• Title/Summary/Keyword: Immature

Search Result 1,580, Processing Time 0.031 seconds

Detrimental Effect of Bovine Serum Albumin in a Maturation Medium on Embryonic Development after Somatic Cell Nuclear Transfer in Pigs

  • Lee, Hanna;Lee, Yongjin;Park, Bola;Elahi, Fazle;Lee, Joohyeong;Choi, Jung Hoon;Lee, Seung Tae;Park, Choon-Keun;Hyun, Sang-Hwan;Lee, Eunsong
    • Journal of Embryo Transfer
    • /
    • v.29 no.4
    • /
    • pp.361-368
    • /
    • 2014
  • This study was designed to evaluate the effect of bovine serum albumin (BSA) in a maturation medium on oocyte maturation and embryonic development in pigs. Immature pig oocytes were matured for 44 h in a medium supplemented with 0.4% (w/v) BSA, 0.1% (w/v) polyvinyl alcohol (PVA), or 10% (v/v) pig follicular fluid (PFF). After IVM, oocytes reached metaphase II stage were activated for parthenogenesis (PA) or used as cytoplasts for somatic cell nuclear transfer (SCNT). Nuclear maturation (89.5%, 90.7% and 91.3% for BSA, PVA and PFF, respectively) and intraoocyte glutathione contents (1.20, 1.16 and 1.00 pixels/oocyte for BSA, PVA and PFF, respectively) were not altered by the macromolecules added to maturation medium. IVM of oocytes in a medium containing BSA (21.4%) and PVA (20.7%) showed significantly lower blastocyst formation after PA than culture in medium with PFF (39.2%). After SCNT, oocytes matured in medium with BSA showed decreased embryonic development to the blastocyst stage (9.2%) compared to those matured in medium with PFF (28.9%), while 23.6% of SCNT oocytes matured in medium with PVA developed to the blastocyst stage. When the effect of BSA in a maturation medium during the first 22 h and the second 22 h of IVM in combination with PFF or PVA was examined, PVA-BSA showed a higher nuclear maturation (94.1%) than BSA-PFF (84.5%). However, there was no significant difference in the blastocyst formation among tested combinations (47.3, 52.2, 50.0, 44.4 and 49.0% for PFF-PFF, PFF-BSA, PVA-BSA, BSA-PVA and BSA-PFF, respectively). Our results demonstrate that BSA and PVA added to maturation medium can support oocyte maturation comparable to PFF-supplemented medium. However, maturation of oocytes in a BSA-containing medium decreases embryonic development after PA and SCNT when compared with the medium supplemented with PFF.

Effects of Fructose in a Chemically Defined Maturation Medium on Oocyte Maturation and Parthenogenetic Embryo Development in Pigs (돼지 난자의 체외성숙에서 합성배양액에 첨가된 과당이 난자의 성숙 및 단위발생 배아의 체외발육에 미치는 영향)

  • Shin, Hyeji;Kim, Minji;Lee, Joohyeong;Lee, Seung Tae;Park, Choon-Keun;Hyun, Sang-Hwan;Lee, Eunsong
    • Journal of Embryo Transfer
    • /
    • v.32 no.3
    • /
    • pp.139-146
    • /
    • 2017
  • The objective of this study was to determine the effect of fructose that was supplemented to a chemically defined in vitro maturation (IVM) medium on oocyte maturation and embryonic development after parthenogenesis in pigs. The base medium for in vitro maturation (IVM) was porcine zygote medium (PZM) that was supplemented with 0.05% (w/v) polyvinyl alcohol (PVA) or 10% (v/v) porcine follicular fluid (pFF). In the first experiment, when immature pig oocytes were matured in a chemically defined medium that was supplemented with 5.5 mM glucose or with 1.5, 3.0 and 5.5 mM fructose, 3.0 mM fructose resulted in a higher nuclear maturation (91.5%) than 1.5 and 5.5 mM fructose (81.9 and 81.9%, respectively) but showed a similar result with 5.5 mM glucose (94.2%). However, there was no significant differences among groups in the embryo cleavage (89.4-92.4%), blastocyst formation (37.5-41.1%), and mean cell number of blastocyst (30.8-34.2 cells). Fructose at the concentration of 3.0 mM (1.08 pixels/oocyte) resulted in a higher intra-oocyte glutathione (GSH) content than 1.5 and 5.5 mM fructose (1.00 and 0.87 pixels/oocytes, respectively) while the cumulus cell expansion was not influenced. In the second experiment, effect of individual and combined supplementation of a chemically defined maturation medium with 5.5 mM glucose or 3.0 mM fructose was examined. No significant effect was found in the nuclear maturation (86.3-92.6%). Embryo cleavage was significantly increased by the combined supplementation with glucose and fructose (95.2%) compared to that with 3.0 mM fructose only (85.7%) while blastocyst formation (37.3-42.8%) and embryonic cell number (33.3-34.1 cells) were not altered. Effect of supplementation of pFF-containing medium with glucose and fructose + glucose was examined in the third experiment. No significant effect by the supplementation with glucose and fructose or glucose alone was observed in the nuclear maturation of oocytes (90.7-94.1%) and blastocyst formation (51.0-56.5%). Our results demonstrate that 3.0 mM fructose was comparable to 5.5 mM glucose in supporting in vitro oocyte maturation and embryonic development after parthenogenesis and could be used as an alternative energy source to glucose for in vitro maturation of pig oocytes.

Detection of circulating antigens in rats experimentally infected with Paragonimus westermani by ELISA (ELISA법을 이용한 폐흡충 감염 백서의 혈청내 항원 검출)

  • 용태순;김동수
    • Parasites, Hosts and Diseases
    • /
    • v.25 no.2
    • /
    • pp.141-148
    • /
    • 1987
  • Circulating antigens in rats experimentally infected with Paragonimus westermani were examined by ELISA. From a total of 22 albino rats, each fed with 25 metacercariae, blood samples were collected until 12 weeks after infection. The specific antibodies against P. westermani in the serum of an infected cat were purified by ammonium sulfate precipitation, DEAE anion-exchange chromatography and affinity chromatography serially. So-called double antibody sandwich ELISA method was used for the detection of circulating antigens. The results were as follows: Mean value of O.D. in control sera was O. 04 (S.D.=0. 04). After infection, mean O.D.(S.D.) values were changed serially: 0.03(0.01) at 0.5 week(3 days), 0.55(0.50) at 1 week, 0.69(0.45) at 1.5 week, O.20 (0.19) at 2 weeks and O.13(0.10) at 2.5 weeks of infection. They returned, thereafter, to the level before infection. When O. 16 (mean+3 S.D.) were considered as cut-off value, those higher than O. 16 were observed only in the sera collected between 1 and 2.5 weeks after infection. Average 8. 4 immature worms (2.2 from the lungs and pleural cavities; 6.2 from muscles) were recovered in a rat at 12 weeks after infection. The fact that circulating antigens were not detected after 3 weeks of infection was considered to the caused by the formation of antigen-antibady complexs.

  • PDF

Epidemio-entomological survey of Japanese encephalitis in Korea (한국에 있어서 일본뇌염의 역학적, 매개동물학적 조사)

  • 백두현;주종윤
    • Parasites, Hosts and Diseases
    • /
    • v.29 no.1
    • /
    • pp.67-86
    • /
    • 1991
  • In order to determine the seasonal prevalence and population dynamics of Culex tritaeniorhynchus in relation to the epidemics of Japanese encephalitis, and ecology of these vector mosquito in Kyungpook Province, Korea, studies were con- ducted during the Period of 7 years from 1984 to 1990. Cx. tritaeniorhynchus first collected in June between 4th and 28th, and trapped in large numbers during the period from mid-August to early September, showed a simple sharply pointed one-peaked curve. There was a gradual decrease from mid-September, with a very small number of them collected until early October in every year. The average number of Cx. tritaeniorhynchus rapidly decreased after 1985, and the number became particularly low in 1989. The highest population density, which was observed in August during the initial three years, was found to be delayed in the following years, accompanied by a decrease in the number of mosquitoes. In the trend of nocturnal activity of Cx. tritaeniorhynchus, with oncoming darkness they become very active, gradually decreasing in activity toward mid night, but slightly increasing toward dawn. The immature stages of Cx. tritaeniorhynchus were first found in rice fields contributing to peak adult densities in mid-July. The highest average densities of Cx. trisaeniorhynchus was 14,900 per m2 on mid-August 19th. The larval Cx. tritaeniorhynchus showed high resistance levels and resistance ratios against 5 organophosphorus compounds. In the adult horisontal life table characteristics of Kyungsan colonies of Cx. tritaeniorhynchus under insectary condi- tions, life expectancy was 28.3 days for males and 59.8 days for females. The net reproductive rate was 7.8 and generation time was 25.6 days.

  • PDF

Effect of exogeneous plant growth regulators on morphogenetic response in vitro by embryo and leaf cultures of Camellia sinensis(L.) O. Kuntze (차나무 잎과 배 배양에 있어서 식물 생장조절물질이 형태형성에 미치는 영향)

  • PARK, Young Goo;AHN, In-Suk;BOZHKOV Peter
    • Korean Journal of Plant Tissue Culture
    • /
    • v.24 no.3
    • /
    • pp.129-135
    • /
    • 1997
  • Morphogenetic responses were investigated by culturing embryo and leaf explants of Korean wild type tea plant, Camellia sinensis (L.) O. Kuntze. Induction of direct somatic embryogenesis as well as adventitious and/or axillary shoots was obtained from mature zygotic embryo cultures on Murashige and Skoog (MS) basal medium having 5 to $20\mu\textrm{M}$cytokinin a lone. Morphogenetic response was decreased dramatically by the addition of auxins tested. One hundred percent of induced and isolated shoots formed roots after four weeks of culture on half-strength MS or quarter-strength Schenk and Hildebrandt (SH) media supplemented with $10\mu\textrm{M}$indole-3-butyric acid (IBA). Immature zygotic embryos were shown to be a suitable explant for embryogenic callus formation in the presence of 2, 4-dichlorophenoxyacetic acid(2, 4-D) in basal medium. Mature zygotic embryo originated leaves were used to test their ability for mophogenesis by incorporating plant growth regulators such as IBA, naphthyl-1-acetic acid (NAA), and 6-benzylaminopurine (BAP). Apparently, the morphogenetic responses of the cultured explant sources on the types and/or levels of plant growth regulators tested were observed visually.

  • PDF

Test turnaround Time for Complete Blood Cell Count using Delta and Panic Value Checks and the Q-flag Limit

  • Koo, Bon-Kyung;Ryu, Kwang-Hyun;Lim, Dae-Jin;Cho, Young-Kuk;Kim, Hee-Jin
    • Korean Journal of Clinical Laboratory Science
    • /
    • v.44 no.2
    • /
    • pp.66-74
    • /
    • 2012
  • Test turnaround time (TAT) is the lead time from reception to reporting. In the complete blood cell count (CBC), 4 units of the XE-2100 (Sysmex Corp., Japan) processed around 80% of quantity, 1 unit of the LH-780 (Beckman-Coulter Incorp., USA) processed around 10% and 1 unit of ADVIA-2120 (Siemens AG, Munich, Germany) processed around 10%. We analyzed the change in the TAT for the CBC for over 7 years, from January of 2005 to December of 2011. The delta check made alterations of delta to WBC, hemoglobin, hematocrit, platelet and metamyelocyte, however, did not made them to band neutrophil, eosinophil, basophil and monocyte. The panic value check made alterations of panic value to hemoglobin, hematocrit, platelet and monocyte. In the criteria of currently slide review, LH-780 and ADVI-2120 analyzers prepared suspect flags of "Blast, Imm NE2, Immature granulocyte, Imm NE1, Left shift, Variant lymphocyte, Atypical lymphocyte, Platelet clumps and NRBC". The New slide review in the XE-2100 analyzer altered the preparations of a smear slide more than a "Platelet clumps flag(${\geq}200unit$), a single flag excluding the "Platelet clumps flag (${\geq}250unit$) and a multiple flag (${\geq}200unit$)". Also, below the 240 unit, medical technologists prepared manual slides selectively according to their evaluations. The automatic reporting rate was 33.4% without alterations, whereas it was 41.0% without alterations, and was thus improved by 7.6%. The slide review rate was 15.2% before using the Q-flag limit, whereas it was 12.1% for a reduce 3.1%. TAT was 45 minutes without the creation alterations of the delta and panic value checks, whereas it was 35 minutes after making alterations of the delta and panic value checks and thus was shortened by 10 minutes. We came to the conclusion that the establishment and operation of delta and panic value checks and slide review criteria suitable for laboratory environment can reduce unnecessary smear slides, re-checking, re-sampling, re-testing, telephone inquiries and concentrated workloads during specific times of the day.

  • PDF

A study on Actinosporeans parasitized in Branchiura sowerbyi (Oligochaeta : Annelida) (아가미지렁이 (Branchiura sowerbyi)에 기생하는 포자충류에 관한 연구)

  • Kim, Young-Gill
    • Journal of fish pathology
    • /
    • v.8 no.1
    • /
    • pp.13-21
    • /
    • 1995
  • Four species of Actinosporeans, Aurantiactinomyxon sp. A, B, C and Neoctinomyxon sp. D were released from Oligochaete, Branchiura sowerbyi captured from three reserviors, where intestinal giant-cystic disease in carp had occured from June to September, 1994. All part of the intestinal epithelial tissue from the gullet to the anus of B. sowerbyi were infected by Actino-sporeans, and many mature Actinosporean were seen more easily at the posterior parts of the body. Just before releasing, mature Actinosporean sporozoites were divied into each individual from the intestinal epithelial tissue of Oligochaete, while immature ones had 6 spores ($20{\times}25{\mu}m$ in size) per each in the oocyst ($60{\times}65{\mu}m$ in size). A total of 1, 762 of B. sowerbyi were investigated in three reservoirs, 86 individuals (4.88%) of them were infected; 0. 74% (13 ind.) of Aurantiactinomyxon sp. A, 2. 27% (40 ind.) of Aurantiactinomyxon sp. B, 1. 59% (28 ind.) of Aurantiactinomyxon sp. C, and 0. 28% (5 ind.) of Neoactinomyxon sp. D. At the room temperature of 22.6-$30.7^{\circ}C$, number of extrusion dates of Actinosporeans from B. sowerbyi for 32 days are 1 day (23.3% of total, 1 time) or 5 days (11.7%. 5 times), and the majority was finished within 15 days, however, 6.7% of total were released for 32 days.

  • PDF

Arthscopic Treatment of Osteochondritis Dissecans in Femoral Condyle (관절경을 이용한 대퇴골과 박리성 골연골염에 대한 치료)

  • Kim Sung-Jung;Kyung Hee-Soo;Ihn Joo-Chul;Lee Seong-Man
    • Journal of the Korean Arthroscopy Society
    • /
    • v.4 no.2
    • /
    • pp.138-143
    • /
    • 2000
  • Purpose : We analyzed clinical and radiological results of the treatment of osteochondritis dissecans in the femoral condyle under arthroscopic guidance. Materials and Methods : The study group consists 19 cases in 17 patients. Average follow up period was 34 months and average age was 16 years. The cases were classified by 4 different groups, using the fellowing system: Group 1-stable lesion and no specific treatment after arthroscopic examination; Group 2-early separation and multiple drilling; Croup 3-unstable lesion and Herbert screw fixation; Croup 4-loose body removal and/or crater curettage. The results were analyzed by the criteria of Hughston which including clinical and radiologic outcomes. Results : There were 14 cases$(74\%)$ of good and excellent results in 19 knees in which, $75\%$(3/4) in Group 1, $75\%$(3/4) in Group 2, $86\%$(7/8) in Group 3 and $33\%$(1/3) in Group 4. The result of Herbert screw fixation group was better than that of other groups with statistically significant differences. Conclusion : In the treatment of osteochondritis dissecans of skekletally immature patients, arthroscopic finding was reliable guidance in decision of treatment method and active fixation was recommended in patients with large, unstable lesion.

  • PDF

Arthroscopic Reduction and Fixation of an Anterior Cruciate Ligament Avulsion Fracture From the Tibial Eminence Using Bioabsorbable Pins - Technical Note - (생흡수성 핀을 이용한 소아 경골 과간부 견열 골절의 관절경적 정복 및 고정 방법 - 수술 술기 -)

  • Lee, Su-Chan;Yang, Il-Soon;Seo, Hee-Soo
    • Journal of the Korean Arthroscopy Society
    • /
    • v.13 no.2
    • /
    • pp.183-187
    • /
    • 2009
  • Purpose: We describe a new and simple technique for arthroscopic fixation of tibial intercondylar eminence avulsion fracture using bioabsorbable pins in skeletally immature patients. Operative Technique: Diagnostic knee arthroscopy is performed using anterolateral and anteromedial portals. Fracture debris and blood clot are debrided to expose the injured site well. The fragment is reduced with the probe and fixed temporarily with a 1.1-mm diameter K-wire that is inserted percutaneously from the anterosuperior aspect of the knee joint. The drill guide is introduced into the joint and the fragment is secured by bioabsorbable, poly-p-dioxanone 1.3-mm pins inserted from different angles. The pins are 40 mm in length. The knee is placed in a long leg cast in extension for 4 weeks to assure that full extension is obtained. Conclusion: Arthroscopic fixation of an tibial intercondylar eminence avulsion fracture using bioabsorbable pins is not a technically demanding, suitable method that ensures fracture healing and restores the stability of the joint.

  • PDF

Regulatory Dendritic Cells Induced by Mesenchymal Stem Cells Ameliorate Dextran Sodium Sulfate-Induced Chronic Colitis in Mice

  • Jo, Hannah;Eom, Young Woo;Kim, Hyun-Soo;Park, Hong Jun;Kim, Hee Man;Cho, Mee-Yon
    • Gut and Liver
    • /
    • v.12 no.6
    • /
    • pp.664-673
    • /
    • 2018
  • Background/Aims: Regulatory dendritic cells (rDCs), which can be induced by mesenchymal stem cells (MSCs), play an important role in inducing and maintaining homeostasis of regulatory T cells and exhibit anti-inflammatory functions. In this study, we investigated whether MSCs could differentiate DCs into rDCs and compared the therapeutic effects of rDCs and MSCs on dextran sodium sulfate (DSS)-induced chronic colitis mice. Methods: Immature DCs (imDCs) and lipopolysaccharide (LPS)-treated mature DCs (mDCs) were co-cultured with MSCs for 48 hours, and then the profiles of surface markers and cytokines and regulatory roles of these DCs for primary splenocytes were analyzed. In addition, the therapeutic effects of MSCs and DCs co-cultured with MSCs were compared in chronic colitis mice. Results: After co-culture of imDCs (MSC-DCs) or LPS-treated mDCs (LPS+MSC-DCs) with MSCs, the expression of CD11c, CD80, CD86, interleukin 6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and interferon-${\gamma}$ (IFN-${\gamma}$), was decreased, but that of CD11b, IL-10, and transforming growth factor-${\beta}$ (TGF-${\beta}$) was increased. Furthermore, MSC-DCs and LPS+MSC-DCs induced the expression of CD4, CD25, and Foxp3 in primary splenocytes isolated from mice. In DSS-induced colitis mice, MSCs and MSC-DCs increased colon length, body weight, and survival rate and induced histological improvement. Moreover, in the colon tissues, the expression of IL-6, TNF-${\alpha}$, and IFN-${\gamma}$ decreased, but that of IL-10, TGF-${\beta}$, and Foxp3 increased in the MSC- and MSC-DC-injected groups. Conclusions: Our data suggest that MSCs differentiate DCs into rDCs, which ameliorate chronic colitis. Thus, rDCs stimulated by MSCs may be therapeutically useful for the treatment of chronic inflammatory diseases.