The killing of male germ cells by radiation and other toxicants has recently been attributed to apoptosis, but a critical evaluation of the presence of the different features of apoptosis in each epithelial stage has not been performed. In this study, mouse testes exposed to radiation were examined by light microscopy and terminal transferase-mediated end labeling (TUNEL) with periodic acid-Schiff (PAS) stains to determine whether the cells were apoptotic according to several criteria. Apoptosis was easily recognized by the presence of peroxidase-stained, entirely apoptotic bodies. In the TUNEL-positive cells or bodies, the stained products correlated precisely with the typical morphologic characteristics of apoptosis as seen at the light microscopic level. The changes that occurred from 0 to 24 hours after exposing the mice to 2 Gy of gamma-rays (2 Gy/min) were examined. The numbers of apoptotic cells reached a peak at 12 hours after irradiation and then declined. The mice that received 0-8 Gy of gamma-rays were examined 8 hours after irradiation. Dose-response relationships were generated for each stage of the epithelial cycle by counting TUNEL-positive cells. The dose-response curves were linear- quadratic [y=(-0.014${\pm}$0.009)$D^{2}$+(0.31${\pm}$0.697)D+0.3575. Where y=the number of apoptotic cells per seminiferous tubule, and D=the irradiation dose in Gy, $r^{2}$=0.9] and there was a significant relationship between the frequency of apoptotic cells and the radiation dose. Although the maximum response was produced by 8 Gy, even 0.5 Gy induced marked changes. These changes were most pronounced in B spermatogonia of stage V and the spermatocyte at the mitotic cells of stage XII.
Journal of The Korean Society of Inherited Metabolic disease
/
v.18
no.1
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pp.30-34
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2018
Niemann Pick type C disease (NPC) is an inherited progressive neurodegenerative disorder, due to defects of intracellular lipid trafficking and storage. Hepatosplenomegaly may prevail, while progressive neurodegenerative symptoms such as cerebellar involvement, dystonia, vertical supranuclear ophthalmoplegia, cataplexy, and eventually seizures starting at juvenile or late infantile period may accompany after normal early development. Here we describe a 3-year-old Korean boy with NPC who presented with splenomegaly at age 3. Liver biopsy showed characteristic foamy cell stained by periodic acid-schiff, and molecular analysis for NPC1 identified the compound heterozygous mutations, novel mutation of c.1631G>A (p.Trp544Ter) and c.2662C>T (p. Pro888Ser) as a known mutation. Filipin was strongly stained with unesterified cellular cholesterol in the patient's skin fibroblasts. The patient has received migulstat since age 3 years and his long-term outcome is needed to be observed.
The ultrastructure of the integumental epidermis of Korean planaria (Dugesia japonica) is studied by light microscope, scanning electron microscope and transmission electron microscope. The planaria has mono-layered integumental epidermis in which most of cells exhibit irregularly columnar shape. The epidermal cells of the integument are classified into six types on the basis of cytochemical and ultrastructural characteristics. 1) Ciliated epithelial cells: These cells have cilia in their free surfaces. The axonemes of cilia exhibits fundamental 9+2 microtubular pattern. 2) Eosinophilic cells: These cells contain a few large eosinophilic granules. The core of eosinophilic granule is consisted of sparsely dispersed fibrillar structures in relatively electron-lucent ground material. 3) Mucous cells: These cells are filled with irregularly shaped, PAS-positive mucous granules which have an average size of $0.8\\times0.3 \\muM$. 4) Rhabdite-forming cells: These cells possess a few strongly-eosinophilic large rhabdite granules. The rhabdite granules are synthesized either in the rhabdite-forming cells which constitute integumental epidermis or in the corresponding cells which are developed in the parenchyma and later transferred to epidermal cells of integumental epidermis through basement membrane. 5) A-type of basophilic granule cells: These granule cells possess round or irregularly-shaped granules which are strongly stained with Alcian blue. These electron-dense granules have an average size of $1.5\\times1.0 \\muM$. This type of cells is derived from parenchymal tissue. 6) B-type of basophilic granule cells: These basophilic granule cells with PAS-positive granules, are found in the epidermis of lateral body wall. The granules, which are about $0.7\\times0.4 \\muM$ in size, occupying most part of this cell type are originated from the parenchyma.
Effect of humic acid (HA), photo-oxidation and adsorption with periodic $N_2$ back-flushing was investigated in hybrid process of ceramic ultrafiltration and photocatalyst for drinking water treatment. It was compared and investigated with the previous result at water back-flushing in viewpoints of membrane fouling resistance ($R_f$), permeate flux (J), and total permeate volume ($V_T$). As decreasing HA, $R_f$ decreased dramatically and J increased, and finally $V_T$ was the highest at HA 2 mg/L. As HA concentration increased from 2 to 10 mg/L, the membrane fouling resistance after 180 mins' operation ($R_{f,180}$) improved 0.8 times more than that of water back-flushing. Therefore, HA concentration should affect on the membrane fouling at $N_2$ back-flushing than water back-flushing. Turbidity treatment efficiencies were almost constant independent of HA concentration, but HA treatment efficiency was the maximum at HA 2 mg/L. This means that adsorption and photo-oxidation of photocatalyst beads could removed HA at HA 2 mg/L, but it was not enough at 4 mg/L. Beyond HA 6 mg/L, UF could effectively treat HA by thick cake layer on membrane surface and severe inner membrane fouling.
Kim, Jae-Hyun;Kim, Gang-Phil;Ryu, Hong-Keun;Suh, Hong-Suk;Lee, Jung-Ho
Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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2008.06a
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pp.241-241
/
2008
Macrofore formation in silicon and other semiconductors using electrochemical etching processes has been, in the last years, a subject of great attention of both theory and practice. Its first reason of concern is new areas of macropore silicone applications arising from microelectromechanical systems processing (MEMS), membrane techniques, solar cells, sensors, photonic crystals, and new technologies like a silicon-on-nothing (SON) technology. Its formation mechanism with a rich variety of controllable microstructures and their many potential applications have been studied extensively recently. Porous silicon is formed by anodic etching of crystalline silicon in hydrofluoric acid. During the etching process holes are required to enable the dissolution of the silicon anode. For p-type silicon, holes are the majority charge carriers, therefore porous silicon can be formed under the action of a positive bias on the silicon anode. For n-type silicon, holes to dissolve silicon is supplied by illuminating n-type silicon with above-band-gap light which allows sufficient generation of holes. To make a desired three-dimensional nano- or micro-structures, pre-structuring the masked surface in KOH solution to form a periodic array of etch pits before electrochemical etching. Due to enhanced electric field, the holes are efficiently collected at the pore tips for etching. The depletion of holes in the space charge region prevents silicon dissolution at the sidewalls, enabling anisotropic etching for the trenches. This is correct theoretical explanation for n-type Si etching. However, there are a few experimental repors in p-type silicon, while a number of theoretical models have been worked out to explain experimental dependence observed. To perform ordered macrofore formaion for p-type silicon, various kinds of mask patterns to make initial KOH etch pits were used. In order to understand the roles played by the kinds of etching solution in the formation of pillar arrays, we have undertaken a systematic study of the solvent effects in mixtures of HF, N-dimethylformamide (DMF), iso-propanol, and mixtures of HF with water on the macrofore structure formation on monocrystalline p-type silicon with a resistivity varying between 10 ~ 0.01 $\Omega$ cm. The etching solution including the iso-propanol produced a best three dimensional pillar structures. The experimental results are discussed on the base of Lehmann's comprehensive model based on SCR width.
Jo, Un-Bock;Jung, Kwun-Soon;Hong, Mal-Sook;Jo, Gi-Jin
Journal of Life Science
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v.10
no.2
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pp.1-8
/
2000
This experiment was performed to study the structure of esophageal mucousa and the histochemical properties of glycosaminoglycans of esophageal mucous cells in four teleostean species, i.e., Agramus agramus, Inimicus japonicus, Epinephelus chlorostigma and Helicolenus dactylopterus. To observe the structure of eosphageal mucosa, hematoxylin-eosin(H-E) staining was used. The glycosaminoglycans was stained with PAS(periodic acid schiff), alcain blue(AB) pH 2.5, AB pH 1.0, aldehyde fuchsin(AF) pH 1.7, AF pH 1.0, AB pH 2.5-PAS, AB pH 1.0-PAS and AF pH 1.7-alcian blue pH 2.5. As for the amount and histochemical composition of glycosaminglycans in Agramus agramus, most of mucous secreting columnar cell and mucous cells contain large and moderate amount of neutral glycosaminoglycans. A few of mucous cells having small amount of neutral glycosamino-glycans and minimal amount of sulfated glycosamino-glycans. In Inimicus japonicus, the esophageal mucous cells were composed of most of medium sized and large mucous cells with moderate amount of neutral glycosaminoglycanonly only, a few of medium sized and large mucous cells and most of small mucous cells with considerable amount of neutral glycosaminoglycans and minimal to small amount of nonsulfated glycosaminoglycans, and a feq of small mucous cells with small amount of neutral glycosaminoglycans and minimal amount of sulfated glycosaminoglycans. In Epinephelus chlorostigma, most of medium sized and large mucous cells were mixed small amount of neutral glycosaminoglycans with sulfated glycosaminoglycans, a few of which were contained with moderate or considerable amount of neutral glycosaminoglycans with sulfated glycosaminoglycans, while most of small mucous cells containing considerable amount of neutral glycosaminoglycans and small to moderate or considerable to minimal amount of nonsulfated glycosaminoglycans(sialomucin) a few of which containing a mixture of considerable amount of neutral glycosaminoglycans and considerable amount of nonsulfated glycosaminoglycans(sialomucin)only. In Helicolenus dactylopterus, most of medium sized and large mucous cells, mixing with moderate to considerable amount of neutral glycosaminoglycans and small to moderate amount of sulfated glycosaminoglycans, while most of small mucous cells with considerable amount of nonsulfated glycosaminoglycans (sialomucin), a few of which having a mixture of considerable amount of neutral glycosaminoglycans and considerable or small amount of nonsulfated glycosaminoglycans(sialomucin).
Park, Mi-Yeon;Moon, Bo-Mi;Gang, Su-Jin;Lee, Jong-Ha;Park, Jin-Woo;Cho, Sung-Woo;Her, Cheol-Ho
Korean Journal of Veterinary Service
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v.44
no.2
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pp.73-83
/
2021
Although many swine farms continuously vaccinated to sow to prevent Porcine epidemic diarrhea(PED), PED has occurred annually in swine herds in Jeonbuk province, Korea. In the present study, the small intestine and feces samples from 17 farms where severe watery diarrhea and death of newborn piglets occurred in 2019 were collected, amplified by RT-PCR and determined the complete nucleotide sequences of the spike (S) glycoprotein genes of nine Jeonbuk PEDV isolates. The spike (S) glycoprotein is an important determinant for molecular characterization and genetic relationship of PEDV. These nine complete S gene isolates were compared with other PEDV reference strains to identify the molecular diversity, phylogenetic relationships and antigenicity analysis. 9 field strains share 98.5~100% homologies with each other at the nucleotide sequence level and 97.3~100% homologies with each other at the amino acid level. The nine Jeonbuk PEDV isolates were classified into G2b group including a genetic specific signal, S-indels (insertion and deletion of S gene). In addition, comparisons the neutralizing epitopes of S gene between 9 field strains and domestic vaccine strains of Korea mutated 12-15 amino acids with SM-98-1 (G1a group) and mutated 0-3 amino acids with QIAP1401 (G2b group). Therefore, the development of G2b-based live vaccines will have to be expedited to ensure effective prevention of endemic PED in Korea. In addition, we will need to be prepared with periodic updates of preventive vaccines based on the PEDV variants for the re-emergence of a virulent strain.
Han, Byung Hyuk;Lee, Hyeon Kyoung;Jang, Se Hoon;Tai, Ai Lin;Yoon, Jung Joo;Kim, Hye Yoom;Lee, Yun Jung;Lee, Ho Sub;Kang, Dae Gill
Herbal Formula Science
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v.29
no.1
/
pp.33-44
/
2021
Renal ischemia-reperfusion injury(IRI), an important cause of acute renal failure (ARF), cause increased renal tubular injury. Jesaeng-sinkihwan (JSH) was recorded in a traditional Chines medical book named "Bangyakhappyeon (方藥合編)". JSH has been used for treatment of diabetes and glomerulonephritis with patients. Here we investigate the effects of Jesaeng-sinkihwan (JSH) in a mouse model of ischemic acute kidney injury. The animals model were divided into four groups at the age of 8 weeks; sham group: C57BL6 male mice (n=9), I/R group: C57BL6 male mice with I/R surgery (n=9), JSH Low group: C57BL6 male mice with surgery + JSH 100 mg/kg/day (n=9) and JSH High group: C57BL6 male mice with surgery + JSH 300 mg/kg/day (n=9). Ischemia was induced by clamping the both renal arteries during 25 min, and reperfusion was followed. Mouse were orally given with JSH (100 and 300 mg/kg/day during 3 days after surgery. Treatment with JSH significantly ameliorates creatinine clearance(Ccr), Creatinine (Cr) and blood urea nitrogen(BUN) in obtained plasma. . Treatment with JSH reduced kidney inflammation markers such as Neutrophil Gelatinase Associated Lipocalin (NGAL) and kidney injury molecule-1 (KIM-1). JSH also reduced the periodic acid schiff (PAS) staining intensity and picro sirius red staining intensity in kidney of I/R group. These findings suggest that JSH ameliorates tubular injury including renal dysfunction in I/R induced ARF mouse.
Ulcerative colitis is a disease that causes inflammation in the mucosal or submucosal layer of the colon. Previous studies have reported that obesity increases the prevalence of ulcerative colitis and aggravates the progression. This study was therefore undertaken to investigate whether curcumin inhibits the progression of ulcerative colitis caused by obesity. Mice were bred on a high-fat diet to induce obesity, and curcumin was administered with the high-fat diet to confirm the anti-inflammatory effect. To induce ulcerative colitis, dextran sulfate sodium (DSS) was administered orally, and clinical symptoms of colitis were subsequently observed. For histological evaluation of curcumin, the colon, liver and abdominal fat tissue samples were prepared and analyzed by hematoxylin and eosin (H&E) and Alcian blue-periodic acid-Schiff (PAS) staining. Our results confirm that consumption of curcumin resulted in decreasing the score of the disease activity index, and inhibited shortening of the colon length. In addition, inflammatory cell infiltration and mucosal damage were inhibited in the colon tissue of ulcerative colitis exacerbated by obesity. We further confirmed that exposure to curcumin significantly reduced the steatosis area of the liver and adipocytes of abdominal fat. In conclusion, we believe that curcumin can be applied as a therapeutic agent to treat ulcerative colitis, by inhibiting the progression of colitis caused by obesity.
Objective: Choline deficiency, one main trigger for nonalcoholic fatty liver disease (NAFLD), is closely related to lipid metabolism disorder. Previous study in a choline-deficient model has largely focused on gene expression rather than gene structure, especially sparse are studies regarding to alternative splicing (AS). In modern life science research, primary hepatocytes culture technology facilitates such studies, which can accurately imitate liver activity in vitro and show unique superiority. Whereas limitations to traditional hepatocytes culture technology exist in terms of efficiency and operability. This study pursued an optimization culture method for duck primary hepatocytes to explore AS in choline-deficient model. Methods: We performed an optimization culture method for duck primary hepatocytes with multi-step digestion procedure from Pekin duck embryos. Subsequently a NAFLD model was constructed with choline-free medium. RNA-seq and further analysis by rMATS were performed to identify AS events alterations in choline-deficency duck primary hepatocytes. Results: The results showed E13 (embryonic day 13) to E15 is suitable to obtain hepatocytes, and the viability reached over 95% by trypan blue exclusion assay. Primary hepatocyte retained their biological function as well identified by Periodic Acid-Schiff staining method and Glucose-6-phosphate dehydrogenase activity assay, respectively. Meanwhile, genes of alb and afp and specific protein of albumin were detected to verify cultured hepatocytes. Immunofluorescence was used to evaluate purity of hepatocytes, presenting up to 90%. On this base, choline-deficient model was constructed and displayed significantly increase of intracellular triglyceride and cholesterol as reported previously. Intriguingly, our data suggested that AS events in choline-deficient model were implicated in pivotal biological processes as an aberrant transcriptional regulator, of which 16 genes were involved in lipid metabolism and highly enriched in glycerophospholipid metabolism. Conclusion: An effective and rapid protocol for obtaining duck primary hepatocytes was established, by which our findings manifested choline deficiency could induce the accumulation of lipid and result in aberrant AS events in hepatocytes, providing a novel insight into various AS in the metabolism role of choline.
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