Immunization of Mice with Gold Nanoparticles Conjugated to Thermostable Cancer Antigens Prevents the Development of Xenografted Tumors.

Gold nanoparticles as part of vaccines greatly increase antigen stability, antigen accumulation in the lymph nodes, and antigen uptake by antigen-presenting cells. The use of such particles as part of anticancer vaccines based on heat shock proteins to increase vaccine effectiveness is timely. We prepared and characterized nanoconjugates based on 15-nm gold nanoparticles and thermostable tumor antigens isolated from MH22a murine hepatoma cells. The whole-cell lysate of MH22a cells contained the main heat shock proteins. BALB/c mice were injected with the conjugates and then received transplants of MH22a cells. The highest titer was produced in mice immunized with the complex of gold nanoparticles + antigen with complete Freund's adjuvant. The immunized mice showed no signs of tumor growth for 24 days. They also showed a decreased production of the INF-γ, IL-6, and IL-1 proinflammatory cytokines compared to the mice immunized through other schemes. This study is the first to show that it is possible in principle to use gold nanoparticles in combination with thermostable tumor antigens for antitumor vaccination. Antitumor vaccines based on thermostable tumor antigens can be largely improved by including gold nanoparticles as additional adjuvants.

Biomedical evaluation of antioxidant properties of lamb meat enriched with iodine and selenium.

The article presents a study of the antioxidant properties of meat from lambs that received organic forms of iodine and selenium during growth. This meat was included in diets of laboratory animals using a model of acute toxic hepatitis. The experiments resulted in developing and testing a technique that was effective in enriching lamb with bioorganic elements of iodine and selenium and contributed to the activation metabolism in the bodies of animals consuming the meat. The purpose of the presented investigation was to compare the roles of bioorganic iodine and selenium and their combination as antioxidants in rat rations using a model of acute toxic hepatitis induced by carbon tetrachloride. The experimental studies have established a hepatoprotective effect of lamb meat enriched with selenium and iodine on rats suffering from toxic xenobiotic effects. This was confirmed by normalized hematological and biochemical measures in the blood of the experimental rats.

Classification of skin cancer using convolutional neural networks analysis of Raman spectra.

BACKGROUND AND OBJECTIVE: Skin cancer is the most common malignancy in whites accounting for about one third of all cancers diagnosed per year. Portable Raman spectroscopy setups for skin cancer "optical biopsy" are utilized to detect tumors based on their spectral features caused by the comparative presence of different chemical components. However, low signal-to-noise ratio in such systems may prevent accurate tumors classification. Thus, there is a challenge to develop methods for efficient skin tumors classification. METHODS: We compare the performance of convolutional neural networks and the projection on latent structures with discriminant analysis for discriminating skin cancer using the analysis of Raman spectra with a high autofluorescence background stimulated by a 785 nm laser. We have registered the spectra of 617 cases of skin neoplasms (615 patients, 70 melanomas, 122 basal cell carcinomas, 12 squamous cell carcinomas and 413 benign tumors) in vivo with a portable Raman setup and created classification models both for convolutional neural networks and projection on latent structures approaches. To check the classification models stability, a 10-fold cross-validation was performed for all created models. To avoid models overfitting, the data was divided into a training set (80% of spectral dataset) and a test set (20% of spectral dataset). RESULTS: The results for different classification tasks demonstrate that the convolutional neural networks significantly (p<0.01) outperforms the projection on latent structures. For the convolutional neural networks implementation we obtained ROC AUCs of 0.96 (0.94 - 0.97; 95% CI), 0.90 (0.85-0.94; 95% CI), and 0.92 (0.87 - 0.97; 95% CI) for classifying a) malignant vs benign tumors, b) melanomas vs pigmented tumors and c) melanomas vs seborrheic keratosis respectively. CONCLUSIONS: The performance of the convolutional neural networks classification of skin tumors based on Raman spectra analysis is higher or comparable to the accuracy provided by trained dermatologists. The increased accuracy with the convolutional neural networks implementation is due to a more precise accounting of low intensity Raman bands in the intense autofluorescence background. The achieved high performance of skin tumors classifications with convolutional neural networks analysis opens a possibility for wide implementation of Raman setups in clinical setting.

In vivo diagnosis of skin cancer with a portable Raman spectroscopic device.

In this study, we performed in vivo diagnosis of skin cancer based on implementation of a portable low-cost spectroscopy setup combining analysis of Raman and autofluorescence spectra in the near-infrared region (800-915 nm). We studied 617 cases of skin neoplasms (615 patients, 70 melanomas, 122 basal cell carcinomas, 12 squamous cell carcinomas and 413 benign tumors) in vivo with a portable setup. The studies considered the patients examined by GPs in local clinics and directed to a specialized Oncology Dispensary with suspected skin cancer. Each sample was histologically examined after excisional biopsy. The spectra were classified with a projection on latent structures and discriminant analysis. To check the classification models stability, a 10-fold cross-validation was performed. We obtained ROC AUCs of 0.75 (0.71-0.79; 95% CI), 0.69 (0.63-0.76; 95% CI) and 0.81 (0.74-0.87; 95% CI) for classification of a) malignant and benign tumors, b) melanomas and pigmented tumors and c) melanomas and seborrhoeic keratosis, respectively. The positive and negative predictive values ranged from 20% to 52% and from 73% to 99%, respectively. The biopsy ratio varied from 0.92:1 to 4.08:1 (at sensitivity levels from 90% to 99%). The accuracy of automatic analysis with the proposed system is higher than the accuracy of GPs and trainees, and is comparable or less to the accuracy of trained dermatologists. The proposed approach may be combined with other optical techniques of skin lesion analysis, such as dermoscopy- and spectroscopy-based computer-assisted diagnosis systems to increase accuracy of neoplasms classification.

Synthesis of silymarin-selenium nanoparticle conjugate and examination of its biological activity in vitro.

Silymarin (Sil) was conjugated to selenium nanoparticles (SeNPs) to increase Sil bioavailability. The conjugates were monodisperse; the average diameter of the native SeNPs was ~ 20-50 ± 1.5 nm, whereas that of the conjugates was 30-50 ± 0.5 nm. The use of SeNPs to increase the bioavailability of Sil was examined with the MH-22a, EPNT-5, HeLa, Hep-2, and SPEV-2 cell lines. The EPNT-5 (glioblastoma) cells were the most sensitive to the conjugates compared to the conjugate-free control. The conjugates increased the activity of cellular dehydrogenases and promoted the penetration of Sil into the intracellular space. Possibly, SeNPs play the main part in Sil penetration of cells and Sil penetration is not associated with phagocytosis. Thus, SeNPs are promising for use as a Sil carrier and as protective antigens.

Single-Center Experience of Surgical Treatment of Primary Retroperitoneal Tumors.

This study is an evaluation of surgical treatment results of primary retroperitoneal tumors. In Samara Regional Clinical Oncology Dispensary, from 2008 to 2015, the treatment of 187 patients (53 men and 134 women) was conducted. One hundred fifteen patients got tumor removal within the healthy tissue (R0), and 61 patients went through complete resection of tumor with wide margins (R0). Complete resection of tumor with wide margins (R0) with preoperative tumor vessel embolization was performed in 11 patients. According to the histological examination, malignant retroperitoneal tumor was detected in 85 patients (48.4%); in most cases it was presented by various forms of sarcoma. A benign tumor was diagnosed in 71 patients (40.3%), fibrolipomas (17.1%), and neurofibromas (12.5%). The diagnosis of 20 patients needs subsequent clarification, as mesenchymal tumor (6.2%) and histiocytoma (5.1%) were diagnosed. Short-term results of surgical treatment for the group, where complete resection of tumor with wide margins was performed: intraoperative blood loss 410.91 + - 113.31(ml), operation time 185.15 + -32.49(min); postoperative complications 10 (16,4%); mortality 3 (4,9%); LOS 23,14 ± 6,31; for removal of the tumor within healthy tissues: intraoperative blood loss 281.33 + -110.94 (ml), operation time 58.33 + -27.14(min) postoperative complications 7 (6,08%); mortality 2 (1,74%); LOS 6,98 ± 4,83; (t = 279, p = 0,015). For patients who went through preoperative tumor feeding vessel embolization, intraoperative blood loss was 121.33 ± 27.94 (ml), time of operation 43.13 ± 16.11 (min), postoperative complication 1 (4.5%), mortality 0 (0%), and length of stay 12.72 ± 1.49. After the complete resection of tumor with wide margins, intraoperative blood loss, operation time, the number of postoperative complications, and postoperative LOS were significantly greater in comparison with the group of patients where the tumor was removed within healthy tissues. The method of preoperative embolization of the tumor feeding vessels can reduce intraoperative blood loss, the time of operation, and the number of postoperative complications.

Portable spectroscopic system for in vivo skin neoplasms diagnostics by Raman and autofluorescence analysis.

The present paper studies the applicability of a portable cost-effective spectroscopic system for the optical screening of skin tumors. in vivo studies of Raman scattering and autofluorescence (AF) of skin tumors with the 785 nm excitation laser in the near-infrared region included malignant melanoma, basal cell carcinoma and various types of benign neoplasms. The efficiency of the portable system was evaluated by comparison with a highly sensitive spectroscopic system and with the diagnosis accuracy of a human oncologist. Partial least square analysis of Raman and AF spectra was performed; specificity and sensitivity of various skin oncological pathologies detection varied from 78.9% to 100%. Hundred percent accuracy of benign and malignant skin tumors differentiation is possible only with a combined analysis of Raman and AF signals.

Prospects for the use of spherical gold nanoparticles in immunization.

Recent years have seen extremely fast development of new viral nanovaccines and diagnostic agents using nanostructures prepared by biological and chemical synthesis. We used spherical gold nanoparticles (average diameter, 15 nm) as a platform for the antigen for swine transmissible gastroenteritis virus (TGEV). The literature data demonstrate that immunization of animals with the TGEV antigen coupled to gold nanoparticles (GNPs) not only activates antigen-presenting cells but also increases the proliferative activity of splenic lymphoid (antibody-forming) cells. The contents of γ-IFN, IL-1ß, and IL-6 in animals immunized with GNP-antigen conjugates were found to be higher than those in intact animals or in animals given the antigen alone. The increased concentration of IL-1ß in the immunized animals directly correlated with the activity of macrophages and stimulated B cells, which produce this cytokine when activated. The increased concentration of IL-6 indicates that the injected preparations are stimulatory to cellular immunity. Immunization with the TGEV antigen conjugated to GNPs as a carrier activates the respiratory activity of lymphoid cells and peritoneal macrophages, which is directly related to their transforming activity and to the activation of antibody generation. Furthermore, the use of this conjugate allows marked improvement of the structure of the animals' immune organs and restores the morphological-functional state of these organs. The microanatomical changes (increased number of follicles) indicate the activation of the B-dependent zone of the spleen and, consequently, the development of a humoral-type immunological reaction. The degradative processes observed in the animals immunized with TGEV antigen alone are evidence of weak resistance to pathogen attack. These results can be used to develop vaccines against this infection by employing TGEV antigen coupled to gold nanoparticles as a carrier.

Combined Raman and autofluorescence ex vivo diagnostics of skin cancer in near-infrared and visible regions.

The differentiation of skin melanomas and basal cell carcinomas (BCCs) was demonstrated based on combined analysis of Raman and autofluorescence spectra stimulated by visible and NIR lasers. It was ex vivo tested on 39 melanomas and 40 BCCs. Six spectroscopic criteria utilizing information about alteration of melanin, porphyrins, flavins, lipids, and collagen content in tumor with a comparison to healthy skin were proposed. The measured correlation between the proposed criteria makes it possible to define weakly correlated criteria groups for discriminant analysis and principal components analysis application. It was shown that the accuracy of cancerous tissues classification reaches 97.3% for a combined 6-criteria multimodal algorithm, while the accuracy determined separately for each modality does not exceed 79%. The combined 6-D method is a rapid and reliable tool for malignant skin detection and classification.

Comparative analysis of combined spectral and optical tomography methods for detection of skin and lung cancers.

Malignant skin tumors of different types were studied in vivo using optical coherence tomography (OCT), backscattering (BS), and Raman spectroscopy (RS). A multimodal method is proposed for early cancer detection based on complex analysis of OCT images by their relative alteration of scattered-radiation spectral intensities between malignant and healthy tissues. An increase in average accuracy of diagnosis was observed for a variety of cancer types (9% sensitivity, 8% specificity) by a multimodal RS-BS-OCT system in comparison with any of the three methods used separately. The proposed approach equalizes the processing rates for all methods and allows for simultaneous imaging and classification of tumors.

The usage of phage mini-antibodies as a means of detecting ferritin concentration in animal blood serum.

Mini-antibodies that have specific ferritin response have been produced for the first time using sheep's phage libraries (Griffin.1, Medical Research Council, Cambridge, UK). Produced phage antibodies were used for the first time for the development of diagnostic test kits for ferritin detection in the blood of cattle. The immunodot assay with secondary biospecific labeling is suggested as means of ferritin detection in cow blood serum (antiferritin phage antibodies and rabbit antiphage antibodies conjugated with different labels). Сolloidal gold, gold nanoshells, and horse reddish peroxidase used as labels have shown a similar response while detecting concentration of ferritin (0.2 mg/mL). It is shown that the method of solid-phase immunoassay with a visual view of the results allows determination of the minimum concentration of ferritin in the blood of cows at 0.225 g/mL.