Műszerügyi és Méréstechnikai Közlemények

UNIDO Workshop a Műszerügyi és Méréstechnikai Szolgálatnál Újszerű lehetőségek a Kutatófilm és Videotechnikai Főosztályon Műszerkölcsönzés Császár László: Üzemeltetési és szerviztapasztalataink (3.) A GOULD gyártmányú digitális oszcilloszkópok Új irányok a műszer- és méréstechnikában Radnai Rudolf: Gyakorlati tanácsok számítógépes mérőrendszerek üzembehelyezéséhez és üzemeltetéséhez Kőfalvi Jenő: Mikrovezetékes analitika az integrált áramkörök mintájára Szaktanácsadás Kőfalvi Jenő: Válogatás az Országos Műszernyilvántartás nagyértékű műszerújdonságaiból Külföldi műszerújdonságok. Összeállította: Csont Tamás - Fekete Gábor - Kőfalvi Jenő Könyvismertetés. Összeállította: Radnai Rudolf - Kőfalvi Jenő Műszerkölcsönzés Görgényi László: A kölcsönműszerpark szaporulata Szolgálatunk életébő

<i>BTG1</i> microdeletion occurrence within cytogenetic subgroups of ALL.

<p><i>BTG1</i> deletion status was determined using MLPA.</p>a<p>Because of missing values, numbers do not always add up to 722 BCP-ALL cases. Data was available for 637 cases on hyperdiploidy; 513 cases for <i>ETV6-RUNX1</i>; 648 cases for <i>BCR-ABL1;</i> 649 cases for <i>MLL</i>-rearranged.</p>b<p>The ‘other’ subgroup encompasses cases negative for <i>ETV6-RUNX1</i> or <i>BCR-ABL1</i> translocations, <i>MLL</i>-rearrangement and/or hyperdiploidy. This group includes 10 cases with <i>E2A-PBX1</i> translocation, of which none harbor a <i>BTG1</i> deletion.</p>c<p>Subgroup unknown includes all cases in which no data is available in one or more cytogenetic classifications.</p>d<p>Fisher's exact test was used when sample groups were small.</p

Expression of <i>BTG1</i> truncated read-through transcripts in BCP-ALL cells with <i>BTG1</i> deletions.

<p>(A) Schematic representation of the wild-type human <i>BTG1</i> gene, existing of two partly coding exons, and five different <i>BTG1</i> transcripts due to <i>BTG1</i> gene deletions. Exons are represented by black (coding) or white (non-coding) bars. Indicated are the RT-PCR primers that were used to detect expression of the wild-type <i>BTG1</i> transcript (primers A and B), or one of the <i>BTG1</i> truncated read-through transcripts for deletion II (primers A and C), deletion III (pimers A and D), deletion IV (primers A and E), deletion V (primers A and F), or deletion VIII (primers A and G). (B) RT-PCR analyses on total RNA isolated from the BCP-ALL cell lines Nalm6 and RS4;11 (<i>BTG1</i> wild-type) and REH, SUP-B15 and 380, each with distinct monoallelic <i>BTG1</i> deletions. (C) RT-PCR analyses on primary BCP-ALL samples in which a single <i>BTG1</i> deletion (Pt1, Pt2, Pt3 and Pt5), multiple <i>BTG1</i> deletions (Pt4 and Pt6) or no <i>BTG1</i> deletions were detected with genomic PCR (Pt7). Type of deletions (III, V, or VIII) and outcome of MLPA (p: deletion-positive; n: deletion-negative) are indicated. <i>BTG1</i> read-through transcripts were verified by sequencing (<a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002533#pgen.1002533.s007" target="_blank">Table S5</a>), except for Pt3-deletion V, which was an unrelated DNA sequence. (D) Quantitative real-time RT-PCR data representing relative expression levels of <i>BTG1</i> measured 5′ (primers exon 1/2) and 3′ of the <i>BTG1</i> breakpoint hotspot (primers exon 2). Expression levels were normalized to <i>HPRT</i> levels, and compared to the expression level in Nalm6, which was set to 1. The data shown represent the average of two independent cDNA reactions and triplicate qRT-PCR reactions.</p

Multiple <i>BTG1</i> deletion-positive clones are present in specific BCP-ALL subtypes.

<p>(A) Recurrence of multiclonal <i>BTG1</i> deletions. A sensitive PCR method was used to screen for eight different deletion breakpoints (deletion I–VIII) in <i>BTG1</i> MLPA deletion positive (+) cases (n = 65), and to screen for the three most frequent deletion breakpoints (deletion III, V and VIII) in <i>BTG1</i> MLPA deletion negative (−) cases (n = 89). (B) <i>BTG1</i> deletion frequency in the two major cytogenetic subgroups of BCP-ALL (Hyperdiploid and <i>ETV6</i>-<i>RUNX1</i>). Presence of a <i>BTG1</i> deletion in the predominant clone was determined by MLPA on the entire cohort of BCP-ALL cases (n = 722), and was compared to deletions detected as a minor clone in MLPA-negative cases (n = 89) by deletion-spanning PCR. Distributions are similar, being depleted from hyperdiploid cases and enriched in <i>ETV6</i>-<i>RUNX1</i>-positive cases as compared to the total group.</p

Increased levels of H3K4me3 at the <i>BTG1</i> locus in BCP-ALL versus T-ALL cell lines.

<p>(A) Quantitative real-time RT-PCR data representing relative expression levels of <i>BTG1</i> in T-ALL cell lines HSB2, Jurkat and KARPAS45, and BCP-ALL cell lines RS4;11, Nalm6 and CCRF-SB (<i>HPRT</i> normalized and related to HSB2 expression levels). Data shown are the average of two independent cDNA reactions and triplicate qRT-PCR reactions. (B and C) Percentage recovery after ChIP performed with H3K4me3 antibody (B) or H3K9/14Ac antibody (C) on T-ALL (HSB2, Jurkat, KARPAS45) and BCP-ALL (RS4;11, Nalm6 and CCRF-SB) cell lines. Real-time quantitative PCR was performed with primers specific for the region 1 kb upstream of the transcription start site (−1 kb prom), directly flanking the transcription start site (prox prom), the second exon near the breakpoint hotspot (exon 2) and towards the end of the 3′untranslated region (3′UTR) at the second (and last) exon of the human <i>BTG1</i> gene. Values represent two independent ChIP experiments. Student's <i>t</i>-test was performed to assess differences between the average recovery of T-ALL versus BCP-ALL samples. Asterisk (*) indicates a p-value<0.05.</p

<i>BTG1</i> deletions in relapsed cases.

a<p>Determined by sequence analysis of breakpoint spanning PCR product.</p>b<p>The deletion breakpoint could not be detected (N.D.) using the eight breakpoint-spanning PCR assays (I–VIII).</p>c<p>The breakpoint does not cluster within <i>BTG1</i> exon 2, but is located 2 kb downstream in the 3′UTR.</p>d<p>Homozygous deletion.</p

Authentic RSSs and candidate RSSs flanking breakpoints of <i>BTG1</i> microdeletions.

<p>Mismatches from consensus are underlined;</p>a<p>RSSs flanking V(D)J gene segments;</p>b<p>Sequence shown is in reverse complement orientation;</p>c<p>Functional cryptic RSSs at proto-oncogene breakpoints <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1002533#pgen.1002533-Marculescu1" target="_blank">[18]</a>.</p

Activating IL7R signaling mutations can confer resistance to steroid treatment.

<p>(A–F) Steroid response curves (from triplicate experiments ± standard deviation) for SUPT1 cells that express (A) JAK1, (B) JAK1^R724H, (C) JAK1^T901A, (D) JAK3, (E) JAK3^M511I, or (F) JAK3^R657Q from doxycycline-inducible lentiviral expression constructs. Steroid response curves are shown for induced (+Dox) and non-induced (−Dox) cells that have been exposed to serial dilutions of prednisolone for 72 h. (G–I) Mean survival of SUPT1 cells (triplicate experiments ± standard deviation) expressing wild-type or mutant IL7R signaling molecules (+Dox: open red bars) following a 72-h exposure to (G) prednisolone (Pred), (H) vincristine (VCR), or (I) <i>L</i>-asparaginase (ASP). Black bars represent the mean survival of all SUPT1 lines under non-induced conditions following exposure to prednisolone, vincristine, or <i>L</i>-asparaginase (−Dox control). The steroid-sensitive panel refers to SUPT1 lines that retain a similarly sensitive steroid response following expression of IL7R, JAK1, JAK3, JAK3^M511I, JAK3^R657Q, or AKT^E17K compared to non-induced control conditions. The steroid-resistant panel refers to lines that acquire steroid resistance following expression of IL7R^RFCPH, JAK1^R724H, JAK1^T901A, NRAS, NRAS^G12D, or AKT. See also <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1002200#pmed.1002200.s005" target="_blank">S5 Fig</a>.</p

MEK and PI3K/AKT pathway inhibitors enhance the steroid response of primary T-ALL patient samples.

<p>MEK and PI3K/AKT pathway inhibitors enhance the steroid response of primary T-ALL patient samples.</p

Mutations/aberrations affecting the IL7R signaling pathway in pediatric T-ALL patients at diagnosis predict diminished steroid response and poor outcome.

<p>Mutations in (A and B) <i>JAK1</i> or (C) <i>KRAS</i> detected by TES in diagnostic samples from 69 T-ALL patients are associated with diminished steroid response and/or poor survival. IL7R signaling mutations in diagnostic samples from 146 T-ALL patients are associated with reduced (D) in vitro steroid sensitivity and (E) relapse-free survival. Patients harboring <i>NR3C1</i> deletion as a consequence of a chromosomal 5q deletion were excluded from these analyses. See also <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1002200#pmed.1002200.s015" target="_blank">S6 Table</a>. ETP-ALL, early thymic progenitor acute lymphoblastic leukemia; T-ALL, T cell acute lymphoblastic leukemia; TES, targeted exome sequencing.</p