Serial quantitation of BCR-ABL IntroductionReverse-transcription real-time quantitative polymerase chain reaction (RQ-PCR) is used routinely to quantify levels of BCR-ABL mRNA in peripheral blood and bone marrow samples from chronic myelogenous leukemia (CML) patients undergoing therapy. The technique can accurately determine response to treatment and is particularly valuable for patients who have achieved a complete cytogenetic response. The National Comprehensive Cancer Network (NCCN) 1 and the European LeukemiaNet (ELN) 2 recommend similar monitoring schedules for patients treated with imatinib and the ELN defines an optimal response as the attainment of a major molecular response (MMR) after 18 months of therapy. Monitoring of BCR-ABL mRNA levels is also useful for gauging therapeutic response for patients with Philadelphia chromosomepositive acute lymphoblastic leukemia (Ph ϩ ALL). The CML meeting at the National Institutes of Health in Bethesda in October 2005 made several recommendations for the harmonization of minimal residual disease (MRD) assessment and proposed an international scale (IS) for BCR-ABL RQ-PCR measurements. 8 Importantly, the IS is essentially identical to that used in the International Randomized Study of Interferon and STI571 (IRIS) study, 9 with the IRIS standardized baseline defined as 100% BCR-ABL IS and MMR (3-log reduction relative to the standardized baseline) defined as 0.1% BCR-ABL IS . The original standards used for the IRIS trial are no longer available, however traceability to the IRIS scale is provided by the extensive quality control data generated by the Adelaide laboratory over a period of several years. 10,11 To enable testing centers to gain access to the IS, the Adelaide laboratory initiated a process to develop and validate laboratoryspecific conversion factors (CFs) that can be used to convert local values to IS values. 11 The strength of this approach is that testing centers can continue to use their existing assay conditions and continue to express results according to local preferences in addition to expressing results on the IS. The concept of the IS is analogous to established procedures for other quantitative assays, for example the International Normalized Ratio (INR) for prothrombin time. 12 Many laboratories with validated CFs have established themselves as national or regional reference laboratories and are in the process of propagating CFs to local centers. 13 While this process has generally worked well, it is apparent that the establishment of CFs is time-consuming, complex, expensive, and open to only a limited number of laboratories at any given time. Furthermore, it is unclear how frequently any individual CF will need to be revalidated. We sought therefore to develop an alternative means for testing laboratories to access the IS by developing calibrated, accredited primary reference reagents for BCR-ABL RQ-PCR analysis. StrategyIdeally, the formulation for primary reference reagents should be as close as possible to the usual analyte, should cove...

BackgroundHypoxia-inducible factor 1 (HIF-1) is a master transcriptional regulator of genes regulating oxygen homeostasis. The HIF-1 protein is composed of two HIF-1α and HIF-1β/aryl hydrocarbon receptor nuclear translocator (ARNT) subunits. The prognostic relevance of HIF-1α protein overexpression has been shown in breast cancer. The impact of HIF-1α alternative splice variant expression on breast cancer prognosis in terms of metastasis risk is not well known.MethodsUsing real-time quantitative reverse transcription PCR assays, we measured mRNA concentrations of total HIF-1α and 4 variants in breast tissue specimens in a series of 29 normal tissues or benign lesions (normal/benign) and 53 primary carcinomas. In breast cancers HIF-1α splice variant levels were compared to clinicopathological parameters including tumour microvessel density and metastasis-free survival.ResultsHIF-1α isoforms containing a three base pairs TAG insertion between exon 1 and exon 2 (designated HIF-1αTAG) and HIF-1α736 mRNAs were found expressed at higher levels in oestrogen receptor (OR)-negative carcinomas compared to normal/benign tissues (P = 0.009 and P = 0.004 respectively). In breast carcinoma specimens, lymph node status was significantly associated with HIF-1αTAG mRNA levels (P = 0.037). Significant statistical association was found between tumour grade and HIF-1αTAG (P = 0.048), and total HIF-1α (P = 0.048) mRNA levels. HIF-1αTAG mRNA levels were also inversely correlated with both oestrogen and progesterone receptor status (P = 0.005 and P = 0.033 respectively). Univariate analysis showed that high HIF-1αTAG mRNA levels correlated with shortened metastasis free survival (P = 0.01).ConclusionsOur results show for the first time that mRNA expression of a HIF-1αTAG splice variant reflects a stage of breast cancer progression and is associated with a worse prognosis.See commentary: http://www.biomedcentral.com/1741-7015/8/45

IntroductionHypoxia-inducible factor-1 (HIF1) controls the expression of genes involved in the cellular response to hypoxia. No information is available on its expression in critically ill patients. Thus, we designed the first clinical study in order to evaluate the role of HIF1α as a prognosis marker in patients suffering from shock.MethodsFifty consecutive adult patients with shock and 11 healthy volunteers were prospectively enrolled in the study. RNA was extracted from whole blood samples and expression of HIF1α was assessed over the first four hours of shock. The primary objective was to assess HIF1α as a prognostic marker in shock. Secondary objectives were to evaluate the role of HIF1α as a diagnostic and follow-up marker. Patient survival was evaluated at day 28.ResultsThe causes of shock were sepsis (78%), hemorrhage (18%), and cardiac dysfunction (4%). HIF1α expression was significantly higher in the shock patients than in the healthy volunteers (121 (range: 72-168) versus 48 (range: 38-54) normalized copies, P <0.01), whatever the measured isoforms. It was similar in non-survivors and survivors (108 (range 84-183) versus 121(range 72-185) normalized copies, P = 0.92), and did not significantly change within the study period.ConclusionsThe present study is the first to demonstrate an increased expression of HIF1α in patients with shock. Further studies are needed to clarify the potential association with outcome. Our findings reinforce the value of monitoring plasma lactate levels to guide the treatment of shock.