The studies were carried out to obtain the basic data for maximizing the protoplast yields from the mycelia of P. ostreatus and P. sajor-caju. Some factors affecting the regeneration of the protoplast of both species and the productivity of their reversion were also examined. The maximum yields of protoplasts were obtained from four days cultured mycelia of both species on cellophan membrane placed on the surface of PSA or MCM media in a petri dish. The optimal concentration of lytic enzyme Novozym 234 for protoplast releasing was 5 mg per ml of 0.5 M phosphate buffer solution with 0.6 M sucrose or 0.6 M $MgSO_4$ at pH 6.0. The greatest number of protoplasts was released 3 hours after incubation of the mycelia of P. ostreatus and after 4 hours for the P. sajor-caju in the lytic enzyme solution. Among the osmotic stabilizer solutions tested 0.6 M sucrose and 0.6 M KCl showed the best regeneration rates of the protoplasts of both species. When 0.75 % agar solution was over-layed on the regeneration media immediately after inoculation of the protoplast the regeneration rates were greatly enhanced. The ampicillin added to the agar solution prevented bacteria from infection. The reverted isolates produced the sporophores and basidial spores just like their parents without any mutations when they were cultivated in a broad mouth bottle with sawdust substrates.
Kim, K.H.;Lee, J.H.;Oh, Y.G.;Kang, S.W.;Lee, S.C.;Park, W.Y.;Ko, Y.D.
Journal of Animal Science and Technology
/
v.47
no.5
/
pp.731-744
/
2005
Ninety Hanwoo steers(initial BW=167.2$\pm$13.4kg) were used to determine the effect of energy concentrations in concentrates and slaughter age on performance, carcass characteristics. Steers were allotted by BW to one of eighteen pens. Eighteen pens were randomly allotted to a low(70-70-71-72% for the growing, the early, the middle or the late fattening periods), medium(70-71-72-73%) or high(70-72- 73-74%) TDN level of concentrates. Five steers for each treatment of energy level were slaughtered every one month from 26 month of age to 31 month of age. Concentrates was fed restrictedly to achieve a predicted gain of 0.7-0.9kg from growing stage to middle fattening stage. All steers were fed orchard grass(Dactylis glomerata L.) hay as roughage during the growing period, fed rice straw gradually substituted for orchard grass hay during the early fattening period, and fed rice straw only thereafter. Overall body weight and feed intake were not affected by TDN levels of concentrates. Average daily gain for all treatments was higher than 0.9kg/d during the 19-21 month of age and decrease thereafter, but sustained above 0.7kg/d. Mean concentrates intake for all treatments was 1.0-1.3% of live BW during the growing period and 1.5% during the early fattening. Thereafter, it decreased up to 1.4% during the middle fattening and 1.0% during late fattening period. Delay of slaughter end point resulted in a gradual increase of rib-eye area, back fat thickness and marbling score, especially after slaughter age of 29 month there was significant increases(P<0.05). The appearance rate of 1+ and 1 grade related to the slaughter ages was 100% at 29, 30 and 31 months of age, whereas those at 26, 27 and 28 months were 93, 86 and 80%, respectively. Dressing rate was significantly(P<0.05) increased and rate of retailed cut weight significantly(P<0.05) decreased when slaughter age increased. In economic analysis, there was pronounced increase in net income up to 32-46% after slaughter age of 29 months. Under the conditions of this study, high TDN intake is not necessarily required for high quality Hanwoo meat production and slaughter age of 29 month might be the optimum for Hanwoo steers.
The objective of this study was to supply excellent genetic resources to livestock farms by transferring embryos produced by genetically superior Korean cows (Hanwoo). Eighty Hanwoo donors were superovulated with gonadotropin ($Folltrpin^(R)\;or\;Antorin^(R)$) for 4 days combined with or without progesterone releasing intravaginal device (CIDR) insertion. The collected fresh or frozen-thawed embryos were transferred to 226 farm recipients. In this study, the effect of CIDR insertion in combination with gonadotropin ($Folltrpin^(R)$) treatments initiated at the random stage of estrous cycle on embryo production was evaluated and compared to conventional superovulation protocol. Moreover, the effect of gonadotropin ($Antorin^(R)$) dose in CIDR-treated Hanwoo donors on the embryo yield was determined. In addition, the effects of embryos (fresh vs. frozen-thawed), embryo transfer person, seasons and farms on the pregnancy rate were evaluated. In Hanwoo donors, CIDR insertion in combination with $Folltrpin^(R)$ treatments regardless of estrous detection resulted in increased numbers of total ova (6.5 vs. 5.8) and transferable embryos (3.9 vs. 3.2) compared to the conventional superovulation protocol (p<0.01). In CIDR-treated Hanwoo donors, the higher dose of $Antorin^(R)$ (36 vs. 28 mg) resulted in the increased number of transferable embryos (8.3 vs. 5.4, p<0.05). The embryos (fresh 43.9% vs. frozen-thawed 23.1%) and embryo transfer person (53.9 vs. $0{\sim}16.7%$) significantly affected the pregnancy rate after embryo transfer (p<0.01). These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos and, multiple ovulation and embryo transfer in Hanwoo might be effectively applied for livestock improvement if pregmancy rate with frozen-thawed embryos and embryo transfer skill would be improved.
The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.
Background : Complicated exudative pleural fluid collections have traditionally been treated by either closed tube thoracostomy drainage or by open surgical drainage. Complete drainage is important in order to control pleural sepsis, restore pulmonary function, and entrapment. Recently intracavitary fibrinolytic therapy has been advocated as a method to facillitate drainage of complicated exudative pleural effusion and to allow enzymatic debridemant of the restrictive fibrinous sheets covering the pleural surface. The purpose of this study is to prospectively evaluate the effects of image-guided catheter drainage with high dose urokinase(UK) instillation in the treatment of complicated pleural effusions. Patients : Twenty complicated pleural effusion patients that poorly respond to image-guided drainage were allocated to receive UK. There were 8 pneumonia and 12 tuberculosis. Methods : Drugs were diluted in 250 mL normal saline and were infused intrapleurally through the chest tube or pig-tail catheter in a daily dose of 250,000 IU of UK. Response was assessed by clinical outcome, fluid drainage, chest radiography, pleural ultrasound and/or computed tomography. Results : The mean UK instillation time was $1.63{\pm}0.10$. The mean volume drained UK instillation was $381.3{\pm}314.4\;mL$, and post-UK was $321.6{\pm}489.5\;mL$. The follow up duration after UK therapy was mean $212.9{\pm}194.5$ days. We had successful results in 19 cases (95.0%). There were 12 pleural thickenings (60.0%), 2 markedly decreased effusions (10.0%) and 5 cases of no thickening or effusion. There was recurrence after treatment in only one patient(5%) with complicated pleural effusiondue to tuberculosis. Conclusions : Image-guided drainage with high dose UK instillation (250,000 U/day) in complicated pleural effusion is a safe and more effective method than closed thoracostomy drainage. And this management, in turn, can obviate surgery in most cases.
Nam Hee Rim;Lim Do Hoon;Ahn Yong Chan;Lee Jung I1;Nam Do-Hyun;Kim Jong Hyun;Hong Seung-Chyul;Lee Jeong Eun;Kang Min Kyu;Park Young Je;Kim Kyung Ju;Park Won;Huh Seung Jae
Radiation Oncology Journal
/
v.22
no.2
/
pp.91-97
/
2004
Propose: To Analyze the survival outcomes and prognostic factors In glioblastoma patients treated with surgery and radiation therapy. Materials and Methods : One hundred twenty glloblastoma patients treated with postoperative radiation therapy from 1994 to 2003 at Samsung Medical Center were retrospectively reviewed. Surgical extents were gross total resection in 22 patients (18$\%$), subtotal resection in 69 (58$\%$), and biopsy only in 29 (24$\%$). The median radiation dose was 50 Gy, ranging from 45 Gy to 72 Gy The median follow-up period was 12 months ranging from 2 to 52 months. Results The overall 1- and 2-year survival rates were 52$\%$ and 14$\%$, respectively, and the median survival duration was 13 months. Favorable prognostic factors by Uunivarlate analyses of prognostic factors on 1-year survival rate were revealed that age under 50 (p<0.01), ECOG performance status 0 or 1 (p=0.03), single lesion (p=0.02), and gross total resection (p=0.04), were the favorable prognostic (actors. and by Mmultlvarlate analyses were revealed that female (p<0.01), age under 50 (p<0.01), ECOG performance status 0 or 1 (p=0.05) and gross total resection (p=0.05) were the favorable prognostic factors. Conclusions : The results of our study were comparable with those previously reported. To Improve treatment outcome, various modifications, Including radiation dose escalation through newer radiation therapy techniques and use of effective chemotherapy regimen, should be further Investigated. Investigated. Also Furthermore, the application of Individualized treatment strategy based on 4he patient's prognostic factors might be needed.
This study was conducted to establish the method for preserving PGCs that enables long-term storage in liquid nitrogen for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of freeze-thaw treatment on viability of PGCs in chickens. PGCs were collected separately from a germinal gonad of an early embryo of 5.5~6 day (stage 28) of Isa brown, Korean Oge (KO), White Leghorn and Commercial breeds. PGCs separated from a germinal gonad of an early embryo of 5.5~6 day (stage 28) are suspended in a freezing medium containing a freezing and protecting agents (e.g. dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The PGCs were then purified using magnetic activated cell sorting (MACS) method. The viability of PGCs after thawing was $87.4{\pm}0.4%$ and $89.4{\pm}0.2%$ with the 10% EG treatments with no significant difference between the Isa brown and Commercial breeds. The viability of PGCs after freeze- thawing was significantly higher for Isa brown ($87.4{\pm}0.4%$) and Commercial breeds ($89.4{\pm}0.2%$) than Korean Oge (KO) ($77.6{\pm}1.1%$) and White Leghorn ($76.2{\pm}0.9%$)(p<0.05) using 10% EG cryoprotectant. This study established a method for pre- serving chicken PGCs that enables systematic storage and labeling of cryopreserved PGCs in liquid ($LN_2$) at agermplasm repository and ease of entry into a data base. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.
The number of thoracic and lumbar vertebrae is known to be an unfixed trait among mammals. This study focused on the relationship between numerical variations of cervical (CER), thoracic (THO), and lumbar (LUM) vertebrae and the total number of vertebrae (TNV) and carcass traits in Jejunative black pigs (JBPs), Landrace pigs, and their intercrossed $F_2$ population. There were no numerical variations in CER vertebrae. On the other hand, the numbers of THO and LUM vertebrae and the TNV varied in all three populations. Of the traits investigated in the three populations, only the means${\pm}$SE of the LUM vertebrae did not show statistical significance (p>0.05). The carcass weights (CW), meat color (MC), marbling score (MS), backfat thickness (BFT), carcass length (CLE), THO vertebrae, and TNV all showed statistical significance (p<0.05). The JBP had 14-16 THO vertebrae, 5-6 LUM vertebrae, and 27-29 TNV. The Landrace pigs had 15-16 THO vertebrae, 5-7 LUM vertebrae, and 28-29 TNV. The $F_2$ population had 14-17 THO vertebrae, 5-7 LUM vertebrae, and 27-30 TNV. In the F2 population, increased numbers of THO vertebrae and TNV were associated with a significant increase in the CW, CLE, and BFT (p<0.05). In particular, the increase in the TNV was caused by an increase in the number of THO rather than LUM vertebrae. Although the animals with a greater number of THO and TNV had thicker backfat, they had a longer CLE and a heavier CW. Both these traits are economically more important than the level of backfat when determining the productivity level. These results suggest that genetic selection to increase the number of vertebrae, especially in Landrace pigs, JBPs, and their related populations, may be an excellent strategy for improving productivity.
Cho Jae Ho;Seong Jinsil;Keum Ki Chang;Kim Gwi Eon;Suh Chang Ok;Roh Jae Kyung;Chung Hyun Cheol;Min Jin Sik;Kim Nam Kyu
Radiation Oncology Journal
/
v.18
no.4
/
pp.293-299
/
2000
Purpose :We conducted a prospective non-randomized clinical study to evaluate the efficacy and toxic of the preoperative concurrent chemoradiotherapy for locally advanced unresectable rectal cancer. Materials and Methods: Between January 1995 and June 1998, 37 conecutive patients with locally unresectable advanced rectal cancer were entered into the study. With 3- or 4- fields technique, a total of 45 Gy radiation was delivered on whole pelvis, followed by 5.4 Gy boost to the primary tumor in some cases. Chemotherapy was done at the first and fifth week of radiation with bolus i.v. 5-Fluorouracil (FU) 370$\~$450 mg/m$^{2}$, days 1$\~$5, plus Leucovorin 20 mg/m$^{2}$, days 1$\~$5. OF 37 patients, 6 patients did not receive all planned treatment course (refusal in 4, disease progression in 1, metastasis to lung in 1). Surgical resection was undergone 4$\~$6 weeks after preoperative concurrent chemoradiotherapy. Results :Complete resection rate with negative margins was 94$\%$ (29/31). Complete response was seen in 7 patients (23$\%$) clinically and 2 patients (6$\%$) pathologically. Down staging of tumor occured in 21 patients (68$\%$). Treatment related toxicity was minimal except grade III & IV leukopenia in 2 patients, respectively. Conclusion : Preoperative concurrent chemoradiotherapy in locally advanced rectal cancer was effective in inducing down staging and complete resection rate. Treatment related toxicity was minimal. Further follow up is on-going to determine long term survival following this treatment.
Purpose: Gastrointestinal stromal tumors (GISTs) are mesenchymal neoplasms of the gastrointestinal tract. GISTs are positive for the expression of c-Kit protein at immunohistochemistry, and their clinical presentations vary. This retrospective study was performed to evaluate the clincopathologic characteristics of GISTs and to define the prognostic factors. Materials and Methods: 40 patients who underwent a complete resection of a GIST during the period $1996\~2003$ at the Department of Surgery, Korea University College of Medicine, were studied. We divided them into low- and high-risk. groups by using tumor size and mitotic count: 23 cases were low risk, and 17 were high risk. Clinicopathologic features, immunohistochemical findings, and prognoses were compared between the low- and the high-risk groups. Results: The mean age of the 40 patients was $61.3\pm11.1$years, and the male-to-female ratio was 1:1.1. There was no significant difference in age and sex between the groups. A comparative analysis revealed tumor size, mitotic count, clinical symptoms, preoperative pathologic diagnosis, ulceration, and necrosis to be variables that had statistically significant differences between the high- and the low-risk groups. In the univariate analysis, tumor size, mitotic count, ulceration, necrosis, and abnormal endoscopic ultrasound findings were associated with disease-free survival, but in the multivariate analysis, mitotic activity was the only independent factor associated with disease-free survival. 8 patients had recurrences during the follow-up period, and four of them were treated with STI-571 (imatinib mesylate, $Gleevec^{(R)}$). The treated patients have survived until now; however, two of non-treated patients died from disease progression. Conclusion: Based on this study, tumor size, ulceration, and necrosis are significant factors affecting survival, and mitotic activity may be a useful prognostic marker. STI-571 may be used in an adjuvant setting because the drug has shown anticancer activity in patients with recurrence or metastasis.
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