Applicability of Gene Expression in Saliva as an Alternative to Blood for Biodosimetry and Prediction of Radiation-induced Health Effects.

As the great majority of gene expression (GE) biodosimetry studies have been performed using blood as the preferred source of tissue, searching for simple and less-invasive sampling methods is important when considering biodosimetry approaches. Knowing that whole saliva contains an ultrafiltrate of blood and white blood cells, it is expected that the findings in blood can also be found in saliva. This human in vivo study aims to examine radiation-induced GE changes in saliva for biodosimetry purposes and to predict radiation-induced disease, which is yet poorly characterized. Furthermore, we examined whether transcriptional biomarkers in blood can also be found equivalently in saliva. Saliva and blood samples were collected in parallel from radiotherapy (RT) treated patients who suffered from head and neck cancer (n = 8) undergoing fractioned partial-body irradiations (1.8 Gy/fraction and 50-70 Gy total dose). Samples were taken 12-24 h before first irradiation and ideally 24 and 48 h, as well as 5 weeks after radiotherapy onset. Due to the low quality and quantity of isolated RNA samples from one patient, they had to be excluded from further analysis, leaving a total of 24 saliva and 24 blood samples from 7 patients eligible for analysis. Using qRT-PCR, 18S rRNA and 16S rRNA (the ratio being a surrogate for the relative human RNA/bacterial burden), four housekeeping genes and nine mRNAs previously identified as radiation responsive in blood-based studies were detected. Significant GE associations with absorbed dose were found for five genes and after the 2nd radiotherapy fraction, shown by, e.g., the increase of CDKN1A (2.0 fold, P = 0.017) and FDXR (1.9 fold increased, P = 0.002). After the 25th radiotherapy fraction, however, all four genes (FDXR, DDB2, POU2AF1, WNT3) predicting ARS (acute radiation syndrome) severity, as well as further genes (including CCNG1 [median-fold change (FC) = 0.3, P = 0.013], and GADD45A (median-FC = 0.3, P = 0.031)) appeared significantly downregulated (FC = 0.3, P = 0.01-0.03). A significant association of CCNG1, POU2AF1, HPRT1, and WNT3 (P = 0.006-0.04) with acute or late radiotoxicity could be shown before the onset of these clinical outcomes. In an established set of four genes predicting acute health effects in blood, the response in saliva samples was similar to the expected up- (FDXR, DDB2) or downregulation (POU2AF1, WNT3) in blood for up to 71% of the measurements. Comparing GE responses (PHPT1, CCNG1, CDKN1A, GADD45A, SESN1) in saliva and blood samples, there was a significant linear association between saliva and blood response of CDKN1A (R2 = 0.60, P = 0.0004). However, the GE pattern of other genes differed between saliva and blood. In summary, the current human in vivo study, (I) reveals significant radiation-induced GE associations of five transcriptional biomarkers in salivary samples, (II) suggests genes predicting diverse clinical outcomes such as acute and late radiotoxicity as well as ARS severity, and (III) supports the view that blood-based GE response can be reflected in saliva samples, indicating that saliva is a "mirror of the body" for certain but not all genes and, thus, studies for each gene of interest in blood are required for saliva.

Examining potential confounding factors in gene expression analysis of human saliva and identifying potential housekeeping genes.

Isolation of RNA from whole saliva, a non-invasive and easily accessible biofluid that is an attractive alternative to blood for high-throughput biodosimetry of radiological/nuclear victims might be of clinical significance for prediction and diagnosis of disease. In a previous analysis of 12 human samples we identified two challenges to measuring gene expression from total RNA: (1) the fraction of human RNA in whole saliva was low and (2) the bacterial contamination was overwhelming. To overcome these challenges, we performed selective cDNA synthesis for human RNA species only by employing poly(A)+-tail primers followed by qRT-PCR. In the current study, this approach was independently validated on 91 samples from 61 healthy donors. Additionally, we used the ratio of human to bacterial RNA to adjust the input RNA to include equal amounts of human RNA across all samples before cDNA synthesis, which then ensured comparable analysis using the same base human input material. Furthermore, we examined relative levels of ten known housekeeping genes, and assessed inter- and intra-individual differences in 61 salivary RNA isolates, while considering effects of demographical factors (e.g. sex, age), epidemiological factors comprising social habits (e.g. alcohol, cigarette consumption), oral hygiene (e.g. flossing, mouthwash), previous radiological diagnostic procedures (e.g. number of CT-scans) and saliva collection time (circadian periodic). Total human RNA amounts appeared significantly associated with age only (P ≤ 0.02). None of the chosen housekeeping genes showed significant circadian periodicity and either did not associate or were weakly associated with the 24 confounders examined, with one exception, 60% of genes were altered by mouthwash. ATP6, ACTB and B2M represented genes with the highest mean baseline expression (Ct-values ≤ 30) and were detected in all samples. Combining these housekeeping genes for normalization purposes did not decrease inter-individual variance, but increased the robustness. In summary, our work addresses critical confounders and provides important information for the successful examination of gene expression in human whole saliva.

Rare Hereditary Burden associated with a Hypercalcemic Small-Cell Carcinoma of Cervix in a Young Female Patient.

BACKGROUND: Oncological diseases have, in most cases, a multifactorial etiology, composed of a combination of external and internal environmental factors. Hereditary tumorous syndromes are mostly autosomal dominant diseases with incomplete but very high penetrance. OBSERVATION: The patient, an 18-year-old virgin female, consulted a gynecologist in June 2018 because of metrorrhagia. Magnetic resonance imaging revealed a cervical tumor with the dimensions 80 × 90 × 80 mm. Histological analysis confirmed the presence of a very rare hypercalcemic type of small-cell carcinoma of the cervix. Further investigation of the germinal exom of the patient showed pathological variations in genes PALB2 and BRCA2, presented with recommendation of detailed examination by medical genetics. CONCLUSION: Clinical experience with this type of tumor is very limited, but it still comes with some useful outcome. Small cell carcinomas of the gynecologic tract are very rare, aggressive diseases, with very poor prognosis, affecting mainly young women. Their origin is most often the ovaries, based on most clinical data, but these tumor also localize to the endometrium, cervix, vagina and vulva. It is an extremely rare type of cancer, for which clinical data is scant due to the extremely low number of reported cases. In this patient, the carcinoma had an unusual genetical mutation burden, which she inherited from her parents. In the light of these findings, we recommend that patients suspected of having a small-cell of the gynecologic tract provide a detailed family history, and that genetic testing be considered in similar cases. This work was supported by MH CR grant 16-33209A and research program of Charles University Progress Q40/06. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 10. 6. 2019 Accepted: 9. 9. 2019.

High-Dose Rate Brachyther-apy in the Treatment of Early Stages of Penile Carcinoma.

BACKGROUND: Interstitial low dose rate brachyther-apy is established organ spar-ing treatment of T1- T2 penile carcinoma. Experience with high-dose rate brachyther-apy is limited in this indication. MATERIALS AND METHODS: Twenty-six patients with early penile carcinoma were treated by high-dose rate brachyther-apy at dose 18 × 3 Gy per fraction twice daily between 2002- 2018 at the Department of Oncology and Radiother-apy, University Hospital in Hradec Kralove. Breast interstitial brachyther-apy template was used for fixation and precise geometry reconstruction of stainless hollow needles. RESULTS: Median follow up was 85 months (range 7- 200 months). Acute reaction usually consisted of grade 2 mucositis that dissolved dur-ing 8 weeks after the treatment. Local recurrence occurred in 6 patients, 5 of them were successfully treated with partial amputation. One patient had a nodal recurrence successfully salvaged by lymphadenectomy. One patient developed necrosis of the glans requir-ing partial amputation. Currently, there are 24 patients alive without signs of dis-ease. One patient died of cardiac comorbidity, one died of duplicate lung cancer. Nineteen patients have a preserved penis (73%), 18 of them sexually active before treatment report satisfactory intercourse. CONCLUSION: Hyperfractionated interstitial high-dose rate brachyther-apy with 18 × 3 Gy per fraction twice daily is a promis-ing method in selected patients with penile carcinoma and deserves further evaluation in a larger prospective study. Key words penile neoplasms -  conservative treatment -  brachyther-apy This work was supported by programm Progres Q40. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 8. 1. 2019 Accepted: 15. 1. 2019.

HPV Status and Mutation Analysis Using Multiparallel Sequencing in Distal Oesophageal and Gastro-oesophageal Junction Adenocarcinomas.

The incidence of adenocarcinoma of oesophagus or gastro-oesophageal junction is increasing in Europe and other regions of the Western world. Research of possible causes has shifted to the molecular level. This study evaluated human papillomavirus (HPV) using real-time PCR and mutational status of selected genes using the multiparallel sequencing method (NGS) in DNA extracted from paraffin-embedded tumour tissue of 56 patients with oesophageal or gastro-oesophageal junction adenocarcinoma. The genetic material was in sufficient quality for the analysis in 37 cases (66 %). No HPV-positive sample was found. NGS revealed higher frequency of mutations in TP53, ARID1A, PIK3CA, SMAD4, ERBB2, MSH6, BRCA2, and RET genes. Association between gene mutations and histological grade, subtype according to Lauren, or primary tumour site was not statistically significant. In conclusion, the study did not confirm any HPV-positive sample of oesophageal and gastro-oesophageal junction adenocarcinoma. The study confirmed the usefulness of NGS analysis of paraffin-embedded tissue of these tumours, and it could be used in clinical studies to evaluate the prognostic and/or predictive value of the tested mutations. The association between gene mutations and histological features should be tested in larger patient cohorts.

Validating Baboon Ex Vivo and In Vivo Radiation-Related Gene Expression with Corresponding Human Data.

The research for high-throughput diagnostic tests for victims of radio/nuclear incidents remains ongoing. In this context, we have previously identified candidate genes that predict risk of late-occurring hematologic acute radiation syndrome (HARS) in a baboon model. The goal of the current study was to validate these genes after radiation exposure in humans. We also examined ex vivo relative to in vivo measurements in both species and describe dose-response relationships. Eighteen baboons were irradiated in vivo to simulate different patterns of partial- or total-body irradiation (TBI), corresponding to an equivalent dose of 2.5 or 5 Sv. Human in vivo blood samples were obtained from patients exposed to different dose ranges: diagnostic computerized tomography (CT; 0.004-0.018 Sv); radiotherapy for prostate cancer (0.25-0.3 Sv); and TBI of leukemia patients (2 × 1.5 or 2 × 2 Sv, five patients each). Peripheral whole blood of another five baboons and human samples from five healthy donors were cultivated ex vivo and irradiated with 0-4 Sv. RNA was isolated pairwise before and 24 h after irradiation and converted into cDNA. Gene expression of six promising candidate genes found previously by us in a baboon model ( WNT3, POU2AF1, CCR7, ARG2, CD177, WLS), as well as three genes commonly used in ex vivo whole blood experiments ( FDXR, PCNA, DDB2) was measured using qRT-PCR. We confirmed the six baboon candidate genes in leukemia patients. However, expression for the candidate gene FDXR showed an inverse relationship, as it was downregulated in baboons and upregulated in human samples. Comparisons among the in vivo and ex vivo experiments revealed the same pattern in both species and indicated peripheral blood cells to represent the radiation-responsive targets causing WNT3 and POU2AF1 gene expression changes. CCR7, ARG2, CD177 and WLS appeared to be altered due to radiation-responsive targets other than the whole blood cells. Linear dose-response relationships of FDXR, WNT3 and POU2AF1 using human ex vivo samples corresponded with human in vivo samples, suggesting that ex vivo models for in vivo dose estimates can be used over a wide dose range (0.001-5 Sv for POU2AF1). In summary, we validated six baboon candidate genes in humans, but the FDXR measurements underscored the importance of independent assessments even when candidates from animal models have striking gene sequence homology to humans. Since whole blood cells represented the same radiation-responsive targets for FDXR, WNT3 and POU2AF1 gene expression changes, ex vivo cell culture models can be utilized for in vivo dose estimates over a dose range covering up to 3.5 log scales. These findings might be a step forward in the development of a gene expression-based high-throughput diagnostic test for populations involved in large-scale radio/nuclear incidents.

Analysis of DNA methylation and microRNA expression in NUT (nuclear protein in testis) midline carcinoma of the sinonasal tract: a clinicopathological, immunohistochemical and molecular genetic study.

The aim of this study was a detailed clinicopathological investigation of sinonasal NUT midline carcinoma (NMC), including analysis of DNA methylation and microRNA (miRNA) expression. Three (5%) cases of NMC were detected among 56 sinonasal carcinomas using immunohistochemical screening and confirmed by fluorescence in situ hybridization. The series comprised 2 males and 1 female, aged 46, 60, and 65 years. Two tumors arose in the nasal cavity and one in the maxillary sinus. The neoplasms were staged pT1, pT3, and pT4a (all cN0M0). All patients were treated by radical resection with adjuvant radiotherapy. Two patients died 3 and 8 months after operation, but one patient (pT1 stage; R0 resection) experienced no evidence of disease at 108 months. Microscopically, all tumors consisted of infiltrating nests of polygonal cells with vesicular nuclei, prominent nucleoli and basophilic cytoplasm. Abrupt keratinization was present in only one case. Immunohistochemically, there was a diffuse expression of cytokeratin (CK) cocktail, CK7, p40, p63, and SMARCB1/INI1. All NMCs tested negative for EBV and HPV infection. Two NMCs showed methylation of RASSF1 gene. All other genes (APC, ATM, BRCA1, BRCA2, CADM1, CASP8, CD44, CDH13, CDKN1B, CDKN2A, CDKN2B, CHFR, DAPK1, ESR1, FHIT, GSTP1, HIC1, KLLN, MLH1a, MLH1b, RARB, TIMP3, and VHL) were unmethylated. All NMCs showed upregulation of miR-9 and downregulation of miR-99a and miR-145 and two cases featured also upregulation of miR-21, miR-143, and miR-484. In summary, we described three cases of sinonasal NMCs with novel findings on DNA methylation and miRNA expression, which might be important for new therapeutic strategies in the future.

Importance of Tumour Suppressor Gene Methylation in Sinonasal Carcinomas.

Epigenetic changes are considered to be a frequent event during tumour development. Hypermethylation of promoter CpG islands represents an alternative mechanism for inactivation of tumour suppressor genes, DNA repair genes, cell cycle regulators and transcription factors. The aim of this study was to investigate promoter methylation of specific genes in samples of sinonasal carcinoma by comparison with normal sinonasal tissue. To search for epigenetic events we used methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) to compare the methylation status of 64 tissue samples of sinonasal carcinomas with 19 control samples. We also compared the human papilloma virus (HPV) status with DNA methylation. Using a 20% cut-off for methylation, we observed significantly higher methylation in RASSF1, CDH13, ESR1 and TP73 genes in the sinonasal cancer group compared with the control group. HPV positivity was found in 15/64 (23.4 %) of all samples in the carcinoma group and in no sample in the control group. No correlation was found between DNA methylation and HPV status. In conclusion, our study showed that there are significant differences in promoter methylation in the RASSF1, ESR 1, TP73 and CDH13 genes between sinonasal carcinoma and normal sinonasal tissue, suggesting the importance of epigenetic changes in these genes in carcinogenesis of the sinonasal area. These findings could be used as prognostic factors and may have implications for future individualised therapies based on epigenetic changes.

[Changes in the CD8+ Density of Tumor Infiltrating Lymphocytes after Neoadjuvant Radiochemotherapy in Patients with Rectal Adenocarcinom]. / Vliv predoperacní radiochemoterapie na hustotu CD8+ tumor infiltrujících lymfocytu adenokarcinomu rekta.

BACKGROUND: The goal of this study is to examine the effect of neoadjuvant radiochemotherapy on the density of CD8(+) tumor infiltrating lymphocytes (TILs) in endoscopical biopsies and resection specimens from patients with rectal adenocarcinoma before and after therapy. PATIENTS AND METHODS: In total, 53 patients with locally advanced rectal cancer were studied. RESULTS: The median density of CD8(+) TILs in pretreatment biopsies was 12 (1- 232) and that in surgical specimens after radiochemotherapy was 18 (1- 319). During radiochemotherapy, the density of CD8(+) TILs increased in 30 patients (57%), decreased in 18 (34%), and did not change in one. It was not possible to assess the dynamics of CD8(+) TILs density in four patients. The increased density of CD8(+) TILs after radiochemotherapy was associated with a median survival rate 2.5 times longer than that associated with no increase in density. CONCLUSION: In the present study, the density of CD8(+) TILs in endoscopical biopsies before radiochemotherapy, the density in resection specimens after radiochemotherapy, or in changes in the density after radiochemotherapy showed no predictive or prognostic significance. However, studying a larger number of patients may show that CD8(+) TILs density is of predictive or prognostic significance.

Cancer in the elderly in the Czech Republic.

Population ageing presents a challenge to oncological care due to the particularities of cancer treatment in this population. We evaluate cancer epidemiology, treatment and survival, in the Czech Republic by age groups. Data published by the Czech National Cancer Registry from the years 2006 to 2010 were used for this study. The following cancer types were evaluated: colorectal, pancreatic, head and neck, lung, skin melanoma, breast, gynaecological, prostate, kidney and stomach cancers. The following data were recorded and analysed: crude incidence by 5-year age group; dynamics of crude incidence rates in the age group ≥70 years; disease stage; percentage of patients treated by surgery, radiotherapy and chemotherapy; and age standardised 1-year mortality and 5-year relative survival according to age group. Patients over age 70 accounted for 41% and 46%, respectively, of the cancer incidence and mortality of the whole population. Anticancer therapies are significantly less common in patients over age 70 (P < 0.050), with the exception of skin melanoma. Survival was markedly worse in older patients (P < 0.050) when radical treatment modalities were significantly underused (P < 0.050). In the Czech Republic, the crude cancer incidence in seniors is increasing. In general, elderly patients are undertreated, with worse treatment results compared with younger patients.

[Influence of preoperative chemoradiotherapy on changes of epidermal growth factor receptor expression in patients treated by preoperative chemoradiotherapy for local advanced rectal carcinoma]. / Vliv predoperacní chemoradioterapie na zmenu exprese receptoru pro epidermální rustový faktor u pacientu lécených predoperacní chemoradioterapií pro lokálne pokrocilý karcinom rekta.

AIM: The aim of this retrospective study was to determine the prognostic impact of expression of epidermal growth factor receptor (EGFR) changes during neoadjuvant chemoradiotherapy in patients with locally advanced rectal adenocarcinoma. MATERIAL AND METHODS: One hundred and three patients with locally advanced rectal adenocarcinoma of stage II and III were evaluated. All patients were administered the total dose of 44 --  50.4 Gy. Concomitantly, the patients received capecitabine in the dose 825 mg/ m² in two daily oral administrations or 5- fluorouracil in the dose 200 mg/ m² in continuous infusion. Surgery was indicated at intervals of 4-8 weeks from chemoradiotherapy completion. EGFR expression in the pretreatment biopsies and in resected specimens was assessed with immunohistochemistry. RESULTS: All of 103 patients received radiotherapy without interruption up to the total planned dose. Downstaging was described in 64 patients. Six patients had complete pathologic remission. Recurrence occurred in 49 patients. Local recurrence was found in 22 patients, generalization of disease was reported in 27 patients. A total of 51 patients died. Increased EGFR expression was found in 26 patients. The statistically significantly shorter overall survival (p < 0.001) and disease-free survival (p < 0.001) was found in patients with increased expression of EGFR compared with patients where no increase in the expression of EGFR was observed during neoadjuvant chemoradiotherapy. CONCLUSIONS: The overexpression of EGFR during neoadjuvant chemoradiotherapy for locally advanced rectal adenocarcinoma is associated with significant shorter overall survival and disease-free survival.

ATM and TGFB1 genes polymorphisms in prediction of late complications of chemoradiotherapy in patients with locally advanced cervical cancer.

The purpose of our study was to evaluate a possible correlation between genetic polymorphisms in ATM and TGFB1 genes and late toxicity of chemoradiotherapy for locally advanced cervical cancer. Fifty five patients with FIGO stage IIB and higher without a disease recurrence with a mean follow up of 6 years were included. Late toxicity was assessed by EORTC/RTOG late toxicity criteria. Univariate and multivariate logistic regression model was used for statistical analysis. Degree of association between polymorphisms and late toxicity of chemotherapy was assessed on the basis of phi-coefficient (φ) as well. We did not find any association between 5557G>A polymorphism in the ATM gene or single TGFB1 polymorphisms and late toxicity. TGFB1 compound homozygosity (-1552delAGG, -509C>T, L10P) was a significant predictive factor of grade III-IV and any grade of complications in both univariate and multivariate logistic regression analyses and statistical significance of association between polymorphisms and late toxicity of chemoradiotherapy was confirmed also by the evaluation of phi-coefficient (φ). We conclude that haplotypes instead of single nucleotide polymorphic sites in the genes may better characterize the individual radiosensitivity.

Quantitative evaluation of the benefit of fiducial image-guidance for prostate cancer intensity modulated radiation therapy using daily dose volume histogram analysis.

To quantitatively evaluate the extent to which fiducial-based image-guidance improves dose coverage of the target volume and sparing of critical organs for prostate cancer patients treated with intensity modulated radiotherapy (IMRT) and determination of planning margins by original approach of detailed daily dose volume histogram (DVH) and patient's position correction analysis. Sixty-two patients divided in two groups (clinical target volume (CTV) → planning target volume (PTV) margin 10 and 7 mm) were treated with IMRT using implanted fiducial markers. Each patient's treatment fraction was recalculated as it would have been treated without fiducial-guided positioning. For both plans (IGRT and non-IGRT), equivalent uniform doses (EUD), maximal and minimal doses for target volumes, normal tissue complication probability (NTCP), maximum and mean doses for organs at risk and the whole DVH differences were assessed. In the group with 10 mm margins, the only significant difference was worse rectal NTCP by 4.5%, but the CTV dose coverage remained at the same level. Recalculated plans with 7 mm margin could not achieve the prescribed target volume coverage, and the EUD decreased by 3.7 and 0.6 Gy for PTV and CTV, respectively. Desired CTV → PTV margin for non-IGRT plans should be no lower than 12 mm to guarantee 95% instances when delivered dose to CTV maintain as planned, for IGRT plans decrease this requirement to 2 mm. Prostate IMRT strategies involving margin reduction below 7 mm require image-guidance to maintain the planned dose coverage. Using fiducial-based image-guidance and large margins seems to be superfluous.

[Radiotherapy in vulvar cancer treatment]. / Radioterapie v lécbe karcinomu vulvy.

OBJECTIVE: To provide an actual review of radiotherapy in the treatment of vulvar carcinoma. DESIGN: A review article. SETTING: Department of Oncology and Radiotherapy, University Hospital in Hradec Králové. METHODS: A review article evaluating the application of ionizing radiation in the treatment of early and advanced vulvar carcinoma, based on the most significant previously published studies. CONCLUSION: Postoperative groin irradiation in patients with positive groin lymph-nodes improves local control, time to progression, and overall survival; especially in 2 positive nodes and in N2/3 initial findings. In case of positive inguinal nodes, radiotherapy of both groins and at least lower pelvic iliac node-chains should follow. Adjuvant irradiation of the primary remains controversial, except for positive resection margins where radiotherapy improves overall survival. Concurrent chemoradiotherapy seems to be appropriate primary treatment of advanced vulvar carcinomas, in attempt to avoid mutilating intervention or exenteration. Chemoradiation should be followed by subsequent surgery, including potential groin dissection in case of lymph-node involvement. Definitive chemoradiotherapy is of limited evidence, and radical dose escalation to the gross tumor is essential for its implementation. Modern radiotherapy techniques, especially with intensity modulation, are convenient for dose escalation.

[Prognostic significance of changes of tumor epidermal growth factor receptor expression after neoadjuvant chemoradiation in patients with rectal adenocarcinoma].

PURPOSE: The aim of the present study was to examine the effect of neoadjuvant chemoradiation on tumor epidermal growth factor receptor (EGFR) expression in patients with locally advanced rectal adenocarcinoma. PATIENTS AND METHODS: A total of 53 patients with rectal adenocarcinoma (clinical stages II and III) were studied. Neoadjuvant treatment consisted of 50.4 Gy/28 fractions external radiation with concomitant continuous 5-fluorouracil. Surgical resection was performed 4-6 weeks after the chemoradiation. EGFR expression in the pretreatment biopsies and in the resected specimens was assessed with immunohistochemistry. RESULTS: Patients with an increase of EGFR expression during chemoradiation had significantly shorter disease-free survival (DFS; p = 0.003) and overall survival (OS; p = 0.005) compared to patients with either no change or decrease in EGFR expression. The 5-year DFS in patients with increased EGFR expression was only 29% compared to 61% in patients without an increase of EGFR expression. Similarly, the 5-year OS of the patients with increased EGFR expression was 29% compared to 66% in patients without an increase of EGFR expression. All recurrences in patients who had an increase of EGFR expression occurred within the first 2 years after the treatment. The increase in EGFR expression was the only significant predictor of DFS (p = 0.007) and OS (p = 0.04) using multivariate Cox regression analysis. CONCLUSION: An increase of EGFR expression during chemoradiation may be associated with significantly shorter DFS and OS. The increase of EGFR could identify a population of patients in whom the effect of the treatment with anti-EGFR therapy should be studied.

Successful radiotherapy treatment of lacrimal gland infiltration in patient with Sjögren΄s syndrome.

OBJECTIVE: To present a single case report on successful radiotherapy treatment of lacrimal gland infiltration in patient with Sjögren΄s syndrome. BACKGROUND: Radiotherapy is occasionally used for the treatment of benign disorders. There is no report on use of radiotherapy for local treatment of the Sjögren΄s syndrome in the literature. METHODS: Female patient with lacrimal gland involvement as a part of Sjögren΄s syndrome with diplopia and visus deterioration was treated by radiotherapy with eye shielding. RESULTS: Regression of the infiltration with full restoration of visus and minimal acute radiation reaction was achieved. CONCLUSION: A case report of successful use of local radiotherapy in the treatment of lacrimal gland affected by Sjögren΄s syndrome is presented (Fig. 3, Ref. 6). Full Text in PDF www.elis.sk.

Molecular predictive factors of outcome of radiotherapy in cervical cancer.

Radical radiotherapy with concurrent cisplatin-based chemotherapy is an established treatment for cervical cancer patients with stage FIGO IIB and higher. The tumor control can be achieved in 40-80% of patients, the treatment is associated with the risk of late postiradiation complications in 10 - 15% of cases. Detection of the factors predictive for tumor control and late morbidity is a possible direction how to individualize radiotherapy dose and technique. The aim of our review is to summarize results of studies inquiring various molecular markers predicting tumor response to radiotherapy and a risk of late complications. A lot of candidate molecules were evaluated in histochemical studies: membrane receptors (EGFR, HER-2), cell cycle regulators (p53, p21), proliferative markers (Ki-67), hypoxia and angiogenetic factors (HIF, VEGF), HPV status, and others (COX-2), with promising results in some of them (HPV, HIF-1α, Ku80, ATM polymorphism). Microarray studies identified decades of genes with different expression in radiosensitive/radioresistant cervical tumors and sets of genes are able to comletely separate responding and nonresponding tumors, but these sets differ across studies. Further well designed studies will be necessary to achieve results matured for use in clinical practice.

[Intensity modulated radiation therapy technique in the treatment of gynecologic malignancies]. / Technika radioterapie s modulovanou intenzitou zárení v lécbe gynekologických malignit.

OBJECTIVE: A description of intensity modulated radiation therapy (IMRT) technique and its implementation in the treatment of gynecologic malignancies. SUBJECT: A review article. SETTING: Department of Oncology and Radiotherapy, University Hospital in Hradec Králové. SUBJECT AND METHOD: General explanation of IMRT priciples, its benefits and limitations, and a review of published data about its utilization in the treatment of endometrial, cervical, and vulvar carcinoma. CONCLUSION: IMRT represents an accessible and highly conformal external beam radiothrapy technique, which enables a significant sparing of healthy tissue with consequent reduction of radiation morbidity in comparison with other conventional and conformal techniques. A feasibilty of dose excalation with preservation of low toxicity is another advandage of IMRT. It can be utilized in the treatment of most frequent gynecologic tumors, especially in endometrial, cervical, and vulvar carcinoma.

High dose rate versus low dose rate brachytherapy in the treatment of tongue carcinoma - a radiobiological study.

UNLABELLED: Low dose rate (LDR) brachytherapy is a well established treatment for the early stages of tongue cancer. High dose rate (HDR) afterloading devices have replaced LDR brachytherapy in many radiotherapy departments, but the effect and safety of HDR brachytherapy in comparison with LDR brachytherapy for interstitial applications is an unresolved question. The aim of our radiobiological study was to utilize dose volume histiograms from patients treated in our institution to simulate the risk of complication of LDR and HDR brachytherapy. Normal tissue complication probabilities (NTCP) of acute mucositis, late mucosal necrosis and osteoradionecrosis of two HDR brachytherapy schedules (18 x 3 Gy bid and 10 x 6 Gy bid) and of LDR brachytherapy with identical tumor control probability were compared using data from 8 brachytherapy applications. A linear quadratic (LQ) model was used to calculate the biologically equivalent doses, the effective volume method of Kutcher and Burman and Lyman's model was used to calculate NTCP. The Student's two-tailed test was used for statistical analysis. For 18 x 3 Gy bid the risk of acute mucositis and of late mucosal necrosis was 1.48 and 1.66 times higher with HDR in comparison with LDR brachytherapy. For 10 x 6 Gy bid the risk of acute mucositis, mucosal necrosis and osteoradionecrosis was 1.3, 3.44 and 13.18 times higher with HDR brachytherapy. All differences were statistically highly significant. Our radiobiological study supported the hypothesis that HDR has a higher risk of complication in comparison with LDR brachytherapy for the same tumor control probability. KEYWORDS: tongue cancer, brachytherapy, low dose rate, high dose rate.