A Novel Technique for Preparation, Staining, and Visualization of Tissue with Metal Implants and Extraskeletal Calcification Areas.

The aim of the study was to evaluate the efficacy of a novel technique for preparation, staining, and visualization of tissues containing extra-skeletal mineralization areas, all-metal implants or their prototypes for their subsequent examination using scanning electron microscopy in the backscattered electron mode. MATERIALS AND METHODS: After fixation in 10% formalin (24 h), the biomaterial (a titanium nickelide plate with the surrounding tissues after subcutaneous implantation, patented titanium alloy plates with the surrounding tissues after cranioplasty, primary and secondary calcified atherosclerotic plaques) were fixed with 1% osmium tetroxide (12 h) and then stained with 2% aqueous solution of osmium tetroxide (48 h). The samples were further stained with 2% alcoholic uranyl acetate (5 h), dehydrated with isopropanol (5 h) and acetone (1 h), impregnated with a mixture of acetone and epoxy resin Epon (1:1, 6 h) and then embedded into a fresh portion of epoxy resin (24 h), which was followed by polymerization at 60°C. After grinding and polishing, epoxy blocks were counterstained with lead citrate (7 min) and sputter-coated with carbon, then the samples were visualized by scanning electron microscopy in the backscattered electron mode. The elemental composition was studied using X-ray microanalysis. RESULTS: The developed technique allows obtaining high-quality images at five thousand-fold magnifications, provides the possibility to identify the shape and structure of intact metal and mineral inclusions, and to type the surrounding cells, distinguishing mesenchymal and immunocompetent cells by shape and cytoplasmic content. Apart from connective tissue capsule thickness and leukocyte infiltration, this technique makes it possible to estimate the number and area of newly formed small-caliber vessels representing a surrogate marker of inflammation. CONCLUSION: The proposed technique provides the possibility to investigate adequately the structure of samples when their sectioning is impossible or significantly complicated, with image quality remarkably higher than that obtained by light microscopy.

Correlation of Metabolic Profiles of Plasma and Cerebrospinal Fluid of High-Grade Glioma Patients.

This work compares the metabolic profiles of plasma and the cerebrospinal fluid (CSF) of the patients with high-grade (III and IV) gliomas and the conditionally healthy controls using the wide-range targeted screening of low molecular metabolites by HPLC-MS/MS. The obtained data were analyzed using robust linear regression with Huber's M-estimates, and a number of metabolites with correlated content in plasma and CSF was identified. The statistical analysis shows a significant correlation of metabolite content in plasma and CSF samples for the majority of metabolites. Several metabolites were shown to have high correlation in the control samples, but not in the glioma patients. This can be due to the specific metabolic processes in the glioma patients or to the damaged integrity of blood-brain barrier. The results of our study may be useful for the understanding of molecular mechanisms underlying the development of gliomas, as well as for the search of potential biomarkers for the minimally invasive diagnostic procedures of gliomas.

Autologous and Pooled Tumor Lysates in Combined Immunotherapy of Patients with Glioblastoma.

Although major progress has been made in the standard treatment for glioblastomas, encompassing the maximal surgical resection, chemotherapy and radiation therapy, it is possible to increase survival rates significantly only in a few patients. Therefore, it is necessary to explore new therapeutic modalities, one of which is immunotherapy. The aim of the study was to evaluate the efficacy of the combined use of autologous and pooled tumor lysates in comprehensive treatment of patients with glioblastoma. MATERIALS AND METHODS: All patients (n=58, including 30 males and 28 females aged 18-70 years) were randomized into three groups, two of which received immunotherapy based on injection of autologous dendritic cells pulsed with autologous tumor lysates (first protocol) or pooled lysates (second protocol) from more than one tumor, in addition to the planned standard treatment. The patients of group 3 (control) received the standard comprehensive treatment encompassing the maximum safe tumor resection followed by radiation therapy and chemotherapy. RESULTS: The tolerability of both applied immunotherapy protocols was good: there were no anaphylactic reactions observed or patients who prematurely discontinued participation in the study. The final analysis of the data revealed no significant differences in median survival values of patients in each of the three groups. However, when analyzing the Karnofsky Performance Status in patients of group 2, it was found that it tended to improve. CONCLUSION: The study shows that the proposed immunotherapy protocols are safe for clinical use and have the potential to improve the patient's life quality. However, these findings should be considered intermediate until the findings of multicenter randomized clinical trials with a larger number of patients are obtained.

[Cerebral actinomycotic granuloma mimicking malignant brain tumor].

We report a clinical case of cerebral actinomycotic granuloma that was preoperatively diagnosed as an intracerebral tumor. On the basis of a histological examination performed after lesion resection, the diagnosis of a cerebral actinomycotic granuloma was made. The article presents long-term outcomes of the surgery. The literature data on cases of various forms of cerebral actinomycosis are provided.

CCL19/CCL21-Dependent Chemotaxis of Dendritic Cells in Healthy Individuals and Patients with Brain Tumors.

We compared migration activities of IFN-α- and IL-4-induced dendritic cells (IFN-DC and IL4-DC) generated from blood monocytes of healthy donors and analyzed migration activity of IFN-DC from patients with brain tumors. In the presence of CCL19 chemokine, donor IFN-DC exhibited higher migration activity than IL4-DC, the expression of chemokine CCR7-receptor being similar in the two cell types. IFN-DC of patients with malignant gliomas were characterized by low chemotaxis in response to CCL19 and CCL21 stimulation despite a trend to higher expression of CCR7 in comparison with donor IFN-DC.