• Title/Summary/Keyword: molecular genetics

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Molecular Cytogenetic Characterization of Supernumerary Marker Chromosomes by Chromosomal Microarray (염색체 마이크로어레이를 이용한 표지염색체의 분자세포유전학적 특성)

  • Bae, Mi-Hyun;Yoo, Han-Wook;Lee, Jin-Ok;Hong, Maria;Seo, Eul-Ju
    • Journal of Genetic Medicine
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    • v.8 no.2
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    • pp.119-124
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    • 2011
  • Purpose: Supernumerary marker chromosome (SMC) could be associated with various phenotypic abnormalities based on the chromosomal origin of SMCs. The present study aimed to determine the genomic contents of SMCs using chromosomal microarray and to analyze molecular cytogenetic characterizations and clinical phenotypes in patients with SMCs. Materials and Methods: Among patients with SMCs detected in routine chromosomal analysis, SMCs originating from chromosome 15 were excluded from the present study. CGH-based oligonucleotide chromosomal microarray was performed in 4 patients. Results: The chromosomal origins of SMCs were identified in 3 patients. Case 1 had a SMC of 16.1 Mb in 1q21.1-q23.3. Case 2 showed 21 Mb gain in 19p13.11-q13.12. Case 3 had a 4.5 Mb-sized SMC rearranged from 2 regions of 2.5 Mb in 22q11.1-q11.21 and 2.0 Mb in 22q11.22-q11.23. Conclusion: Case 1 presented a wide range of phenotypic abnormalities including the phenotype of 1q21.1 duplication syndrome. In case 2, Asperger-like symptoms are apparently related to 19p12-q13.11, hearing problems and strabismus to 19p13.11 and other features to 19q13.12. Compared with cat-eye syndrome type I and 22q11.2 microduplication syndrome, anal atresia in case 3 is likely related to 22q11.1-q11.21 while other features are related to 22q11.22-q11.23. Analyzing SMCs using high-resolution chromosomal microarray can help identify specific gene contents and to offer proper genetic counseling by determining genotype-phenotype correlations.

Investigation of the Condition of the Operation of the Livestock Liquid Manure Bin and Assessment of Malodorant Emissions (축분뇨 액비 저장조의 운영실태 및 악취 물질 발생량 조사)

  • Kim T. I.;Song J. I.;Joung S.;Jeong J. W.;Chung E. S.;Barroga A. J.;Yoo Y. H.;Yang C. B.;Kim M. K.
    • Journal of Animal Environmental Science
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    • v.11 no.3
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    • pp.189-196
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    • 2005
  • This study was carried out to determine the acceptability of liquid manure bin among livestock farmers and agricultural farmers, and quantify its malodorous compounds. The results were as follows; 1. The size of the manure bins owned by $93\%$ of 60 farmers surveyed was 200 M/T and were all in normal operation. Around $57\%$ of the normally operated bins were processed under aerobic condition. 2. Filly percent of the respondents utilized their liquid manure bin twice a year while $64\%$ used commercial microbial products to enhance maturity of their liquid manure and abatement of malodorous emissions. On the other hand, $43\%$ mentioned problems on the labor requirement, mechanical maintenance and lending cost of liquid manure processing and utilization, and the price of the commercial microbial products. 3. Malodorants emitted from livestock liquid manure bins and their boundary bin depended upon the livestock liquid manure processing condition. Within bin under the aerobic processing condition, Iso-valeric and propionic acid were ranged 0.012 to 0.07ppm and 0.17 to 2.85ppm, respectively. Within bin under the anoxic processing condition, n-butyric, n-valeric acid, and acetaldehyde were ranged 1.5 to 2.3ppm, 1.3 to 1.8ppm, and 0.8 to 2.1ppm, respectively. Malodorants emitted from the boundary of livestock liquid manure bins under the anoxic processing condition were detected the range of 0.4 to 0.9 ppm, more than the concentration of law regulation, as an acetaldehyde, but under the aerobic processing condition, they did not any detection.

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Effect of Microbial Inoculant to Pig Liquid Fertilizer on Germination Index of Chinese Cabbage (돈 분뇨 액비에 미생물 첨가가 배추의 발아지수에 미치는 영향)

  • Kim, T. I.;Yoo, Y. H.;Chung, E. S.;Barroga, Antonio J.;Yang, C. B.;Kim, M. K.
    • Journal of Animal Environmental Science
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    • v.11 no.2
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    • pp.135-146
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    • 2005
  • This study was carried out to investigate the effect of Synechocystis sp. KACC 91007 when added to a pig slurry or pig liquid fertilizer (PLF) on germination index (GI) of Chinese cabbage. The preliminary experiment involved the screening of inoculant levels which were; 0.05, 0.1, 0.2, and $0.3\%$, respectively. The $0.05\%$ level of inoculant was selected based on low phytotoxicity and high GI.. The PLF underwent a 107 day aerobic and anoxic processing conditions. The T-N, T-P, $NH_4$, and $NO_3-N$ concentrations of the untreated pig slurry were; 2,873, 753, 1,441.6, and 16.48 ppm, respectively. Using aerobic processing treatment, the fertilizer value of the PLF was 3,672, 164, 183.87, and 21.97 ppm, respectively. In contrast, the fertilizer value of the PLF processed under anoxic condition was reduced to 1,261, 68, 161, and 16.87 ppm. The GI value of the untreated PLF under aerobic and anoxic processing condition was 83 and $40.4^{*}\%$, respectively. With the addition of the $0.05\%$ microbial inoculant, the GI improved by more than 40 and $50\%$ respectively, when the PLF was processed under anoxic and aerobic conditions. The above findings proved that the aerobic processing of PLF for 107 days was better than anoxic and yielded higher T-N which is a macro-nutrient fertilizer material. Consequently, the addition of $0.05\%$ microbial inoculant resulted to a higher GI of the Chinese cabbage specifically under aerobic processing condition.

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Expression Analysis of Glutathione Peroxidase Genes in the Stage-Specific Seminiferous Tubules of Mice Excised by a Laser Capture Microdissection (Laser Capture Microdissection으로 절제된 마우스의 특정 단계별 정세관에서 Glutathione Peroxidase 유전자의 발현 분석)

  • Yon, Jung-Min;Lin, Chun-Mei;Park, Jung-Hoon;Hong, Min-Ki;Jung, A-Young;Kim, Mi-Ra;Baek, In-Jeoung;Lee, Beom-Jun;Nam, Sang-Yoon;Yun, Young-Won
    • Development and Reproduction
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    • v.14 no.2
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    • pp.99-105
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    • 2010
  • The seminiferous epithelium, with its division into 12 spermatogenic stages in the mouse, is a very complex tissue. Glutathione peroxidase (GPx) is a representative antioxidant enzyme that is capable of reducing organic hydroperoxides to their corresponding hydroxyl compounds utilizing glutathione and is related to the mammalian spermatogenesis. In this study, a real-time PCR was performed in the stage-specific seminiferous tubules of mouse testes excised by a laser capture microdissection (LCM) in order to quantitate the expression levels of a series of GPx genes including cytosolic GPx (cGPx), gastrointestinal GPx (GI-GPx), plasma GPx (pGPx), and phospholipid hydroperoxide GPx (PHGPx). Frozen sections (10 ${\mu}m$) were obtained from normal adult mouse testes. LCM was used to capture all the cells that were grouped into stages I-V, VII-VIII, and IX-XI in cross-sections of seminiferous tubules. The expression level of PHGPx mRNA was remarkably higher than those of other GPx mRNAs in mouse testes. During spermatogenesis, the expressions of GI-GPx, pGPx, and PHGPx mRNAs were highest on stages VII-VIII, began to decrease after stage XI, and showed a lowest level on stage I-V. However, the expressions of cGPx mRNA were highest on stages VII-VIII, and showed a lowest level on stage XI-XI. These findings indicate that GPx genes are expressed differentially on mouse spermatogenesis and also LCM can be an useful tool in cellular quantitative analysis of testes.

Phenotype-genotype correlations and the efficacy of growth hormone treatment in Korean children with Prader-Willi syndrome (프래더 윌리 증후군의 유전학적 발병 기전에 따른 표현형 및 성장 호르몬 치료 효과에 관한 연구)

  • Bae, Keun Wook;Ko, Jung Min;Yoo, Han Wook
    • Clinical and Experimental Pediatrics
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    • v.51 no.3
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    • pp.315-322
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    • 2008
  • Purpose : Prader-Willi syndrome (PWS) is a complex genetic disorder, caused by the deletion of the paternally derived 15q11-13 region or the maternal uniparental disomy of chromosome 15 (mUPD(15)). In this study, we compared phenotypic differences between those patients whose disease was caused by microdeletion and those caused by mUPD(15). In addition, a comparison of the efficacy of growth hormone (GH) therapy between these two PWS genotypes was analyzed. Methods : Fifty-three patients were diagnosed as having PWS based on molecular and cytogenetic analyses and clinical features. Data that included maternal age, birth weight, a feeding problem in the neonatal period, cryptorchidism, developmental delay or mental retardation, short stature, hypopigmentation, changes in height, weight, and body mass indexes (BMI) before and after GH treatment were obtained by a retrospective review of medical records. The data from the patients with microdeletion were compared with those from the patients with mUPD(15). Results : Of the 53 patients with genetically confirmed PWS, 39 cases had microdeletion and 14 mUPD(15). Maternal ages were significantly higher in the mUPD(15) group, and hypopigmentation and a feeding problem in the neonatal period were more frequent in the microdeletion group. Growth hormone was administered to 20 patients [14 with microdeletion, 6 with mUPD(15)]. There were no differences between the two groups in height velocity, weight and height SDS, and BMI after GH therapy. Conclusion : Phenotype and genotype correlations were observed in Korean PWS patients, such as more advanced maternal ages in the mUPD(15) group and more feeding problems and hypopigmentations in the microdeletion group. Further long-term prospective studies are needed to correlate other aspects of the phenotypes.

Relationship among porcine lncRNA TCONS_00010987, miR-323, and leptin receptor based on dual luciferase reporter gene assays and expression patterns

  • Ding, Yueyun;Qian, Li;Wang, Li;Wu, Chaodong;Li, DengTao;Zhang, Xiaodong;Yin, Zongjun;Wang, Yuanlang;Zhang, Wei;Wu, Xudong;Ding, Jian;Yang, Min;Zhang, Liang;Shang, Jinnan;Wang, Chonglong;Gao, Yafei
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.2
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    • pp.219-229
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    • 2020
  • Objective: Considering the physiological and clinical importance of leptin receptor (LEPR) in regulating obesity and the fact that porcine LEPR expression is not known to be controlled by lncRNAs and miRNAs, we aim to characterize this gene as a potential target of SSC-miR-323 and the lncRNA TCONS_00010987. Methods: Bioinformatics analyses revealed that lncRNA TCONS_00010987 and LEPR have SSC-miR-323-binding sites and that LEPR might be a target of lncRNA TCONS_00010987 based on cis prediction. Wild-type and mutant TCONS_00010987-target sequence fragments and wild-type and mutant LEPR 3'-UTR fragments were generated and cloned into pmiRRB-REPORTTM-Control vectors to construct respective recombinant plasmids. HEK293T cells were co-transfected with the SSC-miR-323 mimics or a negative control with constructs harboring the corresponding binding sites and relative luciferase activities were determined. Tissue expression patterns of lncRNA TCONS_00010987, SSC-miR-323, and LEPR in Anqing six-end-white (AQ, the obese breed) and Large White (LW, the lean breed) pigs were detected by real-time quantitative polymerase chain reaction; backfat expression of LEPR protein was detected by western blotting. Results: Target gene fragments were successfully cloned, and the four recombinant vectors were constructed. Compared to the negative control, SSC-miR-323 mimics significantly inhibited luciferase activity from the wild-type TCONS_00010987-target sequence and wild-type LEPR-3'-UTR (p<0.01 for both) but not from the mutant TCONS_00010987-target sequence and mutant LEPR-3'-UTR (p>0.05 for both). Backfat expression levels of TCONS_00010987 and LEPR in AQ pigs were significantly higher than those in LW pigs (p<0.01), whereas levels of SSC-miR-323 in AQ pigs were significantly lower than those in LW pigs (p<0.05). LEPR protein levels in the backfat tissues of AQ pigs were markedly higher than those in LW pigs (p<0.01). Conclusion: LEPR is a potential target of SSC-miR-323, and TCONS_00010987 might act as a sponge for SSC-miR-323 to regulate LEPR expression.

Effect of Antioxidants for Porcine Oocytes during In Vitro Maturation, Fertilization and Development (돼지 난포란으로부터 체외수정란의 생산에 있어서 항산화제의 첨가가 배 발달에 미치는 효과)

  • Park H.;Kim J. Y.;Kim J. Y.;Lee J. H.;Park H. D.;Kim J. M.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.245-255
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    • 2004
  • In recent years, an increasing number of studies on pig in vitro maturation(IVM) and in vitro fertilization(IVF) have been separated. the wide range of new technologies, including that in applied molecular genetics, has increased this interest. the production of viable porcine embryos in vitro is a prerequisites for the successful production of transgenic pigs to date. The efficiency of IVM/IVF techniques in the porcine is lower than that obtained in other species such as cattle and mouse. The several problems are generally thought to be the cause of poor results: the low rate of MPN formation derived from inadequate IVM of oocytes, the high incidence of polyspermy after IVF and cell blocking at 4 cell during embryos culture. For there reasons overcoming, many studies have been conducted to improve in vitro embryo-genic competence of oocytes. In the last several years, many maturation culture media have been evaluated and various exogenous factors such as hormones and grows factors have been tested to improve the efficiency of porcine in vitro system. In the study several antioxidants have been examined to improve in vitro fertilization and development of porcine oocytes. In this study, several antioxidants were examined to determine the effects on the development of oocytes to the cleavage, morula and blastocyst stage when added at the maturation(IVM) or in vitro fertilization(IVF) or in vitro culture(IVC) of porcine embryos. Porcine oocytes were matured, fertilized and embryos were cultured in defind conditioned medium in vitro with or without supplementation with the antioxidents of cysteine, catalase and glutathione. 1. Significant improvement of blastocyst rate (27.2% versus 15.4%, p<0.05) were achieved when catalase(500U/$m\ell$) were added to TCM-199 medium and morula rate(72.0% versus 53.9%, p<0.05) were significantly higher when glutathione(1.0mM/$m\ell$) were added to TCM-199 medium than those of control. 2. In mTBM medium for oocytes fertilization, the addition of cysteine, catalase and glutathione had no positive effect on embryonic development. glutathione had no positive effect on embryonic development. In conclusion, this study shows that addition of catalase, gluththione during IVM improved the rate of porcine embryo development.

Effects of Feeding Ferritin Gene Transferred Yeast (Saccharomyces serevisiae) on Performance, Iron Concentration in Organs and Egg of Chickens (Ferritin 유전자 전이 효모(Saccharomyces serevisiae)의 급여가 닭의 생산성, 장기 및 계란의 철분함량에 미치는 영향)

  • Ryu, Byeong-Seon;Park, Jae-Hong;Kim, Dae-Hyeok;Ryu, Kyeong-Seon
    • Korean Journal of Poultry Science
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    • v.30 no.4
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    • pp.245-251
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    • 2003
  • Three experiments were conducted to investigate the effect of feeding yeast accumulated transgenic ferritin(FRT, Saccharomyces cerevisiae) as a probiotic on the performance, iron contents in the liver, spleen, bone and yolk of laying hens and broiler chicks. Effects of feeding FRT were compared with that of feeding wild-type yeast(W0) and yeast grown on 20 mM ferric citrate-added medium (W20). In Expt 1, to investigate the effect of feeding yeast (control, W0 FRT) on performance and iron content of organs of broiler chicks which were fed basal diet supplemented with 75mg/kg iron(Fe75) or not (Fe0), three hundred sixty one-day-old male broiler chicks were fed a corn-sov based diet for five weeks. Weight gain, feed intake and feed conversion were measured weekly. In Expt 2, fifteen 33-week-old ISA Brown laying hens were placed in individual cages and were fed control, W0 and FRT diets for Four weeks. In Expt 3, twenty four 45-week-old ISA Brown laying hens were placed in individual cages and were fed a basal diet for a week. Then, experimental diets (control, W0, W20, FRT) were fed for three weeks. Iron contents in the liver, heart, spleen and tibia were determined at the end of all experiments. Iron content in yolk was measured weekly (expt 2, 3). The level of yeast added and iron concentration of FRT were $1{\times}10^8$cfu/kg diet and 500 mg/kg cell (DM) respectively in Expt 3, yeast was supplemented at $2{\times}10^{10}$cfu/kg diet and the iron content of FRT was 1000mg/kg cell (DM). In Expt 1. birds fed Fe75 showed significantly higher weight gain compared with Fe0 (P<0.05). However, weight gain and feed intake of birds fed FRT was significantly lower than control (P<0.05). In Expt 2, the iron content of the liver was decreased in the FRT treatment (P<0.05). In Expt 3, iron concentration of the liver and spleen tended to be increased by feeding FRt. However, the iron content of the tibia tended to be decreased in the FRT treatment. These results suggest that feeding FRT as a probiotic cannot improve performance and iron content in organs of broiler chicks and laying hens.

Differential Responses to TGF Alpha in between Invasive Squamous Cell Carcinoma Cell Line and Noninvasive One (침투성 상피암세포주와 비침투성 상피암세포주의 TGF alpha에 대한 반응의 차이)

  • Son, Young-Sook;Chey, Myoung-Jae;Fuchs, Elaine;Chung, Myung-Hee;Park, Chan-Woong
    • The Korean Journal of Pharmacology
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    • v.29 no.1
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    • pp.139-148
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    • 1993
  • Both SCC 12 and SCC 13 cell lines were derived from squamous cell carcinoma (SCC) of the skin (Wu and Rheinwald, 1981). In the present study, we compared the inherent invasive activity in their raft cultures where most in vivo characteristics of epidermis can be reproduced by cell culture method. The raft culture of SCC 12 cell line produced many invading colonies within the collagen lattice and basal-like cells in the middle of differentiating cell layers, but no invasive activity was observed in the SCC 13 raft culture. We investigated which factors are implicated in inherent invasive activity of SCC 12 cell line by examining basal levels of type I collagenase, EGF receptor, fibronectin, and its receptor in two cell lines. Among them, only type I collagenase was significantly higher in invasive SCC 12 cells than in non-invasive SCC 13 cells. Furthermore, we tried to investigate mechanisms underlying between SCC 12 cell's inherent invasive activity and its high basal level of type I collagenase. As one of them, discrepancy in TGF alpha mediated responses between two cell lines was observed. In SCC 13 cells, TGF alpha initially stimulated type I collagenase at 12 h after TGF alpha treatment and then its down regulation was followed from 24 h even though TGF alpha was continuously present in the medium. However in SCC 12 cells, TGF alpha continuously stimulated type I collegenase up to 48 h. We propose that defect in EGF receptor's down-regulation may be involved in lack of type I collagenase's down-regulation and its possible connection to invasive activity of SCC 12 cell line.

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Increased Expression of Phospholipase C-$\gamma1$ Activator Protein, AHNAK in Human Lung Cancer Tissues (인체 폐암조직에서 Phospholipase C-$\gamma1$의 활성화 단백, AHNAK의 발현양상)

  • Oh, Yoon-Jung;Park, Chun-Seong;Choi, So-Yeon;Cheong, Seong-Cheoll;Lee, Sun-Min;Hwang, Sung-Chul;Lee, Yi-Hyeong;Hahn, Myung-Ho;Lee, Kyi-Beom;Ryu, Han-Young;Ha, Mahn-Joon;Bae, Yoon-Su;Rhee, Seo-Goo
    • Tuberculosis and Respiratory Diseases
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    • v.47 no.3
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    • pp.347-355
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    • 1999
  • Background: Phospholipase C(PLC) plays a central role in cellular signal transduction and is important in cellular growth, differentiation and transformation. There are currently ten known mammalian isozymes of PLC reported to this date. Hydrolysis of phosphatidylinositol 4,5-bisphosphate($PIP_2$) by PLC produces two important second messengers, inositol 1,4,5-trisphosphate($IP_3$) and diacylglycerol. PLC-${\gamma}1$, previously, was known to be activated mainly through growth factor receptor tyrosine kinase. Other mechanisms of activating PLC-yl have been reported such as activation through tau protein in the presence of arachidonic acid in bovine brain and activation by $IP_3$, phosphatidic acid, etc. Very recently, another PLC-${\gamma}1$ activator protein such as tau has been found in bovine lung tissue, which now is considered to be AHNAK protein. But there has been no report concerning AHNAK and its associated disease to this date. In this study, we examined the expression of the PLC-${\gamma}1$ activator, AHNAK, in lung cancer specimens and their paired normal. Methods: From surgically resected human lung cancer tissues taken from twenty-eight patients and their paired normal counterparts, we evaluated expression level of AHNAK protein using immunoblot analysis of total tissue extract Immunohistochemical stain was performed with primary antibody against AHNAK protein. Results: Twenty-two among twenty-eight lung cancer tissues showed overexpression of AHNAK protein (eight of fourteen squamous cell lung cancers, all of fourteen adenocarcinomas). The resulting bands were multiple ranging from 70 to 200 kDa in molecular weight and each band was indistinct and formed a smear, reflecting mobility shift mainly due to proteolysis during extraction process. On immunohistochemistry, lung cancer tissues showed a very heavy, dense staining with anti-AHNAK protein antibody as compared to the surrounding normal lung tissue, coresponding well with the results of the western blot Conclusion: The overexpression of PLC-${\gamma}1$ activator protein, AHNAK in lung cancer may provide evidence that the AHNAK protein and PLC-${\gamma}1$ act in concerted manner in carcinogenesis.

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