• Title/Summary/Keyword: Cellular metabolism

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Proteome analysis of sorghum leaf and root in response to heavy metal stress

  • Roy, Swapan Kumar;Cho, Seong-Woo;Kwon, Soo Jeong;Kamal, Abu Hena Mostafa;Lee, Dong-Gi;Sarker, Kabita;Lee, Moon-Soon;Xin, Zhanguo;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.24-24
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    • 2017
  • Heavy metals at toxic levels have the capability to interact with several vital cellular biomolecules such as nuclear proteins and DNA, leading to oxidative stress in plants. The present study was performed to explore the metal tolerance mechanism in Sorghum seedling. Morpho-physiological and metal ions uptake changes were observed prominently in the seedlings when the plants were subjected to different concentrations of $CuSO_4$ and $CdCl_2$. The observed morphological changes revealed that the plants treated with Cu and Cd displayed dramatically altered shoot lengths, fresh weights, and relative water content. In addition, the concentration of Cu and Cd was markedly increased by treatment with Cu and Cd, and the amount of interacting ions taken up by the shoots and roots was significantly and directly correlated with the applied level of Cu and Cd. Using the 2-DE method, a total of 24 and 21 differentially expressed protein spots from sorghum leaves and roots respectively, 33 protein spots from sorghum leaves under Cd stress were analyzed using MALDI-TOF/TOF MS. However, the over-expression of GAPDH plays a significant role in assisting Sorghum bicolor to attenuate the adverse effects of oxidative stress caused by Cu, and the proteins involved in resistance to stress helped the sorghum plants to tolerate high levels of Cu. Significant changes were absorbed in the levels of proteins known to be involved in carbohydrate metabolism, transcriptional regulation, translation and stress responses. In addition, the up-regulation of glutathione S-transferase and cytochrome P450 may play a significant role in Cd-related toxicity and stress responses. The results obtained from the present study may provide insights into the tolerance mechanism of seedling leaves and roots in Sorghum under heavy metal stress.

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Identification of Genes Involved in Primordial-primary Follicle Transition by Suppression Subtractive Hybridization

  • Park, Chang-Eun;Yoon, Se-Jin;Jeon, Eun-Hyun;Kim, Young-Hoon;Lee, Sook-Hwan;Lee, Kyung-Ah
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.98-98
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    • 2002
  • Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

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Metabolic Changes in Patients with Parkinson's Disease after Stereotactic Neurosurgery by Follow-up 1H MR Spectroscopy

  • Choe, Bo-Young;Baik, Hyun-Man;Chun, Shin-Soo;Son, Byung-Chul;Kim, Moon-Chan;Kim, Bum-Soo;Lee, Hyoung-Koo;Suh, Tae-Suk
    • Journal of the Korean Magnetic Resonance Society
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    • v.5 no.2
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    • pp.99-109
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    • 2001
  • Authors investigated neuronal changes of local cellular metabolism in the cerebral lesions of Parkinsonian symptomatic side between before and after stereotactic neurosurgery by follow-up 1H magnetic resonance spectroscopy (MRS). Patients with Parkinson's disease (PD) (n = 15) and age-matched normal controls (n = 15) underwen MRS examinations using a stimulated echo acquisition mode (STEAM) pulse sequence that provided 2${\times}$2${\times}$2 ㎤ (8ml) volume of interest in the regions of substantia nigra, thalamus, and lentiform nucleus. Spectral parameters were 20 ms TE, 2000 ms TR, 128 averages,2500 Hz spectral width, and 2048 data points. Raw data were processed by the SAGE data analysis package (GE Medical Systems). Peak areas of N-acetylaspartate (NAA), creatine (Cr), choline-containing compounds (Cho), inositols (Ins), and the sum (Glx) of glutamate and GABA were calculated by means of fitting the spectrum to a summation of Lorentzian curves using Marquardt algorithm. After blindly processed, we evaluated neuronal alterations of observable metabolite ratios between before and after stereotactic neurosurgery using Pearson product-moment analysis (SPSS, Ver. 6.0). A significant reduction of NAA/Cho ratio was observed in the cerebral lesion in substantia nigra of PD patient related to the symptomatic side after neurosurgery (P : 0.03). In thalamus, NAA/Cho ratio was also significantly decreased in the cerebral lesion including the electrode-surgical region (P : 0.03). A significant reduction of NAA/Cho ratio in lentiform nucleus was not oberved, but tended toward significant reduction after neurosurgery (P = 0.08). In particular, remarkable lactate signal was noted from the surgical thalamic lesions of 6 among 8 patients and internal segments of globus pallidus of 6 among 7 patients, respectively. Significant metabolic alterations of NAA/Cho ratio might reflect functional changes of neuropathological processes in the lesion of substantia nigra, thalamus, and lentiform nucleus, and could be a valuable finding fur evaluation of Parkinson's disease after neurosurgery. Increase of lactate signals, being remarkable in surgical lesions, could be consistent with a common consequence of neurosurgical necrosis. Thus, IH MRS could be a useful modality to evaluate the diagnostic and prognostic implications fur Parkinsons disease after functional neurosurgery.

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Ultrastructural Study on the Cerebellar Purkinje Cell of the Head-Irradiated Rat (과량의 방사선 국소조사가 흰쥐 소뇌 Purkinje세포의 미세구조에 미치는 영향)

  • Ahn, E-Tay;Yoon, Kyoo-Tae;Yang, Nam-Gil;Ko, Jeong-Sik;Park, Kyung-Ho;Kim, Jin-Gook
    • Applied Microscopy
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    • v.24 no.2
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    • pp.48-62
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    • 1994
  • The acute irradiation effect on rat Purkinje cell was carried out. Anesthetized rats, weighing 200-250g each, were exposed their heads to the linear accelerator (ML-4MV) with the doses of 3,000 rads or 6,000 rads respectively. Irradiated rats were sacrificed by perfusion fixation under anesthesia, six hours, two days and six days following the irradiations. Rats were perfused with the fixative of 1% glutaraldehyde-1% paraformaldehyde solution (pH 7.4). Small pieces of cerebellar cortices were taken out. Tissue blocks were washed out, and were refixed in the 2% osmium tetroxide solution. After dehydration, tissues were embedded in the araldite mixture. Ultrathin sections stained with uranyl acetate-lead citrate solution, were examined with an electron microscope. The results observed were as follow; 1. Many dark Purkinje cells exhibited most severe cellular alterations on 6 hours. But after the 2 or 6 days, the cells exhibited only some alterations of cytoplasmic organelles. 2. Many granular and agranular endoplasmic reticula exhibited the fusion of cisterns. These reticular alterations were most severe on 6 hours following irradiation. But the alterations were hardly found on 6 days. 3. In the Golgi region, alterations including the adhesion of lamelliform cisterns, enlarged saccules, and increased number of vesicles, etc, were seen on 6 hours. But the Golgi complexes were almost recovered on 6 days. 4. Lysosomes were abundant on 6 hours or 2 days, but some residual bodies were found on 6 days. 5. Mitochondrial changes were also most severe at on hours, and they were recovered thereafter. From the results, it was concluded that the cerebellar Purkinje cells reacted to the high doses of irradiation by hyperactive protein synthesis, autolytic activities and energy metabolism. The reaction was most active in the early stage. It implies that motor-control function of Purkinje cells are severely disturbed in the early stage of irradiation.

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Effects of Mixed Medicinal Herbs on Adipogenesis and Apoptosis in 3T3-L1 Adipocytes (배합된 한약재의 추출물이 3T3-L1 지방세포의 지방생성과 세포사멸에 미치는 영향)

  • Hwang, Su-Jung;Shin, In-Soon;Kim, Mi-Ryeo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.24 no.4
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    • pp.661-667
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    • 2010
  • Astragali Radix (AR) has been used in traditional medicinal herb, which is known to tonify the qi and blood. It has been asserted to be a tonic that can improve the function of the lungs, adrenal glands and the gastrointestinal tract, increase metabolism, promote healing and reduce fatigue. Also, Plantago asiatica (PA) has the anti-inflammatory, antiseptic, antitussive, cardiac, diuretic, expectorant, haemostatic effects, though it is considered to be less powerful than the seed. Recent research is showing that it is promising in lowering cholesterol and controlling diabetes. Obesity is characterized at the cellular level by an increase in the number and size of adipocytes differentiated from fibroblastic preadipocytes in adipose tissue. In the present study, we investigated the effects of 3 to 7 extracts of mixed medicinal herbs (water and ethanol extract of Astragali Radix; ARW and ARE, water and ethanol extract of Plantago asitica; PAW and PAE, and those mixed extracts; ARW+PAE, ARW+PAW and ARE+PAE) on adipogenesis and apoptosis in 3T3-L1 adipocytes by MTT assay, Oil-Red-O staining and TUNEL assay. And then, we determined total contents of phenolic compounds and flavonoid compounds in mixed herbal extracts. These result showed that herbal extracts (ARW, PAE and ARW+PAE) affected on adipogenesis and apoptosis in 3T3-L1 adipocytes among 3 to 7 extracts tested. Especially herbal mixed extracts (ARW+PAE) had the highest inhibition on differentiation of preadipocytes and lipoid accumulation of maturing adipocytes. And it did not affect cell viability of mature adipocytes, but herbal mixed extracts (ARW+PAE) increased apoptosis in mature adipocytes, as proved by highest concentration $200\;{\mu}g/m{\ell}$ using TUNEL assay. In addition, contents of total phenoilc compounds and flavonoids showed the highest level in ARE+PAE and PAE than ARW. These results suggest that this main target for adipocyte functions could be effective in improving the metabolic syndrome including obesity.

Disorders of Potassium Metabolism (칼륨 대사 장애)

  • Lee, Joo-Hoon
    • Childhood Kidney Diseases
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    • v.14 no.2
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    • pp.132-142
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    • 2010
  • Hypokalemia usually reflects total body potassium deficiency, but less commonly results from transcellular potassium redistribution with normal body potassium stores. The differential diagnosis of hypokalemia includes pseudohypokalemia, cellular potassium redistribution, inadequate potassium intake, excessive cutaneous or gastrointestinal potassium loss, and renal potassium wasting. To discriminate excessive renal from extrarenal potassium losses as a cause for hypokalemia, urine potassium concentration or TTKG should be measured. Decreased values are indicative of extrarenal losses or inadequate intake. In contrast, excessive renal potassium losses are expected with increased values. Renal potassium wasting with normal or low blood pressure suggests hypokalemia associated with acidosis, vomiting, tubular disorders or increased renal potassium secretion. In hypokalemia associated with hypertension, plasam renin and aldosterone should be measured to differentiated among hyperreninemic hyperaldosteronism, primary hyperaldosteronism, and mineralocorticoid excess other than aldosterone or target organ activation. Hypokalemia may manifest as weakness, seizure, myalgia, rhabdomyolysis, constipation, ileus, arrhythmia, paresthesias, etc. Therapy for hypokalemia consists of treatment of underlying disease and potassium supplementation. The evaluation of hyperkalemia is also a multistep process. The differential diagnosis of hyperkalemia includes pseudohypokalemia, redistribution, and true hyperkalemia. True hyperkalemia associated with decreased glomerular filtration rate is associated with renal failure or increased body potassium contents. When glomerular filtration rate is above 15 mL/min/$1.73m^2$, plasma renin and aldosterone must be measured to differentiate hyporeninemic hypoaldosteronism, primary aldosteronism, disturbance of aldosterone action or target organ dysfunction. Hyperkalemia can cause arrhythmia, paresthesias, fatigue, etc. Therapy for hyperkalemia consists of administration of calcium gluconate, insulin, beta2 agonist, bicarbonate, furosemide, resin and dialysis. Potassium intake must be restricted and associated drugs should be withdrawn.

Formation and Stability of Nanoemulsion Containing CoQ10 by Mechanical Emulsification (코엔자임 Q10을 함유하는 나노에멀젼의 제조)

  • Yoo, In-Sang
    • Applied Chemistry for Engineering
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    • v.23 no.5
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    • pp.467-473
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    • 2012
  • Coenzyme Q10 (CoQ10) is a natural lipid cofactor with antioxidant and anti-aging properties as cosmetic and food ingredients, involved in cellular energy metabolism. Here, nano-emulsions with CoQ10 were fabricated with lecithin, ethanol, oil, and sorbitan monostearate (Arlacel 60), as major components. Phase inversion emulsion method with ultrasonicator was utilized in producing CoQ10 solution, and stabilization effects from lecithin and ethanol and other diverse perturbation factors were evaluated over time. Physical properties of the emulsion were characterized such as its size, surface charges by zeta-potential, and the overall structures. Optimal concentrations of CoQ10 and Arlacel 60 were 0.8% and 3%, respectively, for producing the smallest sizes of nanoemersions in a 100 nm diameter with best morphology. No notable changes in the size were observed over 7 days from Ostwald ripening, when the concentration of Arlacel 60 was higher than 2%. Even after 270 days at room temperature, the size of nanoemulsions maintained as 115 nm in diameter, revealing only a 10% increase with high degrees of long termed stability and substantiality. In addition, changes in the surface potential occurred possible due to the flocculation effect on the nanoparticles.

Effect of Bisphenol A on Insulin-Mediated Glucose Metabolism In Vivo and In Vitro

  • Ko, Jeong-Hyeon;Kang, Ju-Hee;Park, Chang-Shin;Shin, Dong-Wun;Kim, Ji-Hye;Kim, Hoon;Han, Seung-Baik
    • Molecular & Cellular Toxicology
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    • v.4 no.4
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    • pp.348-354
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    • 2008
  • Bisphenol A (BPA), an environmental endocrine disrupter, enters the human body continuously in food and drink. Young children are likely to be more vulnerable than adults to chemical exposure due to the immaturities of their organ systems, rapid physical development, and higher ventilation, metabolic rates, and activity levels. The direct effect of BPA on peripheral tissue might also be of importance to the development of insulin resistance. However, the influence that BPA has on insulin signaling molecules in skeletal muscle has not been previously investigated. In this study, we examined the effect of BPA on fasting blood glucose (FBG) in post-weaned Wistar rats and on insulin signaling proteins in C2C12 skeletal muscle cells. Subsequently, we investigated the effects of BPA on insulin-mediated Akt phosphorylation in C2C12 myotubes. In rats, BPA treatment (0.1-1,000 ng/mL for 24 hours) resulted in the increase of FBG and plasma insulin levels, and reduced insulin-mediated Akt phosphorylation. Furthermore, the mRNA expression of insulin receptor (IR) was decreased after 24 hours of BPA treatment in C2C12 cells in a dose-dependent manner, whereas the mRNA levels of other insulin signaling proteins, including insulin receptor substrate-1 (IRS-1) and 5'-AMP-dependent protein kinase (AMPK), were unaffected. Treatment with BPA increased GLUT4 expression and protein tyrosine phosphatase 1B (PTP1B) activity in C2C12 myotubes, but not in protein levels. We conclude that exposure to BPA can induce insulin resistance by decreasing IR gene expression, which is followed by a decrease in insulin- mediated Akt activation and increased PTP1B activity.

Comparative Genomics Profiling of Clinical Isolates of Helicobacter pylori in Chinese Populations Using DNA Microarray

  • Han, Yue-Hua;Liu, Wen-Zhong;Shi, Yao-Zhou;Lu, Li-Qiong;Xiao, Shudong;Zhang, Qing-Hua;Zhao, Guo-Ping
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.21-28
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    • 2007
  • In order to search for specific genotypes related to this unique phenotype, we used whole genomic DNA microarray to characterize the genomic diversity of Helicobacter pylori (H. pylori) strains isolated from clinical patients in China. The open reading frame (ORF) fragments on our microarray were generated by PCR using gene-specific primers. Genomic DNA of H. pylori 26695 and J99 were used as templates. Thirty-four H. pylori isolates were obtained from patients in Shanghai. Results were judged based on In(x) transformed and normalized Cy3/Cy5 ratios. Our microarray included 1882 DNA fragments corresponding to 1636 ORFs of both sequenced H. pylori strains. Cluster analysis, revealed two diverse regions in the H. pylori genome that were not present in other isolates. Among the 1636 genes, 1091 (66.7%) were common to all H. pylori strains, representing the functional core of the genome. Most of the genes found in the H. pylori functional core were responsible for metabolism, cellular processes, transcription and biosynthesis of amino acids, functions that are essential to H. pylori's growth and colonization in its host. In contrast, 522 (31.9%) genes were strain-specific genes that were missing from at least one strain of H. pylori. Strain-specific genes primarily included restriction modification system components, transposase genes, hypothetical proteins and outer membrane proteins. These strain-specific genes may aid the bacteria under specific circumstances during their long-term infection in genetically diverse hosts. Our results suggest 34 H. pylori clinical strains have extensive genomic diversity. Core genes and strain-specific genes both play essential roles in H. pylori propagation and pathogenesis. Our microarray experiment may help select relatively significant genes for further research on the pathogenicity of H. pylori and development of a vaccine for H. pylori.

The Relationships between UCP-1 Polymorphism and the Degree of Obesity or Plasma Lipid Profile in Prepubertal Children (소아에서의 UCP-1 다형성과 비만도 및 혈액 지질수치와의 관련성에 관한 연구)

  • Oh, Hyun-Hee;Shin, Eun-Jung;Lee, Myoung-Sook
    • Journal of Nutrition and Health
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    • v.41 no.8
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    • pp.767-775
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    • 2008
  • Uncoupling protein-1 (UCP-1) plays a major role in thermogenesis at brown adipose tissues and has been implicated in the pathogenesis of obesity and metabolic disorders. The purpose of this study was to estimate the effects of A-3826G polymorphism in 117 Korean prepubertal children aged 8-11 years olds. Anthropometry by bioelectrical impedance analysis method, plasma lipid profiles by auto-biochemical analyzer and UCP-1 genotyping by PCR-RFLP were done. The frequencies of UCP-1 genotypes were AA; 17.7%, AG; 57.8%, GG; 26.6%. The frequencies of each G allele (55.5%) was similar to Japanese's (49%) and higher than Caucacian's (25%). No correlation UCP-1 polymorphism and BMI (kg/$m^2$) or the degree of obesity described by the relative percentiles of the standard weight according to height in prepubertal children. However, plasma total- and LDL-cholesterol were significantly increased in G allele when sex, age and weight were adjusted. Our results suggested that G allele of UCP-1 gene was stronger risk factors in hyperLDLcholesterolemia than A allele. This impact might be progressed as the precaution against the revalence of obesity based-metabolic disease.