RPL5 on 1p22.1 is recurrently deleted in multiple myeloma and its expression is linked to bortezomib response.

Chromosomal region 1p22 is deleted in ⩾20% of multiple myeloma (MM) patients, suggesting the presence of an unidentified tumor suppressor. Using high-resolution genomic profiling, we delimit a 58 kb minimal deleted region (MDR) on 1p22.1 encompassing two genes: ectopic viral integration site 5 (EVI5) and ribosomal protein L5 (RPL5). Low mRNA expression of EVI5 and RPL5 was associated with worse survival in diagnostic cases. Patients with 1p22 deletion had lower mRNA expression of EVI5 and RPL5, however, 1p22 deletion status is a bad predictor of RPL5 expression in some cases, suggesting that other mechanisms downregulate RPL5 expression. Interestingly, RPL5 but not EVI5 mRNA levels were significantly lower in relapsed patients responding to bortezomib and; both in newly diagnosed and relapsed patients, bortezomib treatment could overcome their bad prognosis by raising their progression-free survival to equal that of patients with high RPL5 expression. In conclusion, our genetic data restrict the MDR on 1p22 to EVI5 and RPL5 and although the role of these genes in promoting MM progression remains to be determined, we identify RPL5 mRNA expression as a biomarker for initial response to bortezomib in relapsed patients and subsequent survival benefit after long-term treatment in newly diagnosed and relapsed patients.

Computed tomography dose measurements with radiochromic films and a flatbed scanner.

Gafchromic XR-QA films were developed for patient dosimetry in diagnostic radiology. A possible application of these films is the measurement of doses in computed tomography. In this study a method to evaluate the CTDI using Gafchromic XR-QA film and a flatbed scanner was developed and tested. Film samples were cut to dimensions of 6 x 170 mm2 in order to have an integration area similar to that of a pencil ionization chamber, with the possibility of changing the integration length. Prior to exposing these films to a computed tomography beam, the angular dependence of the film dose response was investigated by exposing film strips to a static x-ray beam at different angles in the range 0 degrees-180 degrees. A difference of 49% was found between the response with the axis beam parallel to the film surface (90 degrees) and with the axis beam perpendicular (0 degrees and 180 degrees). Integrating over a 360 degrees exposure like the one in computed tomography, a difference of less than 2% was estimated, which is comparable with the measurement error obtainable with XR-QA film. A calibration with a CT beam in the scout mode was performed and film strips were then exposed to single axial scans and to helical scans both in air and in phantoms. Two different types of flatbed scanners were used to read the film samples, a Microtek ScanMaker 9800XL scanner and an Epson Expression 10000 XL scanner, and the accuracy of the results were compared. For beam collimations above 10 mm differences between CTDI measured by film and CTDI measured by ionization chamber below 9% were found for the Epson scanner, with an average estimated error at 1 sigma level of 5%. For the Microtek scanner and for the same film samples, differences below 11% with an average error at 1 sigma level of 8% were founded. The 1 sigma uncertainty of the measured CTDI was provided by the method for each measurement, and it was shown that about the 95% of the differences between the CTDI measurements with radiochromic films and with the ionization chamber were below the estimated 2 sigma uncertainty, for both scanners. After an accurate calibration procedure and the consideration of the uncertainty associated with the measurement, Gafchromic XR-QA films can be used to evaluate the CTDI.

A novel H208D TP63 mutation in a familial case of ectrodactyly-ectodermal dysplasia-cleft lip/palate syndrome without clefting.

Ectrodactyly-ectodermal dysplasia-cleft lip/palate (EEC) syndrome is an autosomal dominant form of ectodermal dysplasia associated with limb anomalies and orofacial clefting. The TP63 gene has been shown to be the cause of the disease, and some tentative genotype-phenotype correlations have been reported. We describe a familial case of EEC syndrome, diagnosed in two siblings affected by severe ectrodactyly and mild ectodermal dysplasia, without clefting. Moreover, one of the siblings had a history of delayed developmental milestones in the first years of life. Family history revealed mild hand malformations in the father and grandfather, who were not available for clinical evaluation. The TP63 gene molecular study showed in both siblings a heterozygous H208D mutation, which has not been previously reported to our knowledge, suggesting that this molecular lesion is associated with EEC syndrome without orofacial clefting.

Effective dose and image quality evaluations of an automatic CT tube current modulation system with an anthropomorphic phantom.

The purpose of this study was to evaluate the consequences of different choices of acquisition parameters on the actual image noise and on the patient dose with an automatic tube current modulation system. The CT investigated was a GE Lightspeed 16-slice and an anthropomorphic phantom was used to simulate the patient. Several acquisitions were made varying noise index (NI), kilovoltage and pitch values. Tube current values were compared for the different acquisitions. Patient dose was evaluated in terms of volumetric computed tomography dose index (CTDI(vol)) and also as effective dose. The noise actually present in the images was analyzed by a region of interest analysis considering representatively phantom sections in the regions of the shoulders, of the lungs and of the abdomen. The obtained results generally evidenced a good agreement between the noise index and the measured noise for the abdomen sections, whereas for the shoulders and the lungs sections the measured noise was respectively greater and lower of the NI. Varying the kV the automatic current modulation system provided images with a substantially constancy of the actual noise and of the patient dose. An increase of the pitch generally decreased the patient dose, whereas the noise was slightly greater for the lowest pitch and almost constant for the other pitch values. This study outlines some important relationships between an automatic tube current modulation system and other CT acquisition parameters, providing useful informations for the choice requested by radiologists in the task of optimization of the CT acquisition protocols. Unless there are other considerations in place, pixel pitches below 1.375 should be avoided, and kVp settings can be changed with no real impact on dose or image noise.

Patient dose evaluation by means of DICOM images for a direct radiography system.

PURPOSE: The purpose of this work was to evaluate the statistical distribution of patient dose for different examinations by using the data stored in a DICOM image archive of a direct digital radiography system. MATERIALS AND METHODS: An automatic procedure to extract dose data and exposure parameters from the image archives was implemented. During a 4.5-month period, 8,292 images were collected. Exposure parameters and the air kerma area product (P (KA)) stored in the image DICOM header were examined for each image. The entrance surface air kerma (K (a,e)), a quantity comparable to the current diagnostic reference levels (DRL), was estimated considering the P (KA) and the geometric parameters of each examination. RESULTS: P (KA) and K (a,e ) distributions showed highly variable values. The obtained K (a,e ) values were substantially lower than the DRL. DRL were more than six times the mean value of K (a,e ) distribution for the abdomen anteroposterior (AP) and lumbar spine lateral (LAT) projections, whereas the ratio was equal to 2.7 for posteroanterior (PA) chest examinations. CONCLUSIONS: The method adopted proved to be effective for characterising the dose of each examination by means of the statistical analysis of the dose quantities over extensive samples. The dose values obtained and the comparison with the DRL showed that this radiographic device allows substantial dose savings compared with estimations made for nondigital or for phosphor-based systems.

Radiation dose evaluations during radiological contrast studies in patients with morbid obesity.

PURPOSE: The purpose of this study was to evaluate the radiation dose to patients during radiological contrast studies performed after vertical banded gastroplasty (VBG) or Roux-en-Y gastric bypass (RYGBP) surgery in patients with morbid obesity. MATERIALS AND METHODS: Dose evaluations were performed on a sample of 39 patients (32 women and 7 men) with a mean weight of 117 kg (range 68-175 kg) and a mean body mass index (BMI) of 43.7 (range 22.2-54.9). Between the second and seventh postoperative day, patients underwent radiological follow-up after oral administration of approximately 70 ml of water-soluble iodinated contrast material (Gastrografin) and images acquired in anteroposterior, right and left oblique projections with the patient upright and then supine. Exposure conditions, dose-area product (DAP) and entrance skin dose (ESD) were recorded for each procedure. On the basis of these data, the effective dose (ED) was calculated using simulation software based on the Monte Carlo method for determining the absorbed dose to organs. To assess the optimal exposure conditions and the dose contributions of fluoroscopy and radiography, the effective dose rates were also evaluated using Plexiglas phantoms of different thickness to simulate different patient sizes. RESULTS: The phantom measurements showed a fourfold dose increase when passing from normal-sized patients to obese patients. Mean DAP value obtained from in-vivo measurements was 70 Gy cm(2) (range 17-147 Gy cm(2)), and mean effective dose was 21 mSv (range 5-45 mSv). CONCLUSIONS: When performing radiological contrast studies in patients with morbid obesity, every possible precaution should be taken to minimise patient dose. Special care should be taken to evaluate justification of the radiological procedure.

Dose area product evaluations with Gafchromic XR-R films and a flat-bed scanner.

Gafchromic XR-R films are a useful tool to evaluate entrance skin dose in interventional radiology. Another dosimetric quantity of interest in diagnostic and interventional radiology is the dose area product (DAP). In this study, a method to evaluate DAP using Gafchromic XR-R films and a flat-bed scanner was developed and tested. Film samples were exposed to an x-ray beam of 80 kVp over a dose range of 0-10 Gy. DAP measurements with films were obtained from the digitalization of a film sample positioned over the x-ray beam window during the exposure. DAP values obtained with this method were compared for 23 cardiological interventional procedures with DAP values displayed by the equipment. The overall one-sigma dose measurement uncertainty depended on the absorbed dose, with values below 6% for doses above 1 Gy. A maximum discrepancy of 16% was found, which is of the order of the differences in the DAP measurements that may occur with different calibration procedures. Based on the results presented, after an accurate calibration procedure and a thorough inspection of the relationship between the actual dose and the direct measured quantity (net optical density or net pixel value variation), Gafchromic XR-R films can be used to assess the DAP.

Dose and energy dependence of response of Gafchromic XR-QA film for kilovoltage x-ray beams.

There is a growing interest in Gafchromic films for patient dosimetry in radiotherapy and in radiology. A new model (XR-QA) with high sensitivity to low dose was tested in this study. The response of the film to different x-ray beam energies (range 28-145 kVp with various filtrations, dose range 0-100 mGy) and to visible light was investigated, together with the after exposure darkening properties. Exposed films were digitized with a commercially available, optical flatbed scanner. A single functional form for dose versus net pixel value variation has been determined for all the obtained calibration curves, with a unique fit parameter different for each of the used x-ray beams. The film response was dependent on beam energy, with higher colour variations for the beams in the range 80-140 kVp. Different sources of uncertainties in dose measurements, governed by the digitalization process, the film response uniformity and the calibration curve fit procedure, have been considered. The overall one-sigma dose measurement uncertainty depended on the beam energy and decreased with increasing absorbed dose. For doses above 10 mGy and beam energies in the range 80-140 kVp the total uncertainty was less than 5%, whereas for the 28 kVp beam the total uncertainty at 10 mGy was about 10%. The post-exposure colour variation was not negligible in the first 24 h after the exposure, with a consequent increase in the calculated dose of about 10%. Results of the analysis of the sensitivity to visible light indicated that a short exposure of this film to ambient and scanner light during the measurements will not have a significant impact on the radiation dosimetry.

Familial tumoral calcinosis and testicular microlithiasis associated with a new mutation of GALNT3 in a white family.

BACKGROUND: Familial tumoral calcinosis (FTC) is a rare autosomal recessive disease characterised by the development of multiple calcified masses in periarticular soft tissues; GALNT3 gene mutations have recently been described in an African American and in a Druse Arab family with FTC. OBJECTIVE: To report the clinical and histological features caused by a new GALNT3 mutation in a white family. RESULTS: Homozygosity for the nonsense mutation Lys463X was found in both affected siblings, who displayed a classic phenotype, the male also having testicular microlithiasis. He is the first subject described with testicular microlithiasis in FTC. CONCLUSIONS: The high testicular expression of GALNT3 suggests that the gene alteration could act locally by causing deposition of calcium, and the testis may be an underestimated site of calcification in FTC. Autoimmune diseases are present in several members of the family. Although immune disorders have been described in FTC, autoimmunity does not segregate with the GALNT3 mutation in this family.

A method for the inter-calibration of a matrix of sensors.

We present a quick and easy method for the calibration of a matrix of sensors. The algorithm is based on a three-step irradiation procedure which relies only on the constancy of the delivered fluence at each step. With this method the gain of each sensor is derived relative to a reference detector. The algorithm has been applied to a matrix of (32 x 32) ionization chambers. After the calibration coefficients have been applied, by comparing the response of the matrix of chambers to a reference detector over a large field, we determined that the fluence measurement of individual chambers is better than 0.7%. The algorithm solves the cumbersome problem of the relative gain calibration of a matrix of a large number of sensors.

D-IMRT verification with a 2D pixel ionization chamber: dosimetric and clinical results in head and neck cancer.

Dynamic intensity-modulated radiotherapy (D-IMRT) using the sliding-window technique is currently applied for selected treatments of head and neck cancer at Institute for Cancer Research and Treatment of Candiolo (Turin, Italy). In the present work, a PiXel-segmented ionization Chamber (PXC) has been used for the verification of 19 fields used for four different head and neck cancers. The device consists of a 32x32 matrix of 1024 parallel-plate ionization chambers arranged in a square of 24x24 cm2 area. Each chamber has 0.4 cm diameter and 0.55 cm height; a distance of 0.75 cm separates the centre of adjacent chambers. The sensitive volume of each single ionization chamber is 0.07 cm3. Each of the 1024 independent ionization chambers is read out with a custom microelectronics chip.The output factors in water obtained with the PXC at a depth of 10 cm were compared to other detectors and the maximum difference was 1.9% for field sizes down to 3x3 cm2. Beam profiles for different field dimensions were measured with the PXC and two other types of ionization chambers; the maximum distance to agreement (DTA) in the 20-80% penumbra region of a 3x3 cm2 field was 0.09 cm. The leaf speed of the multileaf collimator was varied between 0.07 and 2 cm s-1 and the detector response was constant to better than 0.6%. The behaviour of the PXC was measured while varying the dose rate between 0.21 and 1.21 Gy min-1; the mean difference was 0.50% and the maximum difference was 0.96%. Using fields obtained with an enhanced dynamic wedge and a staircase-like (step) IMRT field, the PXC has been tested for simple 1D modulated beams; comparison with film gave a maximum DTA of 0.12 cm. The PXC was then used to check four different IMRT plans for head and neck cancer treatment: cervical chordoma, parotid, ethmoid and skull base. In the comparison of the PXC versus film and PXC versus treatment planning system, the number of pixels with gamma parameter

Two-dimensional and quasi-three-dimensional dosimetry of hadron and photon beams with the Magic Cube and the Pixel Ionization Chamber.

Two detectors for fast two-dimensional (2D) and quasi-three-dimensional (quasi-3D) verification of the dose delivered by radiotherapy beams have been developed at University and Istituto Nazionale di Fisica Nucleare (INFN) of Torino. The Magic Cube is a stack of strip-segmented ionization chambers interleaved with water-equivalent slabs. The parallel plate ionization chambers have a sensitive area of 24 x 24 cm2, and consist of 0.375 cm wide and 24 cm long strips. There are a total of 64 strips per chamber. The Magic Cube has been tested with the clinical proton beam at Loma Linda University Medical Centre (LLUMC), and was shown to be capable of fast and precise quasi-3D dose verification. The Pixel Ionization Chamber (PXC) is a detector with pixel anode segmentation. It is a 32 x 32 matrix of 1024 cylindrical ionization cells arranged in a square 24 x 24 cm2 area. Each cell has 0.4 cm diameter and 0.55 cm height, at a pitch of 0.75 cm separates the centre of adjacent cells. The sensitive volume of each single ionization cell is 0.07 cm3. The detectors are read out using custom designed front-end microelectronics and a personal computer-based data acquisition system. The PXC has been used to verify dynamic intensity-modulated radiotherapy for head-and-neck and breast cancers.

Dosimetric characterization of a large area pixel-segmented ionization chamber.

A pixel-segmented ionization chamber has been designed and built by Torino University and INFN. The detector features a 24 x 24 cm2 active area divided in 1024 independent cylindrical ionization chambers and can be read out in 500 micros without introducing dead time; the digital charge quantum can be adjusted between 100 fC and 800 fC. The sensitive volume of each single ionization chamber is 0.07 cm3. The purpose of the detector is to ease the two-dimensional (2D) verifications of fields with complex shapes and large gradients. The detector was characterized in a PMMA phantom using 60Co and 6 MV x-ray photon beams. It has shown good signal linearity with respect to dose and dose rate to water. The average sensitivity of a single ionization chamber was 2.1 nC/Gy, constant within 0.5% over one month of daily measurements. Charge collection efficiency was 0.985 at the operating polarization voltage of 400 V and 3.5 Gy/min dose rate. Tissue maximum ratio and output factor have been compared with a Farmer ionization chamber and were found in good agreement. The dose profiles have been compared with the ones obtained with an ionization chamber in water phantom for the field sizes supplied by a 3D-Line dynamic multileaf collimator. These results show that this detector can be used for 2D dosimetry of x-ray photon beams, supplying a good spatial resolution and sensibly reducing the time spent in dosimetric verification of complex radiation fields.

Flt-3 and its ligand are expressed in neural crest-derived tumors and promote survival and proliferation of their cell lines.

Flt-3 ligand (FL) is a cytokine that promotes the survival, proliferation, and differentiation of hematopoietic progenitors in synergy with other growth factors, such as stem cell factor. Previously we have demonstrated that stem cell factor and its receptor c-kit are expressed in neural crest-derived tumor cells and that a c-kit block induces their apoptosis. Here we have evaluated the expression of flt-3 and its ligand in 12 neuroectodermal tumor cell lines from neuroblastoma (NB), neuroepithelioma (NE), Ewing sarcoma (ES), and peripheral neuroectodermal tumor (PNET) and in 38 biopsies: 19 from NB and 19 from ES and PNET. RT-PCR demonstrated the expression of flt-3 and FL in all lines. Coexpression was observed in 42% of NB and in 74% of ES and PNET biopsies. Flow cytometry confirmed the presence of membrane and cytoplasmic flt-3 and membrane FL in all lines, whereas soluble FL protein was not measurable in their supernatants. Microphysiometric demonstration of acidification of the medium provided evidence of the specific response of cell lines to FL stimulation. Specific flt-3 phosphorylation after FL treatment was also demonstrated by Western blotting analysis. In cells growing in RPMI plus 1% fetal calf serum, FL revealed a significant proliferating activity, more evident in NB and NE lines (mean increase of viable cells, 73 +/- 26% after 1 day). Treatment with flt-3 antisense oligonucleotides significantly inhibited cell growth. FL also displayed an antiapoptotic activity: after a 12-hour culture in the presence of 0.1% fetal calf serum, FL caused a 50% reduction of apoptotic cells. These results provide further evidence that neuroectodermal and hematopoietic cells share common regulatory pathways, and could be of interest in the clinical management of neuroectodermal tumors.

Diamond-Blackfan anemia: report of seven further mutations in the RPS19 gene and evidence of mutation heterogeneity in the Italian population.

Diamond-Blackfan anemia (DBA) is a congenital disease characterized by defective erythroid progenitor maturation and physical malformations. Most cases are sporadic, but dominant or, more rarely, recessive inheritance is observed in 10% of patients. Mutations in the gene encoding ribosomal protein (RP) S19 have recently been found in 25% of patients with either the dominant or the sporadic form. DBA is the first human disease due to mutations in a ribosomal structural protein. Families unlinked to this locus have also been reported. In an investigation of 23 individuals, we identified eight different mutations in 9 patients. These include five missense, one frameshift, one splice site defect, and one 4-bp insertion in the regulatory sequence. Seven mutations are new; one has so far been found in 8 patients and is a relatively common de novo event. Two mutations are predicted to generate a truncated protein. We also report the prevalence of RPS 19 mutations in the Italian DBA population, as shown by an analysis of 56 patients. No genotype-phenotype correlation was found between patients with the same mutation. The main clinical applications for molecular analysis are clinical diagnosis of patients with an incomplete form of DBA and testing of siblings of a patient with a severe form so as to avoid using those who carry a mutation and a silent phenotype as allogeneic stem cell donors.

Diamond-Blackfan Anaemia: an overview.

Diamond Blackfan Anaemia (DBA) is a congenital disease characterised by defective erythroid progenitor maturation. It is usually diagnosed during the first year of life. The main clinical sign is profound isolated normochromic or macrocytic anaemia, with normal numbers and function of the other haemopoietic cells. Reticulocyte counts in patients with DBA are very low. Bone marrow reflects the defective erythropoiesis, showing a very low number of erythropoietic precursors and a reduction of erythroid burst-forming unit progenitor cells. The proliferation and differentiation of the other lineages are normal. More than one-third of patients have malformations, most often involving the upper limbs and head, and the urogenital or cardiovascular systems. However, the link between these malformations and defective erythropoiesis is unclear and a defect in a molecule acting on both early embryonic development and haematopoiesis has been proposed. Whereas most cases are sporadic, inheritance is observed in 10% of patients, with a dominant or, more rarely, recessive pattern. One locus on chromosome 19q13.2 encoding ribosomal protein S19 accounts for a quarter of patients with either the dominant or the sporadic form. Families not linked with this locus have also been described. The diagnosis of DBA may be difficult and differential diagnoses include Fanconi's anaemia and acquired erythroid aplasias. Erythrocyte adenosine deaminase levels are generally high in DBA patients, which may help in the diagnosis, but they are not pathognomic. Corticosteroids are the main treatment option in DBA and these agents induce erythropoiesis in over 60% of patients. Some patients achieve complete remission, which may be either corticosteroid-induced or spontaneous. The increased in vitro erythropoiesis occasionally induced by the addition of specific cytokines, namely interleukin (IL)-3 and stem cell factor (SCF), has suggested their use in vivo. However, few patients have responded to IL-3, whereas SCF administration, though interesting in theory, has not yet been attempted. Patients who do not respond to corticosteroids and those who have to discontinue treatment because of adverse events must rely on long term transfusions, and are thus exposed to all of the associated complications. Bone marrow or cord blood transplantation has been performed in some patients. The former approach is burdened with severe complications and high mortality.

Mutations in ribosomal protein S19 gene and diamond blackfan anemia: wide variations in phenotypic expression.

Mutations of the ribosomal protein S19 (RPS19) gene were recently identified in 10 patients with Diamond Blackfan anemia (DBA). To determine the prevalence of mutations in this gene in DBA and to begin to define the molecular basis for the observed variable clinical phenotype of this disorder, the genomic sequence of the 6 exons and the 5' untranslated region of the RPS19 gene was directly assessed in DBA index cases from 172 new families. Mutations affecting the coding sequence of RPS19 or splice sites were found in 34 cases (19.7%), whereas mutations in noncoding regions were found in 8 patients (4.6%). Mutations included nonsense, missense, splice sites, and frameshift mutations. A hot spot for missense mutations was identified between codons 52 and 62 of the RPS19 gene in a new sequence consensus motif W-[YFW]-[YF]-x-R-[AT]-A-[SA]-x-[AL]-R-[HRK]-[ILV]-Y. No correlation between the nature of mutations and the different patterns of clinical expression, including age at presentation, presence of malformations, and therapeutic outcome, could be documented. Moreover, RPS19 mutations were also found in some first-degree relatives presenting only with isolated high erythrocyte adenosine deaminase activity and/or macrocytosis. The lack of a consistent relationship between the nature of the mutations and the clinical phenotype implies that yet unidentified factors modulate the phenotypic expression of the primary genetic defect in families with RPS19 mutations.

Diamond-Blackfan anaemia in the Italian population.

Diamond-Blackfan anaemia (DBA) is a congenital disease characterized by defective erythroid progenitor maturation: 30% of patients have congenital malformations. The link between these malformations and defective erythropoiesis is unclear: a defect in a molecule acting both on embryo development and haemopoiesis has been proposed. Inheritance is autosomal dominant in most familial cases, but recessive families have also been reported. Many cases are sporadic. A DBA locus has been mapped on chromosome 19q13.2 (Gustavsson et al, 1997), but several families unlinked to this locus have also been reported (Gustavsson et al, 1998). This paper presents clinical, epidemiological and molecular data for DBA in the Italian population. Segregation analysis of 19q markers in patients with DBA showed exclusion of this locus in 5/12 families with inherited DBA. There was evidently locus heterogeneity for DBA in this population. A new microdeletion was identified in one patient. Other families, in which DBA segregates concordantly with the 19q critical region, suggest incomplete penetrance and expressivity of the DBA gene.