Transcriptional Regulation of the Human Calcitonin Gene: A Progress Report

We have applied DNA transfer techniques lo study the transcriptional regulation of the calcitonin (CT) gene in a C-cell line (TT) derived from a human medullary thyroid carcinoma. TT cells were transfected with a fusion gene containing the CT gene promoter and 5\u27 -flanking DNA attached to the promoter-less growth hormone gene (reporter). We quantitated the reporter gene product to monitor transcriptional activation by the CT promoter and deletion mutants of the 5\u27 -flanking DNA. We found that the proximal CT promoter which includes the DNA sequence from +1 to -129 bp upstream from the CT transcription start site did not induce transcription in C-cells or in NIH 3T3 cells. The attachment of additional 5\u27-flanking DNA, extending up to -1460 bp enhanced transcription up to twelvefold in TT cells but had no effect on transcription in 3T3 cells. Deletion of a sequence located at -1290 to -820 bp on the CT 5\u27 -flanking DNA abolished the transcription of the reporter gene. Attachment of the DNA sequence located between -1333 to -731 to the fusion gene, containing the CT promoter (+1 to -129) and the reporter gene, restored transcription of the reporter gene in TT cells. We conclude that an enhancer of CT transcription, which is active in C-cells but not in 3T3 cells, is located between -1290 and -820 of the CT 5\u27-flanking DNA

Breast cancer and communication: Monocentric experience of a self-assessment questionnaire

Background: The communication of the diagnosis of breast cancer induces to the patient a strong psychological trauma. Radiologists are at the forefront of communicating, either for a biopsy or the probable diagnosis of malignancy. This is a complex task, which requires the knowledge and application of correct “communicative models”, among which the SPIKES protocol rep-resents a fundamental reference. Design and methods: 110 patients, with a history of breast cancer, filled out a questionnaire consisting of six questions: five aimed at defining communication compliance with the SPIKES protocol, the sixth, consisting of six feelings, aimed at the knowledge of the next emotional state. Results: Regarding compliance with various “strategic points” of the SPIKES protocol, questionnaires show that 70% of patients reported no omissions, while the remaining 30% reported omissions relatively to perception (56%), emotions (23%), setting (13%), knowledge (6%) and invitation (2%). The results showed the existence of a correlation between the final emotional state and the correct application of the SPIKES protocol; in fact, patients who reacted with a positive final emotional state-reported greater adherence to the strategic points of the SPIKES protocol. Conclusions: In healthcare, knowing the communicative compliance of a team in giving “bad news” is fundamental, especially in breast cancer. The SPIKES protocol is recognized by the Literature as a fundamental reference able to affect “positively” the emotional state of patients. The proposed questionnaire is a valid tool to identify the weak points of communication and related criticalities, to improve clinical practice

The INGV Mobile Telemetered Network (Re.Mo.Tel.): long-term monitoring analysis.

Since 2008, the Istituto Nazionale di Geofisica e Vulcanologia (INGV) developed a temporary real-time telemetered seismic network infrastructure (Re.Mo.Tel.) to densify the Italian National Seismic Network in epicentral area, thus improving the location of the aftershocks distribution after a mainshock. This infrastructure consists of various mobile and autonomous seismic stations that in group of three are telemetered to a Very Small Aperture Terminal (VSAT). Using a dedicated bandwidth on UHF, Wi-Fi and satellite frequency, the system is able to stream real-time data to the INGV acquisition centers in Rome and Grottaminarda. The deployment of the seismic network is supported by a geographical information system (GIS) that visualizes, for the epicentral area, information about instrumental seismicity, seismic risk, macroseismic felts and territorial data. The April, 6th, 2009 Mw 6.3 L'Aquila destructive earthquake represented the first real-case where the entire Re.Mo.Tel. infrastructure has been deployed. Less than 6 hours after the earthquake occurrence, a first accelerometric station was streaming data to the INGV seismic monitoring headquarters. A total number of 9 seismic stations was installed within 4 days after the main event, with the aim of recording continuous data to contribute in the aftershocks detection. In detail the first day a number of 3 stations was installed, surrounding the mainshock. Then the rest of the stations was deployed on the third and the fourth day to the south and the north, following the post-seismic evolution. A project funded by the Dipartimento di Protezione Civile (DPC-Civil Defense) gave the opportunity to maintain some of the deployed stations throughout the duration of the project of one year (although the stations are still installed) to assure the long-term monitoring of the triggered seismic sequence. In this work, we investigate: • The impact of the Re.Mo.Tel. on earthquakes detection and real-time monitoring of the sequence evolution; • The location improvement due to the deployment of stations closer to the epicentral area (with installations that followed the early evolution and complexity of fault fractures); • The network reliability in terms of data stored throughout the working period

Telomere length shortening is associated with treatment-free remission in chronic myeloid leukemia patients

We studied telomere length in 32 CML patients who discontinued imatinib after achieving complete molecular remission and 32 age-sex-matched controls. The relative telomere length (RTL) was determined by q-PCR as the telomere to single copy gene (36B4) ratio normalized to a reference sample (K-562 DNA). Age-corrected RTL (acRTL) was also obtained. The 36-month probability of treatment-free remission (TFR) was 59.4 %. TFR patients showed shorter acRTL compared to relapsed (mean ± SD = 0.01 ± 0.14 vs 0.20 ± 0.21; p = 0.01). TFR was significantly higher in CML patients with acRTL ≤0.09 (78.9 vs 30.8 %, p = 0.002). CML stem cells harboring longer telomeres possibly maintain a proliferative potential after treatment discontinuation

Next-generation sequencing for BCR-ABL1 kinase domain mutation testing in patients with chronic myeloid leukemia: A position paper

BCR-ABL1 kinase domain (KD) mutation status is considered to be an important element of clinical decision algorithms for chronic myeloid leukemia (CML) patients who do not achieve an optimal response to tyrosine kinase inhibitors (TKIs). Conventional Sanger sequencing is the method currently recommended to test BCR-ABL1 KD mutations. However, Sanger sequencing has limited sensitivity and cannot always discriminate between polyclonal and compound mutations. The use of next-generation sequencing (NGS) is increasingly widespread in diagnostic laboratories and represents an attractive alternative. Currently available data on the clinical impact of NGS-based mutational testing in CML patients do not allow recommendations with a high grade of evidence to be prepared. This article reports the results of a group discussion among an ad hoc expert panel with the objective of producing recommendations on the appropriateness of clinical decisions about the indication for NGS, the performance characteristics of NGS platforms, and the therapeutic changes that could be applied based on the use of NGS in CML. Overall, these recommendations might be employed to inform clinicians about the practical use of NGS in CML

Next-generation sequencing for BCR-ABL1 kinase domain mutation testing in patients with chronic myeloid leukemia: A position paper

BCR-ABL1 kinase domain (KD) mutation status is considered to be an important element of clinical decision algorithms for chronic myeloid leukemia (CML) patients who do not achieve an optimal response to tyrosine kinase inhibitors (TKIs). Conventional Sanger sequencing is the method currently recommended to test BCR-ABL1 KD mutations. However, Sanger sequencing has limited sensitivity and cannot always discriminate between polyclonal and compound mutations. The use of next-generation sequencing (NGS) is increasingly widespread in diagnostic laboratories and represents an attractive alternative. Currently available data on the clinical impact of NGS-based mutational testing in CML patients do not allow recommendations with a high grade of evidence to be prepared. This article reports the results of a group discussion among an ad hoc expert panel with the objective of producing recommendations on the appropriateness of clinical decisions about the indication for NGS, the performance characteristics of NGS platforms, and the therapeutic changes that could be applied based on the use of NGS in CML. Overall, these recommendations might be employed to inform clinicians about the practical use of NGS in CML

Dihydroorotate dehydrogenase inhibition reveals metabolic vulnerability in chronic myeloid leukemia

The development of different generations of BCR-ABL1 tyrosine kinase inhibitors (TKIs) has led to the high overall survival of chronic myeloid leukemia (CML) patients. However, there are CML patients who show resistance to TKI therapy and are prone to progress to more advanced phases of the disease. So, implementing an alternative approach for targeting TKIs insensitive cells would be of the essence. Dihydroorotate dehydrogenase (DHODH) is an enzyme in the de novo pyrimidine biosynthesis pathway that is located in the inner membrane of mitochondria. Here, we found that CML cells are vulnerable to DHODH inhibition mediated by Meds433, a new and potent DHODH inhibitor recently developed by our group. Meds433 significantly activates the apoptotic pathway and leads to the reduction of amino acids and induction of huge metabolic stress in CML CD34+ cells. Altogether, our study shows that DHODH inhibition is a promising approach for targeting CML stem/progenitor cells and may help more patients discontinue the therapy

Validation and reference values of the EORTC QLQ-CML24 questionnaire to assess health-related quality of life in patients with chronic myeloid leukemia

Health-related quality of life (HRQOL) assessment is important to facilitate decisions in the current treatment landscape of chronic myeloid leukemia (CML). Therefore, the availability of a validated HRQOL questionnaire, specifically developed for CML patients treated with tyrosine kinase inhibitors (TKIs), may enhance quality of research in this area. We performed an international study including 782 CML patients to assess the validity of the EORTC QLQ-CML 24 questionnaire, and to generate HRQOL reference values to facilitate interpretation of results in future studies. Internal consistency, assessed with Cronbach’s alpha coefficients, ranged from 0.66 to 0.83. In the confirmatory factor analysis, all standardized factor loadings exceeded the threshold of 0.40 (range 0.49–0.97), confirming the hypothesized scale structure. Reference values stratified by age and sex were also generated. Our findings support the use of the EORTC QLQ-CML 24, in conjunction with the EORTC QLQ-C30, as a valuable measure to assess HRQOL in CML patients