Volume 49 (2015): Issue 3 (September 2015)

Background. Efficient stem cell differentiation is considered to be the holy grail of regenerative medicine. Pursuing the most productive method of directed differentiation has been the subject of numerous studies, resulting in the development of many effective protocols. However, the necessity for further improvement in differentiation efficiency remains. This review contains a description of molecular processes underlying the response of stem cells to ionizing radiation, indicating its potential application in differentiation procedures. In the first part, the radiation-induced damage response in various types of stem cells is described. Second, the role of the p53 protein in embryonic and adult stem cells is highlighted. Last, the hypothesis on the mitochondrial involvement in stem cell development including its response to ionizing radiation is presented.

Conclusions. In summary, despite the many threats of ionizing radiation concerning genomic instability, subjecting cells to the appropriate dosage of ionizing radiation may become a useful method for enhancing directed differentiation in certain stem cell types.

Background. Gamma-enolase, known also as neuron-specific enolase (NSE), is an enzyme of the glycolytic pathway, which is expressed predominantly in neurons and cells of the neuroendocrine system. As a tumour marker it is used in diagnosis and prognosis of cancer; however, the mechanisms enrolling it in malignant progression remain elusive. As a cytoplasmic enzyme gamma-enolase is involved in increased aerobic glycolysis, the main source of energy in cancer cells, supporting cell proliferation. However, different cellular localisation at pathophysiological conditions, proposes other cellular engagements.

Conclusions. The C-terminal part of the molecule, which is not related to glycolytic pathway, was shown to promote survival of neuronal cells by regulating neuronal growth factor receptor dependent signalling pathways, resulting also in extensive actin cytoskeleton remodelling. This additional function could be important also in cancer cells either to protect cells from stressful conditions and therapeutic agents or to promote tumour cell migration and invasion. Gamma-enolase might therefore have a multifunctional role in cancer progression: it supports increased tumour cell metabolic demands, protects tumour cells from stressful conditions and promotes their invasion and migration.

Background. The aim of the study was to explore the influence of various time-of-flight (TOF) and non-TOF reconstruction algorithms on positron emission tomography/computer tomography (PET/CT) image quality.

Materials and methods. Measurements were performed with a triple line source phantom, consisting of capillaries with internal diameter of ~ 1 mm and standard Jaszczak phantom. Each of the data sets was reconstructed using analytical filtered back projection (FBP) algorithm, iterative ordered subsets expectation maximization (OSEM) algorithm (4 iterations, 24 subsets) and iterative True-X algorithm incorporating a specific point spread function (PSF) correction (4 iterations, 21 subsets). Baseline OSEM (2 iterations, 8 subsets) was included for comparison. Procedures were undertaken following the National Electrical Manufacturers Association (NEMA) NU-2-2001 protocol.

Results. Measurement of spatial resolution in full width at half maximum (FWHM) was 5.2 mm, 4.5 mm and 2.9 mm for FBP, OSEM and True-X; and 5.1 mm, 4.5 mm and 2.9 mm for FBP+TOF, OSEM+TOF and True-X+TOF respectively. Assessment of reconstructed Jaszczak images at different concentration ratios showed that incorporation of TOF information improves cold contrast, while hot contrast only slightly, however the most prominent improvement could be seen in background variability - noise reduction.

Conclusions. On the basis of the results of investigation we concluded, that incorporation of TOF information in reconstruction algorithm mostly affects reduction of the background variability (levels of noise in the image), while the improvement of spatial resolution due to incorporation of TOF information is negligible. Comparison of traditional and modern reconstruction algorithms showed that analytical FBP yields comparable results in some parameter measurements, such as cold contrast and relative count error. Iterative methods show highest levels of hot contrast, when TOF and PSF corrections were applied simultaneously.

Background. Irreversible electroporation (IRE) is a tissue ablation method, which relies on the phenomenon of electroporation. When cells are exposed to a sufficiently electric field, the plasma membrane is disrupted and cells undergo an apoptotic or necrotic cell death. Although heating effects are known IRE is considered as non-thermal ablation technique and is currently applied to treat tumors in locations where thermal ablation techniques are contraindicated.

Materials and methods. The manufacturer of the only commercially available pulse generator for IRE recommends a voltage-to-distance ratio of 1500 to 1700 V/cm for treating tumors in the liver. However, major blood vessels can influence the electric field distribution. We present a method for treatment planning of IRE which takes the influence of blood vessels on the electric field into account; this is illustrated on a treatment of 48-year-old patient with a metastasis near the remaining hepatic vein after a right side hemi-hepatectomy.

Results. Output of the numerical treatment planning method shows that a 19.9 cm³ irreversible electroporation lesion was generated and the whole tumor was covered with at least 900 V/cm. This compares well with the volume of the hypodense lesion seen in contrast enhanced CT images taken after the IRE treatment. A significant temperature raise occurs near the electrodes. However, the hepatic vein remains open after the treatment without evidence of tumor recurrence after 6 months.

Conclusions. Treatment planning using accurate computer models was recognized as important for electrochemotherapy and irreversible electroporation. An important finding of this study was, that the surface of the electrodes heat up significantly. Therefore the clinical user should generally avoid placing the electrodes less than 4 mm away from risk structures when following recommendations of the manufacturer.

Background. The aim of our study was (1) to describe central nervous system (CNS) manifestations in children with Langerhans cell histiocytosis (LCH) based on images sent to a reference center and meeting minimum requirements and (2) to assess the inter-rater agreement of CNS-MRI results, which represents the overall reproducibility of this investigation.

Methods. We retrospectively reviewed brain MRI examinations in children with LCH, for which MRI minimum requirements were met. Abnormalities were rated by two experienced neuroradiologists, and the inter-rater agreement was assessed.

Results. Out of a total of 94 imaging studies, only 31 MRIs met the minimum criteria, which included T2w, FLAIR, T1w images before/after contrast in at least two different section planes, and thin post contrast sagittal slices T1w through the sella. The most common changes were osseous abnormalities, followed by solid enlargement of the pineal gland, thickened enhancing stalk and signal changes of the dentate nucleus. Whereas inter-rater agreement in assessing most of the CNS lesions was relatively high (κ > 0.61), the application of minimum criteria often did not allow to evaluate the posterior pituitary.

Conclusions. The diversity of radiological protocols from different institutions leads to difficulties in the diagnosis of CNS abnormalities in children with LCH. Although the inter-rater agreement between neuroradiologists was high, not all the LCH manifestations could be completely ruled out when using the minimum criteria. Brain MRIs should therefore follow LCH guideline protocols and include T1 pre-gadolinium sagittal images, and be centrally reviewed in order to improve the comparison of clinical trials.

Background. The primary objective of the study was to evaluate the association between the minimum apparent diffusion coefficient (ADC_min) and Ki-67, an index for cellular proliferation, in non-small cell lung cancers. Also, we aimed to assess whether ADC_min values differ between tumour subtypes and tissue sampling method.

Methods. The patients who had diffusion weighted magnetic resonance imaging (DW-MRI) were enrolled retrospectively. The correlation between ADC_min and the Ki-67 index was evaluated.

Results. Ninety three patients, with a mean age 65 ± 11 years, with histopathologically proven adenocarcinoma and squamous cell carcinoma of the lungs and had technically successful DW-MRI were included in the study. The numbers of tumour subtypes were 47 for adenocarcinoma and 46 for squamous cell carcinoma. There was a good negative correlation between ADC_min values and the Ki-67 proliferation index (r = −0.837, p < 0.001). The mean ADCmin value was higher and the mean Ki-67 index was lower in adenocarcinomas compared to squamous cell carcinoma (p < 0.0001). There was no statistical difference between tissue sampling methods.

Conclusions. Because ADC_min shows a good but negative correlation with Ki-67 index, it provides an opportunity to evaluate tumours and their aggressiveness and may be helpful in the differentiation of subtypes non-invasively.

Background.Helicobacter pylori infection is the main cause of gastric cancer. The disease progression is influenced by the host inflammatory responses, and cytokine single nucleotide polymorphisms (SNPs) may have a role in the course of the disease. The aim of our study was to investigate proinflammatory cytokine polymorphisms, previously associated with the development of gastric cancer, in a Slovenian population.

Patients and methods. In total 318 patients and controls were selected for the study and divided into three groups: (i) patients with gastric cancer (n = 58), (ii) patients with chronic gastritis (n = 60) and (iii) healthy control group (n = 200). H. pylori infection in patient groups was determined by serology, histology and culture. Four proinflammatory gene polymorphisms were determined (IL-1β, IL-1rα, TNF-α, TLR-4) in all subjects.

Results. We found a statistically significant difference between males and females for the groups (p = 0.025). Odds ratio (OR) for gastric cancer risk for females was 0.557 (95% confidence interval [CI]: 0.233―1.329) and for chronic gastritis 2.073 (95% CI: 1.005―4.277). IL-1B-511*T/T homozygous allele for cancer group had OR = 2.349 (95% CI: 0.583―9.462), heterozygous IL-1B-511*T had OR = 1.470 (95% CI: 0.583―3.709) and heterozygotes in TNF-A-308 genotype for chronic gastritis had OR = 1.402 (95% CI: 0.626―3.139). Other alleles had OR less than 1.

Conclusions. We could not prove association between gastric cancer and chronic gastritis due to H. pylori in any cytokine SNPs studied in Slovenian population. Other SNPs might be responsible besides infection with H. pylori for the progression from atrophy to neoplastic transformation.

Background. Inflammatory myofibroblastic tumors are rare in the pediatric population. Most common localizations were reported in the lungs. A localization in the pancreas needs differentiation from other tumors and chronic pancreatitis. Treatment is surgical resection, although there are reports of treatment with oral steroids and radiation therapy.

Case report. A 6-month-old child was treated due to a tumor in the head of the pancreas. On admission he was jaundiced with pruritus. US and MRI confirmed pancreatic tumor. Preoperative biopsy wasn’t conclusive regarding the nature of the tumor. Duodenopancreatectomy was performed. Postoperative course was uneventful. Histologic examination confirmed the diagnosis of inflammatory myofibroblastic tumor. On follow up, he remained with no evidence of recurrence.

Conclusions. A literature review revealed 10 cases of pancreatic inflammatory myofibroblastic tumors in the pediatric age group. Our patient is the youngest reported. Despite major resection, there were no complications. However, management of this child might be possible with steroids, but conservative treatment might be insufficient, especially in aggressive forms of tumors.

Background. There is a paradigm that chemotherapy is ineffective in thyroid carcinoma. The aim of our study was to find out whether neoadjuvant chemotherapy before thyroid surgery had an effect on the size of primary tumour in patients with poorly differentiated thyroid carcinoma (PDTC) based on Turin proposal.

Patients and methods. Altogether, 13 patients (8 women, 5 men; median age 61 years) with PDTC based on Turin proposal were treated with neoadjuvant chemotherapy between 1986 and 2005. Tumour diameter was from 4.5 to 18 cm (median 9 cm). Regional and distant metastases were detected in 6 and 9 patients, respectively. Eight patients had pT4 tumour.

Results. Altogether, 29 (range 1–5) cycles of chemotherapy were given. Tumour diameter decreased in all the patients and by more than 30% in 5 patients (= 38%). Two of these five patients had also preoperative external beam irradiation (EBRT). Total thyroidectomy, lobectomy and neck dissection were performed in 10, 3 and 5 cases, respectively. R0 and R1 resection was done in 5 and 8 cases, respectively. Eight patients had postoperative EBRT of the neck and upper mediastinum. The 5-year and 10-year cause-specific survival rates of patients were 66% and 20%, respectively.

Conclusions. After neoadjuvant chemotherapy a partial tumour regression was observed in 38% of patients with PDTC based on Turin proposal.

Background. Fibulin-3 is a new potential biomarker for malignant mesothelioma (MM). This study evaluated the potential applicability of fibulin-3 plasma levels as a biomarker of response to treatment and its prognostic value for progressive disease within 18 months. The potential applicability of fibulin-3 in comparison with or in addition to soluble mesothelin-related peptides (SMRP) was also assessed.

Patients and methods. The study included 78 MM patients treated at the Institute of Oncology Ljubljana between 2007 and 2011. Fibulin-3 levels in plasma samples obtained before treatment and in various responses to treatment were measured with the enzyme-linked immunosorbent assay.

Results. In patients evaluated before the treatment, fibulin-3 levels were not influenced by histopathological sub-types, tumour stages or the presence of metastatic disease. Significantly higher fibulin-3 levels were found in progressive disease as compared to the levels before treatment (Mann-Whitney [U] test = 472.50, p = 0.003), in complete response to treatment (U = 42.00, p = 0.010), and in stable disease (U = 542.00, p = 0.001). Patients with fibulin-3 levels exceeding 34.25 ng/ml before treatment had more than four times higher probability for developing progressive disease within 18 months (odds ratio [OR] = 4.35, 95% confidence interval [CI] 1.56–12.13). Additionally, patients with fibulin-3 levels above 34.25 ng/ml after treatment with complete response or stable disease had increased odds for progressive disease within 18 months (OR = 6.94, 95% CI 0.99–48.55 and OR = 4.39, 95% CI 1.63–11.81, respectively).

Conclusions. Our findings suggest that in addition to SMRP fibulin-3 could also be helpful in detecting the progression of MM.

Background. The present study aimed to analyse potential prognostic factors, with emphasis on tumour volume, in determining progression free survival (PFS) for malignancies of the nasal cavity and the paranasal sinuses.

Patients and methods. Retrospective analysis of 106 patients with primary sinonasal malignancies treated and followed-up between March 2006 and October 2012. Possible predictive parameters for PFS were entered into univariate and multivariate Cox regression analysis. Kaplan-Meier curve analysis included age, sex, baseline tumour volume (based on MR imaging), histology type, TNM stage and prognostic groups according to the American Joint Committee on Cancer (AJCC) classification. Receiver operating characteristic (ROC) curve analysis concerning the predictive value of tumour volume for recurrence was also conducted.

Results. The main histological subgroup consisted of epithelial tumours (77%). The majority of the patients (68%) showed advanced tumour burden (AJCC stage III-IV). Lymph node involvement was present in 18 cases. The mean tumour volume was 26.6 ± 21.2 cm³. The median PFS for all patients was 24.9 months (range: 2.5–84.5 months). The ROC curve analysis for the tumour volume showed 58.1% sensitivity and 75.4% specificity for predicting recurrence. Tumour volume, AJCC staging, T- and N-stage were significant predictors in the univariate analysis. Positive lymph node status and tumour volume remained significant and independent predictors in the multivariate analysis.

Conclusions. Radiological tumour volume proofed to be a statistically reliable predictor of PFS. In the multivariate analysis, T-, N- and overall AJCC staging did not show significant prognostic value.

Background. Volumetric modulated arc therapy (VMAT) has quickly become accepted as standard of care for the treatment of prostate cancer based on studies showing it is able to provide faster delivery with adequate target coverage and reduced monitor units while maintaining organ at risk (OAR) sparing. This study aims to demonstrate the potential to increase dose conformality with increased planner control and OAR sparing using a hybrid treatment technique compared to VMAT.

Methods. Eleven patients having been previously treated for prostate cancer with VMAT techniques were replanned with a hybrid technique on Varian Treatment Planning System. Multiple static IMRT fields (2 to 3) were planned initially based on critical OAR to reduce dose but provide some planning treatment volume (PTV) coverage. This was used as a base dose plan to provide 30-35% coverage for a single arc VMAT plan.

Results. The clinical VMAT plan was used as a control for the purposes of comparison. Average of all OAR sparing between the hybrid technique and VMAT showed the hybrid plan delivering less dose in almost all cases except for V80 of the bladder and maximum dose to right femoral head. PTV coverage was superior with the VMAT technique. Monitor unit differences varied, with the hybrid plan able to deliver fewer units 37% of the time, similar results 18% of the time, and higher units 45% of the time. On average, the hybrid plan delivered 10% more monitor units.

Conclusions. The hybrid plan can be delivered in a single gantry rotation combining aspects of VMAT with regions of dynamic intensity modulated radiation therapy (IMRT) within the treatment arc.

Background. Penumbra characteristics play a significant role in dose delivery accuracy for radiation therapy. For treatment planning, penumbra width and radiation field offset strongly influence target dose conformity and organ at risk sparing.

Methods. In this study, we present an analytical and numerical approach for evaluation of the rounded leaf end effect on penumbra characteristics. Based on the rule of half-value layer, algorithms for leaf position calculation and radiation field offset correction were developed, which were advantageous particularly in dealing with large radius leaf end. Computer simulation was performed based on the Monte Carlo codes of EGSnrc/BEAMnrc, with groups of leaf end radii and source sizes. Data processing technique of curve fitting was employed for deriving penumbra width and radiation field offset.

Results. Results showed that penumbra width increased with source size. Penumbra width curves for large radius leaf end were U-shaped. This observation was probably related to the fact that radiation beams penetrated through the proximal and distal leaf sides. In contrast, source size had negligible impact on radiation field offset. Radiation field offsets were found to be constant both for analytical method and numerical simulation. However, the overall resulting values of radiation field offset obtained by analytical method were slightly smaller compared with Monte Carlo simulation.

Conclusions. The method we proposed could provide insight into the investigation of rounded leaf end effects on penumbra characteristics. Penumbra width and radiation field offset calibration should be carefully performed to commission multileaf collimator for intensity modulated radiotherapy.

Background. This study was designed to compare the quality assurance (QA) results of four dosimetric tools used for intensity modulated radiation therapy (IMRT) and to suggest universal criteria for the passing rate in QA, irrespective of the dosimetric tool used.

Materials and methods. Thirty fields of IMRT plans from five patients were selected, followed by irradiation onto radiochromic film, a diode array (Mapcheck), an ion chamber array (MatriXX) and an electronic portal imaging device (EPID) for patient-specific QA. The measured doses from the four dosimetric tools were compared with the dose calculated by the treatment planning system. The passing rates of the four dosimetric tools were calculated using the gamma index method, using as criteria a dose difference of 3% and a distance-to-agreement of 3 mm.

Results. The QA results based on Mapcheck, MatriXX and EPID showed good agreement, with average passing rates of 99.61%, 99.04% and 99.29%, respectively. However, the average passing rate based on film measurement was significantly lower, 95.88%. The average uncertainty (1 standard deviation) of passing rates for 6 intensity modulated fields was around 0.31 for film measurement, larger than those of the other three dosimetric tools.

Conclusions. QA results and consistencies depend on the choice of dosimetric tool. Universal passing rates should depend on the normalization or inter-comparisons of dosimetric tools if more than one dosimetric tool is used for patient specific QA.