Alignments were sorted and filtered for mapping quality (MAPQ > 10) using SAMtools v1.5 (ref. WT1xIfnrs mice were viable and fertile, with no obvious phenotypes. 55, 53–65 (2019). J. Med. Th~ Vacutain~rsystem !s quicker and simpler to use, with greater versatility, producing blood specimens of higher quality a~dat a ~ostcom­ parable with that by the syringe system. contributed to methodology, investigation, formal analysis and manuscript review. Produktkatalog Kapitel venöse Blutentnahme. Comparison of the fold-changes for these DEGs in Dp16/WT versus Dp162xIfnrs/WT revealed significant attenuation of gene expression changes in Dp162xIfnrs in every tissue examined, albeit to variable degrees (Fig. J. Hum. Genet. When reintroduced to the shock context on day 2, both WT and Dp16 froze at higher baseline rates relative to the beginning of day 1, yet WT mice froze at a substantially higher rate than Dp16. Our findings define a role for the Ifnr locus during heart development. Cervical cancer screening Einmalhalter aus Kunststoff für Röhrchen mit ⌀ 13 und 16 mm Durchmesser. J Hosp Infect. c, Swim path efficiency of mice navigating to the escape platform in an MWM for male and female (M and F), WT (n = 39, 19 male and 20 female), Dp16 (n = 28, 13 male and 15 female) and Dp162xIfnrs (n = 25, 12 male and 13 female). Sample sizes are WT (n = 5, 2 male and 3 female), Dp16 (n = 6, 3 male and 3 female) and Dp162xIfnrs (n = 5, 3 male and 2 female). Each mouse was given two practice sessions at 16 revolutions per minute (RPM), followed by two test sessions at 16, 24 and 32 RPM. Number of animals per group were as follows: rotarod WT (n = 16, 8 male and 8 female), Dp16 (n = 14, 7 male and 7 female) and Dp162xIfnrs (n = 17, 8 male and 9 female); MWM WT (n = 39, 19 male and 20 female), Dp16 (n = 28, 13 male and 15 female) and Dp162xIfnrs (n = 25, 12 male and 13 female); CFC WT (n = 33, 13 male and 20 female), Dp16 (n = 17, 8 male and 9 female) and Dp162xIfnrs (n = 23, 11 male and 12 female). Anesthesia delivery 4f and Extended Data Fig. As for other tissues, triplicated genes were clearly overexpressed in Dp16 and Dp162xIfnrs (Extended Data Fig. During both phases, freezing behavior was measured with FreezeScan v2.00 (Clever Sys). c, GSEA results with Hallmark gene sets for whole-blood gene expression correlations between HSA21- and non-HSA21-encoded genes in individuals with T21 only (n = 304). Using these parameters, we confirmed the presence of the deletion with no other coherent CNVs of similar size. © 5b). Sources supporting this summary overview is provided in Source Data Extended Data Fig. Fixative was washed-out in FACS buffer then cells were analyzed using a Cytek Aurora instrument. Trisomy 21 dysregulates T cell lineages toward an autoimmunity-prone state associated with interferon hyperactivity. Dp16 mice display key phenotypes of DS including hyperactive IFN signaling, a dysregulated antiviral response, increased prevalence of heart defects, developmental delays, cognitive impairments and craniofacial anomalies6,16,17,18,19,20,21. PCR products spanning the deleted region were subjected to Sanger sequencing using a 3730xl DNA Analyzer (Thermo Fisher Scientific) to identify transmission of a single modified allele (oligonucleotides in Supplementary Table 4). Hall, J. C. & Rosen, A. Salient examples include genes involved in synaptogenesis, SNAP receptor (SNARE) signaling and dopamine signaling. Virol. Given that all four Ifnrs employ JAK/STAT signaling, creating the potential for genetic redundancy, we designed a strategy to delete the 192 kb genomic segment encoding all four Ifnrs in wild-type (WT) mice (Methods, Fig. 83). BD Vacutainer® Venous Blood Collection Tube Guide BD Diagnostics Preanalytical Systems 1 Becton Drive Franklin Lakes, NJ 07417 USA * Invert gently, do not shake ** The performance characteristics of these tubes have not been established for infectious disease testing in general; therefore, users must, System, Blood, Tubes, Collection, Diagnostics, Venous, Vacutainer, Bd vacutainer venous blood collection tube, Bd vacutainer, 174 venous blood collection tube, Preanalytical, Bd diagnostics preanalytical systems. e, GSEA results with Hallmark gene sets for the adult mouse brain transcriptome comparison of Dp162xIfnrs to WT mice. Positions are indicated in base pairs (bp) for the GRCm38 assembly of the M. musculus genome. Dev. has provided consulting services for Eli Lilly Co. and Gilead Sciences Inc. and serves in the advisory board of Perha Pharmaceuticals. Ten micrograms of protein lysates were resolved on a 10% polyacrylamide gel and transferred to a PVDF membrane. 4f,g). The CFC20 test of associative learning and memory was performed in CFC boxes (30.5 × 24.1 × 29.2 cm) consisting of a light, recording device and a metal grid on the floor (Med Associates). GSEA was used to test for the enrichment of specific gene sets among upregulated or downregulated mRNAs or among positively or negatively correlated features. Source data are provided with this paper. Xie, C. & Tammi, M. T. CNV-seq, a new method to detect copy number variation using high-throughput sequencing. Dabei werden durch die standardisierten Entnahmebedingungen Fehlerquellen minimiert. Kelly D. Sullivan or Joaquin M. Espinosa. Psychol. Google Scholar. Biol. Natl Med. 213, 2527–2538 (2016). J. Primatol. The majority of BD Vacutainer® Blood Collection Tubes are made of clear latex-free Except for calvaria rounding, which was done on mice 7–10 weeks of age, all qualitative assessments of skulls from decapitated mice were done at 7–8 weeks of age by an expert in craniofacial morphology blinded to genotypes. Sample sizes are WT (n = 6, 2 male and 4 female), Dp16 (n = 5, 2 male and 3 female) and Dp162xIfnrs (n = 7, 4 male and 3 female). In alternative mouse models also harboring triplication of Ifnrs and displaying hyperactive IFN signaling20, developmental delays were rescued by prenatal treatment with the anti-inflammatory compound apigenin74. a, DESeq2 differential expression analysis results for whole-blood transcriptome, comparing individuals with T21 (n = 304, 163 male and 141 female) to euploid controls (D21, n = 96, 44 male and 52 female). 181, 41–46 (1996). Gel image represents example of genotyping PCR used to characterize >50 pups derived from modified zygotes. contributed to investigation and manuscript review. Cell Host Microbe 19, 181–193 (2016). Mice navigate to a hidden platform more efficiently over time (d). sharing sensitive information, make sure you’re on a federal Autoantibodies neutralizing type I IFNs are present in ~4% of uninfected individuals over 70 years old and account for ~20% of COVID-19 deaths. c, DESeq2 differential expression analysis results for adult mouse brain transcriptomes, comparing Dp162xIfnrs to Dp16 mice. Duchon, A. Nandi, S. S. & Mishra, P. K. Harnessing fetal and adult genetic reprograming for therapy of heart disease. Western blot analysis showed elevated phospho-STAT1 in the developing heart tissue of Dp16, but not in Dp162xIfnrs (Extended Data Fig. Dams were checked daily for vaginal plugs; the first morning of visual confirmation was denoted embryonic day (E)0.5. Rueda, N., Florez, J., Dierssen, M. & Martinez-Cue, C. Translational validity and implications of pharmacotherapies in preclinical models of Down syndrome. Vacutainer® Lithium-Heparin, hellgrün, mit Trenngel. e, Kaplan–Meier plot comparing survival across genotypes during chronic stimulation with the TLR3 agonist poly(I:C); significance determined by Mantel–Cox log-rank test. 6d). 5d–f). Ann. Paired-end 150 bp sequencing was carried out on an Illumina NovaSeq 6000 instrument. J. Exp. 364815 Haben Sie Fragen? Notably, correction of Ifnr locus dosage does not fully rescue transcriptome signatures of inflammation and immune dysregulation in Dp16 mice, indicating the presence of additional mechanisms contributing to immune dysregulation in DS. BD, das BD Logo und alle anderen Marken sind – sofern nicht anders angegeben – Eigentum der Becton, Dickinson und Company. Using the package CNV-seq v0.2-8, we confirmed the presence of Ifnr deletion in the F0 male WT1xIfnrs. Globin RNA depletion, poly-A+ RNA enrichment and strand-specific library preparation were carried out using the GLOBINclear Human Kit (Thermo Fisher Scientific), NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) and NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs). Die durchsichtige Hülse der Blutentnahmekanüle mit einer Drehbewegung abziehen. Broom: Convert Statistical Objects into Tidy Tibbles. PubMed  Consistently with this, Dp16 heart tissues show increased IFN-α and IFN-γ signaling concurrent with the elevation of diverse inflammatory pathways, elevated expression of genes involved in epithelial-mesenchymal transition (EMT) and decreased expression of genes associated with cell proliferation (for example, MYC targets and E2F targets; Fig. 6b). e, IFNR protein expression on immune subsets isolated as in (d) from CD45 + WBCs. Disabil. Nair Kesavachandran, C., Haamann, F. & Nienhaus, A. Pups were assessed in a pseudorandom order by blinded investigators in the following order: (1) surface righting 2 d in a row (D3–10); (2) first day both eyes open (D7–21); (3) first day both ears twitch (D7–21) and (4) the first day of auditory startle (D11–21). Transcriptome analysis of adult mouse mesenteric lymph nodes. Cardiac valve involvement in ADAR-related type I interferonopathy. Waugh, K. A., Cox, T., Sullivan, K. D. & Espinosa, J. M. Triplication of the Ifnr locus exacerbates craniofacial anomalies in a mouse model of Down syndrome. Cervical cancer screening 56, 675–688 (2004). All raw scan data were reconstructed to multiplanar slice data using NRecon v1.7.4.6 (Bruker). Love, M. I., Huber, W. & Anders, S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. To gain further insight, we completed transcriptome analysis of neural crest-derived facial mesenchyme at E10.5, an embryonic tissue relevant to craniofacial development. 3f). Multiple genes could contribute to a specific phenotype, and the aneuploidy itself could exert effects independent of gene content3,12. 111, 1420–1435 (2019). 12, 156–175 (2003). 35, 1473–1500 (2014). ), T32CA190216 (to K.A.W. Spezialsysteme für Molekulardiagnostik, Immunologie und Proteomics, Snowden-Pencer™ – Laparoskopische Instrumente, Snowden-Pencer™ – Instrumente für die plastische Chirurgie, Durchsuche alle Produkte für die Chirurgie, Durchsuche alle Produkte zur Infektionsprävention, Durchsuche alle Produkte für die Infusion, Integrierte Analyselösungen - Ressourcenbibliothek, Durchsuche alle Produkte für Interventionsverfahren, Technologien für niedergelassene Apotheken, Durchsuche alle Produkte für das Medikamentenmanagement, Sichere Handhabung gesundheitsgefährdender Medikamente, PhaSeal™ geschlossenes Arzneimitteltransfersystem, Sicherheitsspritzen und Sicherheitskanülen, Durchsuche alle Produkte für das Versorgungsmanagement, BD Direktabnahmeadaption für Blutkulturen, BD Eclipse™ Sicherheitskanüle zur Blutentnahme. Impaired local intrinsic immunity to SARS-CoV-2 infection in severe COVID-19. 10, 4766 (2019). Truett, G. E. et al. Health supervision for children and adolescents with Down syndrome. Nr. J. Phys. 1 and Extended Data Fig. J. Blood contamination during venepuncture and laboratory manipulations of specimen tubes. 3g and Extended Data Fig. Third, the phenotypic differences observed upon correction of Ifnr locus dosage are accompanied by partial attenuation of global gene expression changes, and it is unclear which of the pathways affected by Ifnr triplication contribute to the rescued phenotypes. Science 369, 706–712 (2020). Google Scholar. Peripheral blood was collected in PAXgene RNA Tubes (QIAGEN), and total RNA was purified using the PAXgene Blood RNA Kit (QIAGEN). 8600 Rockville Pike f, Frequency of complete ISS fusion (black) compared between cohorts by pairwise two-sided Fisher’s exact test with significance set at P < 0.05. Once criteria were reached, testing was stopped for that mouse. Heterozygote WT1xIfnr (Ifnr−/+) mutant mice were made in collaboration with Dr. Jennifer Matsuda and James Gross of the Genetics Core Facility at National Jewish Health, CO. DNA was genotyped by PCR34. Article  Für BD Bactec Blutkulturflaschen. 5c,d). 0:00 / 15:20 FamProp Wochenthema B1 - Blutabnahme mit dem Vacutainer™-System WUM - Wiener unabhängige Medizinstudierende 356 subscribers Subscribe 157 Share 24K views 5 years ago Dieses Video. The asterisks indicate q < 0.1 from either GSEA or Spearman correlations with permutation test and Benjamini–Hochberg correction. In the Ts16 mouse strain carrying triplication of essentially all MMU16 genes, including many genes not orthologous to HSA21, reduction of IFN signaling improved some aspects of Ts16 fetal development33. The Vacutainer system is particularly effective when multiple samples are required. Br J Nurs. In a-b, each dot represents an independent biological animal replicate with the mean indicated with a dash. Although our results do not demonstrate that Ifnr triplication is sufficient to cause CHD, they indicate that the Ifnr locus contributes to this trait, likely potentiating the effects of other necessary genes. Hygienisches Einwegprodukt. 57, 475–478 (2020). Neurosci. Assoc. Rev. Sung YH, Hwang MS, Lee JH, Park HD, Ryu KH, Cho MS, Yi YH, Song S. J Korean Acad Nurs. Learn. b, Concentration values (pg ml−1) for IL6 protein, as measured by Meso Scale Discovery assay, in plasma of individuals with T21 (n = 249). PLoS One 12, e0185056 (2017). Blood 127, 3154–3164 (2016). Demographics of human research cohort. Immunol. Bull, M. J. et al. sichtbar, z.T. a, Representative micro-computed tomography (μCT) reconstructions of a wild-type (WT) mouse skull. PCR products were purified and sequenced on an Illumina NovaSeq 6000 instrument. 5i). q-values determined by DESeq2 with significance set at q < 0.1 after Benjamini-Hochberg correction. 6i,j, Extended Data Fig. e, Scatter plots comparing mRNA fold-changes for Dp16 DEGs in mesenteric lymph nodes (top, sample sizes as in c) and brain (bottom, WT (n = 6, 2 male and 4 female), Dp16 (n = 5, 2 male and 3 female), and Dp162xIfnrs (n = 7, 4 male and 3 female)) for Dp16/WT and Dp162xIfnrs/WT, with Ifnrs highlighted in red, Dp16 triplicated genes in blue, nontriplicated Dp16 DEGs in gray, and slope (m) colored accordingly. Dabei durchsticht die Kanüle im Sicherheitsventil den Gummistopfen des Vacutainers. For all assays, mice were allowed to acclimate to the experimental room in their home cages for 30 min before testing began each day at 1200–1600 h. All experiments were performed by experimenters blinded to genotype, but Dp16 mice are somewhat smaller and have craniofacial differences, which may be noticeable to some personnel. Nat. We then employed genome editing to correct the dosage of the Ifnr locus in a mouse model of DS, which revealed that the Ifnr locus contributes to multiple key phenotypes in mice, with potential therapeutic implications for the management of this condition. Coordinates for 30 homologous landmarks were independently collected from each 3D-rendered skull with Drishti v2.6.5 by two investigators blinded to genotype. Written informed consent was obtained from participants who were cognitively able or their guardians. 107, 273–283 (1998). 1986 Sep;8(2):178-83. doi: 10.1016/0195-6701(86)90044-7. Am. Significance was determined by CNV-seq (*p < 0.1). There is a greater chance of trauma with a Vacutainer system. Examples include a shortening of the basisphenoid (BS) bone (distance between landmarks 24 and 27), increase in the sphenofrontal suture width (a proxy for intertemple width; distance between landmarks 21 and 22) and alterations in many mandibular interlandmark distances (for example, landmarks 18–20), all of which are observed in Dp16 but attenuated in Dp162xIfnrs (Fig. J. Neurosci. Hazardous drug safety PubMed Central  Article  Upon quick freeze/thaw at 37 °C, mesenteric lymph nodes were placed in Lysing Matrix D tubes also containing RLT with 2-ME. Three copies of four interferon receptor genes underlie a mild type I interferonopathy in Down syndrome. Number of animals as in b. P values determined by one-way ANOVA with Tukey’s HSD test, with significance set at P < 0.05; horizontal dashes indicate means. These subtle yet significant differences by genotype are in line with previous studies using MWM to investigate Dp16 deficits in memory extinction and relearning16,18,19,20. Patient monitoring and temperature management a, Top—diagram indicating genomic locations of the mouse Ifnr gene cluster on MMU16 and gRNAs (orange arrowheads) employed for genome editing using CRIPSR–Cas9 technology. Hinweis: In Ihrer Region sind möglicherweise nicht alle Produkte, Dienstleistungen oder Funktionen von Produkten und Dienstleistungen verfügbar. Genet 107, 911–931 (2020). Type III interferons disrupt the lung epithelial barrier upon viral recognition. Behav. Data are represented as mean population estimates (black dots) with 95% confidence intervals (lines) colored according to differences versus WT where green represents larger inter-landmark distance, blue represents smaller inter-landmark distance, and gray represents confidence intervals that intersect 1 (that is, no difference). See Source Data and Extended Data Fig. Biol. Das BD Vacutainer® Blutentnahmesystem ist mit allen anderen Blutentnahmesystemen kompatibel. Wound care, Support Type Spearman correlations were defined for HSA21-encoded mRNAs versus all other mRNAs using only trisomy 21 samples and ρ values as the GSEA ranking metric; middle—Spearman correlations between mRNAs encoded on HSA21 versus mRNAs for indicated DEGs encoded elsewhere in the genome among individuals with DS; bottom—Spearman correlations between mRNAs encoded on HSA21 and plasma levels of CRP and IL6 in individuals with DS. In all cases, to adjust for multiple hypothesis testing, q values were calculated from P values using the BH approach. Mouse model of SARS-CoV-2 reveals inflammatory role of type I interferon signaling. Bitte auswählen Res. An algorithm for fast preranked gene set enrichment analysis using cumulative statistic calculation. J. Orthod. The 3–5 mesenteric lymph nodes per animal were flash-frozen together while brain and adult heart samples were first placed in Lysing Matrix D tubes (MP Biomedicals) containing 594 μl of lysis buffer RLT Plus (QIAGEN) and 6 μl of 2-mercaptoethanol (Sigma-Aldrich) before storage at −80 °C. We used IPA (release winter 2022), which employs right-tailed Fisher’s exact tests to define the enrichment of a particular gene set within a gene list, to investigate pathways enriched among DEGs in the adult brain transcriptome study. Maxillary and mandibular processes were manually dissected from the face. Powers, R. K. et al. a, Manhattan plots of human chromosomes 3 and 21 (HSA3 and HSA21) displaying results of whole-blood transcriptome analysis for individuals with trisomy 21 (T21, n = 304, 163 male and 141 female) versus euploid controls (D21, n = 96, 44 male and 52 female). Halter für BD Vacutainer Kanülen und Blutentnahmesets. 5h). Sci. Brain Res. Ann. D21: euploid controls (n = 96, 44 male and 52 female). Despite many research efforts, the mechanisms driving these hallmarks of DS are largely unknown. Get the most important science stories of the day, free in your inbox. 36, 60–90 (2001).