• Title/Summary/Keyword: single cell injection

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Enhanced Efficacy of Human Brain-Derived Neural Stem Cells by Transplantation of Cell Aggregates in a Rat Model of Parkinson's Disease

  • Shin, Eun Sil;Hwang, Onyou;Hwang, Yu-Shik;Suh, Jun-Kyo Francis;Chun, Young Il;Jeon, Sang Ryong
    • Journal of Korean Neurosurgical Society
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    • v.56 no.5
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    • pp.383-389
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    • 2014
  • Objective : Neural tissue transplantation has been a promising strategy for the treatment of Parkinson's disease (PD). However, transplantation has the disadvantages of low-cell survival and/or development of dyskinesia. Transplantation of cell aggregates has the potential to overcome these problems, because the cells can extend their axons into the host brain and establish synaptic connections with host neurons. In this present study, aggregates of human brain-derived neural stem cells (HB-NSC) were transplanted into a PD animal model and compared to previous report on transplantation of single-cell suspensions. Methods : Rats received an injection of 6-OHDA into the right medial forebrain bundle to generate the PD model and followed by injections of PBS only, or HB-NSC aggregates in PBS into the ipsilateral striatum. Behavioral tests, multitracer (2-deoxy-2-[$^{18}F$]-fluoro-D-glucose ([$^{18}F$]-FDG) and [$^{18}F$]-N-(3-fluoropropyl)-2-carbomethoxy-3-(4-iodophenyl)nortropane ([$^{18}F$]-FP-CIT) microPET scans, as well as immunohistochemical (IHC) and immunofluorescent (IF) staining were conducted to evaluate the results. Results : The stepping test showed significant improvement of contralateral forelimb control in the HB-NSC group from 6-10 weeks compared to the control group (p<0.05). [$^{18}F$]-FP-CIT microPET at 10 weeks posttransplantation demonstrated a significant increase in uptake in the HB-NSC group compared to pretransplantation (p<0.05). In IHC and IF staining, tyrosine hydroxylase and human ${\beta}2$ microglobulin (a human cell marker) positive cells were visualized at the transplant site. Conclusion : These results suggest that the HB-NSC aggregates can survive in the striatum and exert therapeutic effects in a PD model by secreting dopamine.

Influences of dietary flavonoid (quercetin) supplementation on growth performance and immune response of growing pigs challenged with Escherichia coli lipopolysaccharide

  • Park, Jae-Hong;Sureshkumar, Shanmugam;Kim, In Ho
    • Journal of Animal Science and Technology
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    • v.62 no.5
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    • pp.605-613
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    • 2020
  • This study was conducted to evaluate the effects of dietary supplementation of plant flavonoid (quercetin) on immune parameters, growth performance, and nutrient digestibility in growing pigs challenged with Escherichia coli lipopolysaccharide (LPS). A total of 40 crossbred ([Landrace × Yorkshire] × Duroc) growing pigs; initial body weight (BW) of 26.95 ± 1.26 kg were used in a six-week experimental trial. Pigs were randomly allocated into one of four treatment groups in a 2 × 2 factorial arrangement with the following factors; without LPS challenge and with LPS challenge (day 21) supplemented with or without 0.1% flavonoid according to BW (2 replicate pens per treatment with 2 gilts and 3 barrows per pen). The single-dose LPS (100 ug / kg BW) injection showed trends tended to be increased in interleukin-6 (IL-6) after 2 h and 6 h of challenge compared with unchallenged pigs. However, other measured immune indices (white blood cell, immunoglobulin G, lymphocyte, and tumor necrosis factor), growth performance, and nutrient digestibility were not significantly different between challenged and non-challenged animals. The supplementation of flavonoid significantly increased (p < 0.05) average daily gain (ADG) during day 0-21, tended to increase dry matter and nitrogen digestibility, significantly reduced IL-6, increased Ig-G and WBC concentrations and increased lymphocytes percentage regardless of LPS challenge.

Fumonisin $B_1$ induced renal toxicity in Sprague-Dawley rats

  • Lim, Chae-Woong;Yoon, Hyun-Sang;Rim, Byung-Moo
    • Korean Journal of Veterinary Pathology
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    • v.3 no.1
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    • pp.7-14
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    • 1999
  • We investigated the effect of a single intravenous dose of Fumonisin $B_1(FB_1$) of rat kidney on the time sequence. Male Sprague-Dawley rats were intravenouslyin jected with FB$_1$at 1.25 mg/kg and were euthanized at 12 hrs, 1, 2, 4, and 6 days after the injection. In $FB_1$ treated rats, serum BUN and creatinine were elevated from 12 hrs. Microscopically, the initial target site was tubules of inner stripe, with mild degenerative and necrotic changes at 12 hrs, but the tubules recovered on day 4. In outer stripe, there were only a few scattered necrotic cells on day 1. These changes became more obvious over the time passed and most severe on day 4. On day 6, regeneration occurred, manifest as hypertrophic, basophilic tubular cells. The dying cells were proved to necrotic cells instead of apoptotic cells by TUNEL. Ultrastructural changes were cytoplasmic vacuole, dilated endoplasmic reticulum, swollen mitochondria, ballooned microvilli of the tubular cell in the outer stripe. These results showed that the renal tubules of outer medulla were the target to $FB_1$-induced nephrotoxicity in the rat. However, initial target was mner stnpe of medulla.

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Invasion of Calponin-positive Glomerular Parietal Epithelial Cells into Glomerular Tuft Is Related to the Development of Glomerulosclerosis

  • Choi, Jae-Youn;Nam, Sun-Ah;Cha, Jung-Ho
    • Applied Microscopy
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    • v.44 no.4
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    • pp.117-122
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    • 2014
  • We previously have reported that periglomerular calponin expression of the glomerulosclerotic glomeruli in the chronic nephropathy. To investigate the role of calponin during glomerulosclerosis, we examined the detailed localization pattern of calponin in chronic nephropathy rat model using serial morphometric analysis. Male Sprague-Dawley rats were used, and chronic nephropathy models were established at 8 and 12 weeks after single intraperitoneal injection of adriamycin (10 mg/kg body weight; n=5). In nephropathy models, 16.3% (8 weeks) and 23.4% (12 weeks) glomeruli showed calponin-positivity at glomerular area. In all these glomeruli, showing various sclerotic changes, calponin-immunoreactivities were present only both the glomerular parietal epithelial cells (PECs) and periglomerular myofibroblasts (PMFs). However, in the glomeruli with weak calponin-positive, immunoreactivity was mostly detected in PECs, suggesting that calponin may be expressed in PECs earlier than in PMFs in the glomerulosclerotic change. Some calponin-positive PECs invaded glomerular tuft with loop-shaped projection, and around this projection, nestin expression of glomerular tuft were much reduced. These results suggested that calponin-positive PECs may play a key role in the development of glomerulosclerosis, and direct contact with PECs and glomerular tuft may be more important to degenerative changes of glomeruli.

Production of Identical Rabbit Offspring by Nuclear Transplantation (토끼 핵이식에 의한 복제산자의 생산효율 향상에 관한 연구)

  • 박충생;윤희준;조성근;노규진;이효종;최상용
    • Journal of Embryo Transfer
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    • v.14 no.3
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    • pp.195-201
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    • 1999
  • This study was carried out to improve a technique of cloned animal prodcution by preactivation of nuclear recipient oocytes with ionomycin and 6-dimethylaminopurine (6-DMAP) in rabbits. The oocytes were collected from the oviduct of superovulated rabbit at 19∼20 hours post hCG injection. The collected oocytes were preactivated and self-enucleated by treating 5 uM ionoycin for 5 min, and 2.0 mM 6-DMAP for two hours. Microsurgical removel of the chromation complex in the second polar bodies was effectively performd and single blastomere separated from 32-cell stage rabbit embryos was injected into the perivitelline space of the enculeated recipient oocyts. Follwoing electrofusion and in vitro culture for 18 hours, the nuclear transplant(NT) embryos were transferred into the uterine horns of naturally mated or synchronized recipient does. When 32 NT embryous reconstituted with preactivated oocytes were transferred to 2 recipient does, one foster doe delivered two offspring (6.3%), while not a offspring was delivered from three foster does which received 17 NT embryos reconstituted with non-preactivated oocytes. A total of 68 NT embryos reconstituted with preactivated oocytes were transferred into the uterine horns of 7 synchronized ecipient does. Among them, two recipients were pregnant and delivered three offspring(5.9%).

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The application of electrochemotherapy in three dogs with inoperable cancers

  • Yeom, Seung Chul;Song, Kun Ho;Seo, Kyoung Won
    • Korean Journal of Veterinary Research
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    • v.61 no.1
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    • pp.9.1-9.6
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    • 2021
  • Factors such as location, volume, and the type of neoplasm complicate achieving tumor control. Electrochemotherapy (ECT) is a supplementary treatment for inoperable neoplasms in veterinary patients. Three dogs were diagnosed with a tumor. Two were squamous cell carcinoma (SCC), and the other was liposarcoma, each with a single tumor with the size range of 1 to 5 cm. The tumor locations were the cervical, oral, and abdominal cavity. ECT was selected as a treatment. Bleomycin was injected intratumorally at the dose of 0.5 to 1.0 mg/㎤. Five minutes after the injection, electric pulses applied in a sequence of eight pulses lasting 100 μsec each, were delivered in 1,000 V/cm. An evaluation was performed after 1 week, and the next session was administered 2 weeks later. In a patient with oral SCC, the tumor was in partial remission after two sessions of ECT. Another patient with SCC on her neck was showed complete remission after 2 weeks of ECT administration. A third patient showed stable disease for 8 weeks. Complications were mild and transient and included skin necrosis, edema, local pain, and gait disturbance. ECT is a valid adjuvant, especially for inoperable, cutaneous, or accessible intra-abdominal tumors.

Protective Effect of Panax ginseng on TCDD-induced Liver Toxicity in Male Guinea Pigs

  • Hwang, Seock-Yeon;Wee, Jae-Joon;Kim, Si-Kwan;Youn, Nae-Young
    • Biomedical Science Letters
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    • v.9 no.1
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    • pp.51-58
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    • 2003
  • This study was carried out to investigate the preventive and therapeutic effect of Panax ginseng water extract (PG-WE) on the toxicity induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), one of the most notorious toxic environmental pollutants belonging to the group of polyhalogenated aromatic hydrocarbons. Normal control (NC) group guinea pigs (180~200 g) received vehicle and saline, and TCDD-treated (TT) group was given TCDD and saline. P100 and P200 group animals received PG-WE for 28 days since 1 week before TCDD exposure at daily doses of 100 mg/kg b.w. and/or 200 mg/kg b.w., respectively. C100 and C200 group received PG-WE for 14 days starting 1 week after TCDD-exposure. Toxicity was induced by a single intraperitoneal injection of TCDD (1 $\mu\textrm{g}$/kg b.w.). Abnormal increase in AST and ALT activities in TT group was significantly improved by the administration of PC-WE. Microscopically, there were mild to moderate swelling of hepatocytes, hyperchromatism of individual cells, acidophilic cytoplasm and cytoplasm vacuolation of some hepatocytes, slight to moderate variations of staining density, occasional single cell necrosis, variable size and shape of some hepatocytes, small groups of degenerating hepatocytes surrounded by mononuclear cells, dilated sinusoids of centrilobular zone and some loss of lobular architecture in TT group liver. From these results, we could find the protective and therapeutic role of PG-WE in TCDD-induced liver toxicity by examining the blood chemical parameters and histopathological observation.

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AN IMMUNOHISTOCHEMICAL STUDY ON DNA SYNTHESIS OF SALIVARY GLAND TISSUE CEllS AND ENDOTHELIAL CELL AFTER IRRADIATION (방사선조사 후 타액선 세포와 혈관 내피세포의 DNA합성에 관한 면역조직학적 연구)

  • Shin Jong-Sup;You Dong-Soo
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.21 no.2
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    • pp.183-197
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    • 1991
  • After single fraction of 2, 5, 10 Gy irradiation on submandibular gland of 40 male rats, weighing 150gm, respectively, these animal were sacrificed two hours after 0.1㎎/g bromodeoxyuridine (Sigma) peritoneal injection in 1, 3, 7, 15 hours, 1, 3, 7 days after irradiation. And excised submandibular gland were fixed in Carnoy's and Bouin's solution for 2 hours. Paraffin sections were stained with H&E, and PAS for the observation of the change of salivary gland tissue, and with Feulgen for the study of the DNA distribution, and immunohistochemically stained with anti-bromodeoxyuridine (Sanbyo Co.) for detection of DNA synthetic cells in order to study the distribution of DNA synthetic cells of salivary gland tissue and endothelium after irradiation in 5 different sites of 6 slides on X 200 high power field. The results were as followings. 1. In PAS staining 3 days after 5Gy irradiation, decreased mucine secretion of serous cells were found, and 7 days after l0Gy irradiation, decreased mucine secretion of mucous cells were found. 2. In histopathologic features, degeneration of serous cells were found in 3 days after 2 Gy irradiation and there was little change in mucous cells and excretory duct cells. 3. In Feugen staining, 3 days after 2 Gy, 5 Gy irradiation, more high percentage of DNA synthetic cells were found in intercalated duct cells, striated duct cells and excretory duct cells than in BrdU staining. 4. In immunohistochemical features, DNA synethsis of serous cells and granular convoluted tubular cells abruptly decreased in early period after irradiation and showed no recovery in 7 days after irradiation but there was an increase in DNA synthesis of intercalated duct cells, striated duct cells and excretory duct cells, which have less S-phase cells comparatively, in 7 days after 2 Gy, 5 Gy irradiation. 5. In immunohistochemical features, the DNA synthesis of endothelial cells was continuously decreased after irradiation but showed slight increase in 7 days after 2 Gy and S Gy irradiation.

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Imperatorin is Transported through Blood-Brain Barrier by Carrier-Mediated Transporters

  • Tun, Temdara;Kang, Young-Sook
    • Biomolecules & Therapeutics
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    • v.25 no.4
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    • pp.441-451
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    • 2017
  • Imperatorin, a major bioactive furanocoumarin with multifunctions, can be used for treating neurodegenerative diseases. In this study, we investigated the characteristics of imperatorin transport in the brain. Experiments of the present study were designed to study imperatorin transport across the blood-brain barrier both in vivo and in vitro. In vivo study was performed in rats using single intravenous injection and in situ carotid artery perfusion technique. Conditionally immortalized rat brain capillary endothelial cells were as an in vitro model of blood-brain barrier to examine the transport mechanism of imperatorin. Brain distribution volume of imperatorin was about 6 fold greater than that of sucrose, suggesting that the transport of imperatorin was through the blood-brain barrier in physiological state. Both in vivo and in vitro imperatorin transport studies demonstrated that imperatorin could be transported in a concentration-dependent manner with high affinity. Imperatorin uptake was dependent on proton gradient in an opposite direction. It was significantly reduced by pretreatment with sodium azide. However, its uptake was not inhibited by replacing extracellular sodium with potassium or N-methylglucamine. The uptake of imperatorin was inhibited by various cationic compounds, but not inhibited by TEA, choline and organic anion substances. Transfection of plasma membrane monoamine transporter, organic cation transporter 2 and organic cation/carnitine transporter 2/1 siRNA failed to alter imperatorin transport in brain capillary endothelial cells. Especially, tramadol, clonidine and pyrilamine inhibited the uptake of [$^3H$]imperatorin competitively. Therefore, imperatorin is actively transported from blood to brain across the blood-brain barrier by passive and carrier-mediated transporter.

Multiple Ovulations and In vitro Fertilization in the Domestic Fowl (Gallus domesticus)

  • Han, Haitang;Zhao, Chen;Li, Zandong
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1652-1656
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    • 2004
  • The aim of this study was to obtain mature ova or embryos at a single cell stage, which can be used in avian transgenesis and nuclear transfer through multiple ovulations, in vitro fertilization and culture. Chicken anterior pituitary extract (CAPE) or acetone-dried chicken anterior pituitary extract (ACAPE) was used to induce multiple ovulations in hens pretreated with pregnant mare' serum gonadotrophin (PMSG). In vitro fertilization of the multiple ovulated ova was performed by inseminating sperm onto the germinal disks in m-Ringer' solution and incubating the ova at 41$^{\circ}C$, 5% $CO_2$ for 10 h in DME-F12 medium containing 20% liquid albumen. The in vitro fertilization process was observed using an environmental scanning electron microscope. When normal laying hens (white Leghorn) were administered daily with PMSG (100 IU), egg laying ceased in most hens within 3 to 8 days. Ovulation began to occur about 7.5 h after injection of CAPE and ACAPE. The number of ovulated ova was 1.00${\pm}$0.00, 2.33${\pm}$0.52 and 2.20${\pm}$0.45, respectively, after receiving 100, 200 and 300 mg CAPE. The number of ovulated ova was 2.00${\pm}$0.00, 2.86${\pm}$0.69 and 3.00${\pm}$1.22, respectively, after receiving 10, 15 and 20 mg ACAPE. The fertilized and cultured ova were able to develop into embryos up to the 32 cell stage. The present experiments demonstrated that multiple ovulations can be induced by CAPE and ACAPE successfully, and the ova resulted from the treatment retained the capability for further fertilization and embryonic development. These data provide new information to support the establishment of an in vitro culture system for future avian transgenesis studies.