• Title/Summary/Keyword: quantitative expression analysis

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Nuclear Akt promotes neurite outgrowth in the early stage of neuritogenesis

  • Park, Ji-Hye;Lee, Sang-Bae;Lee, Kyung-Hoon;Ahn, Jee-Yin
    • BMB Reports
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    • v.45 no.9
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    • pp.521-525
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    • 2012
  • In addition to its pivotal role in neuronal survival, PI3K/Akt signaling is integral to neuronal differentiation and neurite outgrowth. However, the exact role of Akt in neuronal differentiation is still controversial. Here, we found that nuclear expression of CA-Akt resulted in unusual rapid neurite outgrowth and overexpression of KD-Akt caused multiple dendrite growth without specific axon elongation. Moreover, microarray data revealed that the expression of FOXQ1 expression was about 10-fold higher in cells with nuclear, active Akt than in control cells. Quantitative real-time PCR analysis showed that mRNA levels were upregulated in NLS-CA-Akt cells as compared to KD or EV cells. Furthermore, our FACS analysis demonstrated that overexpression of NLS-CA-Akt accumulate cells in the G1 phase within 24 h, fitting with the rapid sprouting of neuritis. Thus, our data implied that at least in this early time frame, the overexpression of nuclear, active Akt forced cells into neurite development through probably FOXQ1regulation.

Expression of Poly (ADP-ribose) Polymerase During Apoptosis Induced by Ultraviolet Radiation in HeLa $S_3$ Cells

  • Chang, Jeong-Hyun;Kwon, Heun-Young
    • Biomedical Science Letters
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    • v.12 no.4
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    • pp.349-354
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    • 2006
  • Induction of apoptosis allows the organism to get rid of abnormal cells and also of tumor cells. Understanding the mechanism involved in Ultraviolet radiation (UV) induced apoptosis may improve its therapeutic efficacy. In this study, we present expression of poly (ADP-ribose) polymerase (PARP) during apoptosis induced by UV in HeLa $S_3$ cells. Four different assays were performed in this study: morphological assessment of apoptotic cells and cell viability, DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, and expression of PARP by the western blot analysis. The percentages of apoptotic HeLa $S_3$ cells irradiated with $75J/m^2$ UV was increased continuously from 3 hrs incubation. DNA ladder pattern was appeared at 6 hrs. The amount of nucleosomal DNA fragments in cells treated UV increased from 3 to 12 hrs incubation and gradually decreased. The cleavage of PARP in HeLa $S_3$ cells irradiated with UV was induced, and the cleavage of PARP was more delayed in the cells pretreated with $5J/m^2$ UV and subsequently irradiated with $75J/m^2$ UV. than that in the cells only irradiated with $75J/m^2$ UV. Thus these data suggest that the cleavage of PARP relates with DNA fragmentation associated with apoptosis.

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Molecular characterization and immunohistochemical localization of a mitogen-activated protein kinase, Accp38b, from Apis cerana cerana

  • Zhang, Liang;Meng, Fei;Li, Yuzhen;Kang, Mingjiang;Guo, Xingqi;Xu, Baohua
    • BMB Reports
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    • v.45 no.5
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    • pp.293-298
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    • 2012
  • The p38 mitogen-activated protein kinase (MAPK) is involved in various processes, including stress responses, development, and differentiation. However, little information on p38 MAPK in insects is available. In this study, a p38 MAPK gene, $Accp38b$, was isolated from $Apis$ $cerana$ $cerana$ and characterized. The quantitative real-time PCR (Q-PCR) analysis revealed that $Accp38b$ was induced by multiple stressors. Notably, the expression of $Accp38b$ was relatively higher in the pupae phase than in other developmental phases. During the pupae phase, Accp38b expression was higher in the thorax than in the head and abdomen and higher in the fat body than in the muscle and midgut. Immunohistochemisty showed significant positive staining of Accp38b in sections from the brain, eyes, fat body, and midgut of $A.$ $cerana$ $cerana$. These results suggest that Accp38b may play a crucial role in stress responses and have multiple aspects function during development.

The Effects of Saussurea Radix and Plantaginis Herba on Cellular Viability, Proliferation, Apoptosis and Expression of Cell Cycle-related Genes in Gastric Cancer Cells (목향(木香)과 차전초(車前草)가 위암세포(胃癌細胞)의 활성(活性), 증식(增殖), 자기살해능(自己殺害能) 및 세포주기관련 유전자 발현에 미치는 영향)

  • Oh, Hee-Rah;Ko, Seong-Gyu
    • THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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    • v.7 no.1
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    • pp.1-18
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    • 2001
  • Objective: This experimental study was carried out to evaluate the effects of Saussurea Radix and Plantaginis Herba on cellular viability, proliferation, apoptosis and expression of the cell cycle-related genes in cultured gastric cancer cells. Method :MTT assay for analysis of cellular toxicity and the effect on suppression of cellular viability, $[^{3}H]$ thymidine incorporation assay for evaluation of the effect on suppression of DNA replication, tryphan blue exclusion assay for measurement of induction of apoptosis and Quantitative RT-PCR for analysis of the effects on expression of cell cycle or apoptosis-related genes were performed. Results: Antitumor activity of Saussurea Radix associated with inhibition of cell cycle progression and promotion of apoptosis caused by transcriptional regulation of p53, p21/Wafl and the other related genes was observed.

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Apoptosis and Anti-proliferaction by Saussurea lappa and Pharbitis nil in AGS Human Gastric Cancer Cell Line

  • Ko Seong-Gyu;Oh Hee-Rah;Lee Sun-Dong;Hwang Gwi-Seo
    • The Journal of Internal Korean Medicine
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    • v.24 no.1
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    • pp.134-143
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    • 2003
  • Objectives : We performed this study to understand the molecular basis of the antitumor effect of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for cancer treatment in Korean traditional medicine. Design: We analyzed, the effect of these medicinal herbs on proliferation and apoptosis of tumor cells and its association with gene expression, We performed semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) analysis of cell cycle- and apoptosis-related genes using a gastric cancer cell line AGS. Results : Cell counting assay and $[^3H]thymidine$ uptake analysis showed that Saussurea lappa and Pharbitis nil strongly inhibit cell proliferation of AGS in a dose-dependent manner. Interestingly, gene espression assay revealed that mRNA espression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly decreased by Saussurea lappa and Pharbitis nil. Furthermore, Saussurea lappa was identified to activate expression of the p53 tumor suppressor and its downstream effector $p21^{Wafl}$, which leads to $G_1$ cell cycle arrest and apoptosis. These observations suggest that the anticancer effect of Saussurea lappa and Pharbitis nil might be associated with their regulatory capability of tumor-related gene expression.

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Gene Expression Analysis of Pregnant Specific Stage in the Miniature Pig Ovary

  • Yun, Seong-Jo;Noh, Won-Gun;Yoon, Jong-Taek;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.33 no.4
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    • pp.249-255
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    • 2009
  • The miniature pig is considered to be a better organ donor breed for xenotransplantation than other pig breeds because the size of the organs of the miniature pig is similar to that of humans. In this study, we aimed at identifying differentially expressed genes in the miniature pig ovary during pregnancy. For this, we used the miniature pig ovary model, annealing control primer-based reverse transcription polymerase chain reaction (PCR), quantitative real-time PCR (qRT-PCR), and northern blotting analysis. We identified 13 genes showing differential expression on the based of pregnancy status and validated 8 genes using qRT-PCR. We also sequenced the full-length cDNA of ephrin receptor A4 (EphA4), which had a significant difference in expression level, and validated it by northern blotting. These genes may provide a better understanding of the cellular and molecular mechanisms during pregnancy in miniature pig ovary.

Correlation between chromosome abnormalities and genomic imprinting in developing human - 1) Frequent biallelic expression of insulin-like growth factor II (IGF2) in gynogenetic Ovarian Teratomas: Uncoupling of H19 and IGF2 imprinting

  • Choi, Bo-Hwa;Lee, In-Hwan;Chun, Hyo-Jin;Kang, Shin-Sung;Chang, Sung-Ik
    • Journal of Genetic Medicine
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    • v.2 no.1
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    • pp.41-47
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    • 1998
  • Human uniparental gestations such as gynogenetic ovarian teratomas provide a model to evaluate the integrity of parent-specific gene expression - i.e. imprinting - in the absence of a complementary parental genetic contribution. The few imprinted genes characterized so far include the insulin-like growth factor-2 gene (IGF2) coding for a fetal growth factor and H19 gene whose normal function is unknown but it is likely to act as an mRNA. IGF2 is expressed by the paternal allele and H19 by the maternal allele. This reciprocal expression is quite interesting because both H19 and IGF2 genes are located close to each other on chromosome 11p15.5. In situ RNA hybridization analysis has shown variable expression of the H19 and IGF2 alleles according to the tissue origin in 11 teratomas. Especially, Skin, derivative of ectoderm, is expressed conspicuously. We examined imprinting of H19 and IGF2 in teratomas using PCR and RT-PCR of exonic polymorphism. H19 and IGF2 transcript could be expressed either biallelically or monoallelically in the teratomas. Biallelic expression (i.e., loss of imprinting) of IGF2 occurred in 5 out of 6 mature teratomas and 1 out of 1 immature teratoma. Biallelic expression of H19 occurred in 4 out of 10 mature teratomas and 1 out of 1 immature teratoma. Expression levels of H19 and IGF2 transcript using the semi-quantitative RT-PCR had no relation between monoallelic and biallelic expression. Moreover, IGF2 biallelic expression did not affect allele-specificity or levels of H19 expression. These results demonstrate that both genes, H19 and IGF2, can be imprinted, expressed and regulated independently and individually of each other in ovarian teratoma.

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Expression of heat shock protein genes in Simmental cattle exposed to heat stress

  • Luis Felipe Guzman;Guillermo Martinez-Velazquez;Fernando Villasenor-Gonzalez;Vicente Eliezer Vega-Murillo;Jose Antonio Palacios-Franquez;Angel Rios-Utrera;Moises Montano-Bermudez
    • Animal Bioscience
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    • v.36 no.5
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    • pp.704-709
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    • 2023
  • Objective: In tropical, subtropical and arid zones, heat stress is the main cause of productivity reduction in cattle. When climate stressors occur, animals become thermal adapted through differential expression of some genes, including heat shock proteins (HSP) family. The aim of this study was to determine levels of expression of HSP60, HSP70, and HSP90 genes in Simmental cattle raised in tropical environments of Mexico. Methods: In this study, expression of HSP60, HSP70, and HSP90 genes was analyzed in 116 Simmental cattle from three farms with tropical climate located in western Mexico. Animals were sampled twice a day, in the morning and noon. Gene expression was evaluated by quantitative polymerase chain reaction using probes marked with fluorescence. The MIXED procedure of SAS with repeated measures was used for all statistical analysis. Results: HSP60 gene expression differences were found for sex (p = 0.0349). HSP70 gene differences were detected for sampling hour (p = 0.0042), farm (p<0.0001), sex (p = 0.0476), and the interaction sampling hour×farm (p = 0.0002). Gene expression differences for HSP90 were observed for farm (p<0.0001) and year (p = 0.0521). HSP70 gene showed to be a better marker of heat stress than HSP60 and HSP90 genes. Conclusion: Expression of HSP70 gene in Simmental herds of the tropical region of western México was different during early morning and noon, but the expression of the HSP60 and HSP90 genes was similar. Identification of resilient animals to heat stress will be useful in the genetic improvement of the Simmental breed.

The Effect of Aerobic Exercise on Brain-Derived Neurotrophic Factor (BDNF) in Individuals with Mild Cognitive Impairment: a Systematic Review and Meta-Analysis of a Randomized Controlled Trials

  • Kim, Hyun-Joong;Lee, DongJin;Lee, YeonSeop
    • Physical Therapy Rehabilitation Science
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    • v.11 no.3
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    • pp.304-310
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    • 2022
  • Objective: Mild cognitive impairment (MCI) is a condition in which cognitive and executive functions are reduced, and older adults with MCI are ten times more likely to develop dementia than healthy older adults. Expression of brain-derived neurotrophic factor (BDNF) through aerobic exercise is associated with increased cognitive and executive functions. in this review, randomized controlled trials (RCTs) on the effects of aerobic exercise on BDNF in individuals with mild cognitive impairment are summarized and qualitatively and quantitatively analyzed to suggest the necessity of aerobic exercise. Design: a systematic review and meta-analysis. Methods: RCTs were searched for changes in BDNF through aerobic exercise using four international databases. Quality assessment and quantitative analysis were performed using RevMan 5.4. Quantitative analysis was quantified with a standardized mean difference (SMD) and presented as a random effect model. Results: Three RCTs evaluated BDNF in 123 patients with MCI. There was a significant improvement in the experimental group that performed aerobic exercise compared to the control group. The results analyzed using the random effects model were SMD = 0.48. Conclusions: In this review, we reported the effects and mechanisms of aerobic exercise in individuals with MCI. As a result of synthesizing RCTs that performed aerobic exercise, a significant increase in BDNF was confirmed.

Polymorphisms and expression levels of TNP2, SYCP3, and AZFa genes in patients with azoospermia

  • Mohammad Ismael Ibrahim Jebur;Narges Dastmalchi;Parisa Banamolaei;Reza Safaralizadeh
    • Clinical and Experimental Reproductive Medicine
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    • v.50 no.4
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    • pp.253-261
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    • 2023
  • Objective: Azoospermia (the total absence of sperm in the ejaculate) affects approximately 10% of infertile males. Despite diagnostic advances, azoospermia remains the most challenging issue associated with infertility treatment. Our study evaluated transition nuclear protein 2 (TNP2) and synaptonemal complex protein 3 (SYCP3) polymorphisms, azoospermia factor a (AZFa) microdeletion, and gene expression levels in 100 patients with azoospermia. Methods: We investigated a TNP2 single-nucleotide polymorphism through polymerase chain reaction (PCR) restriction fragment length polymorphism analysis using a particular endonuclease. An allele-specific PCR assay for SYCP3 was performed utilizing two forward primers and a common reverse primer in two PCR reactions. Based on the European Academy of Andrology guidelines, AZFa microdeletions were evaluated by multiplex PCR. TNP2, SYCP3, and the AZFa region main gene (DEAD-box helicase 3 and Y-linked [DDX3Y]) expression levels were assessed via quantitative PCR, and receiver operating characteristic curve analysis was used to determine the diagnostic capability of these genes. Results: The TNP2 genotyping and allelic frequency in infertile males did not differ significantly from fertile volunteers. In participants with azoospermia, the allelic frequency of the SYCP3 mutant allele (C allele) was significantly altered. Deletion of sY84 and sY86 was discovered in patients with azoospermia and oligozoospermia. Moreover, SYCP3 and DDX3Y showed decreased expression levels in the azoospermia group, and they exhibited potential as biomarkers for diagnosing azoospermia (area under the curve, 0.722 and 0.720, respectively). Conclusion: These results suggest that reduced SYCP3 and DDX3Y mRNA expression profiles in testicular tissue are associated with a higher likelihood of retrieving spermatozoa in individuals with azoospermia. The homozygous genotype TT of the SYCP3 polymorphism was significantly associated with azoospermia.