[Clinical, immunohistochemical, and molecular genetic prognostic factors in adult patients with glioblastoma]. / Klinicheskie, immunogistokhimicheskie i molekulyarno-geneticheskie faktory prognoza u bol'nykh c glioblastomoi.

UNLABELLED: Glioblastoma is the most common primary malignant glial tumor of the brain in adult patients. AIM: to define the prognostic value of isocitrate dehydrogenase-1 (IDH-1) mutation and methylguanine-DNA methyltransferase (MGMT) methylation status in patients with glioblastoma (GB) and to analyze the impact of clinical data (gender, age, and tumor site), histological variants of the tumor structure, and time to development of recurrences on the course of the disease. SUBJECTS AND METHODS: The investigation enrolled 63 GB patients aged 18 to 71 years who had received combined treatment (surgery, chemo- and radiotherapy) at the N.I. Burdenko Research Institute of Neurosurgery, Ministry of Health of the Russian Federation, in the period 2008 to 2011. The investigators performed a morphological examination of all tumor tissue samples and an immunohistochemical examination using anti-IDH-1 R-132 antibody clone («Dianova¼, Germany) and defined MGMT methylation status by a polymerase chain reaction using the CpGenome DNA Modification Kit («Chemicon International¼, USA). The data were statistically processed using a package of Statistica 6.0 programs. RESULTS: Patient age, time to development of recurrent glioblastoma, mutations in the IDH-1 gene and MGMT were found to be prognostic factors for overall survival among adult patients in this category. CONCLUSION: Analysis of clinical findings and identification of molecular genetic aberrations in the tumor cells will be able to elaborate an individual approach to treating patients with glioblastoma in order to increase their survival rates and to improve quality of life.

[Breast carcinoma metastasis to the optic nerve: case report and review of literature].

Metastatic tumours of the optic nerve are extremely rare. The review of literature revealed only 12 cases of breast carcinoma metastasis to the optic nerve. All patients survived less then 6 month after surgical treatment. We describe a case of metastatic breast carcinoma to the optic nerve that occurred 8 years after radical mastectomy followed by chemotherapy. The metastasis manifested with progressive decrease in visual acuity in the right eye during 3 month. CT and MRI demostrated enhancing lesion in the muscle cone apex of the right orbit with an extension to the optic canal. The presumable diagnosis was optic nerve sheath meningioma, and surgical resection was performed. The tumour involved the optic nerve and has been resected togeher with the nerve. Histology report confirmed metastatic tumour. Postoperatively, the patient received additional stereotactic radiotherapy. Patient died of tumour dissemination 2,5 years after surgery. Breast carcinoma metastases to the optic nerve usually have unfavorable prognosis both for survival and for visual acuity. Isolated metastatic tumors of the optic nerve remain a diagnostic challenge because of their clinical and radiological similarities to more common primary tumors of the optic nerve.

State of oncomarker protein B23/nucleophosmin in HeLa cells.

Western blot after SDS-PAGE for protein separation showed two immunoreactive bands corresponding to monomers (38-40 kDa) and oligomers (210-230 kDa) of nucleophosmin in HeLa cell lysates. Decreasing the buffer ionic strength during the incubation of cells and nuclei destabilized these oligomers. We also showed the existence of two B23/nucleophosmin pools in nuclei of HeLa cells with different sensitivity to hypotonic buffer treatment: one extractable from the nucleus and the other non-extractable and tightly bound to the nucleus. A detailed structural analysis of the extractable B23 pool was carried out: two closely related nucleophosmin isoforms (B23.1 and B23.2) were identified as a result of analysis of C-terminal amino acid sequences using carboxypeptidase hydrolysis; the N-termini of both isoforms are blocked by an acetyl group. As a result of sequencing of the deacetylated proteins, it has been established that the N-terminal amino acid sequence of nucleophosmin in these preparations is truncated by nine amino acid residues and the acetylated residue is Ser. The truncated monomer of nucleophosmin (represented only by the extractable part of the protein) on addition of magnesium ions to low ionic strength buffer or increase in buffer ionic strength was shown to form oligomers with molecular weights (210-230 kDa) similar to those revealed in the total cell lysate. It should be noted that the set of oligomers in this case differs from the one in total cell lysate. Our strategy of characterization of B23 forms for HeLa cells can be applied for other tumor cells.

[Antibodies to synthetic fragments of nucleophosmin for the specific detection of its monomeric and oligomeric forms].

Immunoactive fragments corresponding to the N-terminal (19-36) and C-terminal (283-294) regions of the NPM1.1 isoform of nucleophosmin and their shortened fragments were chosen and synthesized. Rabbits were immunized with free full-size peptides and their protein conjugates. Antibodies produced against the 19-36 and 283-294 peptides were purified by affinity chromatography on bromocyanogen-activated sepharose that was preliminary conjugated with the synthetic peptides. An analysis of immunoblots of lysates of the HeLa and Ramos cells demonstrated that the antibodies produced against the 19-36 peptide detected the monomeric form of nucleophosmin, whereas the antibodies against the 283-294 peptide predominantly revealed its oligomeric form. It was established by immunocytochemical analysis that the antibodies induced by the 19-36 peptide stained the nucleoplasm and perinuclear space of the cytoplasm of the HeLa and Ramos cells, but did not stain the nucleoli, while the antibodies against the 283-294 peptide stained only the nucleoli of the same cells. On the basis of these results, one could propose that the monomeric and oligomeric forms of nucleophosmin were located in the nucleoplasm and nucleoli of the examined cells, respectively. Thus, antibodies which can predominantly detect monomeric and oligomeric forms of nucleophosmin were produced for the first time. An analysis of the monomeric-oligomeric state and the location of the nucleophosmin in tumor cells could be performed using these antibodies.