• Title/Summary/Keyword: Incubation volume

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Rapid Determination of Volatile Organic Compounds in Human Whole Blood Using Static Headspace Sampling with Gas Chromatography and Mass Spectrometry

  • Lee, Ji-Young;Kim, Seungki;Lee, Jong-Tae;Choi, Jong-Ho;Lee, Jeongae;Pyo, Heesoo
    • Bulletin of the Korean Chemical Society
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    • v.33 no.12
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    • pp.3963-3970
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    • 2012
  • Headspace (HS) and headspace solid-phase microextraction (HS-SPME) were studied for extracting volatile organic compounds (VOCs) from whole blood, with chemical and instrumental variables being optimized for maximum sensitivity: incubation at $60^{\circ}C$, equilibration for 30 min, pH 11, and 2 mL injection volume. Both techniques provided accurate analyses, with detection limits of 0.05-0.1 ng $mL^{-1}$ and 0.05-0.5 ng $mL^{-1}$. HS showed better sensitivity, reproducibility, and analysis times than HS-SPME. Overall levels of chloroform in whole blood were found to be 0.05-5.84 ng $mL^{-1}$; detected levels of benzene were 0.05-2.20 ng $mL^{-1}$.

Organic-Inorganic Hybrid Nanoflowers as Potent Materials for Biosensing and Biocatalytic Applications

  • Tran, Tai Duc;Kim, Moon Il
    • BioChip Journal
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    • v.12 no.4
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    • pp.268-279
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    • 2018
  • Flower-shaped organic-inorganic hybrid nanostructures, termed nanoflowers, have received considerable recent attention as they possess greatly enhanced activity, stability, durability, and even selectivity of entrapped organic biomolecules, which are much better than those from the conventional methods. They can be synthesized simply via co-incubation of organic and inorganic components in aqueous buffer at room temperature and yield hierarchical nanostructures with large surface-to-volume ratios, allowing for low-cost production by easy scale-up, as well as the high loading capacity of biomolecules without severe mass transfer limitations. Since a pioneering study reported on hybrid nanoflowers prepared with protein and copper sulfate, many other organic and inorganic components, which endow nanoflowers with diverse functionalities, have been employed. Thanks to these features, they have been applied in a diverse range of areas, including biosensors and biocatalysis. To highlight the progress of research on organic-inorganic hybrid nanoflowers, this review discusses their synthetic methods and mechanisms, structural and biological characteristics, as well as recent representative applications. Current challenges and future directions toward the design and development of multi-functional nanoflowers for their widespread utilization in biotechnology are also discussed.

Kinesin-13, a Motor Protein, is Regulated by Polo-like Kinase in Giardia lamblia

  • Park, Eun-Ah;Kim, Juri;Shin, Mee Young;Park, Soon-Jung
    • Parasites, Hosts and Diseases
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    • v.60 no.3
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    • pp.163-172
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    • 2022
  • Kinesin-13 (Kin-13), a depolymerizer of microtubule (MT), has been known to affect the length of Giardia. Giardia Kin-13 (GlKin-13) was localized to axoneme, flagellar tips, and centrosomes, where phosphorylated forms of Giardia polo-like kinase (GlPLK) were distributed. We observed the interaction between GlKin-13 and GlPLK via co-immunoprecipitation using transgenic Giardia cells expressing Myc-tagged GlKin-13, hemagglutinin-tagged GlPLK, and in vitro-synthesized GlKin-13 and GlPLK proteins. In vitro-synthesized GlPLK was demonstrated to auto-phosphorylate and phosphorylate GlKin-13 upon incubation with [γ-32P]ATP. Morpholino-mediated depletion of both GlKin-13 and GlPLK caused an extension of flagella and a decreased volume of median bodies in Giardia trophozoites. Our results suggest that GlPLK plays a pertinent role in formation of flagella and median bodies by modulating MT depolymerizing activity of GlKin-13.

Studies on the postnatal development of the Leydig cell in rabbits (토끼에서 출생 후 고환간질세포의 발생에 관한 연구)

  • Tae, Hyun-Jin;Park, Young-Jae;Kang, Hyung-Sub;Kim, Nam-Soo;Park, Sang-Youel;Yang, Hong-Hyun;Ahn, Dong-Choon;Kim, In-Shik
    • Korean Journal of Veterinary Research
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    • v.45 no.3
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    • pp.325-334
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    • 2005
  • Changes in the rabbit Leydig cell from birth to adulthood were studied in New Zealand white rabbits of 1, 7, 21, 35, 49, 70, 105, 147, 196, and 252 days (n = 8 rabbits per group) of age. The objectives of this study were to understand the fate of the fetal Leydig cells, to determine the changes in serum testosterone levels, and leutenizing hormone-stimulated testosterone production per testis in vitro, and to quantify adult Leydig cells by number and average volume with age. Testes of rabbits were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative (stereological) morphological studies were performed. Testosterone levels in the incubation medium of luteinizing hormone-stimulated (100 ng/ml) testosterone secretion per testis in vitro, and in serum were determined by radioimmunoassay. The average volume of a testis of 1-day-old rabbits was determined as $0.0073cm^3$ and the parameter increased linearly from birth to 252 days ($3.93cm^3$). The volume density of the seminiferous tubules increased with age from 33.76% at day 1 to 88.2% at day 252. The volume density of the interstitium represents 66.24% of the testicular parenchyma at day 1. This proportion progressively diminished during development to reach a value of 11.8% at day 252. The volume density of Leydig cells increased almost linearly from birth (0.001%) to 252 days (2.62%). Leydig cell mass per testis increases from 0.0012 mg to 0.25 mg between days 1 and 35, from 2.66 mg to 44.3 mg between days 49 and 105 and from 65.42 mg and 102.9 mg between days 147 and 252. The absolute numbers of adult Leydig cells per testis increased linearly from birth to 252 days. The average volume of adult Leydig cell on days 1, 7, 21 and 35 was not significantly different; a gradual and continued increase was observed thereafter, reaching a 3-fold increase at 196 and 252 days. Serum testosterone concentrations were not significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Values at days 70 and 105 and days 147, 196, and 252 were not significantly different. LH-stimulated testosterone production per testis in vitro was significantly different at day 1 compared days 7, 21, 35. Significant increases were observed at days 49 and 70. Hormonal values at days 105, 147, 196, and 252 were not significantly different. These data suggested Leydig cell developmental phase can be classified: a neonatal phase (1-7 days), a prepubertal phase (14-49 days) and an adult phase (70-252 days). Immature and mature adult Leydig cells, initially detected at days 7 and 49, respectively, and mature adult Leydig cells were abundant Leydig cell type according to the number and absolute volume per testis form day 49 onwards.

Evaluation of Feed Values of Korean Straws Using Pressure Transducer (Auto-Pressure Transducer를 이용한 국내산 고간류의 사료가치평가)

  • Lee, Sang S.;Ha, J. K.;Chang, M. B.
    • Journal of Animal Science and Technology
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    • v.45 no.2
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    • pp.241-250
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    • 2003
  • Accurate and rapid evaluation of the nutritional quality of Korean straws is important because of the recent increase in the use of these feedstuffs in Korean. The aim of the study was to establish with relationships between ruminal fermentation of Korean straws and in vitro gas production using a pressure transducer. The pressure transducer system includes pressure censors, AD board, LED monitor, and the computer with real-time graphics. Both gas production and DM digestibility data were fitted into the exponential equation P = a + b (1-e-$1-e^{-ct}$). The initial rate of gas production was highest for rice straw, followed by barley straw and wheat straw. The gas production rate of constant (c) in gas production for rice straw, wheat straw, and barley straw were 3.8, 2.5, and 2.5 $%h^{-1}$, respectively. Total VFA concentration (mM) produced after 72h incubation was similar among three Korean straws, even though was variable during the early (12h) fermentation. Volume of gas production was related (P> 0.05: r = 0.76 to 0.83) to DM disappearance and also strongly related (p< 0.05: r = 0.91 to 0.98) to VFA concentration at all incubation times. Linear correlation showed between gas production and DM disappearance and VFA by in vitro will be matched in in vivo digestibility.

BIOLOGICAL EFFECTS OF pH CONCENTRATION ON CULTURED HUMAN PERIODONTAL LIGAMENT CELL ACTIVITY IN VITRO (수소이온 농도의 변화가 배양 인체 치주인대 세포의 활성에 미치는 영향)

  • Kim, Seong-Ho;Park, Kui-Woon;Yoo, Hyung-Keun;Shin, Hyung-Shik
    • Journal of Periodontal and Implant Science
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    • v.25 no.3
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    • pp.539-556
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    • 1995
  • Periodontal therapeutic modalities should be re-establishing and regenerating the periodontal tissue previously lost to the disease. To achieve periodontal regeneration, periodontal ligament cells must selective migrate to the deneded root surface, attached and proliferated it. Local pH concentration is one of the most factors that periodontal regeneration. The aims of this study were to examine on biological effects of pH to the human periodontal ligament cells in vitro, especially on the cell morphology, attachment, activity, vitality and viability. Human periodontal ligament cells were cultured from extracted tooth for non-periodontal reason. Immediately after extraction, any soft tissue adhering to the cervical parts of the roots was carefully removed with a sterile curette. To produce different pH levels in the media, Eagle's MEM was adjusted from pH 6.6 to 8.2 in 0.2 intervals with 1 M NaOH and 1 N HCl. After cultivation, Then, Periodontal ligament cells were cultured at pH ranging from 6.6-8.2. attachment assay was done at 1, 2 day incubation and activity assay was done at 1, 2, 3 day incubation. The experiments were evaluated by scaning electron microscopic techniques (HITACHIX-650 Scaning Electron Microanalyzer, Tokyo, Japan), MTT assay, and the cultured periodontal ligament cells were fixed in neutral formalin for 24 hours and immunohistochemically processed by PCNA for proliferating ability. The surviving cells in the medium showed slightly increased volume and widening intercellular distances at low concentration of pH than control group (pH 7.4), and apparently shrinkage at high concentration of pH than control group (pH 7.4). The results of the statistical analysis from the experiment on attachment, vitality and viability were as follows. Attachment of periodontal ligament cells at 1st and 2nd day, similar attachment rate of low concentration pH compared with control value (pH 7.4). But above pH 8.0, attachment rate were statistically significant decrease from control value(P<0.05). Periodontal ligament cell's activities were maximum at pH 7.6 by MTT assay. Similar with control value at low concentration of pH. But, the activities were statistically significant decrease at high concentraration of pH(P<0.05). Cellular proliferating rate (PCNA index) were statistically significant decrease from control value at low and high concentration of pH(p<0.05). This results suggested that hjgh concentration pH, in other words, alkali pH was cytotoxic effects on human periodontal ligament cells in vitro.

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Variations in Nutrients and CO2 Uptake with Growth of Undaria pinnatifida from the South Coast of Korea (미역 (Undaria pinnatifida)의 생장에 따른 영양염과 CO2 흡수율 변화)

  • Shim, Jeong-Hee;Hwang, Jae-Ran;Lee, Jae-Seong;Kim, Jong-Hyun;Kim, Sung-Soo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.43 no.6
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    • pp.679-686
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    • 2010
  • To investigate the contribution of macroalgae to biogeochemical nutrients and carbon cycles, we measured the uptake rates of nutrients and $CO_2$ by Undaria pinnatifida using an incubation method in an acrylic chamber. From January to March 2010, U. pinnatifida was sampled at Ilkwang, a well-known area of macroalgae culture in Korea. The initial and final concentrations of nutrients, dissolved oxygen, total alkalinity, and pH of the chamber water were measured, and production/uptake rates were calculated using concentration changes, chamber volume, and incubation time. The production rate of dissolved oxygen by U. pinnatifida (n = 32) was about $5.4{\pm}4.0\;{\mu}mol\;g_{fw}^{-1\;}h^{-1}$. The uptake rate of total dissolved inorganic carbon (TDIC), calculated by total alkalinity and pH, was $7.9{\pm}6.5\;{\mu}mol\;g_{fw}^{-1}\;h^{-1}$. Nutrients uptake averaged $141.7{\pm}119.2$ nmol N $g_{fw}^{-1}\;h^{-1}$ and $15.0{\pm}9.1$ nmol P $g_{fw}^{-1}\;h^{-1}$. A positive linear correlation ($r^2$ = 9.6) existed between the production rate of dissolved oxygen and the uptake rate of total dissolved inorganic carbon, suggesting that these two factors serve as good indicators of U. pinnatifida photosynthesis. The relationships between fresh weight and uptake rates of nutrients and $CO_2$ suggested that younger specimens (<~50 g fresh weight) are much more efficient at nutrients and $CO_2$ uptake than are specimens >50 g. The amount of carbon uptake by the total biomass of U. pinnatifida in Korea during the year of 2008 was about 0.001-0.002% of global ocean carbon uptake. Thus, more research should be focused on macroalgae-based biogeochemical cycles to evaluate the roles and contributions of macroalgae to the global carbon cycle.

Quality Characteristics of Rye Mixed Bread Prepared with Substitutions of Naturally Fermented Raisin Extract and Sourdough (건포도 천연 발효액과 Sourdough를 이용한 호밀 혼합빵의 품질 특성)

  • Kim, Mun-Yong;Chun, Soon-Sil
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.1
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    • pp.87-94
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    • 2008
  • In this study, a nanatural fermentation starter formulation was developed for manufacturing bread products by substituting baker's yeast with naturally fermented raisin extract and sourdough. Four experimental groups containing 2.5, 5.0, 7.5, 10% naturally fermented raisin extract per 2,000 g of flour were compared based on quality characteristics, including the fermentation power on dough expansion, specific volume, baking loss, water activity, color, textural characteristics, and internal surface appearance. The activities of the naturally fermented raisin extract were examined in terms of pH changes, total titratable acidity, brix, and viable yeast counts. The raisin extract, which was cultured for 7 days at 30$^{\circ}C$, smelled of alcohol and produced $CO_2$. Yeast were also found in the extract after separation. As the incubation time of the raisin extract and sourdough increased, pH decreased, while total titratable acidity increased. The brix of the raisin extract increased until the $2^{nd}$ day of fermentation, and viable yeast counts increased until the $5^{th}$ day however, these gradually decreased by the $7^{th}$ day. The fermenting power on dough expansion increased in the bread with increasing incubation time. The bread samples containing 7.5% and 10% raisin extract had significantly higher specific volumes than the other samples. Baking loss was minimal with the 2.5% extract substitution. In analyzing the crumb, water activity, redness, and yellowness were highest in the 10.0% raisin extract bread samples, and lightness was maximal in the 5.0% group. In terms of textural characteristics, hardness was lowest with the 2.5% extract substitution. Gumminess, springiness, and chewiness were not significantly different among the bread samples. Cohesiveness was highest at the 7.5% extract substitution level, and resilience was lowest at the 10% level. In conclusion, based on the results, a natural fermentation starter formulated with 2.5% naturally fermented raisin extract (1 part raisins and 1.5 parts water) and 70% sourdough (1 part rye flour and 1 part water) has high potential as a baker's yeast substitute for making naturally fermented bread.

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Effect of Wheat Flour Brew with Bifidobacterium bifidum on Rheological Properties of Wheat Flour Dough (Bifidobacterium bifidum을 이용한 밀가루 brew가 반죽의 이화학적 성질에 미치는 영향)

  • Cho, Nam-Ji;Lee, Si-Kyung;Kim, Sung-Kon;Joo, Hyun-Kyu
    • Korean Journal of Food Science and Technology
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    • v.30 no.4
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    • pp.832-841
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    • 1998
  • In order to economically utilize flour brew with Bifidobacterium bifidum as a bread improver, the effect of flour brew on the rheological properties of dough, growth curve and acid production, and symbiosis with yeast were investigated. Growth of bifidobacteria was not increased more than initial seed volume but was consistent during 24 hours of incubation. pH was decreased and T.T.A was increased up to 12 hours of incubation. Symbiosis between bifidobacteria and yeast was little. Bifidobacteria produced more lactic acid than acetic acid in flour brew and the opposite in skim milk broth. This result was inferred from Lactobacillus sp. inherent in flour. On rheological properties of dough, farinograms of flour showed progressively decreasing baking absorption, mixing time and stability as the amount of flour brew increased. The validation of extensograms showed that R/E ratio linearly increased with increment of flour brew, and nearly doubled in all treatments comparing to that of control, which suggest the reduction of actual fermentation time. On visco/amylograms, malt index increased with addition of flour brew, accordingly showing the decrease in viscosity. Break down and set back value decreased with increment of flour brew, suggesting that staling rate of bread can be delayed.

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Effect of Amino acids on the Oxidation of Linoleic acid (일부 아미노산이 Linoleic acid의 산화반응에 미치는 영향)

  • Lee, Joon Ho;Lee, Eun Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.12 no.4
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    • pp.336-341
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    • 1983
  • A study on the oxidation effect of some amino acids(glutamic acid, phenylalanine, alanine) addition to the linoleic acid emulsion was conducted. Amino acid solutions in concentrations of $10^{-2}$, $10^{-3}$ and $10^{-4}M$ were mixed with equal volume of linoleic acid emulsions. The prepared samples were incubated at $60{\pm}1^{\circ}C$ and the extent of diene conjugation and TBA values were measured by using the UV visible spectrophotometer. The results were as follows: 1) From the extent of diene conjugation, we found that the addition of phenylalanine and alanine prolonged the induction period and the addition of glutamic acid shortened. There was an optimum concentration for each amino acid to act as an antioxidant during the induction period. The optimum concentration of alanine was $10^{-3}M$ and that of phenylalanine was $10^{-2}M$. 2) The results of TBA values showed that three amino acids possesed antioxidant activity after the induction period. There was also an optimum concentration to act as antioxidant after the induction period. The optimum concentrations of glutamic acid, phenylalanine, and alanine were $10^{-2}$, $10^{-3}$, and $10^{-3}M$, respectively.

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