For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.
Kim, Jae-Do;Kim, Ji-Youn;Jang, Su-Jin;Chung, So-Hak;Jung, Gu-Hee
The Journal of the Korean bone and joint tumor society
/
v.16
no.1
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pp.8-13
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2010
Purpose: This study was performed to evaluate the effi ciency of Platelet-rich plasma (PRP) for acceleration of bone healing process on allograft transplantation after curettage in benign bone tumor. Materials and Methods: From December 2007 to February 2009, twenty-one patients who had benign bone tumor and underwent allograft transplantation after curettage were evaluated. Mean follow-up period was 14.6 months (range, 12-26 months). We compared with 13 cases of PRP group and 8 cases of non-PRP group in terms of size of lesion, bone resorption, amount of applied PRP and complications. The mean age at surgery was 23.6 years (range, 4-73 years). The most common diagnosis was simple bone cyst (7) followed by enchondroma (4), giant cell tumor (3), undifferentiated benign bone tumor (3) and so on. Results: The mean size of lesion was 33.5 $cm^3$ (range, 2.3-181.9 $cm^3$) (29.4 $cm^3$ in PRP group and 40.2 $cm^3$ in non-PRP group). The mean volume of injected PRP was 7.4 cc (range, 3-12 cc). Bone union started at 3.0 months (range, 1.5-5.8 months) in PRP group and 5.3 months (range, 4-8 months) in non-PRP group. Three cases for each group were excluded due to recurrence and pathologic fracture. One patient had febrile episode 3 weeks later after surgery which subsided with antibiotics. Conclusion: The PRP could accelerate bone union in allograft transplantation after curettage of benign bone tumor. Furthermore, we expect that PRP can accelerate bone union in fracture or non-union.
Sodium is an essential nutrient with very important functions, including regulation of the extracellular fluid volume and active transport of molecules across the cell membranes. Since high levels of dietary sodium are associated with a high prevalence of hypertension, prehypertension, and other adverse effects on health, many national and international health organizations (WHO, FAO, etc.) recommend that sodium intake should be significantly decreased. In developed nations, cheese products, from among many processed foods, can cause high salt intake. Hence, there is an urgent need to reduce the content of salt in cheese processing, using various substitutes of sodium chloride (NaCl). In general, salt (NaCl) has been used as a food preservative to limit and (or) kill the growth of foodborne pathogens and spoilage organisms by decreasing the water activity, and to improve texture and flavor. To maintain public health, the salt content in cheese should be decreased without influencing the physicochemical properties of cheese. Therefore, the objective of this review is to outline the upcoming technologies used to reduce the salt content in different types of cheese using various substitutes.
Purpose : This study analyzed the prognostic factors affecting the survival rate and evaluated the role of radiation therapy in muscle-invading bladder cancer. Materials and Methods : Twenty eight patients with bladder cancer who completed planned definitive radiotherapy in the Departments of Therapeutic Radiology and Urology, Chonnam National University Hospital between Jan. 1986 to Dec. 1998 were retrospectively analyzed. The reviews were peformed based on the patients' medical records. There were 21 males and 7 females in this study. The median of age was 72 years old ranging from 49 to 84 years. All patients were confirmed as having transitional cell carcinoma with histological grade 1 in one patient, grade 2 in 15, grade 3 in 9, and uninformed in 3. Radiation therapy was peformed using a linear accelerator with 6 or 10 MV X-rays. Radiation was delivered daily with a 1.8 or 2.0 Gy fraction size by 4 ports (anterior-posterior, both lateral, alternatively) or 3 ports (Anterior and both lateral). The median radiation dose delivered to the isocenter of the target volume was 61.24 Gy ranging from 59 to 66.6 Gy. The survival rate was calculated by the Kaplan-Meier method. Multivariate analysis was peformed on the prognostic factors affecting the survival rate. Results : The survival rate was $76\%,\;46\%,\;33\%,\;33\%$ at 1, 2, 3, 5 years, respectively, with 19 months of median survival. The potential factors of age (less than 70 years vs above 70), sex, diabetes mellitus, hypertension, hydronephrosis, 1-stage (T3a vs T3b), TUR, chemotherapy, total duration of radiotherapy, radiation dose (less than 60 Gy vs above 60 Gy), and the treatment response were investigated with uniand multivariate analysis. Un univariate analysis, the T-stage (p=0.078) and radiation dose (p=0.051) were marginally significant, and the treatment response (p=0.011) was a statistically significant factor on the survival rate. Multivariate analysis showed there were no significant prognostic factors affecting the survival rate. Conclusion : The treatment response and radiation dose are suggested as th은 statistically significant factors affecting the survival rate of muscle invasive bladder cancer. A Further prospective randomized study is needed to confirm these prognostic factors.
Objective of this research was to investigate the influence of pre-plant nitrogen levels in root media on plug seedling growth of radish cv. Soksungbommu. To achieve the research purpose, a root medium, the mixture of perlite, coir dust, and peatmoss (volume percentage of 30:35:35) was formulated and the N levels incorporated during mixing were controlled to 0, 100, 250, 500, 750, 1,000, and $1,500mg{\cdot}L^{-1}$. Then, the seeds were sown into 72-cell plug trays in which the root medium was packed. The measurements of growth and analysis of tissue and root media were conducted 2 and 4 weeks after sowing. Elevation of pre-plant N levels raised EC and turned down pH of root media. But, as seedling grew, the pH rose and EC get lowered in all treatments. The EC as well as $NH_4-N$ and $NO_3-N$ concentrations of root media declined gradually until week 2, but those declined sharply between weeks 2 to 4. The seedling growth 2 weeks after sowing showed quadratic response to pre-plant N levels with the highest growth in $250mg{\cdot}L^{-1}$ treatment and lagging growth in the treatments of lower or higher N levels than $250mg{\cdot}L^{-1}$. The seedling growth 4 weeks after sowing showed also quadratic response with the highest growth in $500mg{\cdot}L^{-1}$ treatment. The tissue N contents were get higher and those of K, Ca, and Mg were get lower as pre-plant N levels were elevated. Above results suggest that lower than $250mg{\cdot}L^{-1}$ in pre-plant N levels is optimistic for growth of plug seedling and avoidance of toxic injury in very young stage.
Kim, Jin Kyu;Kim, Yi Sun;Yoo, Hye Soo;Ahn, So Yoon;Seo, Hyun Ju;Choi, Seo Heui;Park, Soo Kyung;Jung, Yu Jin;Kim, Myo Jing;Jeon, Ga Won;Koo, Soo Hyun;Lee, Kyung-Hoon;Chang, Yun Sil;Park, Won Soon
Clinical and Experimental Pediatrics
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v.53
no.2
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pp.167-172
/
2010
Purpose : With improved survival of extremely low birth weight infants (ELBWI), there is an increase in the incidence of necrotizing enterocolitis (NEC) requiring laparotomy, and the risk of morbidity and mortality in these ELBWI is increased. Thus, we determined the prognostic factors in ELBWI who underwent laparotomy for NEC. Methods : We retrospectively reviewed the medical records of 35 ELBWI who underwent laparotomy for NEC from January 2001 to December 2008 at Samsung Medical Center. Results : Of 480 ELBWI, 35 required laparotomy for NEC; the mortality rate was 20% (Alive group n=28, Dead group n=7). The values of preoperative score for neonatal acute physiology-II (P =0.022) and fraction of inspired oxygen (P <0.001) were significantly higher in the dead group and values of base excess (P =0.004) were significantly lower in the dead group. Values of preoperative heart rate, respiration rate, mean blood pressure, pH, $CO_2$, and potassium ion were not significantly different between the study groups. Intraoperative fluid volume was significantly higher in the alive group than in the dead group (P =0.045). Postoperative infusion rate was significantly lower in the alive group than in the dead group (P =0.022). Conclusion : Good preoperative condition, more intraoperative fluid infusion, and stable postoperative hemodynamic condition were factors associated with favorable prognosis of laparotomy for NEC in ELBWI.
Kim, Jon Soo;Choi, Jun Seok;Choi, Doo Young;You, Chur Woo
Clinical and Experimental Pediatrics
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v.51
no.8
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pp.827-833
/
2008
Purpose : Early identification of iron deficiency in young children is essential to prevent damaging long-term consequences. It is often difficult for the pediatrician to know which indices should be used when diagnosing these conditions especially in hospitalized young children. This study investigated the clinical significances of reticulocyte hemoglobin content in young children with acute infection. Methods : We studied 69 young children aged from 6 to 24 months admitted with acute infection in a single center. Venous blood was drawn to determine hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), hemoglobin content (CH), reticulocyte hemoglobin content (CHr), and red blood cell distribution width (RDW) using ADVIA 120 (Bayer Diagnostics, NY, USA). For evaluating iron status, iron, total iron binding capacity, ferritin and transferrin saturation (Tfsat) were determined. Iron deficiency was defined as Tfsat less than 20%, and iron deficiency anemia as Tfsat less than 20% and Hb level less than 11 g/dL. Results : In all, 47 were iron deficient; 17 of these had iron deficiency anemia. CHr was the only significant predictor of iron deficiency (likelihood ratio test=71.25; odds ratio=0.67; P<0.05). Plasma ferritin level had no predictive value (P=0.519). Subjects with CHr less than 27.4 pg had lower Hb level, MCH, CH, Tfsat, and iron levels than those with CHr 27.4 pg or more (P<0.05 for all). Conclusion : CHr level was a sensitive screening tool and the strongest predictor of iron deficiency in hospitalized infants with acute infection; it was cost saving and avoiding additional sampling. However its reference range should be established.
The purpose of this study was to evaluate the viability of periodontal ligament cells of rat teeth after low-temperature preservation under high pressure by means of MTT assay, WST-1 assay. 12 teeth of Sprague-Dawley white female rats of 4 week-old were used for each group. Both side of the first and second maxillary molars were extracted as atraumatically as possible under tiletamine anesthesia. The experimental groups were group 1 (Immediate extraction), group 2 (Slow freezing under pressure of 3 MPa), group 3 (Slow freezing under pressure of 2 MPa), group 4 (Slow freezing under no additional pressure), group 5 (Rapid freezing in liquid nitrogen under pressure of 2 MPa), group 6 (Rapid freezing in liquid nitrogen under no additional pressure), group 7 (low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa), group 8 (low-temperature preservation at $0^{\circ}C$ under no additional pressure), group 9 (low-temperature preservation at $-5^{\circ}C$ under pressure of 90 MPa). F-medium and 10% DMSO were used as preservation medium and cryo-protectant. For cryo-preservation groups, thawing was performed in $37^{\circ}C$ water bath, then MTT assay, WST-1 assay were processed. One way ANOVA and Tukey HSD method were performed at the 95% level of confidence. The values of optical density obtained by MTT assay and WST-1 were divided by the values of eosin staining for tissue volume standardization. In both MTT and WST-1 assay, group 7 ($0^{\circ}C$/2 MPa) showed higher viability of periodontal ligament cells than other group (2-6, 8) and this was statistically significant (p < 0.05), but showed lower viability than group 1, immediate extraction group (no statistical significance). By the results of this study, low-temperature preservation at $0^{\circ}C$ under pressure of 2 MPa suggest the possibility for long term preservation of teeth.
Kim, Ji Young;Yang, Seung Ho;Cha, Sun Hwa;Kim, Ji Yeun;Jang, Young Chae;Park, Kwan Kyu;Kim, Jin Kyung;Chung, Hai Lee;Seo, Eok Su;Kim, Woo Taek
Clinical and Experimental Pediatrics
/
v.50
no.7
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pp.686-693
/
2007
Purpose : Mycophenolic acid (MPA), the active metabolite of mycophenolate mofetil (MMF), is a potent inhibitor of inosine-monophosphate dehydrogenase (IMPDH), a new immunosuppressive drug used. It was reported that MPA protected neurons after excitotoxic injury, induced apoptosis in microglial cells. However, the effects of MPA on hypoxic-ischemic (HI) brain injury has not been yet evaluated. Therefore, we examined whether MPA could be neuroprotective in perinatal HI brain injury using Rice-Vannucci model (in vivo) and in rat brain cortical cell culture induced by hypoxia (in vitro). Methods : Cortical cells were cultured using a 18-day-pregnant Sprague-Dawley (SD) rats and incubated in 1% $O_2$ incubator for hypoxia. MPA ($10{\mu}g/mL$) before or after a HI insult was treated. Seven-day-old SD rat pups were subjected to left carotid occlusion followed by 2 hours of hypoxic exposure (8% $O_2$). MPA (10 mg/kg) before or after a HI insult were administrated intraperitoneally. Apoptosis was measured using western blot and real-time PCR for Bcl-2, Bax, caspase-3. Results : H&E stain revealed increased brain volume in the MPA-treated group in vivo animal model of neonatal HI brain injury. Western blot and real-time PCR showed the expression of caspase-3 and Bax/Bcl-2 were decreased in the MPA-treated group In in vitro and in vivo model of perinatal HI brain injury, Conclusion : These results may suggest that the administration of MPA before HI insult could significantly protect against perinatal HI brain injury via anti-apoptotic mechanisms, which offers the possibility of MPA application for the treatment of neonatal HI encephalopathy.
Filtering rates of two farming ascidians Styela clava and S. plicata, and of a farming mussel Mytilus edulis were experimentally investigated with reference to effects of water temperature and size. Absorptiometric determinations of filtering rates were carried out in a closed system with experimental animals being decreased indicate dyes neutral red. Optical density (OD) of 440 nm in path length 22 mm cell used as the indication of food particles absorption was appeared directly in proportion with the concentration of neutral red dyes. The filtering rate F is calculated by Kim's equation $F\;=\;V(1-e^{-z})$, where V is the water volume ($\ell$) in the experimental jar, and Z is the decreasing coefficient of OD as meaning of instantaneous removal speed as In $C_t\;=\;In\;C_{o}-Z{\cdot}t$, in this formula $C_t$ is OD at the time t. Filtering rate of S. clava increased as exponential function with increasing temperature while not over critical limit, and the critical temperature for filtering rate was assumed to be between $28^{\circ}C$ and $29^{\circ}C$. In case of S. plicata, the critical temperature was to be below $13^{\circ}C$, and through the temperature range $15\~25^{\circ}C$ appeared a little difference in level even though with significant. M. edulis was not appear any significant effects by water temperature less than $29^{\circ}C$. The model formula derived from the results is as below, where F is filtering rate (${\ell}/hr/animal$), T is water temperature ($^{\circ}C$), and DW is dry meat weight (g) of experimental animal. $$S.\;Clava;\;F\;=\;e xp\;(0.119\;T-4.540)\;(DW)^{0.6745},\;T<29^{\circ}C$$) $$S.\;plicata;\;F\;=\;e xp\;(A_t)\;(DW)^{0.5675},\;(13^{\circ}C$$[A_t =-8.56+0.6805\;T-0.0153\;T^2]$$$$M.\;edulis;\;F\;=\;0.3844\;(DW)^{0.4952},\;<29^{\circ}C$$)
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