• Title/Summary/Keyword: Blood culture

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Strategy for enhancing Production of recombinant Protein in tobacco's suspension culture

  • Lee, Dong-Geun;Lee, Jae-Hwa
    • Proceedings of the Korean Society of Life Science Conference
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    • 2002.12a
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    • pp.48-60
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    • 2002
  • Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a cytokine that stimulates the production of granulocytes, macrophages, and white blood cells. The effects of osmotic pressure on secretion of human GM-CSF into the culture medium were investigated in suspension cultures of transgenic tobacco cells. An increase in osmotic pressure caused by the addition of mannitol decreased the cell size index, with the effect being more pronounced when cells were measured wet rather than dry. Increased osmotic pressure enhanced the secretion of hGM-CSF. At 90 g/L mannitol, the maximum concentration tested, hGM-CSF was present in the culture medium at 980 ug/L. As the concentration of mannitol increased, the total amount of protein secreted also increased, but was disproportionately enriched in GM-CSF NaCl, another osmoticum, had very similar effects on cell growth and hGM-CSF production, but did not cause enrichment for hGM-CSF Additionally, protein-stabilizing polymer was added to culture broth to enhance stability of secreted recombinant protein. Finally, above two method were applied together to maximize the productivity.

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Biological Dosimetry of In Vitro Irradiation with Radionuclides : Comparison of Whole Blood, Lymphocyte and Buffy Coat Culture (전혈, 림프구와 백혈구 연층 각각의 방사성 동위원소 체외 조사 후 배양을 이용한 생물학적 선량측정)

  • Kim, Jong-Ho;Lee, Dong-Soo;Choi, Chang-Woon;Chung, June-Key;Lee, Myung-Chul;Koh, Chang-Soon;Kim, Chong-Soon;Kim, Hee-Geun;Kang, Duck-Won;Song, Myung-Jae
    • The Korean Journal of Nuclear Medicine
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    • v.29 no.1
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    • pp.125-132
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    • 1995
  • The purpose of this study was to establish mononuclear cell cultures such as lymphocytes or buffy coat for the biological dosimetry of in vitro Irradiation of the radionuclide Tc-99m in order to exclude the effect of residual doses seen in the cultures of whole blood. Biological do simetry of Tc-99m on cultured mononuclear cells at doses ranging from 0.05 to 6.00 Gy, by scoring unstable chromosomal aberrations(Ydr) observed in cultured lymphocytes, were performed using peripheral venous blood of healthy normal person. The results showed that; (1) In vitro irradiation of radioisotope in separated lymphocyte or buffy coat showed trace amount of residual doses of isotope after washing. Residual doses of isotopes are increased in proportion to exposed time and irradiated dose without difference between I-131 and Tc-99m. (2) We obtained these linear-quadratic dose response equations in lymphocyte and buffy coat culture after in vitro irradiation of Tc-99m, respectively (Ydr = 0.001949 $D^2$ +0.006279D + 0.000185; Ydr= 0.002531 $D^2$-0.003274 D+0.003488). In conclusion, the linear quadratic dose-response equation from in vitro irradiation of Tc-99m with lymphocyte and buffy coat culture was thought to be useful for assessing Tc-99m induced biological effects. And mono-nuclear cell cultures seem to be the most appropriate experimental model for the assessment of biological dosimetry of internal irradiation of radionuclides.

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Diagnostic value of procalcitonin and CRP in critically ill patients admitted with suspected sepsis

  • Joen, Jae-Sik;Ji, Sung-Mi
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.15 no.3
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    • pp.135-140
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    • 2015
  • Background: Identifying early markers of septic complications can aid in the diagnosis and therapeutic management of hospitalized patients. In this study, the utility of procalcitonin (PCT) vs. C-reactive protein (CRP) as early markers of sepsis was compared. Methods: A series of 2,697 consecutive blood samples was collected from hospitalized patients and serum PCT and CRP levels were measured. Patients were categorized by PCT level as follows: < 0.05 ng/ml, 0.05-0.49 ng/ml, 0.5-1.99 ng/ml, 2-9.99 ng/ml, and > 10 ng/ml. Diagnostic utility was analyzed by receiver operating characteristic (ROC) curves. Results: Mean CRP levels varied among the five PCT categories at $0.31{\pm}2.87$, $5.65{\pm}6.26$, $13.78{\pm}8.01$, $12.15{\pm}10.16$, and $17.77{\pm}10.59$, respectively (P < 0.05). PCT and CRP differed between positive and negative blood culture groups (PCT: 15.9 vs. 4.78 mg/dl;CRP: 11.5 ng/ml vs. 9.57 ng/ml;P < 0.05). The areas under the ROC curves (PCT, 95% confidence interval [CI]: 0.743, range: 0.698-0.789 at a threshold of 0.5 ng/ml; CRP, 95% CI: 0.540, range: 0.478-0.602 at a threshold of 8 mg/l) differed for PCT and CRP (P < 0.05). Conclusions: Therefore, PCT is a reliable marker for sepsis diagnosis and is more relevant than CRP in patients with a positive blood culture. These findings can be useful for the treatment of critically ill sepsis patients.

Study on the Correlation between Dietary Vitamin B Intakes and Clinical Indices of Type 2 Diabetes Patients (제2형 당뇨병 환자의 비타민 B 섭취와 임상지표의 상관관계에 대한 연구)

  • Shim, Eugene;Kwon, Ji-young;Chung, Hae-Yun
    • Journal of the Korean Society of Food Culture
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    • v.35 no.5
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    • pp.493-502
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    • 2020
  • The objective of this study was to investigate the effects of dietary vitamin B intake on biomarkers related to lipid metabolism, inflammation and blood glucose control, that are important in the development of type 2 diabetes and its complications. Seventy-six adults (42 males, 34 females) were recruited from a group of diabetes patients who had visited the medical center for treatment. Data on anthropometric characteristics and dietary intake of thiamine, riboflavin, niacin, vitamin B6 and folate were collected using 24-hour diet recall and the CAN Pro 4.0 program. Also, data on clinical indices such as serum lipids, blood pressure, high-sensitivity C-reactive protein (hs-CRP), hemoglobin A1c (HbA1c) and homeostasis model assessment 2-insulin resistance (HOMA2-IR) were collected and analyzed for correlation with dietary vitamin B intake. Results from the dietary intake survey showed that riboflavin and folate intake (in males) and folate intake (in females) were below the Dietary Reference Intake for Koreans. Statistical analysis revealed a negative correlation between hs-CRP and dietary intake of B vitamins. Riboflavin intake was inversely associated with systolic blood pressure after adjustments for age, BMI, smoking, alcohol consumption, exercise, ingestion of diabetes mellitus medication and energy intake (p<0.05). Our results suggest that dietary vitamin B may influence inflammation and consequently may help in better management of type 2 diabetes.

Production of monoclonal antibodies specific to the surface antigens of chicken peripheral blood mononuclear cells (닭의 혈액내 단핵세포 표면항원 특이 단클론성 항체 생산)

  • Choi, Jun-Gu;Sung, Haan-Woo;Kim, Sun-Joong
    • Korean Journal of Veterinary Research
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    • v.42 no.2
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    • pp.209-217
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    • 2002
  • This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group 1 mAbs (IIB3, IIB10, IIE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group I mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group II (IC6, IG2-2 and IID9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immunohistochemical staining. Five mAbs in Group III showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group IV showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (IC3 and IE9) in Group IV reacted with most of PBMC.

In vitro Expansion of Umbilical Cord Blood Derived Mesenchymal Stem Cells (UCB-MSCs) Under Hypoxic Conditions

  • Yang, Jungyun;Kwon, Jihye;Kim, Miyeon;Bae, Yunkyung;Jin, Hyejin;Park, Hohyun;Eom, Young Woo;Rhee, Ki-Jong
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.40-49
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    • 2015
  • Mesenchymal stem cells (MSCs) have the ability to self-renew and differentiate into multi-lineage cells, thus highlighting the feasibility of using umbilical cord blood-derived MSCs (UCB-MSCs) for cell-therapy and tissueengineering. However, the low numbers of UCB-MSC derived from clinical samples requires that an ex vivo expansion step be implemented. As most stem cells reside in low oxygen tension environments (i.e., hypoxia), we cultured the UCBMSCs under 3% $O_2$ or 21% $O_2$ and the following parameters were examined: proliferation, senescence, differentiation and stem cell specific gene expression. UCB-MSCs cultured under hypoxic conditions expanded to significantly higher levels and showed less senescence compared to UCB-MSCs cultured under normoxic conditions. In regards to differentiation potential, UCB-MSCs cultured under hypoxic and normoxic conditions both underwent similar levels of osteogenesis as determined by ALP and von Kossa assay. Furthermore, UCB-MSCs cultured under hypoxic conditions exhibited higher expression of OCT4, NANOG and SOX2 genes. Moreover, cells expanded under hypoxia maintained a stem cell immnunophenotype as determined by flow cytometry. These results demonstrate that the expansion of human UCB-MSCs under a low oxygen tension microenvironment significantly improved cell proliferation and differentiation. These results demonstrate that hypoxic culture can be rapidly and easily implemented into the clinical-scale expansion process in order to maximize UCB-MSCs yield for application in clinical settings and at the same time reduce culture time while maintaining cell product quality.

Effect of Mitogens, Supplement of β-mercaptoethanol and Fetal Bovine Serum Supplementation in Whole Blood Culture Medium for Bovine Chromosome Analysis

  • Takagi, M.;Tsuno, M.;Ohtani, M.;Acosta, T.J.;Miyamoto, A.;Miyazawa, K.;Sato, K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.5
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    • pp.615-621
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    • 2002
  • We investigated the effects of the mitogen supplements of 3 types, pokeweed mitogen (PWM), phytohemagglutinin (PHA) and concanavalin A (ConA), to a whole blood culture system on the number of metaphase spreads obtained in perinatal bovine chromosome analysis. In addition, the supplementation of ${\beta}$-mercaptoethanol (${\beta}$-ME) and FBS was examined in such system. Significant differences (p<0.05) were seen in the number of metaphase spreads with PHA stimulation compared to both PWM and ConA stimulation. When examined the effects of ${\beta}$-ME supplementation, the number of metaphase spreads was significantly (p<0.05) increased at $30{\mu}M$ ${\beta}$-ME compared to control. When evaluated FBS supplementation during PWM stimulation, no significant effect of the supplementation was found. Finally, the effects of the cortisol concentration (10-20, 20-30 and >30 ng/ml) of the blood samples were examined. There was no significant effect of cortisol concentration (p>0.05) among these 3 cortisol concentration groups. The mean percentages of normal metaphase plates (2n=60) from each calf 1) with ${\beta}$-ME, 2) without ${\beta}$-ME and 3) with FBS stimulated with PWM were not significantly different (p>0.05). In conclusion, these findings may be useful in cytogenetic screening programs for not only perinatal calves but also for mature cattle.

Risk factors of a Complex Disease for the Diabetes Mellitus Group and the Hypertension Group in the Geoje Community (거제지역 당뇨병 및 고혈압 환자의 복합질병 위험요인)

  • Park, Phil-Sook;Jeong, Gi-Man;Kim, Jong-Hyun;Paik, Ji-Hyun;Park, Mi-Yeon
    • Journal of the Korean Society of Food Culture
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    • v.18 no.2
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    • pp.123-133
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    • 2003
  • The purpose of this study is to compare the correlation of clinical characteristics and patterns of disease. Subjects of the study were the adults(207) living in Geoje City, the diabetes mellitus and the hypertension patients(166) and the normal people(41). In the diabetes mellitus group and the hypertension group, blood pressure, blood glucose, total cholestero LDL-cholesterol and atherogenic index(AI) were significantly high. As the obesity index was getting higher, the blood pressure of the diabetes mellitus group was high, and the HDL-cholesterol of the hypertension group was low, but AI of it was significantly high. The AI was significantly high as serum lipid index were getting higher in both groups. The rate of the prevalence was very high in the diabetes mellitus group(74.3%) and the hypertension group(73.7%). The pattern in the diabetes mellitus group was in order of the hypertension, the hyperlipidemia, and the obesity but, in the hypertension group was the hyperlipidemia, and the obesity. The obesity index and serum lipid index of complex patient group were higher than single patient group.

Effect of Plums Produced in Gimcheon on Metabolism in Streptozotocin-induced Diabetic Rats (김천산 자두가 Streptozotocin 당뇨 유발 쥐에 미치는 영향)

  • Nam, Ji-Woon;Kim, Kwang-Ok
    • Journal of the Korean Society of Food Culture
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    • v.33 no.3
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    • pp.291-298
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    • 2018
  • This study examined the effects of plums produced in Gimcheon area on the glucose and lipid metabolism in streptozotocin-induced diabetic rats. Male Sprague-Dawley diabetic rats were divided into four groups: control, diabetic control, Diabetes-low plum (containing 10% plum powder), and Diabetes-high plum (containing 20% plum powder). The animals were fed isocaloric experimental diets based on an AIN-76 diet for 6 weeks. Feed efficiency ratio (FER) of the diabetic groups were significantly lower than that of the control. On the other hand, among the diabetic groups, the FER of the high plum intake group was higher than that of the diabetic control. The liver weight per 100 g body weight of each group was similar but the liver weights tended to decrease as the amount of plum intake was increased. Kidney weight per 100 g body weight of the plum intake groups were significantly different compared to that of the diabetic control. The supplementation of plums lowered the fasting blood glucose level of the diabetic groups and improved the glucose tolerance, thereby lowering the glycosylated hemoglobin index. In addition, the supplementation of plum was lowered the blood total cholesterol concentration and increased the HDL-C/TC (%) significantly, thereby lowering the atherosclerotic index (AI) and hepatic peroxide level. A steady diet of plums produced in Gimcheon may be effective in controlling the blood glucose level and preventing chronic diabetes mellitus.

Ovarian and Fat Body Yolk Protein Synthesis in Culex piplens pallens (홍모기(Culex pipiens pallens) 지방체와 난소에서의 난황단백질합성에 관한 연구)

  • 이승훈;박영민;성기창
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.416-424
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    • 1993
  • Ovarian Yolk protein (YP2) synthesis has been investigated in mosquito, Culex pipiens pallens. Yolk protein amount which was syntheized in fat body, accumulated into ovary were analyzed by Rocket immunoelectrophoresis and in vitro organ culture. The result was that yolk protein synthesis began to occur at 6hrs after blood meal, reached at maximum level by 24hrs, and was completed within 48hrs. Yolk protein accmulation into the ovary began to start at 6hrs and coutinued for up to 60hrs after blood meal. Extract from 0, 24, 48, 72hrs ovaries after blood meal were analyzed by electrophoresis and Western blotting. The result was that 24hrs ovary contain one yolk protein(YP1), and 48, 72hrs ovaries contain two kinds of yolk proteins(YPl and YP2). When 48hr ovaries and fat bodies were incubated in $^3$H-leucine contained medium, protein synthesis was not occurred in fat body, but ovary synthesized much protein contained yolk protein (YP2). The result of crossed immunoelectrophoresis represented the same immunity between YPl and YP2. The present data suggest that ovary synthesize yolk protein(YP2) in mosquito, Culex pipiens pallens.

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