scholarly journals Regulation of mdr2 P-glycoprotein expression by bile salts

1997 ◽  
Vol 321 (2) ◽  
pp. 389-395 ◽  
Author(s):  
Charles M. G. FRIJTERS ◽  
Roelof OTTENHOFF ◽  
Michel J. A. van WIJLAND ◽  
Carin M. J. van NIEUWKERK ◽  
Albert K. GROEN ◽  
...  
Keyword(s):  
Bile Salt ◽  
Bile Salts ◽  
Control Diet ◽  
Mrna Level ◽  
Northern Blotting ◽  
Mrna Levels ◽  
Male Mice ◽  
Complete Replacement ◽  
P Glycoprotein ◽  
Mrna Content

The phosphatidyl translocating activity of the mdr2 P-glycoprotein (Pgp) in the canalicular membrane of the mouse hepatocyte is a rate-controlling step in the biliary secretion of phospholipid. Since bile salts also regulate the secretion of biliary lipids, we investigated the influence of the type of bile salt in the circulation on mdr2 Pgp expression and activity. Male mice were fed a purified diet to which either 0.1% (w/w) cholate or 0.5% (w/w) ursodeoxycholate was added. This led to a near-complete replacement of the endogenous bile salt pool (mainly tauromuricholate) by taurocholate or tauroursodeoxycholate respectively. The phospholipid secretion capacity was then determined by infusion of increasing amounts of tauroursodeoxycholate. Cholate feeding resulted in a 55% increase in maximal phospholipid secretion compared with that in mice on the control diet. Northern blotting revealed that cholate feeding increased mdr2 Pgp mRNA levels by 42%. Feeding with ursodeoxycholate did not influence the maximum rate of phospholipid output or the mdr2 mRNA content. Female mice had a higher basal mdr2 Pgp mRNA level than male mice, and this was also correlated with a higher phospholipid secretion capacity. This could be explained by the 4-fold higher basal cholate content in the bile of female compared with male mice. Our results suggest that the type of bile salts in the circulation influences the expression of the mdr2 gene.

scholarly journals Developmental changes in the protein and mRNA content of a p115/transcytosis-associated protein in the bovine mammary gland

2000 ◽  
Vol 166 (2) ◽  
pp. 319-327 ◽  
Author(s):  
A Watanabe ◽  
I Uchida ◽  
K Kadota ◽  
N Katoh
Keyword(s):  
Mammary Gland ◽  
Mammary Epithelial Cells ◽  
Mrna Level ◽  
Immunoblot Analysis ◽  
Northern Blotting ◽  
Mammary Development ◽  
Mammary Epithelial ◽  
Mrna Levels ◽  
Stage Of Lactation ◽  
Mrna Content

We measured the amounts of a vesicular transport factor, p115/transcytosis-associated protein (p115/TAP) and its mRNA, in mammary glands from cows in which lactation was induced hormonally. The highest level of p115/TAP mRNA, determined by Northern blotting, was detected in the developing stage. In contrast to the mRNA level, the amount of protein, determined by immunoblot analysis using anti-p115/TAP antibodies raised against a p115/TAP-derived recombinant fusion protein, was higher during the lactating stages than at other times. Immunohistochemical study showed that p115/TAP was predominantly localized in mammary epithelial cells. The p115/TAP was also detected in tissues other than the mammary gland but, in contrast to the situation in the mammary gland, the protein and its mRNA levels in those tissues were independent of the stage of lactation. The increased level of p115/TAP mRNA during the developing stage and the maintenance of p115/TAP protein during lactation suggest that the synthesis of p115/TAP is regulated during mammary development and differentiation, and also that the protein is involved in a function related to lactation.

scholarly journals Cytolethal Distending Toxin Is Essential for Helicobacter hepaticus Colonization in Outbred Swiss Webster Mice

2005 ◽  
Vol 73 (6) ◽  
pp. 3559-3567 ◽  
Author(s):  
Zhongming Ge ◽  
Yan Feng ◽  
Mark T. Whary ◽  
Prashant R. Nambiar ◽  
Shilu Xu ◽  
...  
Keyword(s):  
Mrna Level ◽  
Interleukin 10 ◽  
Mouse Strains ◽  
Cytolethal Distending Toxin ◽  
Mrna Levels ◽  
Helicobacter Hepaticus ◽  
Male Mice ◽  
Susceptible Mouse ◽  
Female Mice ◽  
Ifn Γ

ABSTRACT Helicobacter hepaticus, which induces chronic hepatitis and typhlocolitis in susceptible mouse strains, produces a cytolethal distending toxin (CDT) consisting of CdtA, CdtB, and CdtC. A cdtB-deficient H. hepaticus isogenic mutant (HhcdtBm7) was generated and characterized for colonization parameters in four intestinal regions (jejunum, ileum, cecum, and colon) of outbred Swiss Webster (SW) mice. Inactivation of the cdtB gene abolished the ability of HhcdtBm7 to colonize female mice at both 8 and 16 weeks postinfection (wpi), whereas HhcdtBm7 colonized all of four intestinal regions of three of five males at 8 wpi and then was eliminated by 16 wpi. Wild-type (WT) H. hepaticus was detected in the corresponding intestinal regions of both male and female mice at 8 and 16 wpi; however, colonization levels of WT H. hepaticus in the cecum and colon of male mice were approximately 1,000-fold higher than in females (P < 0.0079) at 16 wpi. Infection with WT H. hepaticus, but not HhcdtBm7, at 8 wpi was associated with significantly increased mRNA level of ileal and cecal gamma interferon (IFN-γ) in females (P < 0.016 and 0.031 between WT H. hepaticus-infected and sham-dosed females, respectively). In contrast, the mRNA levels of IFN-γ were significantly higher in the colon (P < 0.0079) and trended to be higher in the cecum (P < 0.15) in the HhcdtBm7-colonized male mice versus the sham-dosed controls at 8 wpi. In addition, mRNA levels of ileal IFN-γ were significantly higher in the control females than males at 8 wpi (P < 0.016). There were significantly higher Th1-associated immunoglobulin G2a (IgG2a), Th2-associated IgG1 and mucosal IgA (P < 0.002, 0.002, 0.002, respectively) responses in the mice infected with WT H. hepaticus when compared to HhcdtBm7 at 16 wpi. Colonic interleukin-10 (IL-10) expressions at 16 wpi were significantly lower in both female and male mice colonized by WT H. hepaticus or in males transiently colonized through 8 wpi by HhcdtBm7 versus control mice (P < 0.0159). These lines of evidence indicate that (i) H. hepaticus CDT plays a crucial role in the persistent colonization of H. hepaticus in SW mice; (ii) SW female mice are more resistant to H. hepaticus colonization than male mice; (iii) there was persistent colonization of WT H. hepaticus in cecum, colon, and jejunum but only transient colonization of H. hepaticus in the ileum of female mice; (iv) H. hepaticus colonization was associated with down-regulation of colonic IL-10 production.

scholarly journals Magnesium and calcium deficiencies additively increase zinc concentrations and metallothionein expression in the rat liver

2012 ◽  
Vol 109 (3) ◽  
pp. 425-432 ◽  
Author(s):  
Megumi Kotani ◽  
Ki Hyun Kim ◽  
Natsumi Ishizaki ◽  
Masayuki Funaba ◽  
Tohru Matsui
Keyword(s):  
Rat Liver ◽  
Control Diet ◽  
Mrna Level ◽  
Male Rats ◽  
Mrna Levels ◽  
Deficient Diet ◽  
Mg Deficiency ◽  
Ca Deficiency ◽  
Zn Concentration ◽  
Dietary Treatments

Mg deficiency increases the concentration of Zn in the liver. We investigated the effect of Mg deficiency on the expression of Zn-regulating factors such as Zn transporters and metallothionein (MT) in the rat liver. Because Ca deficiency alleviates some of the effects of Mg deficiency, we also investigated the interactions associated with Ca and Mg deficiencies. Growing male rats were given a control diet, a Mg-deficient diet, a Ca-deficient diet and a Mg- and Ca-deficient diet for 3 weeks. Mg and Ca deficiencies additively increased the mRNA levels of MT-1 and MT-2, the MT protein concentration and the concentration of Zn in the liver. The hepatic mRNA level of Zip14 increased with Mg deficiency but not with Ca deficiency. The dietary treatments did not affect the mRNA levels of other Zn transporters such as Zip1, Zip5, ZnT1, ZnT5 and ZnT6 in the liver. Ca deficiency was found to decrease the amount of femoral Zn and increase serum Zn concentration. This did not occur in the case of Mg deficiency. These results suggest that Mg deficiency enhances hepatic Zn uptake by the up-regulation of Zip14 expression and increases hepatic Zn concentration, leading to the enhancement of MT expression. Ca deficiency causes a transfer of Zn from the bone to the liver, which increases hepatic Zn concentration and, in turn, up-regulates the expression of MT. Because Mg and Ca deficiencies increase hepatic Zn concentration and increase MT expression by different mechanisms, their effects are additive.

181. SIRT6 PROTEIN IS REDUCED IN TESTES AND SPERM FROM OBESE MALE MICE

2010 ◽  
Vol 22 (9) ◽  
pp. 99
Author(s):  
N. O. Palmer ◽  
T. Fullston ◽  
M. Mitchell ◽  
M. Lane
Keyword(s):  
Dna Damage ◽  
Control Diet ◽  
Caloric Intake ◽  
Cell Types ◽  
Mature Sperm ◽  
Mrna Levels ◽  
Male Mice ◽  
Sperm Dna Damage ◽  
Damage Repair ◽  
Sperm Dna

Obesity in males is associated with altered hormone levels, reduced sperm function and increased sperm DNA damage. However, the underlying molecular mechanism has not been identified. Mammalian SIRT6 protein exhibits caloric intake dependant DNA damage repair in other tissue types. However, a possible role for SIRT6 in male obesity and subfertility has not been considered. Therefore, the aim of this study was to assess the effects of male obesity on SIRT6 in testes and mature sperm. Five week old C57BL6 male mice (n =10 per diet) were fed either a control diet (CD) (6% fat) or a high fat diet (HFD) (21% fat) for 16 weeks before collection of sperm and testes. There was no difference in Sirt6 mRNA levels as determined by qPCR in testes from HFD males. Immunohistochemistry showed SIRT6 localised to the nucleus of transitioningspermatids from late round spermatidsuntil early elongating spermatids. SIRT6 relative fluorescence of these positive cell types was significantly decreased by 22% in males fed the HFD compared to CD (P < 0.05). This was confirmed by a decrease in total SIRT6 protein in testes from HFD males as detected by an immunoabsorbance assay (P < 0.05). Surprisingly, SIRT6 was only present in the acrosome of mature sperm. Acrosomal localisation was confirmed by the loss of SIRT6 staining after an induced acrosome reaction. SIRT6 levels in the acrosome of mature sperm was decreased by 11% in males fed the HFD (P < 0.05). This is the first study to show that SIRT6 is located to the acrosome of mature sperm, specific cells within the testes and is reduced in an obese state. Furthermore, this study suggests a possible role for SIRT6 in the acrosomal reaction and therefore potentially fertilisation, processes which are known to be reduced by male obesity.

Inhibitory Effects of Benzaldehyde, Vanillin, Muscone and Borneol on P-Glycoprotein in Caco-2 Cells and Everted Gut Sac

Pharmacology ◽  
2018 ◽  
Vol 101 (5-6) ◽  
pp. 269-277 ◽  
Author(s):  
Shixiang Wang ◽  
Nan Tan ◽  
Cuicui Ma ◽  
Jie Wang ◽  
Pu Jia ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Prescription Drugs ◽  
High Performance ◽  
Mrna Level ◽  
Herbal Medicines ◽  
Mrna Levels ◽  
Inhibitory Effects ◽  
Apparent Permeability ◽  
P Glycoprotein ◽  
Everted Gut Sac

Aims: In clinical practice, herbal medicines have played an important role in the modulation of drug transporters through the combination of conventional prescription drugs, which necessitates the elucidation of herb-drug interactions. The present study was designed to investigate the inhibitory effects and mechanisms of benzaldehyde, vanillin, muscone, and borneol on P-glycoprotein (P-gp). Methods: The effects of the 4 compounds on the intracellular accumulation of rhodamine-123 (Rho-123) in vinblastine-treated Caco-2 (VB-Caco-2) cells were studied by monitoring fluorescence intensity through a flow cytometry assay, and the effects of these compounds on Rho-123 transport through VB-Caco-2 monolayers and Rho-123 intestinal absorption in the rat everted gut sac were investigated by high-performance liquid chromatography. Moreover, P-gp expression in VB-Caco-2 cells was assessed using flow cytometry and Western blot analysis, and the relative ABCB1 mRNA level was determined by Real-time RT-PCR. Key Findings: The results showed that benzaldehyde, vanillin, muscone, and borneol significantly increased Rho-123 uptake in VB-Caco-2 cells, increased the absorption rate and apparent permeability coefficient of Rho-123 in rat jejunum and ileum, and decreased the efflux ratio of Rho-123 from 6.52 to less than 2 during transport across VB-Caco-2 cell monolayers. In addition, these compounds reduced the protein and ABCB1 mRNA levels of P-gp in VB-Caco-2 cells. Conclusions: These data indicate that benzaldehyde, vanillin, muscone and borneol could effectively reverse multidrug resistance via inhibiting the P-gp function and expression pathway. The data provide fodder for further investigation into the interaction between the 4 compounds and other drugs transported by P-gp.

Effect of oestradiol-17β on the expression of oestrogen receptor mRNA in human tonsillar cells

1995 ◽  
Vol 14 (1) ◽  
pp. 13-19 ◽  
Author(s):  
M Evagelatou ◽  
J Farrant
Keyword(s):  
T Cell ◽  
Oestrogen Receptor ◽  
Mrna Level ◽  
Cell System ◽  
Target Cells ◽  
Mrna Levels ◽  
Mrna Content ◽  
Actin Mrna ◽  
Tonsillar Cells

ABSTRACT We have investigated the expression of oestrogen receptor (ER) mRNA in Ficoll-separated tonsillar cells and the changes that occur with the addition of oestradiol (OE2) both in the presence and the absence of the T cell mitogen phytohaemagglutinin (PHA). The amounts of ER mRNA and β-actin mRNA in the samples were determined by slot blotting and hybridization and quantified by densitometry. The levels of ER mRNA were normalized against the β-actin mRNA content. In the presence of OE2 (7×10−8m) after a 10-h culture there was a significant decrease (P<0·05) to about 66% of the control (0-h culture) ER mRNA levels. Stimulating the cultures with PHA (1 μg/ml, without the presence of OE2, had no effect on the expression of ER mRNA. However, when OE2 was present in a 10-h culture of PHA-stimulated cells, the ER mRNA level was significantly decreased (P<0·05) to about 60% of control levels. In 24-h cultures, the presence of OE2 and/or PHA had no effect. When separated T cell preparations from the tonsils were used, no significant effects of OE2 were seen in either the 10-h or 24-h cultures. In conclusion, OE2 downregulates the ER mRNA content in a tonsillar mononuclear cell system in vitro as it does in many primary oestrogen target cells.

scholarly journals Mannan oligosaccharide supplementation in diets of sow and (or) their offspring improved immunity and regulated intestinal bacteria in piglet1

2019 ◽  
Vol 97 (11) ◽  
pp. 4548-4556 ◽  
Author(s):  
Xudong Duan ◽  
Gang Tian ◽  
Daiwen Chen ◽  
Linhui Huang ◽  
Dan Zhang ◽  
...  
Keyword(s):  
Systemic Inflammation ◽  
Control Diet ◽  
Mrna Level ◽  
Intestinal Bacteria ◽  
Interaction Effects ◽  
Diet Group ◽  
Mrna Levels ◽  
Toll Like Receptor ◽  
Mannan Oligosaccharide ◽  
Ifn Γ

Abstract The objectives of the current study were to explore the effects of mannan oligosaccharide (MOS) supplementation in the diets of sow and (or) their offspring on intestinal bacteria, intestinal and systemic inflammation in the piglet. A total of 60 multiparous sows (4 ± 1 parity; Landrace × Yorkshire) were fed either control diet (sCON, n = 30) or a diet containing 400 mg kg−1 MOS (sMOS, n = 30) from day 86 of gestation until weaning (day 20 of postpartum). On day 7 of age, offspring (Duroc × Landrace Yorkshire) were assigned within sow treatments and fed control diet (pCON) or diet containing 800 mg kg−1 MOS (pMOS) for 28 d (end at 35 d of age), resulting in four piglet diet groups (n = 15 litters per diet group): sCON-pCON, sCON-pMOS, sMOS-pCON, and sMOS-pMOS. Results found that piglet diet MOS increased or tend to increase Lactobacillus amount in the ileum digesta (P < 0.01) and jejunum digesta (P = 0.07), respectively; while tend to decrease Escherichia coli amount in jejunum digesta (P =0.06) and cecum digesta (P = 0.08). Both sow and piglet diets add MOS (sMOS-pMOS) increased Lactobacillus amount but decreased E. coli amount in jejunum digesta (P < 0.05) compared with the sCON-pCON diet group. In addition, sow diet MOS (rather than piglet diet MOS) increased sIgA content in piglet jejunum mucosa compared with control (P = 0.04). Sow diet MOS decreased toll-like receptor 2 (TLR2), toll-like receptor 4 (TLR4), and interleukin 8 (IL-8) mRNA levels (P < 0.05) and tended to decrease nuclear factor-κB p65 (NF-κB p65) mRNA level (P = 0.07) in piglet intestinal lymphatic. The interaction effects between sow and piglet diets were found on the mRNA levels of NF- κB p65 (P = 0.03) and IL-8 (P = 0.02) in piglet jejunum. Finally, the sow diet MOS decreased proinflammatory cytokines IL-2 (P < 0.01) and IL-4 (P < 0.01) concentrations in piglet serum. Piglets diet MOS decreased the contents of IL-2 (P = 0.03), IL-4 (P = 0.01) and interferon (IFN)-γ (P < 0.01) while increased anti-inflammatory cytokine IL-10 (P < 0.01) content in serum. The interaction effects between sows and piglet diets on IL-4 (P = 0.02), IL-10 (P < 0.01), and IFN-γ (P = 0.08) were observed. In conclusion, sow and/or piglet diet MOS could improve intestinal microbiota, enhance intestinal mucosal immune competence, and suppress intestinal and systemic inflammation in the piglet.

scholarly journals Enhanced Expression of the Human Multidrug Resistance Protein 3 by Bile Salt in Human Enterocytes

2001 ◽  
Vol 276 (50) ◽  
pp. 46822-46829 ◽  
Author(s):  
Akihiko Inokuchi ◽  
Eiji Hinoshita ◽  
Yukihide Iwamoto ◽  
Kimitoshi Kohno ◽  
Michihiko Kuwano ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Bile Salt ◽  
Bile Salts ◽  
Promoter Activity ◽  
Enterohepatic Circulation ◽  
Basolateral Membrane ◽  
Mrna Levels ◽  
Multidrug Resistance Protein ◽  
Multidrug Resistance Protein 3 ◽  
Resistance Protein

The enterohepatic circulation is essential for the maintenance of bile acids and cholesterol homeostasis. The ileal bile acid transporter on the apical membrane of enterocytes mediates the intestinal uptake of bile salts, but little is known about the bile salt secretion from the basolateral membrane of enterocytes into blood. In the basolateral membrane of enterocytes, an ATP-binding cassette transporter, multidrug resistance protein 3 (MRP3), is expressed, which has the ability to transport bile salts. We hypothesized that MRP3 might play a role in the enterohepatic circulation of bile salts by transporting them from enterocytes into circulating blood through the up-regulation ofMRP3expression, so we investigated the transcriptional control ofMRP3in response to bile salts.MRP3mRNA levels were increased about 3-fold in human colon cells by chenodeoxycholic acid (CDCA), in a dose- and time-dependent manner. In the promoter assay, the promoter activity ofMRP3was increased about 3-fold over the basal promoter activity when treated with CDCA, and the putative bile salt-responsive elements exist in the region −229/−138 including two α-1 fetoprotein transcription factor (FTF)-like elements. Constructs with a specific mutation in the consensus sequence of FTF elements showed no increase in basal transcriptional activity following CDCA treatment. In electrophoretic mobility shift assay with nuclear extracts, specific binding of FTF to FTF-like elements was observed when treated with CDCA. The expression ofFTFmRNA levels were also markedly enhanced in response to CDCA, and overexpression of FTF specifically activated theMRP3promoter activity about 4-fold over the basal promoter activity. FTF thus might play a key role not only in the bile salt synthetic pathway in hepatocytes but also in the bile salt excretion pathway in enterocytes through the regulation ofMRP3expression. MRP3 may contribute as a plausible bile salt-exporting transporter to the enterohepatic circulation of bile salts.

The Effect of Pomegranate Juice on the Expression of Some Murine UDPGlucuronosyltransferases Genes

2020 ◽  
Vol 14 ◽  
Author(s):  
Yacoub M. Irshaid ◽  
Ruba Alani ◽  
Razan Al-Rawashdeh ◽  
Tuqa Al-Ghazawi ◽  
Hiba Hijazi ◽  
...  
Keyword(s):  
Small Intestine ◽  
Mrna Level ◽  
Folk Medicine ◽  
The Body ◽  
Pomegranate Juice ◽  
Mrna Levels ◽  
Male Mice ◽  
Specific Primers ◽  
Metabolizing Enzymes ◽  
Polymerase Chain

Background: Food-drug interactions may lead to suppression or induction drug metabolizing enzymes. Pomegranate is a commonly used fruit in folk medicine all over the world. Data concerning the effect of pomegranate on the activity of UDP-glucuronosyltransferases (UGTs) is scarce. Objective: The purpose of this work was to investigate the effect of pomegranate juice ingestion on the transcription of ugt2b1, ugt2a3, and ugt1a9 in the liver and small intestine of male mice. Methods: Pomegranate juice was administered to 10 male mice for 14 days in drinking bottles instead of water. Ten control mice received water in the drinking bottles. On the 15th day, the mice were sacrificed and the liver and the small intestine were removed. The small intestine was divided into 3 parts. Total mRNA was extracted from samples of these specimens, and cDNA was synthesized by quantitative real-time polymerase chain reaction (RT-PCR) using specific primers for each ugt gene. Results: The ugt1a9 mRNA level was reduced by 2.25-fold in the liver and by 6-, 1.5-, and 3-folds in the first, second and third part of the small intestine, respectively. The ugt2b1 mRNA level in the liver and the third part of the small intestine was not affected, while it was reduced by 3.7- and 3-folds in the first and second parts of the small intestine, respectively. The ugt2a3 mRNA level was not affected in the liver and the 3 parts of the small intestine. Conclusions: Some ugt mRNA levels may be reduced by the ingestion of pomegranate juice, which may reduce the metabolism of their drug substrates. The consequences may be accumulation of such drugs in the body and enhanced toxicity.

scholarly journals Resistance exercise alters MRF and IGF-I mRNA content in human skeletal muscle

2003 ◽  
Vol 95 (3) ◽  
pp. 1038-1044 ◽  
Author(s):  
Niklas Psilander ◽  
Rasmus Damsgaard ◽  
Henriette Pilegaard
Keyword(s):  
Skeletal Muscle ◽  
Resistance Training ◽  
Resistance Exercise ◽  
Human Skeletal Muscle ◽  
Mrna Level ◽  
Vastus Lateralis Muscle ◽  
Mrna Levels ◽  
Igf I ◽  
Mrna Content ◽  
Heavy Resistance Training

Increasing evidence suggests that the myogenic regulatory factors (MRFs) and IGF-I have important roles in the hypertrophy response observed after mechanical loading. We, therefore, hypothesized that a bout of heavy-resistance training would affect the MRF and IGF-I mRNA levels in human skeletal muscle. Six male subjects completed four sets of 6-12 repetitions on a leg press and knee extensor machine separated by 3 min. Myogenin, MRF4, MyoD, IGF-IEabc (isoforms a, b, and c) and IGF-IEbc (isoform b and c) mRNA levels were determined in the vastus lateralis muscle by RT-PCR before exercise, immediately after, and 1, 2, 6, 24, and 48 h postexercise. Myogenin, MyoD, and MRF4 mRNA levels were elevated ( P < 0.005) by 100-400% 0-24 h postexercise. IGF-IEabc mRNA content decreased ( P < 0.005) by ∼44% after 1 and 6 h of recovery. The IGF-IEbc mRNA level was unaffected. The present study shows that myogenin, MyoD, and MRF4 mRNA levels are transiently elevated in human skeletal muscle after a single bout of heavy-resistance training, supporting the idea that the MRFs may be involved in regulating hypertrophy and/or fiber-type transitions. The results also suggest that IGF-IEa expression may be downregulated at the mRNA level during the initial part of recovery from resistance exercise.

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