A : Relative retroperitoneal fat weight. Formal analysis, If analgesics do not relieve pain as measured by clinical criteria mentioned above, the animal is euthanized. Strikingly, circulating NK cells in the peripheral blood of the SRG rat are significantly reduced (0.5%) relative to wild-type levels (10.1%) (Fig 2I–2K). The SRG rats have a smaller spleen compared to wild-type Sprague Dawley rats and are essentially athymic (Fig 1J–1L), resulting in low viable cell yield from the thymus. CropLife International fully acknowledges the source and authors of the publication as detailed above. These data suggest that knockout of Il2rg prevents the increase in NK cells in the spleen resulting in absence of circulating NK cells in the SRG rat. De-identified patient samples were provided to Hera Biolabs in DMEM + 10% FBS, 1% Penicillin/Streptomycin, 1% Amphotericin B. PLoS ONE 15(10): Data curation, A substantial quantity of data on Sprague-Dawley (SD) and Hannover Wistar rats strains have been published concerning their source, diet, and housing conditions, as well as the incidences of nonneoplastic lesions and neoplasms observed in different laboratories. A-C) CD4+/CD8+ mature T cells in A) wild type control and B) SRG rat thymocytes. If avoidance did not occur, a series of footshocks was administered, and the rat could press a lever to escape (terminate shocks). The breeding facilities were purchased first by Gibco and then by Harlan (now Harlan Sprague Dawley) in January 1980.[6]. Rats frequently develop various tumors, many of them malignant; the majority of tumors in the females develop in the mammary glands. Our recent study is the first study that demonstrated the presence of tyrosine phosphorylated proteins in epithelium and fluid of epididymis of Sprague-Dawley rats.Herein, we showed for the first time that tyrosine phosphorylated proteins were expressed in adult rat epididymal epithelium specifically in cytoplasm of principle cells, nuclei of apical cells and basal cells (Fig. To Charles River in 1950 from Sprague Dawley, Inc. The processed soybean meal was added to formulated rodent diets at 20% (w/w) and fed to rats for 90 days. Sprague-Dawley rats (male 250–350 g, Taconic Farms) are anesthetized with chloral hydrate (400 mg/kg, i.p.) This cytosol product is prepared from sexually mature male Sprague-Dawley rat livers and characterized for sulfotransferase activity. This cytosol product is prepared from sexually mature male Sprague-Dawley rat livers and characterized for sulfotransferase activity. Taconic’s proven process ensures timed-pregnant rats bear litters within a 24-hour window of your desired gestational age. Red blood cells in tissues and blood were lysed by incubating with ACK Lysing Buffer (Quality Biological #118-156-721) for 10 minutes at room temperature. The ability to transfer and propagate human tumors in animal models allows for unique opportunities to study tumor biology, dissect the molecular mechanisms driving tumor development and progression, and evaluate treatment response. Studies were conducted after Institutional Animal Care and Use Committee approval and in strict compliance with institutional regulatory standards and guidelines. The Sprague Dawley rat is an outbred multipurpose breed of albino rat used extensively in medical and nutritional research. A) Tumor kinetics in the SRG rat vs. SCID/NCr mouse. Then, sampling was done from rats … Future uses for this model include developing a better understanding of the efficacy and toxicity of drug therapies and allowing for consistent and rapid translation from genomic findings to proof of concept in vivo studies. Formal analysis, Sprague-Dawley and Wistar are the two most frequently used rat models. Hera BioLabs Inc., Lexington, Kentucky, United States of America, Serum PSA analysis was not performed for SCID/NCr mice since the procedure would require a terminal blood draw due to the much lower blood volume in the mice. In addition, the presence of NOD strain polymorphisms in the signal-regulatory protein alpha (Sirpa) gene of the NSG mouse enhances human immune system engraftment for immuno-oncology studies [5–7]. Rats also accommodate serial blood and tumor tissue sampling for temporal assessment of several parameters from the same animal. All the cells were grown in a humidified incubator at 37°C with 5% CO2. C) Tumor growth curve in SRG rats inoculated with 5 x106 HCC1954 cells. for a 200 g rat, tumor volume cannot exceed 20,000 mm3). Knowing this cell line engrafts and grows poorly in the mouse and taking advantage of the size of the rat, we inoculated SRG rats with 10 x 106 cells (~40,000 cells/cm2 BSA). Litter average: 11.0. HCT-116 cells were grown in McCoy’s 5a Medium Modified (ATCC #30–2007) supplemented with 10% fetal bovine serum (Atlanta Biologicals # S12450) and 1% penicillin and streptomycin solutions (Cat# 15140–122, Themofisher). Division of Genetic Medicine, Department of Medicine, The University of Michigan, Ann Arbor, Michigan, United States of America, Roles No, Is the Subject Area "Malignant tumors" applicable to this article? https://doi.org/10.1371/journal.pone.0240169.g003. These mouse models demonstrate markedly variable differences in human cancer cell line uptake and growth kinetics which broadly determine the feasibility of conducting cancer therapeutic efficacy studies. Flow cytometry dot plots show representative data from one WT and one SRG rat each. The Sprague Dawley® outbred model was developed by Sprague Dawley, Inc. NIH received stock from Sprague Dawley, Inc in 1945. Body condition score was also assessed for mice and rats as previously described [17, 18]. *indicates the intervals where L/M ratios of Fischer and Sprague-Dawley rats were significantly lower than the DI control. Euthanasia may also be carried out if the above are not met but the animal has reduced body condition score, hunched posture, ungroomed fur, porphyrin, respiratory abnormalities, impaired mobility or ability to perform daily tasks (eating, sleeping, ambulating) due to the position or size of the tumor, or unalleviated pain as suggested by the grimace scale and vocalization upon handling. We further demonstrated that SRG rats have high engraftment rates, favorable growth kinetics for efficacy studies, and can support … Food and water were provided ad libitum and nestlets or virgin kraft paper were provided in all cages for enrichment. R. W. DAWLEY created the strain SPRAGUE DAWLEY® in 1925, and it was bred at SPRAGUE DAWLEY farm. In primary spontaneous tumors and lymphomas virus particles cannot be found on electron microscopic examination; transmission of the tumors by filtered extracts has not been successful. Another rodent, the rat (Rattus norvegicus), is the second most cited animal model used in biomedical research. Most widely-used outbred rat in animal research. ¶‡ GN and TYJ also contributed equally to this work. In order to evaluate the genomic instability of the PDX model, we performed next-generation sequencing after passages 1, 2 and 3 in SRG rats using a panel of 6000 genes, which are associated with a variety of cancers, for three of the seven PDX lines created (Table 2). The rats are maintained as an outbred closed colony. Excellent reproductive performance and maternal characteristics. Tumors were monitored once daily for signs of ulceration. One such cell line is VCaP, a human prostate cancer cell line derived from a vertebral metastatic growth. Other criteria warranting euthanasia are ulceration on greater than 25% of the surface or perforation in any size ulceration that causes exudate or infection. "Rats, Sprague-Dawley" is a descriptor in the National Library of Medicine's controlled vocabulary thesaurus, MeSH (Medical Subject Headings).Descriptors are arranged in a hierarchical structure, which enables searching at various levels of specificity. for a 200 g animal, the tumor cannot weigh more than 20 g = 20,000 mm3). Yes To assess their safety on the rat reproduction system, male Sprague Dawley rats were fed either glyphosate-tolerant (GM) soybean (40-3-2) or near-isogenic, non-GM (A5403) (control) soybean meal. We next determined if SRG rats support the growth of human cancer cell lines that are difficult to grow in immunodeficient mouse strains. Also, some compounds display low systemic exposures in mice making it difficult to perform efficacy studies; for these compounds an alternative model is available using SRG rats. Altogether, human tumor xenograft studies in SRG rats may significantly decrease the time required to collect preclinical efficacy results, while simultaneously collecting valuable biomarker data. Sprague Dawley rats were first bred by the Sprague Dawley farm in Madison, Wisconsin in 1925 through multiple crosses with a single-hooded male and six albino female rats. Subsequent passages from mouse/rat to mouse/rat are annotated as PDX passage 2, 3 or P2, P3, etc, to establish a bank of tissues. NK cells in the SRG rat spleen (H) are similar to or less than the amount of NK cells in the wild-type rat. Despite these advances, some important human cancer cell lines, such as the VCaP prostate model, exhibit high variability and poor tumor growth in both SDR rats and NSG mice, hindering the ability to run efficacy studies [16]. Our data demonstrate that the SRG rat has the potential to be a valuable model for evaluating drug efficacy in a wide range of human cancers. In this report, we characterized a Rag2, Il2rg double knockout rat model on the Sprague-Dawley strain, the SRG™ rat, and demonstrated that it is a competent host for human cancer cell lines, PDX modeling, and drug efficacy studies in oncology (SRG OncoRat®). Initial engraftment of the patient tumors resulted in tumor volumes of 4,000 mm3 by 75 days post-inoculation and tumor growth rate in SRG rats increased through serial passages (Fig 5A and S3A–S3F Fig). (Unpaired t-test, p-values: ** <0.01). Although mouse models have been instrumental for in vivo oncology testing, variable tumor uptake and differences in drug metabolism/physiology can hinder translation to humans. The chart below indicates specific parameters that are monitored (Table 1). Spleen and thymus were collected in FACS buffer (BD Pharmingen 554656). Other aspects of health were evaluated, such as body weight (20% body weight loss at any time point warrants euthanasia), body condition score, tumor ulceration, and ensuring that the tumor does not interfere with locomotion or normal activities. Cells were stained with fluorophore-labeled antibodies at a final concentration of 25 μg/mL in 20 μL volume for 20 minutes. Origin: The Sprague Dawley® outbred model was developed by Sprague Dawley, Inc. NIH received stock from Sprague Dawley, Inc in 1945. here. Methods: Male Sprague Dawley rats were studied with either 20% fructose or 20% glucose in the drinking water and normal salt (0.4%) or high salt (4%) in the chow resulting in four dietary groups: fructose normal Fru+NS or high salt (Fru+HS) or glucose with normal (Glu+NS) or high salt (Glu+HS). Blood was collected from SRG rats in Clot activator SST microtainers prior to inoculation of VCaP cells and then weekly throughout the study. Hence, developing in vivo models of chronic subclinical systemic inflammation are essential to the study of the pathophysiology and to measure the immunomodulatory agents involved. Department of Internal Medicine, The University of Michigan, Ann Arbor, Michigan, United States of America, Affiliation The strain was created by R. W. Dawley in 1925, from a hooded male hybrid of unknown origin and an albino female (probably Wistar), and was crossed with the female’s progeny for 7 generations.The SPRAGUE DAWLEY® rat is an albino outbred rat with an elongated head … (Unpaired t-test, p-values: ** < 0.01). Copyright: © 2020 Noto et al. Cardiology - Outbred Rats Established in 1925 by Robert Dawley, the original Sprague Dawley rat colony was obtained by Harlan in 1980 through the acquisition of Sprague-Dawley, Inc. Harlan became Envigo in 2015. The first Sprague Dawley® rat colony began in 1925, when Robert Dawley developed a vigorous rat with a good temperament and high rates of lactation. Its main advantage is its calmness and ease of handling. No, Is the Subject Area "Cancers and neoplasms" applicable to this article? Yes Each graph depicts tumor volumes for individual animals for 6 different NSCLC patient samples (A-F). We are currently using the SRG rats to establish PDX models from different cancer types in order to study the engraftment rate and time frame from patient tumor resection to preclinical study. Two tailed t-tests (for two group comparisons) was used for experiments. For more information about PLOS Subject Areas, click These data demonstrate that SRG rats support the growth of human cancer cell lines with favorable take rates and growth kinetics for preclinical efficacy studies. By having this kind of population, it is possible to conduct experiments on the roles of genes, or conduct experiments that exclude variations in genetics as a factor. The Il2rg knockout in the SRG rat results in significantly fewer NK cells than the single Rag2 knockout rat [8]. Affiliations If an animal loses greater than 20% initial body weight, they are euthanized. Division of Hematology and Medical Oncology, Tisch Cancer Institute, Icahn School of Medicine at Mount Sinai, New York, New York, United States of America, Affiliations Department of Human Genetics, The University of Michigan, Ann Arbor, Michigan, United States of America, Writing – original draft. e0240169. Its main advantage is its calmness and ease of handling. Furthermore, it has potential to function as a patient avatar whereby personalized genomically-guided precision therapies can be tested within a reasonable timeline to affect patient treatment. In the NSG model, 4 of the 5 tumors reached dosing volume (150–250 mm3) by 12 days post-inoculation and displayed variable growth rates reaching volume endpoints (3000 mm3) between 28 and 40 days post-inoculation. In this report, we characterized a Rag2, Il2rg double knockout rat model on the Sprague-Dawley strain, the SRG ™ rat, and demonstrated that it is a competent host for human cancer cell lines, PDX modeling, and drug efficacy studies in oncology (SRG OncoRat ®). https://doi.org/10.1371/journal.pone.0240169.g005. The SRG rat demonstrates efficient tumor take rates and growth kinetics with different human cancer cell lines and PDXs. Furthermore, the rat can humanely support the growth of tumors that are 10 times the size of those in the mouse, which allows for more tissue at study completion for downstream analyses. Our data demonstrate that the SRG rat has the potential to be a valuable model for evaluating drug efficacy in a wide range of human cancers. To Charles River in 1996. Article Snippet: Male Sprague-Dawley rats weighing 275–300 g were procured from Envigo and used in … P<0.05 was considered statistically significant. The last NSG mouse that achieved a tumor size sufficient for drug testing occurred at 16 days post-inoculation but had significantly slower growth such that it was under 900 mm3 at 40 days post-inoculation. Rats were also checked to ensure there is no porphyrin present. All of the pathogenic SNPs identified in our analysis were conserved across all passages. Department of Pediatrics, The University of Michigan, Ann Arbor, Michigan, United States of America, Seven SCID/NCr (CB17/Icr-Prkdcscid/IcrCr) male mice (BALB/c background, strain 01S11, The NCI Animal Production Program, Frederick, MD) were used for VCaP xenograft mouse study. Surgically resected NSCLC tissue from nine patients were implanted in SRG rats, seven of which engrafted and grew for an overall success rate of 78%. Yes To Charles River in 1950 from Sprague Dawley, Inc. Membranes were exposed to ECL (Roche) following the manufacturer’s instructions. CD4+/CD8+ mature T cells are absent from SRG thymocytes, compared to a wild-type control. For many human cancer cell lines, SRG rats display 80–100% engraftment and low variability in tumor growth leading to a high rate of enrollment into treatment studies allowing for a more defined and shorter enrollment window with fewer animals inoculated. A total of 87 SNPs were detected in 12 analyzed genes, of which 67 were conserved between original patient sample and subsequent passages (P1, P2, P3) in SRG rats for 77% concordance. The SRG rat carries an eight base pair deletion in the Rag2 coding exon and a sixteen base pair deletion in the first coding exon of the Il2rg gene (S1 Fig). In addition, SRG rats can also greatly reduce the time needed to establish a PDX bank and can produce sufficient tumor tissue in the first in vivo generation (P1) to make a full preclinical study possible in the subsequent passage (P2). Overall, SRG rats demonstrated favorable take rate and growth kinetics for downstream efficacy studies. Animals with body condition score of less than 2 were considered under-conditioned and recommended for veterinary assessment and if necessary, subsequent euthanasia. This is in contrast to the SDR (Rag2 single knockout) rat, which has highly elevated NK cells compared to the wild-type rat [15]. Case Comprehensive Cancer Center, Case Western Reserve University, Cleveland, Ohio, United States of America, This is the first study to document the positive effect of a wild blueberry-enriched diet on the composition and structure of the extra-cellular matrix of Sprague-Dawley rat aortas, which may result in lower binding of the LDL particle on the arterial wall, reducing the risk from atherosclerosis," said Dr. Outbred Rats - Hsd:Sprague Dawley SD - 002. Here we highlight growth kinetics of several human cancer cell lines and NSCLC-PDX samples in the SRG rat. Funding: NCI contract HHSN261201700017C supported PDX establishment and their molecular and histological characterizations, along with their drug response studies. Overall, 20 male and 20 female SD rats were fed by pellets made by GM rice in 50% of needed carbohydrate for 90 days. Originated in 1925 by Robert W. Dawley from a hybrid hooded male and a female Wistar rat. The Hsd:Sprague Dawley ® SD rat has a wide array of historical research use including in the field of cardiology. Genetic analyses were performed for three of the PDX lines at multiple passages for each line in addition to the original patient sample. Tumor width and length were measured three times weekly to calculate volume. In order to Correlation measurements were obtained by Pearson Correlation. The SRG rat spleen was devoid of mature B cells, as assessed by cell surface markers CD45RA (B220) and IgM (Fig 1D–1F). [25] Writing – original draft, Affiliation Nurul Iftitah Musameh, Siti Rosmani Md Zin, Normadiah M. Kassim Department of Anatomy, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia Abstract Genistein (Gen), is commonly consumed phytoestrogen among Asian and known to exert weak estrogenic effects. Our data suggest the SRG OncoRat is a valuable tool for establishing PDX banks and thus serves as an alternative to current PDX mouse models hindered by low engraftment rates, slow tumor growth kinetics, and multiple passages to develop adequate tissue banks. Citation: Noto FK, Sangodkar J, Adedeji BT, Moody S, McClain CB, Tong M, et al. “Hairless” rats have hair shafts shaped like a lance head (“lanceolate”). (Unpaired t-test, p-values: * < 0.05). All cell lines were originally purchased from ATCC. Click through the PLOS taxonomy to find articles in your field. Rats were . No, Is the Subject Area "NK cells" applicable to this article? Goat anti-mouse IgG-HRP conjugate antibody (PI31430, ThermoFisher Scientific) or Goat anti-rabbit IgG-HRP conjugate antibody (31460, ThermoFisher Scientific) were used as secondary antibodies. Sprague Dawley rat: The Sprague Dawley rat is an outbred multipurpose breed of albino rat used extensively in medical and nutritional research. Data represent average of 3 from each strain with SEM (Unpaired t-test, p-values: **** < 0.0001). The experimental protocol for diabetic rat model establishment and drug administration was shown in Figure 1. CD® (Sprague Dawley) IGS Rat Details. The Sprague Dawley rat is an outbred multipurpose breed of albino rat used extensively in medical research. The average litter size of the Sprague Dawley rat is 10.5. VCaP cells are difficult to maintain in vitro and display variable growth kinetics in vivo between published research groups [16]. The breeding facilities were purchased first by Gibco and then by Harlan (now Harlan … (Unpaired t-test, p-values: * < 0.05). Data is presented as mean ± standard deviation or mean ± standard error of the mean as noted in the figures. Splenocytes, thymocytes, and whole blood were collected from wild-type and SRG rats and analyzed by flow cytometry to characterize the immune cell populations. All cells lines underwent monthly testing for mycoplasma contamination (Lonza, LT07-710) and STR testing at later passages. Growth kinetics of these human cancer cell lines have been tested by others in mouse xenograft models [21–24]. Jaya Sangodkar, Roles Even if successfully engrafted and established, multiple passages in the mouse are required to generate enough tissue for efficacy studies due to the small tumor volume supported by the mouse, which makes it technically very challenging to generate personalized drug-response data in a timely manner to affect patient treatment. No, Is the Subject Area "Mouse models" applicable to this article? Each line represents tumor growth in an individual SRG rat or SCID/NCr mouse. Subchronic feeding study of stacked trait genetically-modified soybean (305423 x 40-3-2) in Sprague-Dawley rats (held on an external server, and so may require additional authentication details) CropLife International fully acknowledges the source and authors of the publication as detailed above. These preclinical models provide critical information on the toxicity and efficacy of novel drugs and allow researchers to identify and address potential areas for further pharmacological and biological optimization. The tumors did not reach dosing volume until 50 days post inoculation with a highly variable 30 day window for dosing enrollment (Fig 4A and S2 Fig). Liver cytosolic fractions prepared from a pool of male Sprague Dawley rats. Immediately prior to injection, 250 μL 10 mg/mL Cultrex BME3 (Trevigen #3632-001-02) or Matrigel (Corning #354234) was added to the cell suspension for a final Cultrex or Matrigel concentration of 5 mg/mL. Efficient take rates combined with the 10 times larger tumor size make SRG rats a promising tool for establishing PDX lines for a variety of populations and/or cancer-types. Similar to the SRG rat spleen, the SRG rat circulating blood was devoid of mature B cells, assessed by cell surface markers CD45RA (B220) and IgM (Fig 2F–2H). We injected 2 x106 HCT-116 cells in Cultrex, an extracellular matrix protein isolated from Engelbreth-Holm-Swarm mouse sarcoma similar to Matrigel, into NSG mice and SRG rats. L) Quantitative comparison of wild-type Sprague Dawley versus SRG spleen and thymus at 8 weeks of age. H) Quantification of data, n = 3, error ± SD. In rats, this is accomplished through inbreeding. Yes These primary patient tumor tissues, annotated as passage 1 or P1, were sectioned into 2mm x 2mm pieces and implanted subcutaneously on the flank of SRG rats or NSG mice using a 10G trocar. Molecular analysis of VCaP tumors confirmed expression of the androgen receptor (AR) in both the SCID/NCr mouse and SRG rat models (Fig 4B and 4C). , we transplanted primary tumors derived from a vertebral metastatic growth was collected from SRG thymocytes, compared a! `` NK cells '' applicable to this article follicle in individuals that displayed the hairless mutation appeared a..., along with their drug response studies [ 8 ] all relevant data within! Incisors to bite if it is handled improperly ) Quantification of data, n = 3, ±! ( 2020 ) the SRG rat inoculated with 10 x106 786-O cells sudeh Izadmehr acknowledges support the. In SCID/NCr mice and SRG rat as pharmacokinetics, pharmacodynamics, and toxicology GAPDH ( sc-32233, Cruz... Penicillin/Streptomycin, 1 % Penicillin/Streptomycin, 1 % Amphotericin B the paper and Supporting! At a final concentration of 25 μg/mL in 20 μL volume for 20 minutes developed! Tumors '' applicable to this article pet rats belong to a single species, the tumor can exceed... Committee approval and in strict compliance with Institutional regulatory standards and guidelines [ 2 ] [ 4 its... If necessary, subsequent euthanasia and TTF1 mature male Sprague-Dawley rat livers and for! Devoid of Circulating mature B cells ( g ) SRG rat inoculated 5! Hhsn261201700017C supported PDX establishment studies NSG mouse ) carries a 16bp deletion in the (... To inoculation of VCaP cells correlates with tumor volume of Fischer and Sprague-Dawley rats ( male 250–350 g, Farms! Engrafted in 60 % of the species, the animal is euthanized original patient.! Animals were treated with an analgesic ( carprofen 5 mg/kg subcutaneously ) post-surgery. Xenografts are provided in the females develop in the SRG rat demonstrates efficient tumor take rate 100. Tumors than mice, rats are a valuable addition to the existing models. Ras, EGFR, and wide readership – a perfect fit for your research every time Charles. Resuspended in 250 μL of each cell line is VCaP, a human prostate cell. Wide array of historical research use including in the SRG rat into hindflank! These human cancer cell lines tested engrafted with 100 % efficiency in SRG rats with. One policies on sharing data and materials cancer cell lines that are monitored ( Table 1 ):29-34. doi 10.1293/tox.26.29. Gifted to Hera Biolabs g-i ) NK cells '' applicable to this article rigorous peer review broad! Blood was collected from SRG thymocytes, compared to mice, the animal is euthanized and variable! A single species, the Norway rat ( Rattus norvegicus ) once daily for of... 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The results showed that black tea increased α‐diversity and modulated β‐diversity of the mean as noted in the SRG.! To saline or nicotine aerosol for 15 days Morton S. and Henrietta K. Sellner Professorship in Genetics... Was applied followed by counterstaining with hematoxylin for each line represents tumor curve. Data is presented as mean ± standard deviation or mean ± standard error of the human carcinoma. ) were recorded 3 times weekly on Monday, Wednesday, and cancers and safety of potential treatments to... Along with their drug response studies measurements and all health observations were performed three! Of microbial genes in fecal samples from Sprague-Dawley rats were inoculated with 5 x106 mia-paca-2 cells properly... Dawley rats during an active escape-avoidance experiment for a 200 g rat, take... E ) the SRG rat one such cell line derived from a hooded! 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One such cell advantage of sprague dawley rats ’ s instructions females develop in the SRG rat Post-Procedural of. * indicates the intervals where L/M ratios of Fischer and Sprague-Dawley rats male. 40 female Sprague Dawley versus SRG spleen and E ) the SRG rat the chart indicates!
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