[Is it possible to detect surface antigen CD133 on patient-derived glioblastoma continuous cell cultures using fluorescent aptamers?] / Vozmozhna li detektsiya poverkhnostnogo antigena CD133 na perevivaemykh kul'turakh kletok glioblastomy patsientov s pomoshch'yu fluorestsentnykh aptamerov?

Theranostics combines diagnostics and therapeutic exposure. Regarding glioblastomas, theranostics solves the problem of detecting and destroying tumor stem cells resistant to irradiation and chemotherapy and causing tumor recurrence. Transmembrane surface antigen CD133 is considered as a potential marker of tumor stem cells. OBJECTIVE: To detect CD133 in patient-derived glioblastoma continuous cell cultures using fluorescence microscopy and modified aptamers (molecular recognition elements) anti-CD133. MATERIAL AND METHODS: To detect CD133, we used mousey fluorescence monoclonal antibodies anti-CD133 MA1-219, FAM-modified DNA aptamers anti-CD133 AP-1-M and Cs5. Non-aptamer DNA oligonucleotide NADO was used as a negative control. Detection was performed for three samples of patient-derived glioblastoma continuous cell cultures coded as 1548, 1721 and 1793. RESULTS: MA1-219 antibodies brightly stained cell culture 1548, to a lesser extent - 1721. There was diffuse staining of cell culture 1793. Cs5-FAM aptamer stained cells in a similar way, but much weaker. AP-1-M-FAM aptamer interacted with cells even weaker and diffusely stained only cell culture 1793. Non-aptamer NADO did not stain cell culture 1548 and very weakly diffusely stained cell culture 1793. CONCLUSION: For both molecular recognition elements (MA1-219 antibody and Cs5 aptamer), 3 cell culture samples can be arranged in the following order possibly reflecting CD133 status decrease: strong signal for cell culture 1548, much weaker for 1721, even weaker for 1793. Only cell culture 1548 can be considered CD133 positive with combination of Cs5+ and NADO signals. Cell culture 1793 is CD133 false positive with combination of Cs5+ and NADO+ signals.

Trifluridine/tipiracil plus bevacizumab in patients with untreated metastatic colorectal cancer ineligible for intensive therapy: the randomized TASCO1 study.

BACKGROUND: We designed an open-label, noncomparative phase II study to assess the safety and efficacy of first-line treatment with trifluridine/tipiracil plus bevacizumab (TT-B) and capecitabine plus bevacizumab (C-B) in untreated patients with unresectable metastatic colorectal cancer (mCRC) who were not candidates for combination with cytotoxic chemotherapies. PATIENTS AND METHODS: From 29 April 2016 to 29 March 2017, 153 patients were randomly assigned (1:1) to either TT-B (N = 77) or C-B (N = 76). The primary end point was progression-free survival (PFS). The primary PFS analysis was performed after 100 events (radiological progression or death) were observed. Secondary end points included overall survival (OS), quality of life (QoL; QLQ-C30 and QLQ-CR29 questionnaires), and safety. RESULTS: Median (range) duration of treatment was 7.8 (6.0-9.7) months and 6.2 (4.1-9.1) months in the TT-B and C-B groups, respectively. Median (range) PFS was 9.2 (7.6-11.6) and 7.8 (5.5-10.1) months, respectively. Median (range) OS was 18 (15.2 to NA) and 16.2 (12.5 to NA) months, respectively. QoL questionnaires showed no relevant changes over time for either treatment. Therapies were well tolerated. Patients receiving TT-B had more grade ≥3 neutropenia (47% versus 5% with C-B). Patients receiving C-B had more grade ≥3 hand-foot syndrome (12% versus 0% with TT-B) and grade ≥3 diarrhea (8% versus 1% with TT-B), consistent with the known safety profiles of these agents. CONCLUSION: TT-B treatment showed promising clinical activity in untreated patients with unresectable mCRC ineligible for intensive therapy, with an acceptable safety profile and no clinically relevant changes in QoL. CLINICAL TRIAL INFORMATION: NCT02743221 (ClinicalTrials.gov).

[Thrombosis of the inferior vena cava and right portions of the heart in a relapsing retroperitoneal malignant tumour]. / Tromboz nizhnei poloi veny i pravykh otdelov serdtsa pri retsidivnoi zabriushinnoi zlokachestvennoi opukholi.

Presented herein is a clinical case report regarding successful treatment of a female patient with a relapsing malignant retroperitoneal tumour complicated by disseminated thrombosis of the inferior vena cava, right atrium and right ventricle of the heart. A relapse of the malignant uterine tumour had developed 3 years after the primary operation and was represented by a large-size mass ingrowing into the infrarenal segment of the inferior vena cava, aorta, as well as jejunum. Additional examination revealed the presence of a tumorous thrombus extending from the primary tumorous node in the lumen of the inferior vena cava to the right atrium and ventricle. The procedures performed consisted in removal of the tumour with resection of the inferior vena cava, aorta, and jejunum, followed by thrombectomy from the right portions of the heart under extracorporeal circulation. The postoperative period turned out uneventful, with no complications observed. The woman was discharged on POD 15. Twelve-month postoperative follow up revealed neither relapse nor progression of the disease. Currently, the patient continues undergoing specific treatment (second-line chemotherapy). Also discussed in the article are current challenges concerning both the classification of tumour thrombosis of the inferior vena cava in retroperitoneal sarcomas, and the choice of optimal strategy and policy of treatment.

ARTERIAL CHEMOINFUSION IN PATIENTS WITH LOCALLY ADVANCED AND METASTATIC PANCREAS CANCER.

The chemoinfusions (310) were carried out in celiac trunk in 167 patients with non-removed pancreas cancer at the period from 2000 to 2015. Locally advanced timorous process (stage III, n=79) was revealed in 79 patients and liver metastases (stage IV, n=88) were noted in 88 cases. The celiac axis infusion by Gemcitabine (1000 mg/m²) was applied for patients and GEMOX (Gemcitabine+Oxaliplatin 75 mg/m²) has been using since 2012. Symptomatic improvement such as decrease of pain, growth of body weight was noted in majority of patients. An average lifetime, median and one-year survival consisted of 7,6 months, 5,8 months and 10%. The patients (133) were treated by 1­2 cycles and after that by course of total body chemotherapeutics. There weren't any serious complications. Toxic manifestations of chemotherapy weren't higher than I­II degree and they were arrested by corrective therapy in 92 patients (55%). The celiac axis infusion is safe in patients with locally advanced and inoperable pancreas cancer. Symptomatic improvement showed the most patients. The objective response to the treatment had 20% patients and performance of repeated cycles led to increase of their survival.

[Multiple sclerosis vaccine: possibilities of the use of dendritic cells immunomodulatory potential].

For the first time we carried out a clinical assessment of the safety, tolerability and clinical efficacy course of repeated administration of experimental modified autologous vaccine interleykin (IL-10) dendritic cells in two patients with secondary-progressive multiple sclerosis patient and one with relapsing-remitting multiple sclerosis. In the course of treatment, we carried out clinical and immunological monitoring. It was found out that intradermal dose of 3 x 106 cells applied to spinal area 6-12. times did not cause any serious side effects. After the treatment with dendritic cells, the following results were observed: 1) a significant positive clinical effect in patients with secondary-progressive multiple sclerosis exacerbations; 2) moderate positive clinical effect in patients with relapsing-remitting multiple sclerosis, in a state of remission; 3) a complete absence of any clinical results in patients with secondary-progressive multiple sclerosis without exacerbations. The immune response was characterized by a significant absolute and relative increase of serum T-regulatory cells. Discovered distinct anti-inflammatory properties of dendritic cell therapy allow us to consider it as a promising area of personalized treatment based on an individual vaccination against multiple sclerosis.

[Experience with the use of gefitinib in patients with inoperable non-squamous cell lung cancer with activating EGFR mutations].

Discovery of activating EGFR mutations led to dramatic modification of treatment schemes for nonsquamous lung cancer. 70 patients with activating EGFR mutations were treated by gefitinib being either a part of prospective phase II trial (n = 25) or, subsequently, subjected to routine clinical management (n = 45). Objective response rate approached to 32.7%. Median time to disease progression was 14 months, and median overall survival was 26.1 months. Subgroup analysis revealed statistically longer time to disease progression (p < 0,0001) and overall survival (p = 0,001) in latter vs. former group, despite the lower rate of objective response (22% vs 48%). Possible explanations include more relaxed standards for routine gefitinib use, i.e. inclusion of the patients with non-measurable tumor lumps, continuation of gefitinib uptake upon slow disease progression, and increasing availability and quality of radiosurgery for brain metastases.

[Pharmacoeconomic analysis of gefitinib therapy in patients with non-cell cell lung cancer].

We performed a treatment efficacy analysis for non-small cell lung cancer (NSCLC) patients' population with EGFR mutation aimed at optimization of pharmacoeconomic factors. The employment of gefitinib leads to an increase in patients' life expectancy for a median of 1.05 years. The average cost-effectiveness of this therapy is 934.8 thousand rubles per additional year (903.9-1100.5 thousand rubles for each year). If gefitinib therapy is given only to patients with proved EGFR mutation it can decrease the average expenses by 211.6-251.8 thousand rubles per patient in comparison to undiagnosed patients's population receiving gefitinib without a decrease in clinical effect. Comparison of selective gefitinib administration with isolated chemotherapy (CT) yields an incremental cost-effectiveness ratio of 960.7 to 1010.0 thousand rubles per additional year. Therefore, the strategy of EGFR gene mutations testing in patients with inoperable NSCLC with consequent gefitinib therapy administration in patients positive for mutation lead to an increase in life expectancy and is characterized by acceptable cost-effectiveness.

[Phase II clinical trial of autologous dendritic cell vaccine with immunologic adjuvant in cutaneous melanoma patients].

This paper describes the clinical results and immunologic changes in cutaneous melanoma patients receiving active specific immunotherapy with autologous dendritic cell vaccine (DCV) in combination with cyclophosphamide used as immunologic adjuvant. Twenty eight patients with morphologically verified stage III-IV cutaneous melanoma receiving therapy in N. N. Petrov Research Institute of Oncology between 2008 and 2011 were included in the study. All patients signed an informed consent form. Nineteen patients (67,9%) received DCV in therapeutic setting, 9 (32,1%) received it in adjuvant setting. DCV therapy was well tolerated. No serious adverse events were registered. Frequent adverse events included Grade 1-2 unspecific symptoms (fever, fatigue, flu-like symptoms) observed in 22% patients after 3,5% of vaccinations. In therapeutic settings the use DCV lead to clinical effect (PR+SD) in 36,6% of patients. PR was observed in 5% of (95% CI 0-15%) patients, SD in 31,6% (95% CI 13-56%). Duration of the objective responses was 168-965+days. Addition of immunologic adjuvant (cyclophosphamide 300 mg/m2 IV 2 hours) 3 days before vaccination increased its efficacy. In this patients group (n=12) the therapy lead to clinical benefit in 42% (95% CI 17-69%) of cases, median time to progression was 91 (95% CI 55-126) days. This regimen was selected for adjuvant therapy. In the adjuvant therapy group (n=9) the median time to progression was 112 (95% CI 58-166) days. Immunologic monitoring showed correlation ofT- and B-cell immune response with DCV clinical efficacy (p<0,05), no correlation with delayed hypersensivity reaction was observed (p>0,1). DCV is well tolerated and shows immunological and clinical response in stage III-IV skin melanoma patients.

Factors predictive of obliteration after arteriovenous malformation radiosurgery.

OBJECTIVE: To investigate predictive factors of complete obliteration following treatment with linac-based stereotactic radiosurgery for intracerebral arteriovenous malformations. METHODS: Archived plans for 48 patients treated at the British Columbia Cancer Agency and who underwent post-treatment digital subtraction angiography to assess obliteration were studied. Actuarial estimates of obliteration were calculated using the Kaplan-Meier method. Univariate and multivariate Cox proportional hazards models were used for analysis of incidence of obliteration. Log-rank test was used to search for parameters associated with obliteration. RESULTS: Complete nidus obliteration was achieved in 38/48 patients (79.2%). Actuarial rate of obliteration was 75.9% at 4 years (95% confidence interval 63.1%-88.6%). On univariate analysis, prescribed dose to the margin (p=0.002) and dose to isocentre (p=0.022) showed statistical significance. No parameters were significant in a multivariate model. According to the log-rank test, prescribed dose to the margin of >20 Gy (p=0.004) and dose to the isocentre of >25 Gy (p=0.004) were associated with obliteration. CONCLUSION: Reported series in the literature suggest a number of different factors are predictive of complete obliteration of arteriovenous malformations following radiosurgery. However, differing definitions of volume and complete obliteration makes direct comparison between series difficult. This study demonstrates that complete obliteration of the nidus following linear accelerator-based stereotactic radiosurgery for arteriovenous malformations appears to be most closely related to the prescribed marginal dose. In particular, a marginal dose of >20 Gy is strongly associated with obtaining complete obliteration of the nidus.

[Expression of Ki-67, p53, bcl-2, estrogen receptors alpha in patients with clear cell renal carcinoma and epidermal growth factor receptor mutation].

Three hundred and thirty-six cases of clear-cell renal carcinoma (CCRC) were examined for epidermal growth factor receptor (EGFR) mutation: exon 19 deletion and L858R mutation in exon 21 of the EGFR gene. The expression of Ki-67, bcl-2, p53, and estrogen receptors alpha was studied in CCRC with EGFR mutation. There were 4 cases of CCRC with EGFR exon 19 deletion. The frequency of EGFR gene mutations was 1.2%. L858R missense mutations in exon 21 of the EGFR gene were absent. In CCRC, EGFR gene mutation (exon 19 deletion) was detected in 3 men and 1 woman with an age range of 50 to 60 years and Fuhrman differentiation grade 2 or 3. The Ki-67 index varied from 4 to 23%. The expression of bcl-2 and p53 was negative. A moderate estrogen receptor alpha expression was revealed in 1 of 4 cases.

[Investigation of hepatoprotector Metrop GP influence on tumor growth or antitumor action of doxorubicin].

An experimental model of solid tumor of Ehrlich failed to reveal any influence of the hepatoprotector Metrop GP on tumor growth or affect antitumor action of doxorubicin.

[Preclinical evaluation of the effect of Metrop GP on acute toxicity and hepatotoxicity of chemotherapy].

Our paper presents the results of preclinical evaluation of the mitigating effect of Metrop GP on the acute toxicity and hepatotoxicity induced by paclitaxel and doxorubicin treatment using functional samples and biochemical and morphological techniques.

[Laboratory evaluation of TGFbeta1, IL-10, VEGF levels in vivo and in vitro in patients with solid tumors].

Tumor cells can acquire the mechanisms of immune response evasion. One of these mechanisms is synthesis and secretion in the microenvironment of immunosuppressive factors able to block immune cells maturation and function. We investigated plasma levels of TGFbeta1 and IL0 in 10 healthy volunteers and 114 patients with solid tumors (breast cancer--24, gastrointestinal tumors--27, renal cell carcinoma--15, lung cancer--9, ovarian cancer--13, cutaneous melanoma--18, primary multiple tumors--2, prostate cancer--6). TGFbeta and IL10 concentration in supernatants of 37 primary cultures (cutaneous melanoma, renal cell carcinoma and prostate cancer) and 10 cultures of melanoma during cultivation were also studied. VEGF was determined by immunocytochemistry staining in 15 melanoma culture specimens. The lowest level of TGFbeta1 was documented in rectal cancer patients, 30.05 +/- 12.30 ng/ml (p < 0.05), the highest in renal cell cancer and pancreatic cancer patients, 145.61 +/- 11.32 and 146.15 +/- 30.56 ng/ml (p < 0.001), respectively. Primary cultures of tumor cells can synthesize traceable amounts of TGFbeta1 and IL10. Cultures of cutaneous melanoma, renal cell carcinoma and prostate cancer cells did not change expression level of IL-10 after several passages. VEGF was expressed in 20% of cutaneous melanoma cultures. We suppose that tests for TGFbeta1, IL10 and VEGF in culture supernatants from tumor cell lines, on the surface of the cells purposed for cancer vaccines and in serum of patients to be vaccinated are potentially useful.

[Immunochemical assay of MIC A production by tumor cells in vitro and in vivo as a component of antitumor vaccine development].

Due to immunological monitoring, most of aggressive tumor cells are selected capable of producing soluble factors of immunosuppression in their microenvironment. A ligand for NK- and T-cells receptor (MICA/B) activation is one of them. We investigated MICA concentrations in serum of 10 healthy volunteers and 114 patients with solid tumors (breast cancer - 24, gastrointestinal tumors - 27, renal cell carcinoma - 15, lung cancer - 9, ovarian carcinoma - 13, cutaneous melanoma - 18, primary multiple tumors - 2, prostate cancer - 6). The lowest level of MICA was reported in ovarian carcinoma (91.19 +/- 29.42 pg/ml), the highest - in rectal cancer (311.13 +/- 50.11 pg/ml) while mean concentration in healthy volunteers was (19.38 +/- 5.91 pg/ml). Cells of melanoma, renal cell carcinoma and prostate cancer became capable of producing MICA during cultivation and antitumor vaccine preparation. It is suggested that culture supernatants be tested for MICA contents as well as patients screened for vaccination. Selection of patients should be carried out on the basis of MICA molecules detection in blood flow.

[Frequency of expression of markers predictive of sensitivity to cytostatics in skin melanoma].

At present, choice of treatment for skin melanoma depends on empirical data on efficacy of medication. Individual treatment may be promoted if certain markers of sensitivity to chemotherapy are evaluated. We studied frequencies of expression of marker sensitivity to fluoropyrimidin [(FPd) (-1)], TS(-)0 and TS(-). TS(-/+) was reported in 36.1%. Sensitivity to platinum drugs [EPCC1(-) - 64.4%], taxotere drugs- [beta = tubulin(-) -72.7%], cyclooxygenase inhibitors [COX2(+)] - 8.9%, mutant tyrosokinase inhibitors [c = kit(+)] - 21.4%, PDGFR = beta(+) -35.5%. Marker expression in tumor tissue was heterogenous. Data on molecular-genetic characteristics of tumor may be used to individualize choice of drugs, dosage and to lower risk of toxicity. Use of cytostatics should be evaluated in clinic for their efficacy in skin melanoma with due consideration of prognostic markers.

[Effectiveness of gefitinib (Iressa) as first-line therapy for inoperable non-small-cell lung cancer with mutated EGFR gene (phase II study)].

Tumor regression was reported in 20-30% of patients with inoperable non-small-cell lung cancer (NSLC) following standard first-line chemotherapy. Clinical trials with second-line gefitinib (Iressa) showed a strikingly high response in patients with mutated EGFR. However, clinical experience with gefitinib as first-line therapy had been limited to small-scale trials mostly among subjects of Asian origin. Our study was not associated with the drug manufacturer and included 25 chemotherapy-naive patients with mutated EGFR inoperable lung adenocarcinoma. Standard dose was 250 mg/day. Complete response was observed in 1 patient (4%), partial--11 (44%), sustained stabilization--13 (52%); median time until tumor progression--186 days. Median overall survival failed to be registered within the duration of the study. Among most frequent side-effects were skin rash (19; 76%) and diarrhea (14; 56%): marked side-effect -toxicity grade III (4; 16%). Gefitinib appeared highly efficient and tolerable and may be recommended as first-line treatment of mutated EGFR inoperable NSLC.

Volumetric modulated Arc therapy and conventional intensity-modulated radiotherapy for simultaneous maximal intraprostatic boost: a planning comparison study.

AIMS: Volumetric modulated arc therapy (VMAT) is a novel extension of intensity-modulated radiotherapy (IMRT) where an optimised three-dimensional dose distribution may be delivered in a single gantry rotation. This optimisation algorithm is the predecessor to Varian's RapidArc. The aim of this study was to compare the ability of conventional static nine-field IMRT (cIMRT) and VMAT to boost as much of the clinical target volume (CTV) as possible to 88.8Gy without exceeding organ at risk (OAR) dose-volume constraints. MATERIALS AND METHODS: Optimal cIMRT and VMAT radiotherapy plans were produced for 10 patients with localised prostate cancer using common planning objectives: (1) Treat >or=98% of the planning target volume (PTV) to >or=95% of the prescription dose (74Gy in 37 fractions); (2) keep OAR doses within predefined limits; (3) treat as much of prostate CTV (minus urethra) as possible to >or=120% of prescription dose (=88.8Gy); (4) keep within maximum dose limits in and out of target volumes; (5) conformality index (volume of 95% isodose/volume of PTV)or=120% of the prescription dose (P=0.002). All dose constraints were kept within predefined limits. VMAT and cIMRT required an average of 949 and 1819 monitor units and 3.7 and 9.6min, respectively, to deliver a single radiation fraction. CONCLUSIONS: VMAT is able to boost more of the CTV to >or=120% than cIMRT without contravening OAR dose constraints, and uses 48% fewer monitor units. Treatment times were 61% less than with cIMRT.

[The specific features of the expression of molecular biological markers for proliferation and apoptosis in patients with lung cancer with epidermal growth factor receptor mutation].

Two hundred and two lung adenocarcinomas were examined for epidermal growth factor receptor (EGFR) mutation: exon 19 deletion and L85R mutation in exon 21. The expression of EGFR, Ki-67, bcl-2, p53 in the lung adenocarcinomas was studied in relation to the type of EGFR mutation. Lung adenocarcinomas harboring the EGFR L858R mutation in exon 21 have a less proliferative activity than those with EGFR mutation in exon 19 and in those without EGFR mutation. The p53 protein expression was higher in a group of adenocarcinomas having the EGFR L858R mutation than in that of adenocarcinomas with EGFR mutation in exon 19. There was no bcl-2 expression in adenocarcinomas showing the EGFR mutation.

[A clinico-experimental study in metronomic therapy of disseminated solid tumors].

Although medication of malignancies made a considerable progress and the rates of apparent response grew markedly, those of survival among patients with disseminated solid tumors increased insignificantly. Metronomic therapy with cytostatics is a new modality of treatment via tumor vessels. Dose-dense regimen uses low dosage administration of drugs at regular short intervals (hours, days, weeks). Cumulative dose may be much lower than that used in standard therapy, thus reducing toxicity as well as amount of therapeutic support. A pre-clinical study established decrease in size of tumors which initially showed resistance to treatment. However, further research in randomized investigation is needed in view of certain inconstinence of our data.