• Title/Summary/Keyword: Sodium pyruvate

Search Result 58, Processing Time 0.026 seconds

Effects of Amino Acids, Carbohydrates and Phosphorus Sources on Growth and Alkaline Phosphatase Activity of the Marine Cyanobacterium Anabaena sp. Strain CA

  • Singh, Jeet Bahadur;Vyas, Deepak;Kumar, Har Darshan
    • Journal of Microbiology and Biotechnology
    • /
    • v.7 no.2
    • /
    • pp.127-131
    • /
    • 1997
  • Alkaline phosphatase (APase) was found to be inducible in Anabaena sp. strain CA Growth was less than control in presence of most amino acids except glycine and serine, but most amino acids enhanced APase activity. Highest APase activity was recorded in tyrosine supplemented culture followed by hydroxyproline, cystein, valine and glutamic acid. Threonine supplemented material showed lowest APase level (1.8 nmol/mg protein/min). Lactose, glucose, sodium pyruvate and succinate stimulated growth but not APase activity. APase activity was high in the presence of sucrose, mellibiose, mannitol, arabinose, maltose and sorbose, even though the growth in these supplements was less than in control. Organic phosphate sources supported good growth of the organism. Best growth occurred in presence of inorganic phosphate, adenosine diphosphate, fructose 1,6-diphosphate or ribulose 1,5-diphosphate, followed by other phosphorus sources tested. APase activity in presence of any of the organic phosphate sources was 3 to 5 fold low as compared to phosphate limited culture. Also, there was no APase activity in cultures grown on inorganic phosphate. These data indicate that most amino acids and a few carbohydrates (sucrose, mellibiose, arabinose and sorbose) are suitable for APase production. Lactose, glucose, pyruvate or succinate may be used as a carbon source during photoheterotrophic growth of the cyanobacterium. Glycine and serine are preferred nitrogen sources for its growth. Phosphate repressible APase activity has been found in Anabaena sp. strain CA.

  • PDF

Maintenance and Differentiation of Pluripotential Embryonic Cell Lines from Mouse Blastocysts (BCF1 생쥐 배반포기 유래 배아간세포 작성에 관한 연구)

  • 이재원;이훈택;정길생
    • Korean Journal of Animal Reproduction
    • /
    • v.18 no.4
    • /
    • pp.235-244
    • /
    • 1995
  • The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.

  • PDF

Improvement of the Detection Technique of Listeria monocytogenes through Modification of the Enrichment Medium and DNA Extraction Buffer (증균배지 및 DNA 추출법 개량을 통한 Listeria monocytogenes의 검출기법 개선 연구)

  • Lee, Jeeyeon;Seo, Yeongeun;Yoon, Yohan
    • Journal of Food Hygiene and Safety
    • /
    • v.35 no.4
    • /
    • pp.334-340
    • /
    • 2020
  • In this study we developed an enrichment medium and lysis buffers to detect Listeria monocytogenes in meat and processed meat products under various lysis conditions. The enrichment efficiency of L. monocytogenes medium listed in the Food Standards was compared, and thus, Listeria Enrichment Broth (LEB) was modified by adding supplements such as carbon source and minerals. The lysis buffers were developed to extract L. monocytogenes DNA quickly and efficiently under various lysis conditions. L. monocytogenes was most rapidly grown in LEB containing 0.1% pyruvate and 0.1% ferric citrate. A lysis buffer mixed with 0.5% or 1% N-lauroylasrcosine sodium salt, 0.5 N NaOH and 0.5 M EDTA for 30 min at room temperature was found to be the best in terms of DNA purity and yield. These results indicate that developed enrichment medium and lysis buffer can be used to detect L. monocytogenes in meat and processed meat products rapidly and efficiently.

Dichloroacetate Inhibits the Proliferation of a Human Anaplastic Thyroid Cancer Cell Line via a p53-independent Pathway (Dichloroacetate의 p53 비의존적 경로를 통한 인간 역분화 갑상선 암세포주의 성장억제 효과)

  • KC, Yam Bahadur;Poudel, Sunil;Jeon, Eon Ju;Shon, Ho Sang;Byun, Sung June;Jeoung, Nam Ho
    • Journal of Life Science
    • /
    • v.28 no.12
    • /
    • pp.1469-1476
    • /
    • 2018
  • Occurrence of the Warburg effect in solid tumors causes resistance to cancer chemotherapy, and targeting energy metabolisms such as aerobic glycolysis is a potential strategy for alternative treatment. Dichloroacetate (DCA), an inhibitor of pyruvate dehydrogenase kinase (PDK), shifts glucose metabolism from aerobic glycolysis to oxidative phosphorylation (OxPhos) in many cancers. In this study, we investigated the anticancer effect of DCA on a human anaplastic thyroid cancer (ATC) cell line, 8505C. We found that DCA selectively inhibits cell proliferation of the 8505C line but not of a normal thyroid line. In 8505C, the cell cycle was arrested at the G1/S phase with DCA treatment as a result of decreased antiapoptotic proteins such as $HIF1{\alpha}$, PDK1, and Bcl-2 and increased proapoptotic proteins such as Bax and p21. DCA treatment enhanced the production of reactive oxygen species which consequently induced cell cycle arrest and apoptosis. Interestingly, DCA treatment not only reduced lactate production but also increased the expression of sodium-iodine symporter, indicating that it restores the OxPhos of glucose metabolism and the iodine metabolism of the ATC. Taken together, our findings suggest that PDK inhibitors such as DCA could be useful anticancer drugs for the treatment of ATC and may also be helpful in combination with chemotherapy and radiotherapy.

Gilt Transfer of Cultured Freezing Embryos by Open Pulled Straw(OPS) Methods (Open Pulled Straw(OPS) 방법에 의한 체외 배양 동결 수정란의 미경산돈 이식)

  • Kim, In-Doc;Seok, Ho-Bong
    • Journal of Embryo Transfer
    • /
    • v.23 no.3
    • /
    • pp.217-222
    • /
    • 2008
  • In previous studies, we reported that sow which was transferred OPS-freezing embryos not able to deliver a piglet (Kim et al, 2004). This study was conducted to investigate a possibility of gilt as recipients which produce piglets after transfer of OPS-freezing embryos. All transferred embryos were prepared by in vitro production (IVP) system. In vitro culture (IVC) medium used glucose-free NCSU23 supplemented with 5mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at $39^{\circ}C$. From day 3 of IVC, 10% fetal bovine serum albumin was added to the culture medium. In preparing of freezing embryos, embryos were treated with 7.5 $\mu g/ml$ cytochalasin-B for 30 min and centrifuged at $13,000{\times}g$ for 13 min. And then, embryos were exposed sequentially to an ethylene glycol (EG) solution, aspirated into open pulled straw (OPS), and plunged or thawed into the liquid nitrogen. In embryo transfer (ET), we used two kinds of type (surgical method vs. non-surgical method). In surgical method of embryo transfer, $55\sim65$ embryo were transferred in both uterine horn of two recipient gilts by plastic straw. Non-surgical method which is like artificial insemination was performed on three gilts. Each 140 frozen embryos were transferred to two gilts and 40 fresh embryos to one gilt. Pregnancy establishment was shown one recipient at 45 days after ET. However, the one recipient was also aborted at 58 days after ET. These results suggest that gilts can be considered as a candidate of recipients for OPS-freezing embryo transfer.

Sow Transfer of Cultured Embryos : Embryo Recovery, Oocyte Collection, IVM-IVF-IVC Treatment, Vitrification/Thaw, and Surgical and Nonsurgical Transfer

  • Kim, In-Doc;Ahn, Mi-Hyun;Hur, Tae-Young;Son, Dong-Soo;Hong, Moon-Pyo;Seok, Ho-Bong
    • Proceedings of the Korean Society of Developmental Biology Conference
    • /
    • 2003.10a
    • /
    • pp.128-128
    • /
    • 2003
  • The aims of this study are 1) to test oocytes and embryos collected from in-vivo and in-vitro to achieving the valuable protocol by culturing, vitrifying and thawing of oocytes/embryos, and 2) to transfer them to recipient, and finally have resulted in pregnancies from recipient females after surgical or nonsurgical transfer. In vitro maturation and fertilization were performed according to the procedures of Funahashi et al. Fertilized oocytes were cultured in glucose-free NCSU 23 supplemented with 5 mM sodium pyruvate, 0.5 mM sodium lactate and 4 mg/ml bovine serum albumin for 2 days at 39$^{\circ}C$, and 10% fetal bovine serum was added to the culture medium thereafter. Embryos were treated with 7.5$\square$g/ml cytochalasin-B for 30 min, centrifuged at 13,000 ${\times}$ g for 13 min and then exposed sequentially to an ethylene glycol (EG) vitrification solution, aspirated into OPSs, and plunged/thawed into/from liquid nitrogen. In vivo embryos were surgically collected from three donors after Al. Forty-six embryos (18, 9 and 19 embryos, respectively) were washed 3 times in mPBS+10%FBS, followed treatments : cultured, centrifuged, vitrified, recovered and transferred to recipients as in vitro prepared embryos. Three recipients received surgically 34(control), 188 and 184 embryos (derived from abattoir), respectively. Another three recipients were received nonsurgically 150, 100 and 150 embryos, respectively. All recipient sows exhibited delayed returns to estrus. To our knowledge, these results suggest that required an improved techniques, more vigorous embryos preparation and cleaner uterous condition(use gilt).

  • PDF

Nuclear Progression through In Vitro Meiotic Maturation of Bovine Oocytes (우 난포란의 체외성숙시 핵의 발달과정)

  • 주영국;공일근;정미경;강대진;박충생
    • Journal of Embryo Transfer
    • /
    • v.7 no.2
    • /
    • pp.125-131
    • /
    • 1992
  • The nudear changes of bovine oocytes during 24 hrs. of culture for mejotic maturation were examined. Bovine oocytes were collected from small(<2 mm), medium(2~6 mm) and large(>6mm) follicles and classified into three grades by their morphological characteristics. A total of 242 oocytes collected were obtained:from 184 small, 157 medium and 1 large follicles, respectively and were classified into 95 grade I, 155 grade H and 92 grade III oocytes. All the bovine oocytes collected and graded were washed with a basal medium and incubated in groups of 10 for 24 hrs in 5% $CO_2$ and 39$^{\circ}C$. The basal medium used was composed of TCM-199 supplemented with sodium bicarbonate, sodium pyruvate, streptomycin, penicillin G and 10% FCS. The oocytes were cultured in drops of 50,$\mu$l basal medium supplemented with 35$\mu$g /ml FSH, 10$\mu$g /ml LH and 1$\mu$g /ml estradiol-17$\beta$. The oocytes were fixed and examined on their chromosomal status by 1% acetorcein staining in the interval of 3 hrs. Most of the grade I oocytes developed to germinal vesicule stage at 0 to 3 hrs., germinal vesicle breakdown at 6 hrs., metaphase I at 9 to 15 hrs., anaphase I and telophase I at 18 hrs., and metaphase II and the first polar body at 24 hrs. after culture for meiotic maturation. However, it was found that compared to grade I oocytes, grade H and W oocytes reached earlier to germinal vesicle breakdown and most of them developed earlier to M II stage at 21 hrs. after culture.

  • PDF

The Effect of Yindong on Gout of Rats induced by Monosodium Urate (인동(忍冬)이 Monosodium Urate로 유발(誘發)된 백서(白鼠)의 통풍(痛風)에 미치는 영향(影響))

  • Lee, Hong-Jin;Cho, Chung-Sik;Kim, Chul-Jung
    • Journal of Haehwa Medicine
    • /
    • v.17 no.1
    • /
    • pp.83-93
    • /
    • 2008
  • In order to study the Effects of yindong(LC) on the variation of blood and joint value the gout induced by microcrystalline sodium urate in rats, for LC is one of the important medicine on gout. After pretreatment of LC(50, 500mg/kg) for 5days, the Effects of LC was evaluated on Serum albumin, Serum globulin, glutamate dxalacetate transminase(AST), glutanate pyruvate transminase(ALT), blood urea nitrogen(BUN), Serum creatitine, Serum uric acid, xanthine oxidase activity, Erythrocyte Sedimentation Rate(ESR), WBC, platelet were measured. The results were obtained as follows : Joint value Increase ratio was not significantly decreased in all LC taken groups compared with the control group. Serum albumin was significantly different in all LC taken groups compared with the control group and Serum globulin was significantly dicreased in 500mg/kgLC taken group compared with the control group. Serum AST, ALT were significantly dicreased in 500mg/kgLC taken group compared with the control group. Serum BUN was significantly decreased in all LC taken groups and Serum creatinine was significantly decreased in 500mg/kgLC taken group compared with the control group. Serum uric acid was significantly different in 500mg/kgLC taken group, and changes in xanthine oxidase activity was significantly decreased in 500mg/kg, 50mg/kgLC taken group. ESR was significantly decreased in all LC taken groups compared with the control group. WBC, platelet count were significantly decreased in 500mg/kgLC taken group compared with the control group. From above results it may be concluded that Yindong can be used for treatment and preventive medcine of gout induced by microcrystalline sodium urate in clinic.

  • PDF

Metabolomic profiling of embryo culture media in patients with repeated implantation failure during assisted reproductive technology cycles

  • Sheyda Nami;Azam Govahi;Nabaa Najjar;Samaneh Ghasemi;Farshid Rezaei;Fatemehsadat Amjadi;Gholamreza Taheripak
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.51 no.3
    • /
    • pp.260-267
    • /
    • 2024
  • Objective: This study investigated the metabolic status of the spent culture media from embryos of patients with repeated implantation failure (RIF) undergoing in vitro fertilization-intracytoplasmic sperm injection cycles in comparison with the embryos from healthy fertile women. Methods: Metabolite levels in spent culture media were assessed and compared between embryos from RIF patients (n=35) and oocyte donors as controls (n=15). Protein levels of insulin-like growth factor 1 (IGF-1) were determined using Western blotting. Concentrations of glucose, pyruvate, and lactate were measured using spectrophotometry. Ionic colorimetric assay kits were utilized to analyze the concentrations of sodium, chloride, calcium, and magnesium ions. High-performance liquid chromatography was employed to measure the concentrations of glutamic acid, aspartic acid, methionine, phenylalanine, and histidine. Results: Glucose consumption and lactate secretion were higher in the control group than in the RIF group. The magnesium concentration was significantly higher in the control group than in the RIF group, but glutamic acid and aspartic acid concentrations were lower in the control group than in the RIF patients (p<0.05). The levels of IGF-1, sodium, calcium, chloride, methionine, histidine, and phenylalanine did not show statistically significant differences between the two groups. Conclusion: The metabolic profile of the culture medium of the embryos in the RIF group differed from that of the control group. These findings suggest potential factors that may affect implantation capacity in RIF patients and provide a new perspective on embryo selection.

Develpment of Low-serum Medium(LSM) for Mouse-mouse Hybridoma Part II. Development of Low Serum Medium by Screening for Serum Replacement (Hybridoma 배양을 위한 저혈청 배지의 개발 제2부: 혈청 대체 물질 선정을 통한 저혈청 배지 제조)

  • 제훈성;최차용
    • KSBB Journal
    • /
    • v.7 no.2
    • /
    • pp.96-101
    • /
    • 1992
  • A low serum medium(LSM) suitable for the growth of a self-constructed hybridoma cell line, KA112, was established by selecting ingredients to replace serum. Insulin and sodium pyruvate was important for the growth of cell line KA112. Various basal media were tested and DMEM gave the most favorable result. Low serum medium (LSM) developed in this work showed cell line stability in the culture for more than 6 months and exhibited cell growth equivalent to that carried out in medium supplemented with 7% FBS. LSM was found to be applicable to the suspension culture of KA112. The reduction of serum level down to 1%(V/V) FBS in LSM resulted in a substantial saving in the cost of media preparation for large scale culture.

  • PDF