Search

Objective — To investigate the apoptotic activity of Gratiola officinalis L. extract on human tumor cell lines by flow cytofluorometry. Material and Methods — The extract of Gratiola officinalis L. was manufactured via our original methodology. Studies were performed on human tumor cell lines: HeLa – cervical carcinoma, Jurkat – T-cell lymphoblastic leukemia, MCF-7 – breast adenocarcinoma, A549 – lung carcinoma, PC-3 – prostate carcinoma, HCT-116 – colon carcinoma, A498 – renal carcinoma, and SK-BR-3 – human breast carcinoma. Induction of apoptosis was studied after incubating cell lines with Gratiola officinalis L. extract at a concentration of 0.9 mg/mL using the Annexin V-FITC Apoptosis Kit. Caspase-dependent apoptosis was examined on a flow cytometer using anti-caspase-3-FITC (BD) kit on the Jurkat cell line. Morphological studies of HeLa cervical carcinoma cells in the alive and dead test were performed using two stains, acridine orange and propidium iodide, at different concentrations of the extract. The statistical data processing was performed using Microsoft Office Excel software. Results — One day after their exposure to Gratiola officinalis L. extract at a concentration of 0.9 mg/mL, tumor cells were mostly in late apoptosis stage. Cytotoxic activity of Gratiola officinalis L. extract was established for all investigated tumor cell cultures but their sensitivities to the extract were different. Mechanisms of antitumor action of Gratiola officinalis L. extract were identified: we established that the extract induced caspase-dependent apoptosis in tumor cells. Conclusion — The identified mechanisms of apoptotic activity of Gratiola officinalis L. extract confirmed the prospects of bioflavonoids as new-generation antitumor agents.

Optical clearing of the lung tissue aims to make it more transparent to light by minimizing light scattering, thus allowing reconstruction of the three-dimensional structure of the tissue with a much better resolution. This is of great importance for monitoring of viral infection impact on the alveolar structure of the tissue and oxygen transport. Optical clearing agents (OCAs) can provide not only lesser light scattering of tissue components but also may influence the molecular transport function of the alveolar membrane. Air-filled lungs present significant challenges for optical imaging including optical coherence tomography (OCT), confocal and two-photon microscopy, and Raman spectroscopy, because of the large refractive-index mismatch between alveoli walls and the enclosed air-filled region. During OCT imaging, the light is strongly backscattered at each air-tissue interface, such that image reconstruction is typically limited to a single alveolus. At the same time, the filling of these cavities with an OCA, to which water (physiological solution) can also be attributed since its refractive index is much higher than that of air will lead to much better tissue optical transmittance. This review presents general principles and advances in the field of tissue optical clearing (TOC) technology, OCA delivery mechanisms in lung tissue, studies of the impact of microbial and viral infections on tissue response, and antimicrobial and antiviral photodynamic therapies using methylene blue (MB) and indocyanine green (ICG) dyes as photosensitizers.

Using fluorescence methods for visualization of human tumor cell apoptosis and necrosis in vitro, the antitumor effects of flavonoid-containing Gratiola officinalis L. and green tea extracts at ultralow concentrations on A498 human kidney carcinoma cells after 24 and 48 h were compared. Fluorescent dyes (propidium iodide and acridine orange) were used in the “living and dead” test, which made it possible to identify the total number of cells that died by necrosis and apoptosis, as well as cells in which apoptosis has started and apoptotic bodies or pycnosis of the nucleus were detected. Gratiola officinalis L. extract had the most pronounced cytostatic activity 48 h after exposure; the number of cells per field of view decreased by a factor of 1.5 compared with the control. Green tea extract at a concentration of 0.0288 mg/mL had the most pronounced cytotoxic activity; its manifestation was growing 48 h after exposure. Gratiola officinalis L. extract exhibited cytotoxic activity at lower concentrations (0.0036 mg/mL).

The purpose of this study was to investigate the prognostic factors of plasmonic photothermal therapy (PPT) efficiency in rats with transplanted liver cancer on the basis of assessment of the degree of tumor vascularization. Before any exposure, rats with transplanted liver cancer PC-1 were subjected to Doppler ultrasonography to assess the degree of vascularization of transplanted tumors. Half of the animals with the transplanted tumors were withdrawn from the experiment after Doppler ultrasonography to determine the content of vascular factors in blood serum by enzyme immunoassay and in tumor tissue by immunohistochemical method. After three intravenous injections of polyethylene glycol-coated gold nanorods (GNRs) at a dose of 0.4 mq/mL, the transplanted tumors were irradiated percutaneously with infrared laser radiation at a wavelength of 808 nm, followed by thermography of local tumor heating. The animals were withdrawn from the experiment 24 h later, and tumor tissue samples were taken for histological examination. It is found that the accumulation of gold in the tumor tissue and the PPT efficiency at repeated intravenous GNR injections are determined by the presence of formed vasculature in the tumor.

The quercetin content in aqueous EtOH extracts of covering leaves of anthocyanin Purple Saratov diploid variety of Zea mays L. grown under dry conditions reached 39.36 ± 0.41 mg/100 g of dry extract and significantly exceeded the value (26.03 ± 0.36 mg/100 g of dry extract) for maize grown under normal wetting conditions. The large amounts of polyphenolic compounds in anthocyanin Purple Saratov diploid variety showed good prospects for using it as a source of them.

Objective — To compare the expression of the LC3B autophagy marker before and after treatment with high-intensity focused ultrasound therapy (HIFU) and combined therapies: androgenic deprivation AD + HIFU, AD + radiation therapy in patients with and without recurrent prostate adenocarcinoma. Material and Methods — The objects of our study were biopsy specimens of prostates obtained from 70 patients with prostate adenocarcinoma before and after treatment. The first group consisted of patients without signs of relapse after treatment, as evidenced by a normal blood PSA level, the results of ultrasound procedure and biopsy. The second group included patients who had a recurrence of adenocarcinoma after treatment. Patients received the following cancer treatments: ultrasound ablation (HIFU), combination therapy: androgenic deprivation AD + HIFU, AD + radiation therapy. An immunohistochemical staining in two groups was performed with an anti-LC3B antibody. Results — In the group of patients without relapse, the expression of the autophagy marker sharply decreased in the tumor tissue after treatment, but it increased in the stroma of muscle connective tissue and the vascular wall. In the group of patients with relapse, the expression of anti-LC3B antibody significantly increased in tumor cells, in stroma of muscle connective tissue, and in vascular wall after treatment. In both groups, the magnitude of the autophagy marker expression did not depend on the treatment method. Conclusion — The autophagy is one of the factors contributing to the survival of prostate adenocarcinoma cells after treatment; the increase in autophagic protein expression after treatment in patients with prostate adenocarcinoma can be used to assess the risk of relapse.

The present work demonstrates the assessment of cytotoxicity upconversion nanoparticles (UCNPs) coated by SiO2 on different normal and cancer murine cell lines in vitro. The cell viability is scored for cytotoxic effects of UCNPs at dark conditions. UCNPs coated by silica shells provide a dose-dependent cytotoxic effect on all studied cell lines which was most pronounced for the Raw264.7 cell line. It is probably caused by the high phagocytic activity of macrophages. The less sensitive cell line was 4T1. The statistically significant differences in cell viability after 24 and 48 h of incubation of cells with particles were observed just for the macrophage cell line. It is worth notifying that after 48 h of incubation the cytotoxic effect on Raw 264.7 cell line increased which shows a possible negative effect on some subpopulations on blood cells. The obtained results confirm a high sensitivity of the UCNPs to the concentration variations within cells. Carriers based on UCNPs and dyes are promising alternatives to photosensitizer for traditional photodynamic therapy and possess prominent potentials in biological and clinical applications.

Optical clearing (OC) of adipose tissue has not been studied enough, although it can be promising in medical applications, including surgery and cosmetology, for example, to visualize blood vessels or increase the permeability of tissues to laser beams. The main objective of this work is to develop technology for OC of abdominal adipose tissue in vivo using hyperosmotic optical clearing agents (OCAs). The maximum OC effect (77%) was observed for ex vivo rat adipose tissue samples exposed to OCA on fructose basis for 90 minutes. For in vivo studies, the maximum effect of OC (65%) was observed when using OCA based on diatrizoic acid and dimethylsulfoxide for 120 minutes. Histological analysis showed that in vivo application of OCAs may induce a limited local necrosis of fat cells. The efficiency of OC correlated with local tissue damage through cell necrosis due to accompanied cell lipolysis.

Background and aim — Gratiola officinalis extract has antimicrobial properties, but its wound healing activity has not been described. Material and Methods — The aqueous solution of dry Gratiola officinalis extract was used for treatment of skin wounds with different routes of administration. In the experiment 40 white laboratory male rats with weight of 150±50 grams were subjected to the incised model of flat skin wounds in the interscapular region under the general anesthesia. A morphological study was carried out to assess the effectiveness of the therapy administered. Results — The most effective method of treatment was found to be local application of the extract, which reduced wound healing time by 8 days. Intramuscular and oral administrations reduce healing time by 6 days. Morphological signs of regeneration, which developed under the influence of Gratiola officinalis extract, allowed to estimate the wound healing activity. Clear differentiation and thickness of layers of the newly formed epidermis, a large number of thin-walled vessels and actively proliferating fibroblasts indicate an active regeneration process. Conclusion — This extract may be interesting to specialists in regenerative medicine as a remedy for treatment of wound processes.

Cancer remains one of the leading causes of death in the world. For a number of neoplasms, the efficiency of conventional chemo- and radiation therapies is insufficient because of drug resistance and marked toxicity. Plasmonic photothermal therapy (PPT) using local hyperthermia induced by gold nanoparticles (AuNPs) has recently been extensively explored in tumor treatment. However, despite attractive promises, the current PPT status is limited by laboratory experiments, academic papers, and only a few preclinical studies. Unfortunately, most nanoformulations still share a similar fate: great laboratory promises and fair preclinical trials. This review discusses the current challenges and prospects of plasmonic nanomedicine based on PPT and photodynamic therapy (PDT). We start with consideration of the fundamental principles underlying plasmonic properties of AuNPs to tune their plasmon resonance for the desired NIR-I, NIR-2, and SWIR optical windows. The basic principles for simulation of optical cross-sections and plasmonic heating under CW and pulsed irradiation are discussed. Then, we consider the state-of-the-art methods for wet chemical synthesis of the most popular PPPT AuNPs such as silica/gold nanoshells, Au nanostars, nanorods, and nanocages. The photothermal efficiencies of these nanoparticles are compared, and their applications to current nanomedicine are shortly discussed. In a separate section, we discuss the fabrication of gold and other nanoparticles by the pulsed laser ablation in liquid method. The second part of the review is devoted to our recent experimental results on laser-activated interaction of AuNPs with tumor and healthy tissues and current achievements of other research groups in this application area. The unresolved issues of PPT are the significant accumulation of AuNPs in the organs of the mononuclear phagocyte system, causing potential toxic effects of nanoparticles, and the possibility of tumor recurrence due to the presence of survived tumor cells. The prospective ways of solving these problems are discussed, including developing combined antitumor therapy based on combined PPT and PDT. In the conclusion section, we summarize the most urgent needs of current PPT-based nanomedicine.

The deposition of amyloid-β (Aβ) in the brain is a risk factor for Alzheimer’s disease (AD). Therefore, new strategies for the stimulation of Aβ clearance from the brain can be useful in preventing AD. Transcranial photostimulation (PS) is considered a promising method for AD therapy. In our previous studies, we clearly demonstrated the PS-mediated stimulation of lymphatic clearing functions, including Aβ removal from the brain. There is increasing evidence that sleep plays an important role in Aβ clearance. Here, we tested our hypothesis that PS at night can stimulate Aβ clearance from the brain more effectively than PS during the day. Our results on healthy mice show that Aβ clearance from the brain occurs faster at night than during wakefulness. The PS course at night improves memory and reduces Aβ accumulation in the brain of AD mice more effectively than the PS course during the day. Our results suggest that night PS is a more promising candidate as an effective method in preventing AD than daytime PS. These data are an important informative platform for the development of new noninvasive and nonpharmacological technologies for AD therapy as well as for preventing Aβ accumulation in the brain of people with disorder of Aβ metabolism, sleep deficit, elderly age, and jet lag.

We report here a pilot identification of the presence of the lumenized Lyve-1/Prox-1-expressing vessels with distinct walls composed of a single endothelial layer in the unaffected brain and with intraventricular hemorrhages. These lymphatic vessels (LVs) have valves and an undulating shape in the distal region that is the classical characteristic of lymphatic precollectors. Furthermore, we identified Lyve-1/Prox-1-expressing lymphatic elements in the enlarged perivascular spaces. These pioneering results might be a revolutionary step in our understanding of anatomy and physiology of the cerebral lymphatics and stimulate a reassessment of basic assumptions in the aetiology of brain diseases associated with lymphatic dysfunction. The discovery of the cerebral LVs is a crucial step in the development of breakthrough technologies of modulations of lymphatic removing of toxins and blood from the brain.One Sentence SummaryLyve1/Prox1-expressing lymphatic vessels and elements in the unaffected and affected human brain.

The influence of long-term peroral injection of gold nanoparticles with various sizes on the liver, spleen, and lymph nodes of adult laboratory rats and their progeny was studied. The morphometric analysis was conducted using a µVizo-103 medical digital microviser. The conglomerated gold nanoparticles were detected in tissues via the dark-field microscopy. Gold nanoparticles with sizes of 2 and 50 nm were found to cause the mostly pronounced morphological alterations in organs that testifies to their size-dependent impact. The liver in a group receiving 50-nm gold particles exhibit noticeable dystrophy of hepatocytes and activation of a monocyte macrophage system. The spleen and lymph nodes reveal the activation of proliferation and differentiation of immunocompetent cells and macrophages. Peroral injection of gold particles during gestation of females led to an increase in a number of hepatocytes with necrobiotic changes as well as to elevated amount of non-parenchymatic elements in the liver of the progeny, especially in groups containing the 2- and 50-nm gold nanoparticles.

The possibilities of using fluorescence-based analytical methods and their advantages for visualization and identification of the type of programmed cell death in human tumor cells exposed to flavonoids have been analyzed in experiments in vitro. HeLa cervical cancer cells and A498 kidney carcinoma cells were used as the objects of the study, and exposure to the flavonoid-containing extract of common hedgehyssop (Gratiola officinalis L.) was tested as the experimental treatment. The following fluorescence-based techniques were used: the “live and dead” test with double staining by propidium iodide and acridine orange and double staining with annexin and propidium iodide. Autophagy induction was confirmed by fluorescence-based tests implemented in a Muse cell analyzer with a Muse Autophagy LC3-Antibody Based Kit. The use of double staining with acridine orange and propidium iodide fluorescent dyes in the “live and dead” test and comparison with phase contrast microscopy enables the visualization of apoptotic body and autophagosome formation processes in the cells and can, therefore, be used as a method of screening assessment of the efficiency of various chemotherapy drugs.

Our previous study has revealed that specific ornithosis antigens extracted from Сhlamydia psittaci (CP) could possess some modulatory activity on the growth of malignant solid tumors in Wistar rats. Here, in the same model we studied dose-dependent dynamics of tumor growth under the influence of CP antigen derived from CP AMK-16 strain, in doses ranged from 0.025 mg/kg to 1.25 mg/kg given as a single subcutaneous injection. Velocity of tumor growth has been estimated by t-LASCA technique.

The goal of our research was to assess the vascularization degree of tumor to predict the efficiency of plasmonic photothermal therapy (PPT) and photodynamic therapy (PDT) in tumor-bearing rats. Before any treatment, 3D Doppler ultrasound imaging was used for assessment of the vascularization degree of transplanted rat cholangiocarcinoma. For PPT, the gold nanorods with aspect ratio of 4:1, functionalized with thiolated polyethylene glycol, were used. After multiple fractional intravenous (IV) injections in rats with cholangiocarcinoma, the tumours were irradiated through the skin by an 808-nm NIR diode laser at a power density of 2.3 W/cm2 for 15 min. For PDT, galactose – Luphthalocyanine, as a photosensitizer, was applied by intratumoural injection, in a dose of 2 mg/kg. Then, tumors were irradiated through the skin by 670 nm-diode laser source with power density 200 mW/cm2 applied for 1000 sec in three non-overlapping zones covering whole tumour surface to obtain a total irradiation dose of 200 J/cm2 for each lesion. The withdrawal of the animals from the experiment and sampling of tissues for morphological study were performed before and 72 hrs after PPT and PDT. The vascular microdensity in tumors was assessed on histological sections as vessel counts or vessel area per unit of assessed tumor area. It was shown that efficiency of PPT and PDT therapy was mostly due to the sufficient accumulation of photothermosensitizers in the tumor, therefore preliminary assessment of tumor vascularization degree was necessary before starting a therapy.

The experimental study of ultrasonic modes was performed for ex vivo skin samples of white Wistar rats. The ultrasound was provided by a Dynatron 125 US system (Dynatronics, USA) after applying an optical clearing agent aqueousalcohol- fructose (20%-30%-50%) solution to the sample surface. Three different ultrasonic modes were used to found the most effective and fastest optical clearing. The optical coherence tomography system, Spectral Radar OCT System OCP930SR 022 (Thorlabs Inc., USA) with a wavelength of 930 nm, was used for obtaining the quantitative data for effectiveness and rate of skin optical clearing. The best mode had a frequency of 3 MHz and a power of 1.5 W in continuous wave mode for 3 minutes.

Polyelectrolyte microcapsules and other targeted drug delivery systems could substantially reduce the side effects of drug and overall toxicity. At the same time, the cardiovascular system is a unique transport avenue that can deliver drug carriers to any tissue and organ. However, one of the most important potential problems of drug carrier systemic administration in clinical practice is that the carriers might cause circulatory disorders, the development of pulmonary embolism, ischemia, and tissue necrosis due to the blockage of small capillaries. Thus, the presented work aims to find out the processes occurring in the bloodstream after the systemic injection of polyelectrolyte capsules that are 5 μm in size. It was shown that 1 min after injection, the number of circulating capsules decreases several times, and after 15 min less than 1% of the injected dose is registered in the blood. By this time, most capsules accumulate in the lungs, liver, and kidneys. However, magnetic field action could slightly increase the accumulation of capsules in the region-of-interest. For the first time, we have investigated the real-time blood flow changes in vital organs in vivo after intravenous injection of microcapsules using a laser speckle contrast imaging system. We have demonstrated that the organism can adapt to the emergence of drug carriers in the blood and their accumulation in the vessels of vital organs. Additionally, we have evaluated the safety of the intravenous administration of various doses of microcapsules.

The blood-brain barrier (BBB) poses a significant challenge for drug delivery to the brain. The limitations of our knowledge about the nature of BBB explains the slow progress in the therapy of brain diseases and absence of methods for drug delivery to the brain in clinical practice. Here we show that the BBB opens for high molecular weight compounds after exposure to loud sound (100 dB 370 Hz) in rats. The role of stress induced by loud sound and the systemic and molecular mechanisms behind it are discussed in the framework of the BBB opening as an informative platform for a novel fundamental knowledge about the nature of BBB and for the development of a non-invasive brain drug delivery technology.

The extract of G. officinalis was separated by five major fractions and the antitumor activity of hydrophobic fractions has been established. For the most active fractions spectral investigations of the main component were performed using UV-Vis electronic spectra, circular dichroism (CD), Fourier-transform infrared spectroscopy (FTIR) and nuclear magnetic resonance (NMR) spectroscopy. On the basis of spectral characteristics, it was demonstrated that the main components of the fractions 4 and 5 of G. officinalis providing its biological activity, highly likely, are of flavonoid nature. Based on the NMR data we propose the structure of the aglycon moiety of the main component from fraction 4 as 4,2’,5’-trihydroxychalcone. The fraction 5 is the most active, the antitumor effect of which is implemented through cytotoxic, cytostatic, apoptotic and autophagosomal activity in respect to human kidney cancer tumor cells.

Intraventricular hemorrhage (IVH) is the most fatal form of brain injury, yet a therapy directed at ameliorating intraventricular clot is very limited. There is accumulating evidence that an augmentation of the meningeal lymphatic (MLVs) functions might be a promising therapeutic target for IVH. In particular, the photostimulation (PS) of MLVs could be promising for non-invasive therapy of IVH via PS of clearance of red blood cells (RBCs) from the brain via MLVs. Indeed, we uncover that PS has therapeutic effects on IVH in mice reducing the mortality, improving the emotional status, accelerating the RBCs evacuation from the ventricles and increasing the ICP recovery. Our findings strongly suggest that the PS-mediated stimulation of drainage and clearing functions of MLVs can be a novel bedside, readily applicable and commercially viable technologies for treatment of IVH. These pilot results open new horizons in a non-invasive therapy of IVH via PS stimulation of regenerative lymphatic mechanisms.

The tumor vascularization in rats with transplanted cholangiocarcinoma was assessed by US power Doppler before plasmonic photothermal therapy (PPT). For PPT, the multiple intravenous injections of PEG-coated gold nanorods and irradiation by 808-nm NIR diode laser were used. We have shown that gold nanoparticles accumulation in tumor and efficiency of PPT are associated with the degree of vascularization of a tumor.

Early detection of a tumor makes it more probable that the patient will, finally, beat cancer and recover. The main goal of broadly defined cancer diagnostics is to determine whether a patient has a tumor, where it is located, and its histological type and severity. The major characteristic of the cancer affected tissue is the presence of the glioma cells in the sample. The current approach in diagnosis focuses mainly on microbiological, immunological, and pathological aspects rather than on the “metamaterial geometry” of the diseases. The determination of the effective properties of the biological tissue samples and treating them as disordered metamaterial media has become possible with the development of effective medium approximation techniques. Their advantage lies in their capability to treat the biological tissue samples as metamaterial structures, possessing the well-studied properties. Here, we present, for the first time to our knowledge, the studies on metamaterial properties of biological tissues to identify healthy and cancerous areas in the brain tissue. The results show that the metamaterial properties strongly differ depending on the tissue type, if it is healthy or unhealthy. The obtained effective permittivity values were dependent on various factors, like the amount of different cell types in the sample and their distribution. Based on these findings, the identification of the cancer affected areas based on their effective medium properties was performed. These results prove the metamaterial model capability in recognition of the cancer affected areas. The presented approach can have a significant impact on the development of methodological approaches toward precise identification of pathological tissues and would allow for more effective detection of cancer-related changes.

The early stages of gastric cancer are not recognized due to its asymptomatic development. Detection of patho- logical changes can be achieved using the photodynamic diagnostics, but this approach provides rather low tumor selectivity. Additional opportunities to improve the detection of precancerous stages may be provided by ana- lyzing blood circulation in the vessels of the stomach. We consider the possibilities of multiresolution analysis in recognizing changes in blood circulation in rats with different levels of the fluorescent signal in the stomach associated with precancerous stages.

When administered into the bloodstream, upconversion nanoparticles used in therapeutic / diagnostic purposes can affect red blood cells. In this work, using light optical microscopy, we show that the removal of citrate groups from the surface of NaYF4: Er3+ , Yb3+ nanoparticles synthesized by the hydrothermal method prevents the formation of large clod-like aggregates from a suspension of washed rat erythrocytes incubated with the nanoparticles. However, the tendency toward a change in cell shape with an increase in nanoparticles concentration is still manifested.

The new generations of photoactive compounds, which are derivatives of already well-established groups of photosensitizers, together with the modern light sources in the spectral region of their activation and in the PDT window range, are emerging as a reliable phototherapy approach for tumour treatment with higher efficiency. The substitution of a highly hydrophobic phthalocyanine with galactose units could improve the solubility and to decrease significant aggregation in polar solvents. The control group of rats with transplanted cholangiocarcinoma without any treatment was used for comparison of PDT effectiveness. In the second group the white outbred male rats with cholangiocarcinoma lesions were treated using Zn-phthalocyanine, as a typical representative of phthalocyanine photosensitizers with known photodynamic properties and the third group of animals was treated with galactose – Lu-phthalocyanine, as a novel derivative from the same family with two specific differences – presence of galactose and replacing of Zn(II) with Lu(III) ion. To compare the PDT effectiveness of both compounds the same drug doses were applied - 2 mg/kg, applied by intratumoural injection. Diode laser source at 670 nm, on 50 mW output power with power density applied 200 mW/cm2 was applied for 1000 sec to obtain total irradiation dose of 200 J/cm2 for each lesion. 3 days after treatment, the animals were withdrawn from the experiment. Tissue from the central zone and periphery of the tumor was taken for morphological examination and fixed in 10% formalin. Serial paraffin sections were stained with hematoxilin-eosin and by immunohistochemical staining with antibodies to proliferation marker Ki-67 and apoptosis marker BAX. After photodynamic therapy, pronounced necrobiotic changes in combination with an inflammatory reaction were developed at the central zone of tumors. The dystrophic changes, reduction of proliferation and development of apoptosis in tumor cells were observed on tumor periphery as well.

Combined spectroscopic and biochemical measurements were used to improve diagnostic accuracy and to evaluate gastrointestinal tract (GIT) neoplasia development parameters noninvasively. Experiments were performed in mongrel male rats divided into 2 groups – control and experimental. To induce gastric cancer, the rats underwent to chronic stress (overpopulation during 9 months) and diet including the daily using of mtoluidine (25 μg/kg weight) in food and water with a solution of sodium nitrite (0.2%) for 9 months. We studied effectiveness of 5-ALA/PpIX fluorescent analysis of gastric carcinoma and biochemical stress-corresponding indices detection for early diagnosis of primary gastric tumours and their metastatic spreading in liver. Affected by precancerous and cancerous alterations mucosa reveal red fluorescence, related to the accumulation of 5- ALA/PpIX. Liver tissues investigated also presented increase of the red fluorescence, which was used as an indicator for possible pathologic process detection there. The histological examination revealed liver metastases in 67.8 % of the rats with gastric cancer. Biochemical indicators detected malignant alterations presence in GIT, and fluorescent observation addressed the exact area and borders of neoplastic lesions. 5-ALA/PpIX fluorescence detection allow to find and precisely map premalignant and malignant areas of gastric mucosa and liver metastases of stress-induced gastric heterogeneous adenocarcinoma and biochemical evaluation of stressrelated compounds increased the efficiency of such diagnosis and reveal information about the dynamics of lesions development. Diagnostic accuracy achieved using fluorescent detection reaches 93% for gastric carcinoma, and 87% for pre-cancerous mucosa alterations observed.

Background The analysis of recent studies on plasmonic photothermal therapy (PPT) after intravenous administration of gold nanorods (GNRs) has demonstrated that the effectiveness of nanoparticle-assisted laser hyperthermia depends on a correct dosage strategy of nanoparticle administration. Accumulation of GNRs in tumor tissue dramatically increases the local heating of the tumor without damage to healthy tissues. However, the optimal doses of GNR intravenous injections (IVIs) for effective accumulation in tumors, and optimal protocols of PPT are not designed yet. The current study aims to improve the efficacy of PPT in tumor-bearing rats using multiple fractional intravenous administration of GNRs. Materials and methods For PPT experiments, the GNRs with aspect ratio of 4.1 were functionalized with thiolated polyethylene glycol (PEG) and their suspensions were used for multiple fractional intravenous administration in outbred albino male rats with experimental model of rat liver cancer (cholangiocarcinoma line PC-1). Doppler ultrasonography was performed to characterize the vascularity of transplanted rat tumors before any treatment. After a final injection of GNRs, tumor was irradiated during 15 minutes by 808-nm NIR diode laser at a power density 2.3 W/cm2 . The animals were withdrawn from the experiment and sampling of tissues for morphological study and gold accumulation was performed 24 hours and 3 weeks after PPT. Results The multiple IVIs of gold nanorods and further PPT of transplanted cholangiocarcinoma provided significant damage to tumor tissue resulting in pronounced necrotic mass and retardation of the tumor growth. More importantly, the proposed PPT protocol had low toxicity as evidenced by histological examination of internal organs. The efficiency of PPT depends on the presence of newly formed vasculature as revealed by the Doppler ultrasound investigation. Conclusion The repeatable IVIs promote greater of GNR accumulation within the tumor thus resulting in higher PPT efficacy. Accompanying ultrasonography can be useful for prognosis and monitoring of treatment. Lasers Surg. Med. 50:1025-1033, 2018. © 2018 Wiley Periodicals, Inc.

Objective of the study: We used fluorescence imaging methods of apoptosis and necrosis in human renal carcinoma A498 tumor cells in vitro to reveal the indicated forms of cell death under the combined effect of flavonoid-containing extract of Gratiola officinalis and cytostatic (cyclophosphamide). Materials and methods: The dyes were propidium iodide and acridine orange, which were used in the “alive and dead” test. This test helped us to identify the total number of dead cells in the forms of necrosis and apoptosis and the number of cells in which apoptosis had started, it was characterized by the appearance of apoptotic bodies or nucleus pyknosis. Results: We found the most pronounced cytotoxic activity at the ratio of extract of Gratiola officinalis and cyclophosphamide concentrations of 1:1. The number of living cells decreased when exposed to the ratio of extract and cytostatic concentrations of 2:1. When the ratio of concentration of the extract relative to the cytostatic increased to 3:1, the cytostatic activity of the extract began to appear, the total number of tumor cells decreased. The number of cells with nucleus pyknosis and the number of cells with apoptosis signs significantly increased at a 3:1 ratio of extract and cytostatic concentrations, which confirms the presence of pro-apoptotic activity of the studied combination. This trend indicates the dependence of a certain form of cell death (apoptosis, necrosis) on the ratio of extract and cytostatic doses, and it also demonstrates the cytostatic and cytotoxic effects of this combination. Conclusion: Fluorescence methods of investigation in the “alive and dead” test allowed us to visualize the forms of cell death of human kidney carcinoma A498 by combined exposure to the flavonoid-containing extract of Gratiola officinalis and cytostatic (cyclophosphamide) 24 h after exposure. We found that the combination with a concentration ratio of the extract and cyclophosphamide of 3:1 has the greatest effectiveness due to stimulation of the cytostatic effect and cytotoxic effect.

This seems to be the time to gain new knowledge about the meningeal lymphatic system and a deeper understanding of its anatomy and physiology. Although it is known that the meningeal lymphatics present in the layers of the brain, limited information is available about the role of this system in brain function. Here, for the first time we clearly demonstrate that the meningeal lymphatic pathway is involved in brain clearing from the blood after intracranial hemorrhage associated with hypoxia and forms a connective bridge between interstitial, cerebral spinal fluid and peripheral lymphatics. We also show that the development of methods to stimulate meningeal lymph flow after hemorrhagic evidence in the brain might be a neuroprotective strategy for effective recovery of the brain after a cerebrovascular catastrophe.

The ability of diagnostics of melanoma and nevus based on spectral analysis of paraffin-embedded tissues in the 0.3-1.5 terahertz (THz) range has been carried out. The principal component analysis was applied to reduce the dimension of the feature space. A comparison of these spectra shows evident differences between samples. The possibility of applying the optical clearing to paraffin-embedded tissue for improving the visualization of the internal structure of tissue for diagnostic and research purposes is shown. In the studies, a sufficiently strong effect of optical clearing of paraffin-embedded muscle was obtained (63%).

We present studies on the effective properties of mice biological tissues samples treating them as disordered metamaterial media aiming to identify cancerous areas in the mice brain tissue biopsies.