early diagnosis and intensive care, the five-year actuarial patient and kidney survival rates are 65% and 55%. We experienced a case in 7-year-old girl of microscopic polyangiitis presenting with rapidly progressive glomerulonephritis which was confirmed by renal biopsy and positive serum perinuclear antineutrophil cytoplasmic autoantibodies(p-ANCA). The diagnosis of patients first renal biopsy was MPA, p-ANCA-associated crescentic glomerulonephritis. The patients second renal biopsy was done 5 years 6 months later since first renal biopsy, and pathologic diagnosis was chronic sclerosing glomerulonephritis, advanced, due to MPA. We began methylprednisolone pulse therapy, combined with a low dose of cyclophosphamide and plasmapheresis therapy. ACE inhibitor, angiotensin II receptor blocker, and cyclophosphamide were used until now and the patients current age is 14 years old. On admission, the patients laboratory findings showed BUN 117 mg/dL and Cr 2.3 mg/dL, while on the hospital day BUN and Cr values fell to 20.8 mg/dL and 1.6 mg/dL. But renal function was progressed to chronic failure with latest laboratory data BUN 51.7 mg/dL and Cr 3.2 mg/dL. ACE inhibitor, angiotensin II receptor blocker and small dose of immunosuppressant with close observation is the key to maintain the patient survival.

• Research in Plant Disease
• /
• v.8 no.2
• /
• pp.92-100
• /
• 2002

For the survey of viruses infected in peanut cultivated in Korea, peanut seeds and leaves showing viral symptoms were collected from their growing areas. Typical symptoms on virus infected peanut leaves including mosaic, mottle with necrosis, yellowing, stripe or vein banding and stunts were observed. Two viruses isolated from the naturally infected peanuts were identified as Bean common mosaic virus(BCMV-PSt) and Peanut mottle virus(PeMoV) by their host range, immunosorbent elcetron microscopy(ISEM), direct immuno staining assay(DISA), RT-PCR, and intracellural symptoms. Direct negative staining method by electron microscope showed filamentous particles of about 780 m in length as well as inclusion bodies. In ultrathin sections of BCMV-PSt and PeMoV infected tissues, cytoplasmic cylindrical inclusions as well as filamentous virus particles were observed in the cytoplasm of parenchyma cells. ISEM revealed filamentous particles strongly decorated with antiserums of BCMV-PSt and PeMoV Peanut seeds were stained with BCMV-PSt and PeMoV antisera indicating the possibility of seed transmission far these viruses. Seedlings germinated from peanut seeds which reacted with antiserums of BCMV-PSt by DISA showed mild mottle or stripe symptoms while mosaic and necrotic mottle symptoms were observed for PeMoV-positive seedlings. Filamentous particles were strongly decorated with each antiserum under ISEM observation. BCMV-PSt coat protein gene of about 1.2 Kbp was amplified by RT-PCR. Altogether these results indicate that BCMV-PSt is the most prevalent virus infecting peanut in Korea.

• Natural Product Sciences
• /
• v.19 no.1
• /
• pp.36-48
• /
• 2013

The aim of the present study was to investigate the effects of several fractions obtained from methylene chloride ($CH_2Cl_2$) extract of self-fermented pine needle (SFPNE) on the acetylcholine (ACh)-evoked CA release from the isolated perfused model of the rat adrenal medulla and to establish the mechanism of the most active fraction (Fr.)-induced inhibitory action on the CA release. We obtained 6 fractions from $CH_2Cl_2$ extract of self-fermented pine needle. For the ACh (5.32 mM)-evoked CA release, the following rank order of inhibitory potency was obtained: Fr.4-5 > Fr.8-11 ${\gg}$ Fr.3 > Fr.6 = Fr.7 > Fr.1-2. Fr. 4 - 5 (60 ${\mu}g/mL$) perfused into an adrenal vein for 90 min produced relatively time-dependent inhibition of the CA secretory responses to ACh (5.32 mM), DMPP (100 ${\mu}M$), McN-A-343 (100 ${\mu}M$) and high $K^+$ (56 mM). Fr. 4 - 5 itself did not affect basal CA secretion. Also, in the presence of Fr. 4 - 5 (60 ${\mu}g/mL$), the CA secretory responses to angiotensin II (AngII, 0.1 ${\mu}M$), veratridine (50 ${\mu}M$), Bay-K-8644 (10 ${\mu}M$), and cyclopiazonic acid (10 ${\mu}M$) were significantly reduced, respectively. In the simultaneous presence of Fr. 4 - 5 (60 ${\mu}g/mL$) and L-NAME (30 ${\mu}M$), the inhibitory responses of Fr. 4 - 5 on the CA secretion evoked by ACh, DMPP, high $K^+$, AngII, Bay-K-8644 and veratridine were considerably recovered to the extent of the corresponding control secretion compared with that of Fr. 4 - 5-treatment alone. The level of NO released from adrenal medulla after the treatment of Fr. 4 - 5 (60 ${\mu}g/mL$) was greatly elevated compared with the basal level. Taken together, these results demonstrate that Fr. 4 - 5 inhibits the CA secretion from the isolated perfused rat adrenal medulla evoked by stimulation of cholinergic receptors as well as by direct membrane-depolarization. It seems that this inhibitory effect of Fr. 4 - 5 is mediated by blocking the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells as well as by inhibition of $Ca^{2+}$ release from the cytoplasmic calcium store, which is evoked at least partly through the increased NO production due to the activation of NO synthase. Based on these results, it is also thought that Fr. 4 - 5 isolated from $CH_2Cl_2$ extract of pine needle may contain beneficial antihypertensive components to prevent or treat hypertension.

• Journal of Embryo Transfer
• /
• v.15 no.1
• /
• pp.67-75
• /
• 2000

This study was conducted to establish the optimal conditions for in vitro embryo production using oocytes derived from follicles of slaughter-house ovaries. The ovaries of Hanwoo were obtained from a local slaughter-house. The oocytes were aspirated from visible follicles of 2~7mm in diameter. The recovered oocytes which were completely surrounded by at least 2 layers of cumulus cells and a homogeneous cytoplasmic pigmentation were used. The selected oocytes were washed 3 or 4 times with D-PBS containing 10% bovine calf serum (BCS) and matured in vitor (IVM) in Ham's F-10 supplemented with 10% BCS or 0.01 $\mu\textrm{g}$/ml epidermal growth factor(EGF) at 39$^{\circ}C$ under 5% CO2 in air for 24 hours. They were fertilizqed in vitro (IVF) with fresh sperm separated by Percoll density gradient or swim-up in TALP media. The zygotes were cultrued with or without bovine oviductal epitherial cells(BOEC) in media(HECM-6 supplemented with 11 amino acid and / or TCM-199 supplemented with 10% BCS) for 7 to 10 days. The results obtained were as follow: The cleavage rate and developmental rate to blastocyst after maturation and IVF were not significantly different between Ham's F-10 with EGF(76.0% vs. 44.0%) and BCS(75.9% vs. 43.6%)(P<0.05). The cleavage rate and development rate to blastocyst after fertilizing by swim-up or Percoll method were not signifciantly(P<0.05) different between swim-up (80.2% vs. 29.2%) and Percoll(81.9% vs. 26.5%) (P<0.05). The cleavage rate in TCM 199(80.5) was signficiantly higher than that in HECM-6 (72.0%) (P<0.05). However, developmental rate to blastocyst using TCM 199 following HECM-6 for 3 or 4 days (42.2%) was significantly higher than that in TCM-199 alone(26.7%)(P<0.05). The cleavage rate and development rate of embryos produced in vitro by exchange timing for HECM-6 media were not significantly different between in day 3(78.6% vs. 45.5%) and day 4(75.0% vs. 43.2%)(P<0.05). The cleavage rate and developmental rate to blastocysts according to co-culture system were not significantly different between with (74.2% vs. 41.4%) and without BOEC(73.95 vs. 43.5%) (P<0.05). The number of blastomere in blastocyst stage after co-culture with or without BOEC was not significantly different (106.7$\pm$5.1 and 96.6$\pm$4.0). In conclusion, the most transferable IVP embryos could be produced from Ham's F-10 medium for IVM, Percoll density gradient method for IVF sperm separation and in vitro culture in HECM-6 until day 3 or day 4, and then transferred into TCM-199 until day 9 within adequate embryo density in culture droplets after insemination.

• Korean Journal of Food Science and Technology
• /
• v.35 no.6
• /
• pp.1182-1187
• /
• 2003

This study was performed to evaluate the potentiality of Reynoutria elliptica Migo., being used as a folk remedy and a herb medicine for urethritis, cystitis, etc., on growth inhibition of Helicobacter pylori which is known as the ulcerogenic pathogen. The minimum inhibitory concentration (MIC) value of methanol extract from Reynoutria elliptica Migo, was determined to be 120 ppm for H. pylori and urease activity derived from H. pylori was inhibited over 80% by the extract at 2 mg/mL in urea broth. Among various solvent fraction of the methanol extract, the hexane fraction showed a significant inhibitory effect on the growth of H. pylori reducing both its growth and urease activity. Scanning and transmission electron micrographs of H. pylori treated with the methanol extract at 2 mg/mL for 3 hr showed that the cell walls and membranes were disrupted so that the cytoplasmic components were leaked from the body. These results suggest that Reynoutria elliptica Migo. possesses a therapeutic potential on the gastric disease caused by H. pylori.

• Journal of radiological science and technology
• /
• v.39 no.3
• /
• pp.451-459
• /
• 2016

The development of radioprotector is being actively conducted in order to reduce the damage from over radiation exposure at radiation accident or radiation therapy. So this study was confirmed for radiation protective effects using the Cynanchi wilfordii Radix that has been known to be effective for antioxidant activity, anti-cancer, immune enhancing effects. The method of this study was administered orally Cynanchi wilfordii Radix ethanol extracts to Sprague Dawley Rat(SD Rat) for 14 days once a day, while measuring changed blood cell, spleen index, liver and uterus tissue along the change in time of 1, 4, 7 and 21 days after X-ray beam of 7 Gy irradiation. As the result of the experiment, the experimental group's rats which are administered with Cynanchi Wilfordii Radix ethanol extracts showed a rapid recovery in white blood cell count(p < 0.05) and spleen index(p < 0.05). In addition, condensation of nuclei, cytoplasmic swelling, and inflammatory cell infiltration in experimental group's liver cell was decreased more than in irradiation group's component. Further, experimental group's Uterine gland decreased the apoptosis more than irradiation group's components did. It is expected that Cynanchi Wilfordii Radix extracts will be useful as a new radioprotector. With above in mind, this paper may provide appropriate implications with the field of emergency management such as radiation accident.

• Journal of Veterinary Clinics
• /
• v.33 no.1
• /
• pp.74-79
• /
• 2016

A 7-year-old female Pointer dog with multiple masses in the axilla, mammary gland, and bladder was submitted to the Pathology Department of the College of Veterinary Medicine in the Jeju National University. Grossly, mass between right axilla and 1st mammary gland, $15{\times}10cm$ in size, was well delineated and firm, slightly soft center, oval shape. And masses in right 1st, 3rd and 5th mammary gland were well delineated and sulphur yellow in color on the cut-surface. Numerous round to oval shaped masses, 0.3 to 2 cm in diameter were existed in the lung. Urinary bladder mucosa had rough and thick and round to oval papillary masses, 0.1 to 2 cm in diameter, on surface. Microscopically, masses in right axilla, 1st mammary gland, lung and axillary lymph node were composed of poorly differentiated tubules originated from apocrine gland. Lining neoplastic epithelium showed high mitotic figures, typical apical secretory blebs, and PAS-positive diastase-resistant cytoplasmic granules. Masses in 3rd and 5th mammary gland were confirmed as mammary complex adenoma and simple adenoma respectively. The masses in the urinary bladder were covered with stratified transitional epithelium with marked cellular atypia and high mitotic figures. Some neoplastic cells showed focal invasion into substantia propria of bladder. Immunohistochemaically, neoplastic transitional epithelium demonstrated positive reactions for cytokeratin 7, AE1/AE3, and MNF116. Based on the gross, histopathologic and immunohistochemical characteristics, this dog was diagnosed as apocrine carcinoma, mammary gland tumor including simple adenoma and complex adenoma and bladder transitional cell carcinoma. And distant metastases of apocrine carcinoma in right axilla were observed in axillary lymph node and lungs. This is the first report for concurrent occurrence of apocrine carcinoma, mammary gland tumor, and transitional cell carcinoma in a same dog.