Navigating challenges: optimising methods for primary cell culture isolation.

Primary cell lines are invaluable for exploring cancer biology and investigating novel treatments. Despite their numerous advantages, primary cultures are laborious to obtain and maintain in culture. Hence, established cell lines are still more common. This study aimed to evaluate a range of techniques for isolating primary breast cancer cultures, employing distinct enzymatic compositions, incubation durations, and mechanical approaches, including filtration. Out of several protocols, we opted for a highly effective method (Method 5) that gave rise to a primary cell culture (BC160). This method combines mechanical disaggregation and enzymatic digestion with hyaluronidase and collagenase. Moreover, the paper addresses common issues in isolating primary cultures, shedding light on the struggle against fibroblasts overgrowing cancer cell populations. To make primary cell lines a preferred model, it is essential to elaborate and categorise isolation methods, develop approaches to separate heterogeneous cultures and investigate factors influencing the establishment of primary cell lines.

Primary cancer-associated fibroblasts exhibit high heterogeneity among breast cancer subtypes.

Background: Cancer-associated fibroblasts (CAFs) are a diverse subset of cells, that is recently gaining in popularity and have the potential to become a new target for breast cancer (BC) therapy; however, broader research is required to understand their mechanisms and interactions with breast cancer cells. The goal of the study was to isolate CAFs from breast cancer tumour and characterise isolated cell lines. We concentrated on numerous CAF biomarkers that would enable their differentiation. Materials and methods: Flow cytometry, immunofluorescence, and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) were used to phenotype the primary CAFs. Results/Conclusions: According to our findings, there was no significant pattern in the classification of cancer-associated fibroblasts. The results of biomarkers expression were heterogeneous, thus no specific subtypes were identified. Furthermore, a comparison of cancer-associated fibroblasts derived from different BC subtypes (luminal A and B, triple-negative, HER2 positive) did not reveal any clear trend of expression.

IROCA-TES: Improving Quality in Radiation Oncology through Clinical Audits - Training and Education for Standardization.

Background: Clinical audits are an important tool to objectively assess clinical protocols, procedures, and processes and to detect deviations from good clinical practice. The main aim of this project is to determine adherence to a core set of consensus- based quality indicators and then to compare the institutions in order to identify best practices. Materials and methods: We conduct a multicentre, international clinical audit of six comprehensive cancer centres in Poland, Spain, Italy, Portugal, France, and Romania as a part of the project, known as IROCATES (Improving Quality in Radiation Oncology through Clinical Audits - Training and Education for Standardization). Results: Radiotherapy practice varies from country to country, in part due to historical, economic, linguistic, and cultural differences. The institutions developed their own processes to suit their existing clinical practice. Conclusions: We believe that this study will contribute to establishing the value of routinely performing multi-institutional clinical audits and will lead to improvement of radiotherapy practice at the participating centres.

Development of a quasi-humanoid phantom to perform dosimetric and radiobiological measurements for out-of-field doses from external beam radiation therapy.

Our understanding of low dose, out-of-field radiation and their radiobiological effects are limited, in part due to the rapid technological advances in external beam radiotherapy, especially for non-coplanar and dynamic techniques. Reliable comparisons of out-of-field doses produced by advanced radiotherapy techniques are difficult due to the limitations of commercially available phantoms. There is a clear need for a functional phantom to accurately measure the dosimetric and radiobiological characteristics of out-of-field doses, which would in turn allow clinicians and medical physicists to optimize treatment parameters. We designed, manufactured, and tested the performance of a quasi-humanoid (Q-H) adult phantom. To test the physics parameters, we used computed tomography (CT) scans of assembled Q-H phantom. Static open field and dynamic techniques were measured both in- and out-of-field with ionization chambers and radiochromic films for two configurations (full solid and with water-filled containers). In the areas simulating soft tissues, lung, and bones, median Hounsfield units and densities were, respectively: 129.8, -738.7, 920.8 HU and 1.110, 0.215, 1.669 g/cm3 . Comparison of the measured to treatment planning systems (TPS) in-field dose values for the sample volumetric arc therapy (VMAT) (6 MV flattening filter-free (FFF)) plan, 96.4% of analyzed points passed the gamma evaluation criteria (L2%/2 mm, threshold (TH) 10%) and less than 1.50% for point dose verification. In the two phantom configurations: full poly(methyl) methacrylate (PMMA) and with water container, the off-axis median doses for open field, relative to the central axis of the beam (CAX) were similar, respectively: 0.900% versus 0.907% (15 cm distance to CAX); 0.096% versus 0.120% (35 cm); 0.018% versus 0.018% (52 cm); 0.009% versus 0.008% (74 cm). For VMAT 6 MV FFF, doses relative the CAX were, respectively: 0.667% (15 cm), 0.062% (35 cm), 0.019% (52 cm), 0.016% (74 cm). The Q-H phantom meets the International Commission on Radiation Units and Measurements (ICRU) and American Association of Physicists in Medicine (AAPM) recommended phantom criteria, providing medical physicists with a reliable, comprehensive system to perform dose calculation and measurements and to assess the impact on radiobiological response and on the risk of secondary tumor induction.

Treatment-integrated imaging, radiomics, and personalised radiotherapy: the future is at hand.

Since the introduction of computed tomography for planning purposes in the 1970s, we have been observing a continuous development of different imaging methods in radiotherapy. The current achievements of imaging technologies in radiotherapy enable more than just improvement of accuracy on the planning stage. Through integrating imaging with treatment machines, they allow advanced control methods of dose delivery during the treatment. This article reviews how the integration of existing and novel forms of imaging changes radiotherapy and how these advances can allow a more individualised approach to cancer therapy. We believe that the significant challenge for the next decade is the continued integration of a range of different imaging devices into linear accelerators. These imaging modalities should show intra-fraction changes in body morphology and inter-fraction metabolic changes. As the use of these more advanced, integrated machines grows, radiotherapy delivery will become more accurate, thus resulting in better clinical outcomes: higher cure rates with fewer side effects.

FLASH radiotherapy: an emerging approach in radiation therapy.

FLASH radiotherapy (RT) is a technique involving the delivery of ultra-high dose rate radiation to the target. FLASH-RT has been shown to reduce radiation-induced toxicity in healthy tissues without compromising the anti-cancer effects of treatment compared to conventional radiation therapy. In the present article, we review the published data on FLASH-RT and discuss the current state of knowledge of this novel approach. We also highlight the technological constraints and complexity of FLASH-RT and describe the physics underlying this modality, particularly how technology supports energy transfer by ionising radiation (e.g., beam on/off sequence, pulse-energy load, intervals). We emphasise that current preclinical experience is mostly based on FLASH electrons and that clinical application of FLASH-RT is very limited. The incorporation of FLASH-RT into routine clinical radiotherapy will require the development of devices capable of producing FLASH photon beams.

The impact of different optimization strategies on the agreement between planned and delivered doses during volumetric modulated arc therapy for total marrow irradiation.

AIM OF THE STUDY: To evaluate the agreement between planned and delivered doses and its potential correlation with the plans' complexity subjected to dosimetric verification. MATERIAL AND METHODS: Four isocentre volumetric modulated arc therapy for total marrow irradiation plans optimized simultaneously with (P1) and without (P2) MU reduction were evaluated dosimetrically by γ method performed in a global mode for 4 combinations of γ-index criteria (2%/2 mm, 2%/3 mm, 3%/2 mm, and 3%/3 mm). The evaluation was conducted for 4 regions (head and neck, chest, abdomen and upper pelvis, and lower pelvis and thighs) that were determined geometrically by the isocentres. The Wilcoxon test was used to detect significant differences between γ passing rate (GPR) analysis results for the P1 and P2 plans. The Pearson correlation was used to check the relationship between GPR and the plans' complexity. RESULTS: Except for the head and neck region, the P2 plans had better GPRs than the P1 plans. Only for hard combinations of γ-index criteria (i.e. 2%/3 mm, 2%/2 mm) were the GPRs differences between P1 and P2 clinically meaningful, and they were detected in the chest, abdomen and upper pelvis, and lower pelvis and thighs regions. The highest correlations between GPR and the indices describing the plans' complexity were found for the chest region. No correlation was found for the head and neck region. CONCLUSIONS: The P2 plans showed better agreement between planned and delivered doses compared to the P1 plans. The GPR and the plans' complexity depend on the anatomy region and are most important for the chest region.

Differences between TNM classification and 2-[18F]FDG PET parameters of primary tumor in NSCLC patients.

BACKGROUND: The aim of the study was to compare the TNM classification with 2-[18F]FDG PE T biological parameters of primary tumor in patients with NSCLC. MATERIALS AND METHODS: Retrospective analysis was performed on a group of 79 newly diagnosed NSCLC patients. PET scans were acquired on Gemini TF PET/CT scanner 60-70 min after injection of 2-[18F]FDG with the mean activity of 364 ± 75 MBq, with the area being examined from the vertex to mid-thigh. The reconstructed PET images were evaluated using MIM 7.0 Software for SUVmax, MTV and TLG values. RESULTS: The analysis of the cancer stage according to TNM 8th edition showed stage IA2 in 8 patients, stage IA3 - 6 patients, stage IB - 4 patients, IIA - 3 patients, 15 patients with stage IIB, stage IIIA - 17 patients, IIIB - 5, IIIC - 5, IVA in 7 patients and stage IVB in 9 patients. The lowest TLG values of primary tumor were observed in stage IA2 (11.31 ± 15.27) and the highest in stage IIIC (1003.20 ± 953.59). The lowest value of primary tumor in SUVmax and MTV were found in stage IA2 (6.8 ± 3.8 and 1.37 ± 0.42, respectively), while the highest SUVmax of primary tumor was found in stage IIA (13.4 ± 11.4) and MTV in stage IIIC (108.15 ± 127.24). CONCLUSION: TNM stages are characterized by different primary tumor 2-[18F]FDG PET parameters, which might complement patient outcome.

Verification of electron beam parameters in an intraoperative linear accelerator using dosimetric and radiobiological response methods.

BACKGROUND: The availability of linear accelerators (linac) for research purposes is often limited and therefore alternative radiation sources are needed to conduct radiobiological research. The National Centre for Radiation Research in Poland recently developed an intraoperative mobile linac that enables electron irradiation at energies ranging from 4 to 12 MeV and dose rates of 5 or 10 Gy/min. The present study was conducted to evaluate the electron beam parameters of this intraoperative linac and to verify the set-up to evaluate out-of-field doses in a water phantom, which were determined through dosimetric and biological response measurements. MATERIALS AND METHODS: The distribution of radiation doses along and across the radiation beam were measured in a water phantom using a semiconductor detector and absolute doses using an ionisation chamber. Two luminal breast cancer cell lines (T-47D and HER2 positive SK-BR-3) were placed in the phantom to study radiation response at doses ranging from 2 to 10 Gy. Cell response was measured by clonogenic assays. RESULTS AND CONCLUSION: The electron beam properties, including depth doses and profiles, were within expected range for the stated energies. These results confirm the viability of this device and set-up as a source of megavoltage electrons to evaluate the radiobiological response of tumour cells.

Cancer incidence in the Greater Poland region as compared to Europe.

Greater Poland is a region with a high risk of cancer. In terms of age-standardised incidence rate, it is ranked 2nd for men and 3rd for women out of Poland's 16 provinces. Incidence structure in the region of Greater Poland is similar to that in other West European countries. The most common cancers in men are lung, prostate and colorectal, in women: breast, colorectal and lung. In 2016, nearly every third cancer-related death in the region was caused by lung cancer. In women, it was cause no. one. The incidence of chronic diseases, including cancer, is expected to further grow in view of the global ageing of the population. This means that malignancies will remain to be a major challenge for public health care.in the Greater Poland region.

Impact of different optimization strategies on the compatibility between planned and delivered doses during radiation therapy of cervical cancer.

PURPOSE: To analyse the impact of different optimization strategies on the compatibility between planned and delivered doses during radiotherapy of cervical cancer. MATERIAL/METHODS: Four treatment plans differing in optimisation strategies were prepared for ten cervical cancer cases. These were: volumetric modulated arc therapy with (_OPT) and without optimization of the doses in the bone marrow and for two sets of margins applied to the clinical target volume that arose from image guidance based on the bones (IG(B)) and soft tissues (IG(ST)). The plans were subjected to dosimetric verification by using the ArcCHECK system and 3DVH software. The planned dose distributions were compared with the corresponding measured dose distributions in the light of complexity of the plans and its deliverability. RESULTS: The clinically significant impact of the plans complexity on their deliverability is visible only for the gamma passing rates analysis performed in a local mode and directly in the organs. While more general analyses show statistically significant differences, the clinical relevance of them has not been confirmed. The analysis showed that IG(ST)_OPT and IG(B)_OPT significantly differ from IG(ST) and IG(B). The clinical acceptance of IG(ST)_OPT obtained for hard combinations of gamma acceptance criteria (2%/2 mm) confirm its satisfactory deliverability. In turn, for IG(B)_OPT in the case of the rectum, the combination of 2%/2 mm did not meet the criteria of acceptance. CONCLUSION: Despite the complexity of the IG(ST)_OPT, the results of analysis confirm the acceptance of its deliverability when 2%/2 mm gamma acceptance criteria are used during the analysis.

Calculation and measurement of doses in the surface layers of a phantom when using Tomotherapy.

BACKGROUND: The calculation and measurement on the surface of the skin presents a significant dosimetric problem because of numerous factors which have an influence on the dose distribution in this region. AIM: The overall aim of this study was to check the agreement between doses measured with thermoluminescent detectors (TLD) during tomotherapy photon beam irradiation of the skin area of a solid water cylindrical phantom with doses calculated with Hi-Art treatment planning system (TPS). MATERIAL AND METHOD: The measurements of the dose were made with the use of a solid water cylindrical phantom - Cheese Phantom. Two bolus phantoms were used: 5 mm and 10 mm Six different planning treatments were generated. The doses were measured using TL detectors. RESULTS: In the case of a tumor located near the surface of the skin, the mean dose for 0.5 cm bolus was - 1.94 Gy, and for 1 cm bolus - 2.03 Gy. For the tumor located inside the phantom and organ at risk on the same side that TL detectors, for a 0.5 cm bolus, mean dose was 0.658 Gy, and for a 1 cm bolus, 0.62 Gy. CONCLUSION: The analysis of results showed that the relative percentage difference between measured and planned dose in the field of irradiation was less than 10%, while the largest differences were on the board of the field of radiation and outside of the field of irradiation, where the dose was 0.08 Gy to 1 Gy.

Carcinogenesis Induced by Low-dose Radiation.

BACKGROUND: Although the effects of high dose radiation on human cells and tissues are relatively well defined, there is no consensus regarding the effects of low and very low radiation doses on the organism. Ionizing radiation has been shown to induce gene mutations and chromosome aberrations which are known to be involved in the process of carcinogenesis. The induction of secondary cancers is a challenging long-term side effect in oncologic patients treated with radiation. Medical sources of radiation like intensity modulated radiotherapy used in cancer treatment and computed tomography used in diagnostics, deliver very low doses of radiation to large volumes of healthy tissue, which might contribute to increased cancer rates in long surviving patients and in the general population. Research shows that because of the phenomena characteristic for low dose radiation the risk of cancer induction from exposure of healthy tissues to low dose radiation can be greater than the risk calculated from linear no-threshold model. Epidemiological data collected from radiation workers and atomic bomb survivors confirms that exposure to low dose radiation can contribute to increased cancer risk and also that the risk might correlate with the age at exposure. CONCLUSIONS: Understanding the molecular mechanisms of response to low dose radiation is crucial for the proper evaluation of risks and benefits that stem from these exposures and should be considered in the radiotherapy treatment planning and in determining the allowed occupational exposures.

Improving radiation oncology through clinical audits: Introducing the IROCA project.

As radiotherapy practice and processes become more complex, the need to assure quality control becomes ever greater. At present, no international consensus exists with regards to the optimal quality control indicators for radiotherapy; moreover, few clinical audits have been conducted in the field of radiotherapy. The present article describes the aims and current status of the international IROCA "Improving Radiation Oncology Through Clinical Audits" project. The project has several important aims, including the selection of key quality indicators, the design and implementation of an international audit, and the harmonization of key aspects of radiotherapy processes among participating institutions. The primary aim is to improve the processes that directly impact clinical outcomes for patients. The experience gained from this initiative may serve as the basis for an internationally accepted clinical audit model for radiotherapy.

Medical physics in radiotherapy: The importance of preserving clinical responsibilities and expanding the profession's role in research, education, and quality control.

Medical physicists have long had an integral role in radiotherapy. In recent decades, medical physicists have slowly but surely stepped back from direct clinical responsibilities in planning radiotherapy treatments while medical dosimetrists have assumed more responsibility. In this article, I argue against this gradual withdrawal from routine therapy planning. It is essential that physicists be involved, at least to some extent, in treatment planning and clinical dosimetry for each and every patient; otherwise, physicists can no longer be considered clinical specialists. More importantly, this withdrawal could negatively impact treatment quality and patient safety. Medical physicists must have a sound understanding of human anatomy and physiology in order to be competent partners to radiation oncologists. In addition, they must possess a thorough knowledge of the physics of radiation as it interacts with body tissues, and also understand the limitations of the algorithms used in radiotherapy. Medical physicists should also take the lead in evaluating emerging challenges in quality and safety of radiotherapy. In this sense, the input of physicists in clinical audits and risk assessment is crucial. The way forward is to proactively take the necessary steps to maintain and advance our important role in clinical medicine.

Challenges in organizing effective oncology service: inter-European variability in the example of head and neck cancers.

The increasing worldwide burden of cancer makes it imperative that every country develop a comprehensive cancer control programme. In the past, cancer control in Central and Eastern Europe was inadequate, particularly when compared to many wealthier Western European countries. We analyse interregional differences in Europe to the approach to comprehensive cancer care, with a focus on head and neck squamous cell carcinoma using the case of Poland as a representative example. Due to national plans major improvements have been achieved in the field of prevention and in radiotherapy delivery having a measurable and positive impact on treatment outcomes. In head and neck cancers a notable move towards multidisciplinary approach has been made, combining surgery, radiotherapy and chemotherapy accompanied by rehabilitation and social support. In Poland and several other Eastern and Central European countries a shortage of physicians in the field of oncology was noted. The main conclusion is that the special plans are needed in Central and Eastern Europe or those existing must be extended for another decade to fulfil the EU requirement of providing all European citizens with equal access to quality cancer care.

Preoperative radiotherapy for rectal cancer: a comparative study of quality control adherence at two cancer hospitals in Spain and Poland.

BACKGROUND: We performed a clinical audit of preoperative rectal cancer treatment at two European radiotherapy centres (Poland and Spain). The aim was to independently verify adherence to a selection of indicators of treatment quality and to identify any notable inter-institutional differences. METHODS: A total of 162 patients, in Catalan Institute of Oncology (ICO) 68 and in Greater Poland Cancer Centre (GPCC) 94, diagnosed with locally advanced rectal cancer and treated with preoperative radiotherapy or radio-chemotherapy were included in retrospective study. A total of 7 quality control measures were evaluated: waiting time, multidisciplinary treatment approach, portal verification, in vivo dosimetry, informed consent, guidelines for diagnostics and therapy, and patient monitoring during treatment. RESULTS: Several differences were observed. Waiting time from pathomorphological diagnosis to initial consultation was 31 (ICO) vs. 8 (GPCC) days. Waiting time from the first visit to the beginning of the treatment was twice as long at the ICO. At the ICO, 82% of patient experienced treatment interruptions. The protocol for portal verification was the same at both institutions. In vivo dosimetry is not used for this treatment localization at the ICO. The ICO utilizes locally-developed guidelines for diagnostics and therapy, while the GPCC is currently developing its own guidelines. CONCLUSIONS: An independent external clinical audit is an excellent approach to identifying and resolving deficiencies in quality control procedures. We identified several procedures amenable to improvement. Both institutions have since implemented changes to improve quality standards. We believe that all radiotherapy centres should perform a comprehensive clinical audit to identify and rectify deficiencies.

Oral cavity and oropharyngeal squamous cell carcinoma in young adults: a review of the literature.

BACKGROUND: Head and neck squamous cell carcinoma (HNSCC) is a disease of middle-aged to elderly adults. However, an increased incidence of HNSCC in young people under 45 years of age has been reported recently. In the present review, we focused on the epidemiology and aetiology of HNSCC in adults under 45 years of age. METHODS: We reviewed literature related to HNSCC in adult patients less than 45 years of age and discussed current treatment options and prognosis. RESULTS: HNSCC in young adults is associated with a higher incidence rate in nonsmokers, lower female-to-male ratio, a higher percentage of oral cavity and oropharynx tumours, and fewer second primary tumours. However, aside from traditional risk factors of tobacco and alcohol exposure, the causes of these cancers in young adults remain unclear. Agents that might contribute to risk include infection with high-risk human papillomavirus subtypes as well as genetic factors or immunodeficiency status. The expected increase in incidence and mortality of the young with HNSCC may become a major public health concern if current trends persist, particularly lifestyle habits that may contribute to this disease. CONCLUSIONS: Given the younger age and potential long-term adverse sequelae of traditional HNSCC treatments, young adults should be treated on a case-by-case basis and post-therapy quality of life must be considered in any treatment-decision making process.

Cancer incidence and mortality in the Greater Poland Region-Analysis of the year 2010 and future trends.

BACKGROUND AND AIM: The Greater Poland Region is one of the most industrialised areas of Poland, with a high rate of cancer incidence and mortality. The present report estimated incidence and mortality data for Greater Poland in the year 2010. METHODS: Statistical reports in this study include absolute number of cases and crude incidence rates. The derived age-, sex-, and site specific rates were age-standardised (ASRs per 100,000 person-years) using the European (ASRE) standard population. RESULTS: In 2010, a total 13,581 new cancer cases were reported to the Greater Poland Cancer Registry. The number of new cases increased by 24% compared to 2001. Greater Poland has the second-highest ASR for both females and males among the 16 regions in Poland. The most common cancers are similar to those in other Western European countries. Among men, the most common cancers are lung (C34), colorectal (C18-C21), and prostate (C61) cancer. In women, breast cancer is the most common (C50), followed by colon (C18-C21) and lung (C34) cancer. Lung cancer in males accounts for more than one-third of all cancer-related deaths in Greater Poland. As in 2009, lung cancer is the leading cause of death in women. CONCLUSIONS: Given the ageing of the population, the incidence of chronic diseases, including cancer, is expected to grow. These data indicate that cancer will continue to represent an important challenge both to local health authorities and the National Health Fund, which will need to meet the growing demand for cancer care.

Wax boluses and accuracy of EBT and RTQA radiochromic film detectors in radiotherapy with the JINR Phasotron proton beam.

AIM: To present the results obtained using radiochromic films EBT and RTQA 1010P for the reconstruction the dose distributions for targets irradiated by proton beam and modified by wax boluses. BACKGROUND: In Medico-Technical Complex at the Joint Institute for Nuclear Research in Dubna implemented technology of wax boluses. MATERIALS AND METHODS: Wax boluses are easier to make and they give better dose distributions than boluses made from modeling clay previously used at our center. We irradiated two imaginary targets, one shaped as a cylinder and the other one as two cuboids. The evaluated calibration curve was used for calculation of the dose distributions measured by the EBT and RTQA radiochromic film. In both cases, the measured dose distributions were compared to the dose distributions calculated by the treatment planning system (TPS). We also compared dose distributions using three different conformity indices at a 95% isodose. RESULTS: Better target coverage and better compliance of measurements (semiconductor detectors and radiochromic films) with calculated doses was obtained for cylindrical target than for cuboidal target. The 95% isodose covered well the tumor for both target shapes, while for cuboidal target larger volume around the target received therapeutic dose, due to the complicated target shape. The use wax boluses provided to be effective tool in modifying proton beam to achieve appropriate shape of isodose distribution. CONCLUSION: EBT film yielded the best visual matching. Both EBT and RTQA films confirmed good conformity between calculated and measured doses, thus confirming that wax boluses used to modify the proton beam resulted in good dose distributions.