scholarly journals β-Catenin/Tcf signaling in murine oocytes identifies nonovulatory follicles

Reproduction ◽  
2012 ◽  
Vol 144 (6) ◽  
pp. 669-676 ◽  
Author(s):  
Macalister Usongo ◽  
Aida Rizk ◽  
Riaz Farookhi
Keyword(s):  
Specific Gene ◽  
Reporter Construct ◽  
Lacz Expression ◽  
Multifunctional Protein ◽  
T Cell Factor ◽  
Antral Follicles ◽  
Follicular Maturation ◽  
Stage Of Development ◽  
Secondary Stage ◽  
Wnt Signal

WNTS are secreted glycoprotein molecules that signal through one of three signaling pathways. The best-characterized pathway involves stabilization of the multifunctional protein β-catenin, which in concert with members of the T-cell factor (Tcf) family activates specific gene transcription. We have examined putative Wnt/β-catenin in the murine ovary using transgenic mice harboring a reporter construct that activates β-galactosidase (lacZ) expression in response to β-catenin/Tcf binding (TopGal mice). Primordial and primary follicles did not stain for lacZ, and the proportion of β-catenin/Tcf signaling oocytes was lower than that of nonsignaling oocytes throughout estrous cycle. β-Catenin/Tcf signaling oocytes were observed in follicles from the secondary stage of development and their proportion increased with follicular maturation (secondary follicles, 20%; early antral and antral follicles, 70%). In contrast, the majority (>90%) of ovulated oocytes did not stain for lacZ. As the oocyte possesses components for WNT signal transduction, our data suggest that β-catenin/Tcf signaling is involved in the development of follicular ovulatory capability and identifies nonovulatory follicles.

scholarly journals Correction to: Forkhead box (FOX) G1 promotes hepatocellular carcinoma epithelial-Mesenchymal transition by activating Wnt signal through forming T-cell factor-4/Betacatenin/FOXG1 complex

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Xingrong Zheng ◽  
Jiaxin Lin ◽  
Hewei Wu ◽  
Zhishuo Mo ◽  
Yunwen Lian ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
T Cell ◽  
Epithelial Mesenchymal Transition ◽  
Mesenchymal Transition ◽  
T Cell Factor ◽  
Forkhead Box ◽  
Wnt Signal

An amendment to this paper has been published and can be accessed via the original article.

scholarly journals Experiments on skeletal growth and development in vitro in relation to the problem of avian phokomelia

1935 ◽  
Vol 118 (807) ◽  
pp. 133-154 ◽  
Keyword(s):  
Bone Formation ◽  
Bone Growth ◽  
Histological Study ◽  
Detailed Comparison ◽  
Specific Gene ◽  
Body Parts ◽  
Continue Development ◽  
Hatching Time ◽  
Stage Of Development

The experiments to be reported in the following pages were suggested by observations made by one of us on the so-called Creeper fowl. Creeper chickens are characterized by a disproportionate shortness of the long bones of the extremities. Histological study has shown that Creeper chickens belong in the same category as the disproportionate dwarfism of mammals known as chondrodystrophy or achondroplasia (Landauer, 1931) . The Creeper characters are inherited as a Mendelian dominant and are lethal in homozygous condition (Landauer and Dunn, 1930). Homozygous Creeper embryos generally die after about 72 hours of incubation, but in rare cases they survive beyond this stage and continue development up to nearly hatching time. These late stages of homozygous Creeper embryos exhibit striking malformations of the extremities which are known as phokomelia (Landauer, 1933). A study of the early embryonic development of homozygous Creeper embryos (Landauer, 1932) led to the conclusion that the effects of the Creeper mutation are not brought about by specific gene action on those body parts which later show deformities, but by a general retardation of body growth at a definite stage of development. This conclusion was strengthened by a detailed comparison of embryonic and post-natal bone growth in heterozygous Creeper and normal chickens (Landauer, 1934). All evidence which so far has been obtained in this work points to the conclusion that the characteristic traits of heterozygous as well as homozygous Creeper chicks are produced by an unspecific retardation of development at a time when formation of the buds of the extremities (and of the head which in homozygous embryos also shows deformities later on) are proceeding at a particularly rapid rate, thereby causing specific disturbances in the differentiation of these parts. It seemed to us that it should be possible to put these conclusions to an experimental test. The most promising way of approach appeared to be an attempt to produce in vitro the extreme abnormalities of bone formation shown by the extremities of phokomelic homozygous Creeper embryos. These abnormalities chiefly consist in (1) a general retardation of cartilage differentiation; (2) lack of bone formation; and (3) frequent partial fusion of ulna and radius on the one hand, tibia and fibula on the other, or presence of only one bone in these segments instead of two.

Follicular dynamics and secretion of inhibin and oestradiol-17β during the oestrous cycle of the hamster

1995 ◽  
Vol 146 (1) ◽  
pp. 169-176 ◽  
Author(s):  
H Kishi ◽  
K Taya ◽  
G Watanabe ◽  
S Sasamoto
Keyword(s):  
Plasma Levels ◽  
Marked Decrease ◽  
Plasma Concentrations ◽  
Follicular Development ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Oestrous Cycle ◽  
Antral Follicles ◽  
Follicular Maturation ◽  
Follicular Dynamics

Abstract Plasma and ovarian levels of inhibin were determined by a radioimmunoassay (RIA) at 3-h intervals throughout the 4-day oestrous cycle of hamsters. Plasma concentrations of FSH, LH, progesterone, testosterone and oestradiol-17β were also determined by RIAs. In addition, hamsters were injected at various times with human chorionic gonadotrophin (hCG) to determine the follicular development. The changes in plasma concentrations of FSH after injection of antisera to oestradiol-17β (oestradiol-AS) and inhibin (inhibin-AS) on the morning of day 2 (day 1=day of ovulation) were also determined. Plasma concentrations of inhibin showed a marked increase on the afternoon of day 1, remained at plateau levels until the morning of day 4, then increased abruptly on the afternoon of day 4 when preovulatory LH and FSH surges were initiated. A marked decrease in plasma concentrations of inhibin occurred during the process of ovulation after the preovulatory gonadotrophin surges. An inverse relationship between plasma levels of FSH and inhibin was observed when the secondary surge of FSH was in progress during the periovulatory period. Plasma concentrations of oestradiol-17β showed three increase phases and these changes differed from those of inhibin. Changes in plasma concentrations of oestradiol-17β correlated well with the maturation and regression of large antral follicles. Follicles capable of ovulating following hCG administration were first noted at 2300 h on day 1. The number of follicles capable of ovulating reached a maximum on the morning of day 3 (24·8± 0·6), and decreased by 0500 h on day 4 (15·0 ± 1·1), corresponding to the number of normal spontaneous ovulations. Plasma concentrations of FSH were dramatically increased within 6 h after inhibin-AS, though no increase in FSH levels was observed after oestradiol-AS. These findings suggest that changes in the plasma levels of inhibin during the oestrous cycle provide a precise indicator of follicular recruitment, and that the changes in plasma concentrations of oestradiol-17β are associated with follicular maturation. These findings also suggest that inhibin may play a major role in the inhibition of FSH secretion during the oestrous cycle of the hamster. Journal of Endocrinology (1995) 146, 169–176

233 ANTI-MÜLLERIAN HORMONE AS A PREDICTIVE ENDOCRINE MARKER FOR SUPEROVULATORY RESPONSE AND EMBRYO PRODUCTION IN BEEF CATTLE

2016 ◽  
Vol 28 (2) ◽  
pp. 248
Author(s):  
K. Center ◽  
D. Dixon ◽  
R. Rorie
Keyword(s):  
Beef Cattle ◽  
Blood Sample ◽  
Beef Cows ◽  
Corpora Lutea ◽  
Embryo Production ◽  
Antral Follicles ◽  
Stage Of Development ◽  
The Mean ◽  
Donor Cows

A study investigated the use of anti-Müllerian hormone (AMH) and/or follicle counts as a predictor of subsequent superovulatory response and embryo production in 79 beef cows. Before initiation of superovulation, ultrasonography was used to scan the ovaries of each donor cow to record the number of 3 to 5 mm follicles present, and a blood sample was collected for measure of serum AMH. At the time of embryo collection, the ovaries of donor cows were palpated to estimate the number of corpora lutea (CL) present on each ovary. Recovered embryos were evaluated for stage of development and morphological quality. Across cows, serum AMH ranged from 0.013 to 0.898 ng mL–1, with a mean of 0.293 ng mL–1. The distribution of AMH concentrations was divided into quartiles (AMH Q1 through Q4, with Q1 the lowest and Q4 the highest, ng mL–1) for analysis (ANOVA). Number of donors/collections in AMH Q1 through Q4 was 26, 22, 24, and 24, respectively. Donor cows in AMH Q4 had a greater (P < 0.001) number of 3- to 5-mm follicles at the start of superovulation than did donors in either Q1 or Q2. At embryo collection, cows in AMH Q3 and 4 had more (P < 0.001) palpable CL than cows in AMH Q1. The mean number of embryos recovered from donor cows in AMH Q4 (20.1 ± 1.8) was greater (P < 0.001) than those recovered from cows in either AMH Q1 (9.8 ± 1.8) or 2 (9.4 ± 1.9), but similar to that of AMH Q3 (15.5 ± 1.8). Percentages of recovered embryos classified as transferrable, degenerate, or unfertilized were similar (P ≥ 0.275) across AMH quartiles. Analysis indicated that AMH was positively correlated (P < 0.001) with mean follicles (r = 0.458), CL (r = 0.452), and embryos recovered (r = 0.430). To determine if follicle counts at the start of superovulation are predictive of superovulatory response, the distribution of follicle counts was divided into quartiles (F Q1 through Q4, with Q1 the lowest and Q4 the highest) for analysis. Donor cows with higher follicle counts (F Q3 and 4) at the start of superovulation had more (P < 0.001) palpable CL at embryo collection than donor cows in F Q1 or 2. More (P < 0.001) embryos (20.0 ± 2.2 v. 9.6 ± 1.8 and 11.6 ± 1.6) were recovered from cows with the highest follicle counts (F Q4) compared with cows having lower (F Q1 and 2) follicle counts, respectively. The percentage of transferable embryos and unfertilized ova were similar (P ≥ 0.688) across follicle count quartiles. As was noted for AMH, mean number of follicles at the start of superovulation was positively correlated (P < 0.001) with mean CL (r = 0.556) and mean embryos (r = 0.423) but not percentages of viable or degenerate embryos or unfertilized oocytes (P ≥ 0.153). Results confirm that relative AMH concentration was positively correlated with number of small antral follicles in the ovaries of cows and might be used to either predict superovulatory response or possibly adjust superovulatory regimen to improve superovulatory response. Further study is needed to determine the effectiveness of using either AMH concentration or follicle counts to adjust superovulatory regimens.

scholarly journals Sp5 and Sp8 recruit β-catenin and Tcf1-Lef1 to select enhancers to activate Wnt target gene transcription

2016 ◽  
Vol 113 (13) ◽  
pp. 3545-3550 ◽  
Author(s):  
Mark W. Kennedy ◽  
Ravindra B. Chalamalasetty ◽  
Sara Thomas ◽  
Robert J. Garriock ◽  
Parthav Jailwala ◽  
...  
Keyword(s):  
Cell Fate ◽  
Target Gene ◽  
Target Genes ◽  
Wnt Signaling Pathway ◽  
Embryonic Stem ◽  
T Cell Factor ◽  
Core Components ◽  
Specificity Protein ◽  
Wnt Signal ◽  
Target Gene Transcription

The ancient, highly conserved, Wnt signaling pathway regulates cell fate in all metazoans. We have previously shown that combined null mutations of the specificity protein (Sp) 1/Klf-like zinc-finger transcription factors Sp5 and Sp8 (i.e., Sp5/8) result in an embryonic phenotype identical to that observed when core components of the Wnt/β-catenin pathway are mutated; however, their role in Wnt signal transduction is unknown. Here, we show in mouse embryos and differentiating embryonic stem cells that Sp5/8 are gene-specific transcriptional coactivators in the Wnt/β-catenin pathway. Sp5/8 bind directly to GC boxes in Wnt target gene enhancers and to adjacent, or distally positioned, chromatin-bound T-cell factor (Tcf) 1/lymphoid enhancer factor (Lef) 1 to facilitate recruitment of β-catenin to target gene enhancers. Because Sp5 is itself directly activated by Wnt signals, we propose that Sp5 is a Wnt/β-catenin pathway-specific transcripton factor that functions in a feed-forward loop to robustly activate select Wnt target genes.

1 DIFFERENCES IN RESUMPTION OF OOCYTE MATURATION IN YOUNG AND OLD MARES

2008 ◽  
Vol 20 (1) ◽  
pp. 81 ◽  
Author(s):  
L. F. Campos-Chillon ◽  
C. M. Clay ◽  
J. L. Altermatt ◽  
G. J. Bouma ◽  
E. M. Carnevale
Keyword(s):  
Granulosa Cells ◽  
Expression Patterns ◽  
Cell Number ◽  
Specific Gene ◽  
Meiotic Arrest ◽  
Temporal Differences ◽  
Time Points ◽  
Follicular Maturation ◽  
Copy Numbers

The decline in fertility of aged mares is linked with declining oocyte quality. Oocyte viability is dependent on the ability of oocytes to remain in meiotic arrest until the initiation of maturation. We hypothesize that aging is associated with quantitative and temporal differences in meiotic arrest and resumption in oocytes, ultimately resulting in a dissociation of oocyte and follicular maturation. The objectives of this study were to determine temporal differences in the mRNA content of amphiregulin and epiregulin in granulosa cells; PDE4 in cumulus and granulosa cells; and PDE3A, GPR3, GDF9, and BMP15 in oocytes during in vivo maturation in young (3–12 years) v. old (>20 years) mares. Oocytes and follicular cells were collected by transvaginal follicular aspiration. Maturation was induced in estrous mares with a follicle >30 mm by injection of 750 �g of recombinant equine LH. Aspirations were attempted at 0, 6, 9, and 12 h after LH administration. Six oocytes and follicular cell samples from each age group and time point were collected and stored immediately after aspiration. Total RNA was isolated from single denuded oocytes and lysed cumulus and granulosa cells. A fraction of the total lysate was used to determine cell numbers from the DNA copy number of the equine CG� subunit gene. DsRED RNA was added to each RNA isolate to serve as an exogenous standard. Quantitative RT-PCR was performed from cDNA with equine primer pairs. Copy numbers were calculated with an intra assay standard curve of plasmid containing the specific gene and corrected with the exogenous DsRED RNA and cell number. For each gene, mean mRNA copy numbers for time points and age groups were compared by ANOVA and Tukey's HSD test. Expression of PDE4D in cumulus cells was similar between young and old mares and time points. However, PDE4D peaked (P < 0.05) at 6 h in granulosa cells from young, but not old mares. Amphiregulin expression in granulosa cells of young mares peaked (P < 0.05) at 9 h and did not increase in the old mares. Epiregulin expression in granulosa cells peaked (P < 0.05) at 9 h and 6 h in young and old mares, respectively. The pattern of expression of PDE3A for oocytes of young and old mares was similar with an increase (P < 0.05) at 9 h. There was an interaction (P < 0.05) in the expression of GPR3 for age � time. Expression peaked at 9 and 6 h in young and old mares, respectively. Pattern of expression of GDF9 was similar between young and old mares except for a decrease (P < 0.05) in expression in old mares at 9 h. There was an interaction (P < 0.05) in the expression of BMP15 for age � time. Expression in young mares peaked at 9 h while that in old mares peaked at 6 h and decreased at 9 h. These results suggest that key gene expression patterns involved in oocyte and follicular maturation cascades are asynchronous for young versus old mares and could explain some aspects of the age-associated decline in fertility.

185 PREVALENCE AND PROLIFERATIVE ACTIVITY OF MULTIOOCYTE FOLLICLES IN BITCHES: PRELIMINARY RESULTS

2015 ◽  
Vol 27 (1) ◽  
pp. 183
Author(s):  
R. C. Justino ◽  
N. T. Lunardon ◽  
K. C. Silva-Santos ◽  
R. L. Oliveira ◽  
M. M. Seneda ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Granulosa Cells ◽  
Proliferative Activity ◽  
Nuclear Antigen ◽  
Oocyte Nucleus ◽  
Follicular Growth ◽  
Stage Of Development ◽  
Secondary Stage ◽  
Cell Proliferation Activity ◽  
Proliferation Activity

Multioocyte follicles (MOF) are follicles that enclose two or more oocytes. They have been described in many mammalian species, but there is no evidence about their activity in the ovaries. The aim was to estimate the prevalence of MOF and to compare the cell proliferation activity between follicles containing one or more oocytes in the ovaries of prepubertal and adult bitches. Eighty ovaries from prepubertal (n = 20) and adult bitches (n = 20) were obtained by elective ovariohysterectomy (OHE). Immediately after OHE, ovaries were immersed in Bouin's fixative for histological processing. 5 µm thick sections were mounted on histological slides and stained with periodic acid-Schiff (PAS) and hematoxylin. Cell proliferation was evaluated by immunohistochemistry using the proliferating cell nuclear antigen (PCNA). Monoclonal antibody PCNA (clone PC1O, 1 : 200 dilution, Biocare, Concord, CA, USA) was used according to manufacturer's instructions and an antibody diluent was used as a negative control. Slides were counterstained with hematoxylin and examined at 200× to 400× magnification under light microscope. Only cells showing PCNA signal exclusively in the nucleus were considered positive. The prevalence of MOF in the ovaries was compared using a Fisher's exact test (P < 0.05). In all females, the prevalence of MOF was 55% (22/40). MOF containing two or three oocytes were more abundant; however, multioocyte follicles with up to 12 oocytes were observed. The prevalence of MOF at the primordial stage was higher for prepubertal bitches (47 v. 28%) but adult bitches exhibited a higher frequency of secondary MOF (49 v. 25%; P < 0.05). There was no difference in the prevalence of MOF at primary stage between prepubertal and adult bitches (28 v. 23%; P > 0.05). Regarding the cell proliferation activity, PCNA immunoreactivity was detected in oocyte nucleus and granulosa cells of multioocyte follicles at different stages of development. Similarly to what was observed for follicles containing only one oocyte, all nuclei of oocytes within multioocyte follicles exhibited PCNA immunoreactivity and there was a gradual increasing of immunoreactivity in granulosa cells according to the stage of follicular growth. Expression of PCNA by granulosa cells of multioocyte follicles was higher in the secondary and antral stage of development; however, some primordial and primary follicles also exhibited some PCNA-positive cells. In conclusion, the prevalence of MOF at the primordial stage of development was higher in prepubertal bitches, whereas MOF at the secondary stage were more frequent in adult bitches. The PCNA expression pattern by the oocyte nucleus of multioocyte follicles was similar to that observed in follicles containing only one oocyte, which is suggestive of similar activity between these follicles. Furthermore, the presence of proliferative activity in granulosa cells of multioocyte follicles suggests an association of the PCNA expression with more advanced stages of follicular growth.

A transgene containing lacZ is expressed in primary sensory neurons in zebrafish

Development ◽  
1992 ◽  
Vol 115 (2) ◽  
pp. 421-426 ◽  
Author(s):  
T.A. Bayer ◽  
J.A. Campos-Ortega
Keyword(s):  
Trigeminal Ganglion ◽  
Ganglion Cells ◽  
Transgenic Fish ◽  
Cell Types ◽  
Single Copy ◽  
Reporter Construct ◽  
Lacz Expression ◽  
Promoter Sequences ◽  
Rous Sarcoma ◽  
Ganglion Neurons

In order to screen for developmentally active chromosomal domains during zebrafish embryogenesis, we generated transgenic fish by microinjecting two different lacZ reporter constructs into fertilized eggs. Transgenic fish were screened among the progeny of injected fish (F0) crossed to non-injected fish. Groups of 15 to 20 progeny of each cross were tested for lacZ expression and/or transmission of injected sequences using PCR and Southern hybridizations. Progeny from 2 of 102 fish injected with supercoiled constructs containing Rous sarcoma virus promoter sequences showed apparently spatially regulated beta-galactosidase (beta-Gal) activity. However, we were not able to detect this reporter construct in DNA from fins of F1 fish. Injections of a linear reporter construct containing mouse heat-shock promoter sequences revealed transmission of injected sequences to F1 progeny in about 6% of cases (8 of 129 fish, tested with PCR). We found one lacZ-expressing line that showed a spatially and temporally restricted expression of lacZ and, therefore, features typical characteristics of “enhancer trap” lines. In this line, lacZ expression starts at 16 hours post-fertilization in trigeminal ganglion cells. At about 24 hours lacZ expression can be detected in trigeminal ganglion neurons and Rohon-Beard neurons, indicating that the development of these two cell types shows common features. The reporter gene has integrated as a single copy. The founder fish was mosaic: 19% of its offspring (3 of 16 tested animals) carried the reporter construct in their fins; about 51% (13 of 27 tested animals) of the progeny of F1 fish were beta-Gal positive indicating full hemizygosity.(ABSTRACT TRUNCATED AT 250 WORDS)

SERUM GONADOTROPHINS AND FOLLICULAR DEVELOPMENT IN IMMATURE RATS AFTER EARLY ANDROGEN ADMINISTRATION

1976 ◽  
Vol 68 (3) ◽  
pp. 461-468 ◽  
Author(s):  
J. TH. J. UILENBROEK ◽  
E. ARENDSEN DE WOLFF-EXALTO ◽  
M. A. BLANKENSTEIN
Keyword(s):  
Testosterone Propionate ◽  
Follicle Stimulating Hormone ◽  
Follicular Development ◽  
Female Rats ◽  
Vaginal Opening ◽  
Follicular Growth ◽  
Antral Follicles ◽  
Follicular Maturation ◽  
Immature Rats ◽  
Plasma Oestradiol

SUMMARY Follicular development and serum gonadotrophin levels were studied in female rats after neonatal androgen administration. After injection of 1250 μg testosterone propionate (TP) on day 5 after birth the composition of the follicular population was altered: at nearly all ages the number of pre-antral follicles (follicular volume 2–20 × 105 μm3) was lower than in oil-treated rats, in some cases the number of small antral follicles (21–249 × 105 μm3) was also lower. Furthermore levels of serum follicle-stimulating hormone and luteinizing hormone were decreased from day 7 to day 20 suggesting that the high gonadotrophin levels before day 20 are of importance for normal follicular development. In contrast, final follicular maturation in TP-treated rats was enhanced; at day 35 more large antral follicles (follicular volume ≥ 500 × 105μm3) were present in TP-treated rats than in oil-treated rats. The presence of more large antral follicles was accompanied by higher plasma oestradiol concentrations, higher uterine weights and advanced vaginal opening. These results demonstrate an inhibition of normal follicular growth and an acceleration of ovarian maturation after neonatal androgen administration.

scholarly journals Skin Basement Membrane: The Foundation of Epidermal Integrity—BM Functions and Diverse Roles of Bridging Molecules Nidogen and Perlecan

2013 ◽  
Vol 2013 ◽  
pp. 1-16 ◽  
Author(s):  
Dirk Breitkreutz ◽  
Isabell Koxholt ◽  
Kathrin Thiemann ◽  
Roswitha Nischt
Keyword(s):  
Basement Membrane ◽  
De Novo ◽  
Molecular Assembly ◽  
Specific Gene ◽  
Stage Of Development ◽  
Molecular Aspects ◽  
And Function ◽  
Molecular Adaptors ◽  
Nidogen 1

The epidermis functions in skin as first defense line or barrier against environmental impacts, resting on extracellular matrix (ECM) of the dermis underneath. Both compartments are connected by the basement membrane (BM), composed of a set of distinct glycoproteins and proteoglycans. Herein we are reviewing molecular aspects of BM structure, composition, and function regarding not only (i) the dermoepidermal interface but also (ii) the resident microvasculature, primarily focusing on theper senonscaffold forming components perlecan and nidogen-1 and nidogen-2. Depletion or functional deficiencies of any BM component are lethal at some stage of development or around birth, though BM defects vary between organs and tissues. Lethality problems were overcome by developmental stage- and skin-specific gene targeting or by cell grafting and organotypic (3D) cocultures of normal or defective cells, which allows recapitulating BM formationde novo. Thus, evidence is accumulating that BM assembly and turnover rely on mechanical properties and composition of the adjacent ECM and the dynamics of molecular assembly, including further “minor” local components, nidogens largely functioning as catalysts or molecular adaptors and perlecan as bridging stabilizer. Collectively, orchestration of BM assembly, remodeling, and the role of individual players herein are determined by the developmental, tissue-specific, or functional context.

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