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A study conducted in Saint Louis, Missouri analyzed a sample of illicit drug specimens and paraphernalia voluntarily submitted to a syringe service program. The study used gas chromatography-mass spectrometry and liquid chromatography quadrupole time-of-flight mass spectrometry to identify and analyze the substances present. The results showed that fentanyl was the most frequent substance detected, followed by cocaine, metamfetamine, and ketamine. The study also found that many specimens contained complex combinations of multiple drugs. These findings suggest changing patterns of adulteration in illicit substances. [Extracted from the article]

The article focuses on a research which investigates role of digital quantitative real-time polymerase chain reaction (RQ-PCR) assays to quantify rare nucleophosmin (NPM1) mutations (NPM1mut). Topics discussed include mRNA isolation from mononuclear cells, test applicability for minimal residual disease (MRD) monitoring and sensitivity and specificity of the new digital PCR assays.

Discusses a study which reported that G protein-couple receptor kinases (GRK) can inhibit receptor signaling under nonphosphorylating conditions. Materials and methods of the study; Results; Discussion of the results.

Presents a letter to the editor replying to the article "Prognostic Significance of a Short Sequence Insertion of the MCL-1 Promoter in Chronic Lymphocytic Leukemia," published in the 2004 issue of the "Journal of the National Cancer Institute."

14q-deletions have been repeatedly described in mature B-cell neoplasms, but not yet characterized in a larger cohort. Based on chromosome banding analysis, the present study identified 47 del(14q) cases in 3054 mature B-cell neoplasms (1·5%) (chronic lymphocytic leukaemia [CLL]: 1·9%; CLL/prolymphocytic leukaemia [PL]: 9·0%; others: 0·2%). Interphase fluorescence in situ hybridization was performed with probes for 14q22.1, 14q24.1, 14q32.33, and IGH@ (14q32.3). The del(14q) had heteregeneous size but showed a breakpoint cluster at the centromeric site in 14q24.1 (62% of cases). At the telomeric side, the most frequent breakpoint was within the IGH@ locus (14q32.3) between IGH@ 3′-flanking and IGHV ( IgVH) probes (45%). In 16 cases (34%), breakpoints occurred within 14q24.1 and 14q32.3. Eighty-one percent of del(14q) cases showed 1–3 additional cytogenetic alterations (in 45%, +12), and 56% were IGHV-unmutated. In all cases (16/16) with breakpoints in 14q24.1 and 14q32.3, a B-CLL immunophenotype was found. Clinical follow-up in 32 del(14q) patients was compared to 383 CLL and CLL/PL patients without del(14q). While 3-year-overall survival did not differ significantly, time to treatment was significantly shorter in the del(14q) cohort (21·0 months vs. 80·1 months, P = 0·015). In conclusion, the del(14q) is a rare recurrent alteration in diverse mature B-cell neoplasms, shows variable size but distinct clustering of breakpoints, and is associated with short time to treatment. [ABSTRACT FROM AUTHOR]

External quality assurance (EQA) programs are vital to ensure high quality and standardized results in molecular diagnostics. It is important that EQA for quantitative analysis takes into account the variation in methodology. Results cannot be expected to be more accurate than limits of the technology used, and it is essential to recognize factors causing substantial outlier results. The present study aimed to identify parameters of specific importance for JAK2 V617F quantification by quantitative PCR, using different starting materials, assays, and technical platforms. Sixteen samples were issued to participating laboratories in two EQA rounds. In the first round, 19 laboratories from 11 European countries analyzing JAK2 V617F as part of their routine diagnostics returned results from in-house assays. In the second round, 25 laboratories from 17 countries participated. Despite variations in starting material, assay set-up and instrumentation the laboratories were generally well aligned in the EQA program. However, EQA based on a single technology appears to be a valuable tool to achieve standardization of the quantification of JAK2 V617F allelic burden. [ABSTRACT FROM AUTHOR]

The article presents a study on role of negative prognostic markers in prognosis of chronic lymphocytic leukemia (CLL). It states that the study analyzed a group of CLL patients without prior treatments to determine the affect of prognostic markers in the disease. The authors mentions correlation between immunoglobulin heavy (IGHV) mutational status and biochemical markers. They state that several markers such as SF3B1mut were found worsening the course of prognosis in CLL patients.

Multiparameter flow cytometry ( MFC) identifies rare cases of biclonal disease in chronic lymphocytic leukaemia ( CLL). By MFC, we identified 76 patients with biclonal disease in a cohort of 5523 CLL patients (1·4%). Fluorescence in situ hybridization and chromosome banding analysis revealed five and six cases, respectively, with two different cytogenetic aberrations due to clonal evolution. Two different B-cell receptor rearrangements and IGHV subtypes were more frequent in biclonal than in monoclonal CLL by MFC (37·1% vs. 2·7%; P < 0·001). Patients with biclonal CLL by MFC showed a trend to a shorter time to treatment than monoclonal CLL ( P = 0·080). [ABSTRACT FROM AUTHOR]

We analyzed the mutational hotspot region of SRSF2 (Pro95) in 275 cases with chronic myelomonocytic leukemia (CMML). In addition, ASXL1, CBL, EZH2, JAK2V617F, KRAS, NRAS, RUNX1, and TET2 mutations were investigated in sub-cohorts. Mutations in SRSF2 (SRSF2mut) were detected in 47% (129 of 275) of all cases. In detail, 120 cases had a missense mutation at Pro95, leading to a change to Pro95His, Pro95Leu, Pro95Arg, Pro95Ala, or Pro95Thr. In 9 cases, 3 new in/del mutations were observed: 7 cases with a 24-bp deletion, 1 case with a 3-bp duplication, and 1 case with a 24-bp duplication. In silico analyses predicted a damaging character for the protein structure of SRSF2 for all mutations. SRSF2mut was correlated with higher age, less pronounced anemia, and normal karyotype. SflSF2mut and EZH2mut were mutually exclusive, but SRSF2mut was associated with 7ET2mut. In the total cohort, no effect of SRSF2mut on survival was observed. However, in the RUNXIvnut subcohort, SRSF2 Pro95His had a favorable effect on overall survival. This comprehensive mutation analysis found that 93% of all patients with CMML carried at least 1 somatic mutation in 9 recurrently mutated genes. In conclusion, these data show the importance of SRSF2mut as new diagnostic marker in CMML. [ABSTRACT FROM AUTHOR]

The karyotype is so far the most impor-tant prognostic parameter in acute my-eloid leukemia (AML). Molecular muta-tions have been analyzed to subdivide AML with normal karyotype into prognos-tic subsets. The aim of this study was to develop a prognostic model for the entire AML cohort solely based on molecular markers. One thousand patients with cy-togenetic data were investigated for the following molecular alterations: PML-RARA, RUNX1-RUNX1T1, CBFB-MYH11,FLT3-ITD, and MLL-PTD, as well as muta-tions in NPM1, CEPBA, RUNX1, ASXL1, and TP53. Clinical data were available in 841 patients. Based on Cox regression and Kaplan-Meier analyses, 5 distinct prognostic subgroups were identified: (1) very favorable: PML-RARA rearrange-ment (n = 29) or CEPBA double muta-tions (n = 42; overall survival [OS] at 3 years: 82.9%); (2) favorable: RUNX1-RUNX1T1 (n = 35), CBFB-MYH11 (n = 31), or NPM1 mutation without FLT3-HD(n = 186; OS at 3 years: 62.6%); (3) inter-mediate: none of the mutations leading to assignment into groups 1, 2, 4, or 5 (n = 235; OS at 3 years: 44.2%); (4) unfa-vorable: MLL-PTD and/or RUNX1 muta-tion and/or ASXL1 mutation (n = 203; OS at 3 years: 21.9%); and (5) very unfavor-able: TP53 mutation (n = 80; OS at 3 years: 0%; P < .001). This comprehensive molecu-lar characterization provides a more power-ful model for prognostication than cytoge-netics. [ABSTRACT FROM AUTHOR]

Summary The World Health Organization classification uses a cut-off point of 5·0 × 109/l cells with a chronic lymphocytic leukaemia (CLL)-phenotype in peripheral blood to discriminate between monoclonal B-lymphocytosis (MBL) and B-CLL. This study analysed 298 MBL patients by multi-parameter flow cytometry, chromosome banding analysis (CBA)/fluorescence in situ hybridization (FISH), and IGHV mutation status and compared them with 356 CLL patients. In MBL, CBA more frequently revealed a normal karyotype and FISH identified less frequently del(6q), del(13q) (as sole alterations), and del(17)(p13). Within the MBL cohort, a shorter time to treatment (TTT) was found for ZAP-70-positivity, 14q32/ IGH-translocations (CBA), del(11)(q22·3) (FISH) and unmutated IGHV status. Higher CD38 and ZAP-70 expression, del(11)(q22·3) (FISH), trisomy 12 (FISH), and 14q32/ IGH-translocations (CBA) were correlated with a shorter TTT in the combined cohort (MBL + CLL); a sole del(13)(q14) (FISH) correlated with longer TTT. Regarding overall survival, unmutated IGHV status and 'other' alterations (CBA) had an adverse impact. There was no correlation between the concentration of CLL-cells and TTT or overall survival. Multivariate analysis confirmed a negative impact on TTT for del(11)(q22·3)/ ATM, trisomy 12 (both by FISH), and 14q32/ IGH-translocations by CBA. These data emphasize a close relationship between MBL and CLL regarding clinically relevant parameters and provide no evidence to strictly separate these entities by a distinct threshold of clonal B-cells. [ABSTRACT FROM AUTHOR]

The BRAFM600E mutation was recently detected in hairy cell leukemia (HCL) by whole exome sequencing. To make use of this new marker for diagnosis and follow-up of HCL, we developed a SRAFVeoOEmut-specific quantitative real-time PCR assay and validated it in 117 HCL patients and 102 non-HCUBRAFwt patients. The cut-off level to discriminate SR/\FV600E-positive/-negative cases was set at 0.023% BRAFV600EI BRAFwt. A total of 115 of 117 HCL (98.3%) demonstrated percentage BRAFV600EI BRAFvji above the cut-off (mean, 29.6 ±41.1). The remaining 2 of 117 HCL with lower percentage BRARJGOOE/BRAFvjt ratios were also BRAFwt by deep-sequencing technology. Sixteen HCL-variant patients showed percentage BRAFVSOOE/BRAFvrt. values corresponding to "non-HCL." Follow-up studies in 19 HCL cases demonstrated a decrease of percentage BRAFVSOOE/BRAFvjt during therapy. The log-reductions as determined by RT-PCR and immunophenotyping correlated significantly (P < .001). In conclusion, we confirmed BRAFmut as a useful marker in HCL, its absence in HCL variant, and developed an RT-PCR-based assay to monitor minimal residual disease in HCL. [ABSTRACT FROM AUTHOR]

Abstract: A new myeloid leukemia cell line (CG-SH) with normal cytogenetics was established from a patient with acute myelogenous leukemia (AML) following myelodysplastic syndrome (MDS). The cells of CG-SH are immature blasts and have an immature myeloid phenotype (positive for myeloperoxidase, CD7, CD34, CD38, CD117, HLA-DR, negative for CD10, CD19, CD20, CD41, CD42). A partial expression of CD13, CD15, CD65 and a weak expression of CD33 and CD133 was noted. The cells are negative for EBER. By molecular analysis, a mutation of NRAS and heterozygous mutations of RUNX1 were detected. No mutations were detected in FLT3-ITD, MLL-PTD or NPM1. By real-time PCR, a series of 19 microRNAs was identified which are strongly expressed in CG-SH. In conclusion, a new cell line was established which will be useful for the study of AML with normal cytogenetics and mutations in NRAS and/or RUNX1. [Copyright &y& Elsevier]

Abstract: Chromosomal aberrations detected by fluorescence in situ hybridization (FISH) on interphase nuclei are important prognostic markers in chronic lymphocytic leukemia (CLL). Deletions in 13q14 and in 11q22.3 are two of the most frequent aberrations in this disease entity (55 and 18%, respectively) and are usually effected by interstitial deletions. Here, we report on the case of a 66-year-old woman with CLL who was analyzed by conventional metaphase cytogenetics as well as fluorescence in situ hybridization. Deletion-specific probes detected a homozygous loss of two anonymous loci in chromosomal band 13q14 in parallel with a heterozygous loss of the ATM gene located in chromosomal band 11q22.3. Karyotype analysis indicated reciprocal but unbalanced translocations involving chromosomes 3, 11, and 13. Deleted sites on 13q14 appeared to be located within the breakpoint regions of the translocations. We show that mechanisms other than interstitial deletions may lead to loss of critical chromosomal regions in CLL. [Copyright &y& Elsevier]