A novel assay to detect calreticulin mutations in myeloproliferative neoplasms.

The myeloproliferative neoplasms are chronic myeloid cancers divided in Philadelphia positive (Ph+), chronic myeloid leukemia, or negative: polycythemia vera (PV) essential thrombocythemia (ET), and primary myelofibrosis (PMF). Most Ph negative cases have an activating JAK2 or MPL mutation. Recently, somatic mutations in the calreticulin gene (CALR) were detected in 56-88% of JAK2/MPL-negative patients affected by ET or PMF. The most frequent mutations in CARL gene are type-1 and 2. Currently, CALR mutations are evaluated by sanger sequencing. The evaluation of CARL mutations increases the diagnostic accuracy in patients without other molecular markers and could represent a new therapeutic target for molecular drugs.We developed a novel detection assay in order to identify type-1 and 2 CALR mutations by PNA directed PCR clamping. Seventy-five patients affected by myeloproliferative neoplasms and seven controls were examined by direct DNA sequencing and by PNA directed PCR clamping. The assay resulted to be more sensitive, specific and cheaper than sanger sequencing and it could be applied even in laboratory not equipped for more sophisticated analysis. Interestingly, we report here a case carrying both type 1 and type2 mutations in CALR gene.

Variable but consistent pattern of Meningioma 1 gene (MN1) expression in different genetic subsets of acute myelogenous leukaemia and its potential use as a marker for minimal residual disease detection.

Meningioma 1 (MN1) gene overexpression has been reported in acute myeloid leukaemia (AML) patients and identified as a negative prognostic factor. In order to characterize patients presenting gene overexpression and to verify if MN1 transcript could be a useful marker for minimal residual disease detection, MN1 was quantified in 136 AML patients with different cytogenetic risk and in 50 normal controls. In 20 patients bearing a fusion gene transcript suitable for minimal residual disease quantitative assessment and in 8 patients with NPM1 mutation, we performed a simultaneous analysis of MN1 and the fusion-gene transcript or NPM1 mutation during follow-up. Sequential MN1 and WT1 analysis was also performed in 13 AML patients lacking other molecular markers. The data obtained show that normal cells consistently express low levels of MN1 transcript. In contrast, high levels of MN1 expression are present in 47% of patients with normal karyotype and in all cases with inv(16). MN1 levels during follow-up were found to follow the pattern of other molecular markers (fusion gene transcripts, NPM1 and WT1). Increased MN1 expression in the BM during follow up was always found to be predictive of an impending hematological relapse.

The Wilms' tumor (WT1) gene expression correlates with the International Prognostic Scoring System (IPSS) score in patients with myelofibrosis and it is a marker of response to therapy.

The Wilms tumor gene WT1 is a useful marker of clonal hematopoiesis and it has been shown to be a good marker of residual disease and it reflects the response to therapy. Although myelofibrosis is characterized by mutations of JAK2 and calreticulin (CALR), these mutations are not useful to monitor response to therapy. In this study we demonstrated that in patients affected by myelofibrosis WT1 correlates with the International Prognostic Scoring System (IPSS) score at diagnosis. Furthermore WT1 is a good marker of response to JAK2 inhibitors especially for patients without blasts and for patients who develop anemia or thrombocytopenia not for progression but as therapy related toxicity. Finally, WT1 transcript reduction can mirror a benefit of therapy on the disease burden. This study demonstrated that WT1 is a good marker for monitoring the response to therapy in patients affected by myelofibrosis.

Effectiveness of interprofessional education by on-field training for medical students, with a pre-post design.

BACKGROUND: Interprofessional Education (IPE) implies how to achieve successful teamwork, and is based on collaborative practice which enhance occasions for relationships between two or more healthcare professions. This study evaluates the effectiveness of IPE in changing attitudes after a training recently introduced to medical education for second-year students at the University of Padova, Italy. METHODS: All medical students following a new program for IPE were enrolled in this study. The Interdisciplinary Education Perception Scale (IEPS) was administered before and after training, according to observation-based and practice-based learning. Data were analysed with Student's paired t-test and Wilcoxon's signed rank test. RESULTS: 277 medical students completed both questionnaires. Statistically significant improvements were found in students' overall attitudes as measured by the IEPS and four subscale scores. Gender-stratified analyses showed that improvements were observed only in female students in subscale 4 ("Understanding Others' Values"). Students who had a physician and/or health worker in their family did not show any improvement in subscales 2 ("Perceived need for cooperation") or 4 ("Understanding Others' Values"). CONCLUSIONS: Our results indicate that IPE training has a positive influence on students' understanding of collaboration and better attitudes in interprofessional teamwork. More research is needed to explore other factors which may influence specific perceptions among medical students.

Trends in RCT nursing research over 20 years: mind the gap.

Randomised controlled trial (RCT) literature plays a fundamental role in informing evidence-based medicine and nursing. This paper aims to track geographical and temporal trends in the publication of RCTs in nursing over the past 20 years by means of a bibliometric analysis. The PubMed database was searched for articles published from 1 January 1991 to 31 October 2011 and satisfying this search strategy: nursing [MeSH Terms] AND (RCT OR trial* OR 'experimental study' OR randomised OR randomisation) AND (English[lang]). Abstracts were reviewed to assess whether they met the criteria for an RCT. A manual search of information on country of origin was carried out and Journal Citation Reports® was used to allocate journals to subject areas. RCT methodology is increasingly drawing the attention of nursing researchers worldwide. However, there is a large disparity in research productivity, at least in terms of number of published RCTs in the English language and listed on PubMed, between the most productive continents, North America and Europe, and the others.