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Background. Gemistocytic astrocytomas (GA) are a variant of diffuse astrocytomas GII (WHO, 2016), which is characterized by a shorter overall survival of patients than diffuse astrocytoma. The role of vascular endothelial growth factor (VEGF) gene expression and its association with the index of proliferative activity (Ki-67) in GA has not been studied Objective. To estimate the correlation of VEGF gene expression on Ki-67 in GA. Design and methods . A retrospective and prospective analysis of morphological and molecular-genetic features of tumors was performed on 9 patients with the diagnosis of GA. Patients over 18 years of age with supratentorial tumor localization were included in the study. The patients were aged from 27 to 37 years, the median age was 32 years. There were 4 men. Fragments of tumor tissue obtained during surgery were subjected to histological, immunohistochemical (Ki-67 and GFAP) and molecular genetic studies. By real-time PCR method was used to determine the level of the VEGF gene expression and the presence of a mutation in IDH1/IDH2 genes. Histological diagnosis was made in accordance with the classification of tumors of the CNS WHO (2016) and the AFIP atlas (2007). Results. The tumor most often affected several lobes of the brain (44,4%). The first clinical manifestations of the disease in the vast majority of cases were epileptic and hypertensive syndromes (55,6%). For the proliferative activity index Ki-67 >5% (71,4% of cases) the moderate proliferation of vascular endothelium was noted in 85,7% of cases and the middle/high level of VEGF gene expression (except one case). In patients with low Ki-67 (≤5%), the low level of the VEGF gene expression was determined in 100% cases, the proliferation of the vascular endothelium was absent. Conclusion. High Ki-67 (>5%), the proliferation of the vascular endothelium, middle/high level of VEGF gene expression are typical attributes of anaplastic astrocytomas. That’s why some GA have features of astrocytomas GIII.

Primary extraspinal myxopapillary ependymoma (MPE) is an exceptionally rare lesion that is mainly located in the subcutaneous sacrococcygeal region. We describe the first case of MPE that presented as an intramuscular tumor mass located in the lumbar area. Absence of the visible connection with the spinal cord and lack of any other tumors in the reported case argue for the primary ectopic origin of the MPE. The differential diagnosis of MPE is discussed. Additionally, we evaluated the expression level of molecular biomarkers that have a prognostic value in central nervous system tumors.

BACKGROUND Although the molecular analysis of epidermal growth factor receptor (EGFR) and anaplastic lymphoma kinase (ALK) in archived lung cancer tissues is relatively well established, the genetic testing of cytological material has not yet become a routine. METHODS The current study used cell samples that were obtained by bronchial brushing, transthoracic needle aspiration, or biopsy imprint preparation between 1993 and 2008. Islets of malignant cells were visually located on the archived cytological slides, lysed in situ by a drop of sodium dodecyl sulfate-containing buffer, and subjected to the standard DNA and RNA extraction. Examination of paraffin-embedded tissue blocks (resection specimens or biopsy material) from the same patients was performed in parallel. RESULTS A total of 75 cytological/histological lung adenocarcinoma sample pairs underwent polymerase chain reaction analysis for the EGFR mutation. Two cytological samples and 1 morphological sample failed to produce DNA. Concordance for the wild-type and mutation status was observed in 54 of 72 and 14 of 72 informative pairs, respectively; 3 pairs and 1 pair, respectively, had mutation only in the cytological or histological material. The discrepancies could be explained by the failure to ensure a high percentage of lung cancer cells in the analyzed samples or, alternatively, by the genuine intratumoral molecular heterogeneity of some neoplasms. RNA extraction followed by reverse transcriptase-polymerase chain reaction analysis for the EML4-ALK translocation was performed for 44 EGFR mutation-negative sample pairs; failures were observed for 2 cytological and 6 histological specimens. All informative pairs were concordant either for the norm (32 of 36 pairs) or for the presence of EML4-ALK gene fusion (4 of 36 pairs). CONCLUSIONS Archived cytological slides appear to be well suited both for EGFR and ALK analysis. Cancer (Cancer Cytopathol) 2013;121:370–376. © 2013 American Cancer Society.

The BLM gene belongs to the RecQ helicase family and has been implicated in the maintenance of genomic stability. Its homozygous germline inactivation causes Bloom syndrome, a severe genetic disorder characterized by growth retardation, impaired fertility and highly elevated cancer risk. We hypothesized that BLM is a candidate gene for breast cancer (BC) predisposition. Sequencing of its entire coding region in 95 genetically enriched Russian BC patients identified two heterozygous carriers of the c.1642 C>T (Q548X) mutation. The extended study revealed this allele in 17/1,498 (1.1%) BC cases vs. 2/1,093 (0.2%) healthy women (p = 0.004). There was a suggestion that BLM mutations were more common in patients reporting first-degree family history of BC (6/251 (2.4%) vs. 11/1,247 (0.9%), p = 0.05), early-onset cases (12/762 (1.6%) vs. 5/736 (0.7%), p = 0.14) and women with bilateral appearance of the disease (2/122 (1.6%) vs. 15/1376 (1.1%), p = 0.64). None of the BLM-associated BC exhibited somatic loss of heterozygosity at the BLM gene locus. This study demonstrates that BLM Q548X allele is recurrent in Slavic subjects and may be associated with BC risk.

Non-small cell lung carcinomas (NSCLC) carrying a mutation in the epidermal growth factor receptor (EGFR) gene show unprecedented sensitivity to gefitinib or erlotinib.We present the follow-up data of 2 EGFR mutation-positive stage IV NSCLC patients who received upfront 250 mg gefitinib daily, then underwent potentially curative surgery, and resumed gefitinib therapy in the adjuvant setting. Patient 1 was diagnosed with a rightsided adenocarcinoma of the upper lobe with multiple metastases to the middle lobe. After 2 months of gefitinib treatment, only a small primary lesion was seen, and a bilobectomy with lymph node dissection was performed. The patient remained disease-free during a scheduled 12-month adjuvant therapy but relapsed 9 weeks after cessation of this treatment. Patient 2 presented with adenocarcinoma of the lower lobe of the right lung and a single metastasis to the left adrenal. After 3 months of receiving gefitinib, the metastasis was no longer detectable, and the primary tumor was significantly reduced. Surgery included lobectomy and adrenalectomy. This patient relapsed with metastasis in the remaining adrenal 4 months after the start of adjuvant therapy.Gefitinib can be used preoperatively for the management of advanced EGFR mutation-positive NSCLC. It remains to be established whether surgical intervention indeed renders survival advantage for this category of patients.

BRCA1 5382insC variant was repeatedly detected in Jewish breast cancer (BC) families residing in USA and Israel as well as in non-Jewish familial BC patients from Poland, Latvia, Hungary, Russia and some other European countries. However, the distribution of BRCA1 5382insC mutation in unselected BC cases vs. controls has been systematically investigated mainly in Ashkenazi Jews. Here we applied a case-control study design in order to evaluate the impact of BRCA1 5382insC allele on BC incidence in St Petersburg, Russia. High frequency of the BRCA1 5382insC allele was detected in a group of bilateral breast cancer patients (10.4%; 15/144). Randomly selected unilateral BC cases demonstrated noticeable occurrence of BRCA1 5382insC mutation as well (3.7%; 32/857), with evident excess of the carriers in the early-onset (40 years) category (6.1%; 6/99) and in patients reporting breast and/or ovarian tumours in first-degree relatives (11.3%; 11/97). Strikingly, none of 478 middle-aged controls and 344 elderly tumour-free women carried the 5382insC variant. The presented data confirm a noticeable contribution of BRCA1 5382insC mutation in BC development in Russia, that may justify an extended BRCA1 5382insC testing within this population.

This study was aimed to assess the role of CHEK2 1100delC mutation in breast cancer (BC) predisposition in Russia. The 1100delC allele was detected in 14/660 (2.1%) unilateral BC cases and in 8/155 (5.2%) patients with the bilateral form of the disease, but only in 1/448 (0.2%) middle-aged control females and in none of 373 elderly tumor-free women. The obtained data point at potentially high clinical relevance of CHEK2 1100delC testing in females of Russian origin and warrant similar case-control studies in ethnically and geographically related regions, especially in Ukraine, Belarus and Baltic countries.

The gene for Nijmegen chromosomal breakage syndrome (NBS1) plays a role in a variety of processes protecting chromosomal stability. Recently, it was suggested in a Polish case-control study that the founder hypomorphic mutation in NBS1, 657del5, which occurs in approximately 0.5% of Slavic subjects, may be associated with an increased risk of breast cancer (BC). We attempted to validate these findings in Russian subjects, who are also of Slavic descent. Heterozygous carriers for the 657del5 mutation were detected in 2 of 173 (1.16%) bilateral breast cancer cases, 5 of 700 (0.71%) unilateral breast cancer patients, 2 of 348 (0.57%) healthy middle-aged females and in 0 of 344 elderly tumor-free women. The difference between the “extreme” cohorts, i.e., biBC patients vs. elderly controls, approached the formal limit of statistic significance (p = 0.046). LOH at NBS1 was detected in only 3 of 5 available breast tumors from NBS1 657del5-carriers. In 2 of these tumors, the loss involved the mutant NBS1-allele. Overall, our data suggest that the NBS1 657del5 allele may contribute only to a limited fraction of breast cancer cases in Russia. © 2004 Wiley-Liss, Inc.

Breast carcinomas caused by inheritance of cancer-predisposing germ-line mutations have specific bioclinical features. This study aimed to analyze the efficacy of conventional cytotoxic treatment in BRCA1 and CHEK2 mutation carriers and non-carriers. The study included 415 Russian breast cancer patients aged 50 years or younger, who were subjected to various standard schemes of neoadjuvant therapy. The choice of therapy was done without the knowledge of the mutations status, because DNA testing was performed retrospectively using the archival tissue samples. 19 BRCA1 (4.6%) and 8 CHEK2 (1.9%) heterozygous genotypes were identified. BRCA1 mutation carriers achieved pathological complete response more frequently than non-carriers [6/19 (31.6%) vs. 46/388 (11.9%), p = 0.024]; this effect was limited to women treated by anthracycline-based therapy without taxanes [5/9 (55.6%) vs. 28/247 (11.3%), p = 0.002] and was not observed in any of 7 BRCA1 carriers receiving taxane-containing regimens. CHEK2 heterozygotes did not experience pathological complete response and showed lower frequency of objective clinical responses as compared to mutation non-carriers [4/8 (50%) vs. 333/388 (85.5%), p = 0.020]; the efficacy of neoadjuvant therapy was particularly poor in CHEK2 carriers receiving anthracyclines without taxanes. This study provides evidence for distinct sensitivity of BRCA1 and CHEK2 mutation-driven breast carcinomas to standard chemotherapeutic schemes.

An analysis of modern methods of diagnostics such as morphological, immunohistochemical and spectral, which included the bronchoscopy and spectrometry by using reflectance and autofluorescent regime, was made. The data involved the results of prospective follow-up study of 167 patients (620 biopsies). An obligatory spectrometry of suspicious area was carried out before the forceps biopsy. The microslides, which met the requirements of criteria of one of the carcinogen steps (n=201), were subjected to the in-depth morphological and immunohistochemical investigations. The tendency of angiogenesis (CD31 and CD34), proliferative activity (Ki-67), level of apoptosis (P53), EGFR expression were estimated. The sensitivity of combined endoscopic method was 94,74% by specificity 79,95% and high prognostic value of negative endoscopic diagnosis - 99,4%.

In 20 primary patients with focal abnormalities on conventional CT we evaluated diagnostic properties of simultaneous double-tracer SPECT. Scintigraphy was performed as a single examination with simultaneous registration of 67Ga and 99mTc-MIBI. Image acquisition was started 48-74 hours after IV injection of 130-175 MBq 67Ga-citrate and immediately after IV injection of 500-740 MBq of 99mTc-MIBI. All images for each agent were classified as positive and negative for primary tumor, N1 and N2 lymph-nodes (LN). According to histology 18 of 20 evaluated patients had non-small cell lung cancer (NSCLC), the other two patients had tuberculosis and nonspecific inflammation. SPECT with 99mTc-MIBI correctly visualized tumor in 18, 67Ga allowed correct visualization in 16 cases. Both tracers were truly negative in a patient with tuberculosis and false positive in a patient with nonspecific inflammation. Double-tracer SPECT was slightly more specific than CT in primary lesions. In 18 patients histological verification of LN status was obtained: NO was revealed in 9 cases, N1 in 4 and N2 in 5 cases. Both tracers correctly discriminated LN-positive and LN-negative cases with 94% specificity. On the contrary, CT was false-positive in 3 and false-negative in another 5 patients. Differentiation between N1 and N2 LN involvement is crucial for therapy planning. 99mTc MIBI and 67Ga revealed N1 in 2 cases and N2 in 4 cases, the diagnosis was later verified by postoperative morphology. In 2 patients SPECT overestimated extent of LN involvement and LN status was changed after surgery from N2 to N1. In 18 patients results of 99mTc-MIBI and 67Ga augmented each other. Accuracy of LN staging by SPECT with 99mTc-MIBI and 67Ga was 83%. CT accurately determined LN stage only in 7 patients, it was overestimated in 7 and underestimated in 4 cases. SPECT with 99mTc-MIBI and 67Ga demonstrated high overall accuracy in diagnostics of regional LN invasion for patients with NSCLC. Diagnostic value of conventional CT was significantly lower. Correct level of LN involvement was determined by SPECT in 83% of cases.

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.

A single institution series of 48 mucosal melanomas (MMs) has been analyzed for the presence of KIT mutations using high-resolution melting and sequencing of abnormally melted DNA fragments. The analysis of exons 9, 11, 13, and 17 has revealed eight of 48 (17%) nonsynonymous alterations, including zero of seven head and neck, six of 24 anorectal, one of 15 genitourinary, one of one gastric, and zero of one mediastinal MMs. Seven of these mutations were potentially associated with the tumor sensitivity to KIT tyrosine kinase inhibitors. One tumor harbored somatically acquired silent nucleotide substitution c.1383A>G (T461T). This study adds to the evidence that a substantial portion of MMs carry a therapeutically relevant mutation in the KIT oncogene.

Several Asian studies demonstrated feasibility of front-line administration of gefitinib for the treatment of non-small cell lung carcinomas (NSCLCs) harboring intragenic epidermal growth factor receptor (EGFR) mutations. The experience of the use of this EGFR tyrosine kinase inhibitor (TKI) in non-Asian subjects remains limited.The study included lung adenocarcinoma (AC) patients treated at the N.N. Petrov Institute of Oncology (Russia).DNA analysis of 192 consecutive AC revealed 38 (20%) TKI-sensitizing mutations. Presence of the exon 19 deletion (del19) or L858R was strongly correlated with nonsmoking status (smokers: 8/98 (8%); non-smokers: 30/94 (32%); p = 0.00004). The efficacy of first-line gefitinib therapy was evaluated in 25 patients with EGFR-mutated advanced AC. Twelve (48%) cases demonstrated tumor response (1 (4%) complete response, 11 (44%) partial responses; 10/17 (59%) patients with del19 mutation vs. 2/8 (25%) cases with L858R substitution, p = 0.11). The remaining 13 (52%) patients experienced disease stabilization. Median progression-free survival was 8.0 months. Grade 3 toxicity was the maximal adverse event, being observed only in 4 (16%) cases.Gefitinib may be considered as an upfront treatment option for EGFR-mutated NSCLC.

This case report describes a 35-year-old woman who was diagnosed with mixed epithelial/mesenchymal metaplastic carcinoma (carcinosarcoma) of the breast. Genetic analysis of blood DNA revealed a common founder mutation, BRCA1 5382insC. Examination of microdissected tumor samples determined that both epithelial and mesenchymal components contained deletion of the wild-type BRCA1 allele. This report exemplifies that even very uncommon breast tumor types may develop through biallelic inactivation of BRCA1 gene, that has to be considered in the genetic testing settings.

Measurement of intratumoral expression of dihydropyrimidine dehydrogenase (DPD) and thymidylate synthase (TS) may have some value in predicting the response to fluoropyrimidine-containing therapy. Patients and Methods: We attempted to validate this association in a series of Russian metastatic colorectal cancer cases. While replicating already published protocols, we unexpectedly found that the use of commonly utilized gene referees, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin, may lead to artifacts due to pseudogene-driven amplification from the genomic DNA template. We have developed a real-time PCR protocol which amplifies short PCR fragments, thus allowing efficient analysis of archival formalin-fixed paraffin-embedded tumor samples, and relies on succinate dehydrogenase (SDHA) as a gene referee, therefore avoiding amplification from genomic DNA. Results: Low content of DPD transcripts was observed in 13/20 (65%) patients with disease control (tumor response or disease stabilization) as compared to only 3/9 (33%) subjects with progressive disease (p = 0.11). Despite the low number of patients, this association reached the level of statistical significance when similar analysis was done for TS expression (11/20 (55%) vs. 1/9 (11%); p = 0.03). Conclusions: Our data confirm that low DPD and TS expressors have higher chances of success of fluoropyrimidine-containing regimens.

OBJECTIVE AND IMPORTANCE: A rare observation of double saccular aneurysms of the meningeal artery is presented. CLINICAL PRESENTATION: This 22-year-old man was referred to the Neurosurgical Institute with a suspicion of an aneurysm of the anterior communicating artery. Bilateral angiography of the carotid arteries was performed 1 week after the subarachnoid hemorrhage, but the aneurysms were not visualized. Routine angiography of both carotid arteries and selective studies of the left vertebral artery were performed again, and angiography of the right carotid artery revealed an aneurysm. The patient's neurological state at the time of admission was normal. Fundoscopic examination revealed papilledema and conjunctival injection of the left eye. INTERVENTION: The patient was treated using a right pterional approach. One aneurysm had caused spontaneous subarachnoid hemorrhage. The aneurysms were removed using a direct approach, with histological examination of dura matter fragment containing both aneurysms. The results of the patient's 2-week follow-up examination were normal. Follow-up angiography of the right carotid artery showed absence of the aneurysm with a clip on the branch of meningeal artery. CONCLUSION: Saccular aneurysms of the meningeal artery can be manifested by subarachnoid hemorrhage, and intradural arterial aneurysms are similar to saccular cerebral vessel lesions structurally.

Amplification of HER2, C-MYC and CCND1 oncogenes is a hallmark of breast cancer (BC); however, its involvement in the bilateral form of this disease has not been investigated yet. In this study, 50 bilateral BC (biBC) pairs (100 tumors) and 72 control unilateral BC were examined using real-time PCR analysis of microdissected archival tissues. In biBC, the frequency of >3-fold oncogene amplification was 6/100 (6%) for HER2, 6/100 (6%) for C-MYC and 7/100 (7%) for CCND1. Altogether, 18/100 (18%) biBC tumors had increased gene dosage of at least one oncogene. Tumors forming synchronous biBC pairs had amplification in 11/46 cases (24%). In 3 of 8 patients with amplification-positive carcinomas, the amplification was detected in both neoplasms: 2 biBC had concordant activation of the same oncogene (HER2 and CCND1, respectively), and in the remaining case distinct oncogenes were affected (HER2 and C-MYC). In contrast, amplifications in metachronous biBC were strongly discordant: none of 27 first carcinomas carried this abnormality, while the frequency of amplification in second tumors (7/27; 26%) was similar to the one observed in unilateral BC (20/72; 28%). The trend toward concordance of oncogene amplification status in synchronous but not in metachronous biBC pairs can be explained by the nearly identical natural history of the disease in simultaneously arising tumors. The skewed pattern of amplifications in metachronous biBC might be attributed to their association with adverse BC prognosis; it appears that only patients with amplification-negative first BC have sufficient chances to survive until the development of the contralateral carcinoma.

was obtained by bronchoscopy, and the morphological analysis classified the tumor as adenocarcinoma. A subsequent EGFR mutation test revealed a canonical TKI-sensitizing deletion located within exon 19 of the gene. Given the poor performance status of the patient, the extensive local tumor spread and the presence of a highly predictive EGFR mutation, immediate surgical intervention was considered unreasonable. Instead, neoadjuvant therapy with gefitinib (250 mg daily) was initiated. In addition, hemostatic treatment with etamsylate (250 mg by intramuscular injection, 3 times daily) was performed for 3 days. The prescribed therapy led to rapid symptomatic relief: by day 4, the ECOG status reached 1, and no signs of hemoptysis or paraneoplastic edema were observed. X-ray and CT examinations were repeated on day 12 of gefitinib therapy. A dramatic reduction of the primary tumor and hilar lymph nodes as well as disappearance of the bifurcational lesions were detected (fig. 1); given the significant downstaging of the disease and good performance status of the patient, a right pneumonectomy with systematic ipsilateral mediastinal lymph node dissection was performed 2 days later. Both preand post-treatment tissue samples contained a sufficient number of neoplastic cells, therefore it was possible to microdissect tumor tissues, isolate RNA, and perform reverse transcription polymerase chain reaction (RT-PCR) expression analysis of a few candidate genes. In addition to the above mentioned EGFR-inducible inflammatory molecules (IL6, IL8, ICAM1), we also considered the TNF-alpha and IL1-beta genes. Both these cytokines contribute to the mobilization of immune cells. TNF-alpha is involved in the formation of peritumoral pleural effusion [7]. A specific haplotype of the IL1-beta was shown to be associated with increased gene expression and elevated lung cancer risk [8]. As a control, we deliberately included in the study 2 genes, which are not related to inflammatory reactions, DPD and ERCC1.

The paper evaluates the available data as well as our own on use of autofluorescence bronchoscopy in conjunction with spectrometric examination. We used qualitative and quantitative assessment of images obtained by conventional and autofluorescence (ClearVu Elite) means in real time. Our double-stage study evaluated sensitivity and specificity of autofluorescence bronchoscopy in diagnosing lung cancer as well as constructed spectrometric curves (ROC) and areas under them (AUC). Endoscopy was used in 171 patients with central lung cancer. Autofluorescence bronchoscopy established high sensitivity--94.74% (95%CI: 80.9-99%) and sufficient specificity--79.95% (95%CI: 75.8-83.6%). Application of a wide range of spectrometric coefficients contributed to high specificity thus reducing the number of biopsies as well as the injury from the treatment. The AUC for a best predictive index was 0.89 (99%: 0.83-0.95).