• Title/Summary/Keyword: Medium Optimization

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Reduction of Polyspermy in Porcine in vitro Fertilization by Modified Swim-UP Method

  • Park, C.H.;B.S. Koo;Kim, M.G.;J.I. Yun;H.Y Son;Lee, S.G.;Lee, C.K.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.110-110
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    • 2003
  • The high incidence of polyspermic fertilization is one of the major causes lowering the overall efficiency of porcine IVF. The common procedure for IVF involves the co-culture of both gametes in the medium drop, which increases sperm concentration and incidence of polyspermy. Therefore, the present study was carried out to increase the efficiency of porcine IVF by reducing polyspermy using a modified swim-up method. This method modifies conventional swim-up washing by placing oocytes directly at the time of washing. Sperm pellet was prepared in the tube and mature oocytes were placed on cell strainer with $70 \mu m$ pore size (Falcon 2350) at the top of the tube. After insemination, the oocytes were stained for examination. Also, the developmental potential of fertilized embryos was measured to evaluate for the feasibility of this method. While having similar penetration rates in both methods ($86.67 \pm 2.36% to 83.33 \pm 1.36%$), there was a significant reduction of polyspermy in modified swim-up method ($17.50 \pm 1.60%$) compare to the control ($44.1 \pm 3.70%$ (p<0.05). Subsequent culture showed higher rate of blastocyst formation in modified swim-up method (20.44$\pm$0.99%) than the control ($15.73 \pm 3.26%$) (P<0.05), even though there was no significant difference. These results suggest that, by controlling the number of spermatozoa reaching the oocytes, porcine oocytes might be protected from polyspermy in vitro. Also, the developmental potential of the fertilized embryos using this method could be improved by increasing the pool of spermatozoa with better quality. Further optimization of the procedure required to implicate this method in routine porcine IVF.

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The Characteristics related to Pulmonary Rehabilitation in Patients with Chronic Obstructive Pulmonary Disease: A Cross-sectional Study, Data from the Korea National Health and Nutrition Examination Survey 2015-2019.

  • Kyeongbong Lee
    • Physical Therapy Rehabilitation Science
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    • v.12 no.3
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    • pp.229-239
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    • 2023
  • Objective: Patients with chronic obstructive pulmonary disease (COPD) may experience reduced physical activity and quality of life (QoL) due to decreased pulmonary function. The purpose of this study was to investigate the level of pulmonary function, physical activity, and QoL of COPD patients. Design: Cross-sectional observational study. Methods: This study examined the published data of the Korea National Health and Nutrition Examination Survey in 2015-2019. Among 39,759 subjects who participated for 5 years, data from 151 patients diagnosed with COPD were analyzed separately. For the pulmonary function, the results of forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), FEV6, forced expiratory flow 25-75%, and peak expiratory flow were observed. Physical activity was identified as frequency and duration. For the QoL, EQ-5D-3L evaluation results were examined, and the frequency and index of the Korean version were investigated. Results: In pulmonary function, all variables were found to be lower than age and weighted matched normal values. COPD patients showed to perform very low levels of high/medium physical activity and sitting time was confirmed to be more than 8 hours a day. In QoL, it was found that the highest reporting rate of some problems was the "pain and discomfort" and "mobility". Conclusions: It was found that COPD patients showed that the prevalence of circulatory disease was relatively high, lowered pulmonary function, and QoL. These can be improved through regular physical activity, and it is thought that this can be achieved through optimization of pulmonary rehabilitation.

Optimizing hormonal and amino acid combinations for enhanced cell proliferation and cell cycle progression in bovine mammary epithelial cells

  • Hyuk Cheol Kwon;Hyun Su Jung;Do Hyun Kim;Jong Hyeon Han;Seo Gu Han;Dong Hyun Keum;Seong Joon Hong;Sung Gu Han
    • Animal Bioscience
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    • v.36 no.11
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    • pp.1757-1768
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    • 2023
  • Objective: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. Methods: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17β-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. Results: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17β-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. Conclusion: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

Numerical and statistical analysis of Newtonian/non-Newtonian traits of MoS2-C2H6O2 nanofluids with variable fluid properties

  • Manoj C Kumar;Jasmine A Benazir
    • Advances in nano research
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    • v.16 no.4
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    • pp.341-352
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    • 2024
  • This study investigates the heat and mass transfer characteristics of a MoS2 nanoparticle suspension in ethylene glycol over a porous stretching sheet. MoS2 nanoparticles are known for their exceptional thermal and chemical stability which makes it convenient for enhancing the energy and mass transport properties of base fluids. Ethylene glycol, a common coolant in various industrial applications is utilized as the suspending medium due to its superior heat transfer properties. The effects of variable thermal conductivity, variable mass diffusivity, thermal radiation and thermophoresis which are crucial parameters in affecting the transport phenomena of nanofluids are taken into consideration. The governing partial differential equations representing the conservation of momentum, energy, and concentration are reduced to a set of nonlinear ordinary differential equations using appropriate similarity transformations. R software and MATLAB-bvp5c are used to compute the solutions. The impact of key parameters, including the nanoparticle volume fraction, magnetic field, Prandtl number, and thermophoresis parameter on the flow, heat and mass transfer rates is systematically examined. The study reveals that the presence of MoS2 nanoparticles curbs the friction between the fluid and the solid boundary. Moreover, the variable thermal conductivity controls the rate of heat transfer and variable mass diffusivity regulates the rate of mass transfer. The numerical and statistical results computed are mutually justified via tables. The results obtained from this investigation provide valuable insights into the design and optimization of systems involving nanofluid-based heat and mass transfer processes, such as solar collectors, chemical reactors, and heat exchangers. Furthermore, the findings contribute to a deeper understanding of stretching sheet systems, such as in manufacturing processes involving continuous casting or polymer film production. The incorporation of MoS2-C2H6O2 nanofluids can potentially optimize temperature distribution and fluid dynamics.

Optimization of Culture Medium for the Production of an Exopolysaccharide (p-CY02) with Cryoprotective Activity by Pseudoalteromonas sp. RosPo-2 from the Antarctic Sea

  • Pilsung Kang;Sung Jin Kim;Ha Ju Park;Il Chan Kim;Se Jong Han;Joung Han Yim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.5
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    • pp.1135-1145
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    • 2024
  • When cells are exposed to freezing temperatures, high concentrations of cryoprotective agents (CPA) prevent ice crystal formation, thus enhancing cell survival. However, high concentrations of CPAs can also cause cell toxicity. Exopolysaccharides (EPSs) from polar marine environments exhibit lower toxicity and display effects similar to traditional CPA. In this study, we sought to address these issues by i) selecting strains that produce EPS with novel cryoprotective activity, and ii) optimizing culture conditions for EPS production. Sixty-six bacteria producing mucous substances were isolated from the Ross Sea (Antarctic Ocean) using solid marine agar plates. Among them, Pseudoalteromonas sp. RosPo-2 was ultimately selected based on the rheological properties of the produced EPS (p-CY02). Cryoprotective activity experiments demonstrated that p-CY02 exhibited significantly cryoprotective activity at a concentration of 0.8% (w/v) on mammalian cells (HaCaT). This activity was further improved when combined with various concentrations of dimethyl sulfoxide (DMSO) compared to using DMSO alone. Moreover, the survival rate of HaCaT cells treated with 5% (v/v) DMSO and 0.8% (w/v) p-CY02 was measured at 87.9 ± 2.8% after freezing treatment. This suggests that p-CY02 may be developed as a more effective, less toxic, and novel non-permeating CPA. To enhance the production of EPS with cryoprotective activity, Response Surface Methodology (RSM) was implemented, resulting in a 1.64-fold increase in production of EPS with cryoprotective activity.

Overproduction of Xanthophyll Pigment in Flavobacterium sp. JSWR-1 under Optimized Culture Conditions

  • Jegadeesh Raman;Young-Joon Ko;Jeong-Seon Kim;Da-Hye Kim;Soo-Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.34 no.3
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    • pp.710-724
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    • 2024
  • Flavobacterium can synthesize xanthophyll, particularly the pigment zeaxanthin, which has significant economic value in nutrition and pharmaceuticals. Recently, the use of carotenoid biosynthesis by bacteria and yeast fermentation technology has shown to be very efficient and offers significant advantages in large-scale production, cost-effectiveness, and safety. In the present study, JSWR-1 strain capable of producing xanthophyll pigment was isolated from a freshwater reservoir in Wanju-gun, Republic of Korea. Based on the morphological, physiological, and molecular characteristics, JSWR-1 classified as belonging to the Flavobacterium species. The bacterium is strictly aerobic, Gram-negative, rod-shaped, and psychrophilic. The completed genome sequence of the strain Flavobacterium sp. JSWR-1 is predicted to be a single circular 3,425,829-bp chromosome with a G+C content of 35.2% and 2,941 protein-coding genes. The optimization of carotenoid production was achieved by small-scale cultivation, resulting in zeaxanthin being identified as the predominant carotenoid pigment. The enhancement of zeaxanthin biosynthesis by applying different light-irradiation, variations in pH and temperature, and adding carbon and nitrogen supplies to the growth medium. A significant increase in intracellular zeaxanthin concentrations was also recorded during fed-batch fermentation achieving a maximum of 16.69 ± 0.71 mg/l, corresponding to a product yield of 4.05 ± 0.15 mg zeaxanthin per gram cell dry weight. Batch and fed-batch culture extracts exhibit significant antioxidant activity. The results demonstrated that the JSWR-1 strain can potentially serve as a source for zeaxanthin biosynthesis.

Rapid Statistical Optimization of Cultural Conditions for Mass Production of Carboxymethylcellulase by a Newly Isolated Marine Bacterium, Bacillus velezensis A-68 from Rice Hulls (통계학적 방법을 사용한 해양미생물 Bacillus velezensis A-68균주의 섬유소 분해효소 생산 조건 최적화)

  • Kim, Bo-Kyung;Kim, Hye-Jin;Lee, Jin-Woo
    • Journal of Life Science
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    • v.23 no.6
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    • pp.757-769
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    • 2013
  • A microorganism producing carboxymethylcellulase (CMCase) was isolated from seawater, identified as Bacillus velezensis by analyses of 16S rDNA and partial sequences of the gyrA, and designated as B. velezensis A-68. The optimal conditions for production of CMCase by B. velezensis A-68 were established using response surface methodology (RSM). The optimal concentrations of rice hulls and yeast extract, and initial pH of the medium for cell growth were 60.2 g/l, 7.38 g/l, and 7.18, respectively, whereas those for production of CMCase were 50.0 g/l, 5.00 g/l, and 7.30. The analysis of variance (ANOVA) implied that the most significant factor for cell growth as well as production of CMCase was yeast extract. The optimal concentrations of $K_2HPO_4$, NaCl, $MgSO_4{\cdot}7H_2O$, and $(NH_4)_2SO_4$ in the medium for cell growth were 7.50, 1.00, 0.10, and 0.80 g/l, respectively, which were the same as those for production of CMCase. The optimal temperatures for cell growth and production of CMCase were 30 and $35^{\circ}C$, respectively. The maximal production of CMCase under optimized conditions was 83.8 U/ml, which was 3.3 times higher than that before optimization. In this study, rice hulls, agro-byproduct, were developed as a substrate for production of CMCase and time for production of CMCase was reduced to 3 days using a newly isolated marine bacterium.

Characterization of Protease Produced by Elizabethkingia meningoseptica CS2-1 and Optimization of Cultural Conditions for Amino Acid Production (닭 우모 분해세균 Elizabethkingia meningoseptica CS2-1이 생산하는 단백질분해효소의 특성 및 아미노산 생산을 위한 배양조건)

  • Kim, Se-Jong;Cho, Chun-Hwi;Whang, Kyung-Sook
    • Journal of Applied Biological Chemistry
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    • v.54 no.2
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    • pp.135-142
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    • 2011
  • A feather-degrading bacterium Elizabethkingia meningoseptica CS2-1 was isolated from compost in a chicken farm. Cultured on a basal medium containing 2% chicken feather, the bacterium showed 729.7 ${\mu}mol/mL$ of amino acid. Optimal culture conditions for feather degradation by E. meningoseptica CS2-1 were $25^{\circ}C$, pH 7.5, and 180 rpm. The optimal pH and temperature for protease activity were 8.0 and $40^{\circ}C$, respectively. The composition of an optimal medium for amino acid production was 0.05% NH4Cl, 0.05% NaCl, 0.03% $K_2HPO_4$, 0.03% $KH_2PO_4$, 0.01% $MgCl_2{\cdot}6H_2O$, 0.1% urea, and 2% chicken feather. Characteristics of amino acids extracted from the optimal medium under the optimal culture conditions of E. meningoseptica CS2-1 were analyzed. The total amino acid content of strain CS2-1 was 1063 ${\mu}mol/mL$, which was 46% higher compared to the basal condition (729.7 ${\mu}mol/mL$). The essential amino acid content in the total amino acid was 315.9 ${\mu}mol/mL$, which was 44% higher than that of the basal condition. Major amino acids were proline (14%), aspartic acid (12%), glutamic acid (11%), serine (10%), alanine (10%), glycine (9%), and tyrosine (7%) by strain CS2-1. These results suggest that strain CS2-1 can be used as a potential microbial resource for the production of amino acid using chicken feathers.

Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • 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.

Optimization of Mycelial Growth of Entomogenous fungi of the Genus Cordyceps (동충하초속균의 균사생장최적화)

  • Hong, In-Pyo;Nam, Sung-Hee;Jung, I-Yeon;Sung, Gyoo-Byung;Nam, Hack-Woo;Kang, Seok-Woo;Hur, Hyeon;Lee, Min-Woong;Guo, Shun-Xing
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.149-156
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    • 2004
  • This study was carried out to obtain basic data on physiological characteristics for an artificial cultivation of fruiting body of Cordyceps. Specimens such as Cordyceps longissima, C. militaris and C. pruinosa were collected at Mt. Halla of Cheju island in July, 2003. Among four different culture media which have been used for culture of mushrooms, MCM medium was selected for the favorable culture medium of the Cordyceps tested. The initial pH of solid medium for mycelial growth of Cordyceps was good in the range of pH 5.0~7.0 lower than 8.0. The mycelial growth of C. longissima was most favorable on culture media supplemented with glucose, one of monosaccharides. In C. militaris, nine carbon sources were favorable to the mycelial growth as compared with control among 11 carbon sources. Six nitrogen sources were favorable to the mycelial growth of C. longissima as compared with control among 9 carbon sources; namely, the mycelial growth of C. longissima was most favorable on culture media contained potassium nitrate, and followed in order by ammonium citrate and sodium nitrate in 4 weeks incubation.

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