• Title, Summary, Keyword: Centrifugation

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Comparison of the methods for platelet rich plasma preparation in horses

  • Lee, Eun-bee;Kim, Jung-Won;Seo, Jong-pil
    • Journal of Animal Science and Technology
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    • v.60 no.8
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    • pp.20.1-20.4
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    • 2018
  • Platelet rich plasma (PRP) is popularly used in the horse industry to enhance regeneration of tissue injury that has limitation of blood supply. This study aimed to compare the methods for platelet rich plasma preparation since they has not been established yet. Blood was collected from six horses and platelets were concentrated by three different methods (2-step centrifugation, separated centrifugation and separated centrifugation using histopaque). Concentrated blood was analyzed using Advia hematology systems. In the result, separated centrifugation with histopaque showed the significantly lower number of red blood cells than other groups. The 2-step centrifugation showed the significantly higher number of white blood cells than other groups, while it contained the highest concentration of red blood cells among three groups. In the 2-step centrifugation, separated centrifugation and separated centrifugation with histopaque, platelets were concentrated 4.5, 5.3 and 5.6 times, respectively. And no significant difference of the platelet concentration between the three groups was found. This study demonstrated that separated centrifugation using histopaque was the best method for platelet rich plasma preparation because of the proper amount of platelets and the separation of red blood cells from platelet rich plasma.

Proper Condition of Centrifugation for the Fat Cell Viability in the Autologous Fat Injection (자가지방주입에서 지방세포의 생존을 위한 적절한 원심분리 조건)

  • Jang, Kyung Min;Kim, Jong Yeop;Yang, Jung Duk;Chung, Ho Yun;Park, Jae Woo;Cho, Byung Chae
    • Archives of Plastic Surgery
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    • v.33 no.4
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    • pp.423-426
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    • 2006
  • Purpose: In the autologous fat injection, the centrifugation is useful for the refinement of harvested fat. As it can be an injury to the fat cell, we studied the fat cell viability with the change of centrifugation velocity and centrifugation time in order to get the limits of centrifugation velocity and centrifugation time. Methods: We used the Colman System in 8 patients. We handled the control group with no centrifugation, group I with the centrifugation with 1500rpm for 1 minute, group II with 1500 rpm for 3 minutes, group III with 1500rpm for 5 minutes, group IV with 3000rpm for 1 minute, group V with 3000rpm for 3 minutes, group VI with 3000rpm for 5 minutes, group VII with 5000rpm for 1 minute, group VIII with 5000rpm for 3 minutes, group IX with 5000rpm for 5 minutes. We used the collagenase to separate the fat tissue. We had evaluated the fat cell viability by checking survival cell counts. Results: There was no significance in group I, II, IV, V, but there was significant difference in group III, VI, VII, VIII, IX. Conclusion: The centrifugation with 3000rpm for 3 minutes is recommendable.

Effect of centrifugation on tryptic protein digestion

  • Kim, Soohwan;Kim, Yeoseon;Lee, Dabin;Kim, Inyoung;Paek, Jihyun;Shin, Dongwon;Kim, Jeongkwon
    • Analytical Science and Technology
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    • v.30 no.2
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    • pp.96-101
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    • 2017
  • This study investigated the effect of centrifugation on tryptic digestion. This was done by applying different centrifugation speeds (6,000, 8,000, 10,000, 20,000, and $30,000{\times}g$) over various durations (0, 10, 20, 30, 40, 50, and 60 min) to digest two model proteins - cytochrome c and myoglobin. The intact proteins and resulting peptides were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Centrifugation greatly improved the tryptic digestion efficiency of cytochrome c, where either an increase in centrifugation speed or in digestion duration significantly improved the digestion of cytochrome c. However, centrifugation did not noticeably improve the digestion of myoglobin; 16 h of centrifuge-assisted tryptic digestion at $30,000{\times}g$ barely removed the myoglobin protein peak. Similar results were also obtained when using conventional tryptic digestion with gentle mixing. When acetonitrile (ACN) was added to make 10% ACN buffer solutions, the myoglobin protein peak disappeared after 6 h of digestion using both centrifuge-assisted and conventional tryptic digestions.

Effect of Temperature and Pre-treatment for Elutriated Acidogenic Fermentation of Piggery Waste (돈사폐수의 세정산발효시 온도와 전처리의 영향)

  • Bae, Jin-Yeon;Min, Kyung-Sok
    • Journal of Korean Society on Water Environment
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    • v.21 no.1
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    • pp.34-39
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    • 2005
  • The performance of elutriated acid fermentation with slurry-type piggery waste was investigated, especially to evaluate the effects of temperature and pre-treatment. In the first phase, the acid elutriation reactor with piggery waste after centrifugation operated at both mesophilic and thermophilic conditions to evaluate the effect of temperature. Solubilization yield($gVFAs/gSCOD_{prod.}$) and acidification rate($gVFAs/gSCOD_{prod.}$) in the thermophilic digestion were 0.45 and 0.55, which were higher than those of the mesophilic digestion, 0.25 and 0.45. In addition, the acid elutriation reactor at thermophilic temperature is more effective in removing e-coli. In the second phase, the acid elutriation reactor was fed with piggery waste before centrifugation. With piggery wastes before centrifugation, the solubilization yield and the acidificaton rate were 0.40 and 0.80, respectively, which were higher than the rates using piggery waste after centrifugation at both mesophilic and thermophilic conditions. The higher sludge volume reduction of 80% benefits sludge management. Furthermore, economical advantages can be achieved by removing the pre-treatment process, such as centrifugation. Consequently, the treatment with piggery waste before centrifugation proved to be effective. Also, the optimum temperature condition was estimated at mesophilic or thermophilic conditions, considering solubilization yields and acidification rates, though the system should be heated.

The Relative Centrifugation Force Permits Visualization of the Germinal Vesicle in Pig Oocytes

  • Hsieh, Chang-Hsing;Lee, Stone;Jaw, Si-Ning;Tseng, Jung-Kai;Tang, Pin-Chi;Chang, Lan-Hwa;Ju, Jyh-Cherng
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.9
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    • pp.1227-1231
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    • 2004
  • Pig oocytes contain high levels of lipids in the ooplasm, which reduces the visibility of the germinal vesicle (GV) under microscopic examination. Therefore, the purposes of this study were to investigate the effects of relative centrifugation force (RCF) on the visibility and maturation rates of the GV stage oocytes after centrifugation. In Experiment 1, cumulus-oocyte-complexes (COCs) were collected from slaughterhouse ovaries and randomly allocated to different RCFs (3,000 rpm: 970 g; 6,000 rpm: 3,900 g; or 10,000 rpm: 10,840 g) for 10 or 20 min. Percentages of visible GV were 76-79% in the oocytes centrifuged with 10,000 rpm, which were significantly higher (p<0.01) than those with 3,000 and 6,000 rpm. No significant differences in GV visibility were observed among oocytes with different lengths of centrifugation (p<0.05) regardless of the RCFs. In esperiment 2, the maturation rate of the oocyte was found significantly lower in the 20 min than in the 10 min group received 10,840 g of RCF (30 vs. 75%, p<0.05). In conclusion, the GV of porcine oocytes can be clearly visible by centrifugation at 10,840 g for 10 min without compromising their subsequent maturation rates and a longer centrifugation time (20 min) had no beneficial influence on the visibility of GV stage pig oocytes.

Effect of virus infectivity titer following centrifugation and filtration during virus extraction from fish samples

  • Kim, Wi-Sik;Kim, Jong-Oh;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.28 no.2
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    • pp.113-116
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    • 2015
  • A $0.45-{\mu}m$ membrane filter is generally used to remove bacterial contamination during virus extraction from fish samples. However, the number of fish viruses is drastically reduced after filtration with a $0.45{\mu}m$ filter. In this study, we investigated the effect of filters on virus infectivity titer and the change in virus titer and bacterial number following different centrifugation conditions to determine a suitable procedure for virus extraction from fish samples. $10^{4.05}$ and $10^{5.05}TCID_{50}/ml$ of infectious hematopoietic necrosis virus (IHNV) and $10^{4.05}$ and $10^{4.55}TCID_{50}/ml$ of Oncorhynchus masou virus (OMV) were not detectable after filtration with two types of $0.45-{\mu}m$ filters, except the IHNV titer was reduced by about 10 fold after filter use (company A). No significant difference was found in the virus titer following centrifugation at $880{\times}g$ (30 min) or $3,500{\times}g$ (30 min), whereas IHNV and OMV titers were reduced by about 10 and 10-1000 fold by centrifugation at $14,000{\times}g$ (30 min) and $14,000{\times}g$ (10 and 30 min), respectively. A total of 97.7-99.9% Escherichia coli were eliminated by centrifugation at $880 {\times}g$ (30 min) and $3,500{\times}g$ (30 min). These results show that fish viruses were affected by filtering, even though the effect differed by virus species and filter type. Therefore, centrifugation at $3,500{\times}g$ (30 min) and use of medium with antibiotics may be useful for virus extraction along with a reduction in bacteria.

Separation of X-and Y-Bearing Spermatozoa IV. Separtion of bull spermatozoa by the combination of density Gradient Centrifugation and Sephadex Gel filtration (X-정자와 Y-정자의 분류에 관한 연구 IV. Sephadex Gel여과법과 Percoll중층원심분류법의 병용에 의한 우 정자의 분류)

  • 이주영;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.12 no.3
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    • pp.141-147
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    • 1988
  • These experiments were carried out to develop new techniques for in vitro separation of x-and Y-bearing spermatozoa. The results obtained in these experiments were summarized as follows: 1. Following centrifugation of discontinuous percoll density gradient, populatin of spermatozoa increased progressively from low to high density. The highest concentration of spermatozoa was observed at the 4th fraction which included 36.6% of spermatozoa. 2. As increasing percoll concentration, the higher motility index was obtained and the highest motility index(74.2) was obtained at the 5th fraction. 3. The percentage of B-body bearing spermatozoa following percoll density gradient centrifugation was decreased from 39.7% to 25.6%. 4. The sperm population following chromatography by sephadex gel and percoll density gradient centrifugation was decreased in 1st, 5th and 6th fractions but the reverse was turn for 2nd, 3rd and 7th fractions, and the highest sperm concentration was observed at the 7th fraction which included 37.4% of spermatozoa. 5. Motility index of spermatozoa was increased from 77.6 to 79.4 after the sephadex gel filtration, however it was decreased at all fractions after percoll density gradient centrifugation. The lowest motility index(33.2) was obtained from the 7th fraction. 6. The rate of B-body bearing spermatozoa was shown the trend to decrease by the sephadex gel filtration and the trend was accelerated by the percoll density gradient centrifugation. The lowest percentage of B-body bearing spermatozoa, 12.0% was obtained from the 5th fraction.

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STUDY ON PLATELET RICH PLASMA CONCENTRATION ACCORDING TO PROCESSING METHOD (처리방법에 따른 혈소판 혈장의 농축도에 관한 연구)

  • Min, Seung-Ki;Kim, Hyung-Ju;Cha, Soo-Ryen
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.27 no.1
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    • pp.24-31
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    • 2005
  • Recently, Platelet rich plasma(PRP) is commonly used because it is now well known that platelets have many functions beyond that of simple hemostasis in aspect of containing autogenous source of several growth factors. It could be responsible for increasing cell mitosis, increasing collagen production, recruiting other cells to the site of injury, initiating vascular ingrowth, and inducing cell differentiation, enhancing bone formation capacity and easily handling to clinician. However, in spite of these clinical advantages, still the theory behind the use of PRP is compelling. This study was to determine preparation techniques used to increase the concentration of platelets and growth factors are all crucial steps in early wound healing of bone graft which may lead to a more rapid and denser bone regenerate. 200 volunteers were sampled and PRP were prepared according to each evaluation item in this study. Higher concentration of platelets have been gained in double centrifugation. 2000 and 2500 rpm showed proper concentration of platelets at first centrifugation and 5000 rpm in second. Timing for 2 minutes was showed good concentration of platelets in high and low centrifugation speed. It was better concentration of platelets in 20 or 30 ml volume during centrifugation. In histomorphologic findings, degrnulated and high concentraion of platelets were found in low centrifugation speed.

Development of an Improved Endotoxin Detection Method Using Centrifugation (원심분리법을 이용한 혈청 내 내독소의 개선된 측정방법 연구)

  • Choi, Hyeong Jwa;Lim, Yoo Jung;Lee, Eun Hee;Park, Jin Yeon;Prabagar, Miglena G.;Park, Hyung Soon;Kang, Young Sun
    • Microbiology and Biotechnology Letters
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    • v.41 no.2
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    • pp.242-248
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    • 2013
  • Endotoxins are part of the outer membrane of the cell wall of gram-negative bacteria and are continuously released during bacterial growth. Endotoxins typically induce severe sepsis and septic shock, which cause more than 50% of mortalities. Endotoxins are easily measured in the serum by the limulus amebocyte lysate (LAL) test. However, a nonspecific result is obtained, because the high concentration of serum proteins disturbs the enzyme reaction of the LAL test. In order to solve this problem, the LAL test was performed in this study after the centrifugation of the boiled serum samples to remove the impurities. As a result, among the various conditions examined, endotoxin measurement with the LAL test was the most accurate and repeatable after centrifugation of the boiled serum at $100^{\circ}C$. Moreover, the endotoxin was accurately and repeatedly measured from the prepared sera of mice that had been administered an intraperitoneal injection of purified lipopolysaccharides (LPS) or E. coli. Therefore, the application of centrifugation to remove impurities from boiled serum gives an accurate measurement of endotoxins in the sera of normal subjects or patients, and this will lead to the improved diagnosis and prevention of diseases caused by endotoxins. In addition, the centrifugation of boiled serum samples should be considered and included in the development of endotoxin test kits.

Removing Lipemia in Serum/Plasma Samples: A Multicenter Study

  • Castro-Castro, Maria-Jose;Candas-Estebanez, Beatriz;Esteban-Salan, Margarita;Calmarza, Pilar;Arrobas-Velilla, Teresa;Romero-Roman, Carlos;Pocovi-Mieras, Miguel;Aguilar-Doreste, Jose-Angel;Commission on Lipoprotein and Vascular Diseases, Sociedad Espanola de Quimica Clinica
    • Annals of Laboratory Medicine
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    • v.38 no.6
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    • pp.518-523
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    • 2018
  • Background: Lipemia, a significant source of analytical errors in clinical laboratory settings, should be removed prior to measuring biochemical parameters. We investigated whether lipemia in serum/plasma samples can be removed using a method that is easier and more practicable than ultracentrifugation, the current reference method. Methods: Seven hospital laboratories in Spain participated in this study. We first compared the effectiveness of ultracentrifugation ($108,200{\times}g$) and high-speed centrifugation ($10,000{\times}g$ for 15 minutes) in removing lipemia. Second, we compared high-speed centrifugation with two liquid-liquid extraction methods-LipoClear (StatSpin, Norwood, USA), and 1,1,2-trichlorotrifluoroethane (Merck, Darmstadt, Germany). We assessed 14 biochemical parameters: serum/plasma concentrations of sodium ion, potassium ion, chloride ion, glucose, total protein, albumin, creatinine, urea, alkaline phosphatase, gamma-glutamyl transferase, alanine aminotransferase, aspartate-aminotransferase, calcium, and bilirubin. We analyzed whether the differences between lipemia removal methods exceeded the limit for clinically significant interference (LCSI). Results: When ultracentrifugation and high-speed centrifugation were compared, no parameter had a difference that exceeded the LCSI. When high-speed centrifugation was compared with the two liquid-liquid extraction methods, we found differences exceeding the LCSI in protein, calcium, and aspartate aminotransferase in the comparison with 1,1,2-trichlorotrifluoroethane, and in protein, albumin, and calcium in the comparison with LipoClear. Differences in other parameters did not exceed the LCSI. Conclusions: High-speed centrifugation ($10,000{\times}g$ for 15 minutes) can be used instead of ultracentrifugation to remove lipemia in serum/plasma samples. LipoClear and 1,1,2-trichlorotrifluoroethane are unsuitable as they interfere with the measurement of certain parameters.