Light-Induced Functional Pinealectomy. Effect on the Thymus of C57BL/6 Mice.

Light-induced functional pinealectomy was simulated in C57BL/6 mice by 14-day exposure to constant lighting. Immunophenotyping of CD3hi and CD3low thymocytes was performed by staining with CD3-APC antibodies followed by flow cytofluorometry. To study the cell cycle distribution of thymus cells, the content of intracellular DNA was measured by the level PI inclusion. In animals with light-induced functional pinealectomy, blood leukocyte content, the relative number of CD3low and CD3hi T cells in the thymus, and the ratio of CD3low/CD3hi thymocytes decreased. The number of G0/G1-phase thymus cells (non-dividing cells) increased and the content of S-phase cells (division phase) decreased. Continuous lighting stimulated the development of thymocyte apoptosis. The results obtained indicate that prolonged 24-h illumination inhibits differentiation and maturation of young CD3low thymocytes into mature CD3hi forms and leads to the development of T-cell apoptosis in the thymus and, as a consequence, to leukopenia.

Effects of Boron Neutron Capture Therapy on the Growth of Subcutaneous Xenografts of Human Colorectal Adenocarcinoma SW-620 in Immunodeficient Mice.

Boron neutron capture therapy (BNCT) can become an instrument for patients with malignant neoplasms of the rectum and colon. Here we evaluate the effectiveness of BNCT performed at the accelerator based epithermal neutron source at G. I. Budker Institute of Nuclear Physics, Siberian Division of Russian Academy of Sciences, in relation to subcutaneous xenografts of human colon adenocarcinoma SW-620 in SCID mice. Utilization of BNCT with boronоphenylalanine (BPA) and sodium borocaptate (BSH), which were injected intravenously into the retroorbital sinus, resulted in a significant decrease in tumor volumes compared to the control group (no radiation).

Manganese Oxide Nanoparticles Inhibit the Growth of Subcutaneous U-87MG Glioblastoma Xenografts in Immunodeficient Mouse.

Our previous study demonstrated that manganese oxide nanoparticles (MnO NP) selectively destroyed U-87MG and U251 human glioblastoma cells in vitro. MnO NP were synthesized and studied by electron microscopy. Their antitumor properties were studied in vivo on the model of immunodeficient SCID mice with subcutaneous xenografts of U-87MG human glioblastoma. The mice were injected subcutaneously with MnO NP in doses of 0.96 and 1.92 mg/kg (calculated for Mn) 3 days a week over 3 weeks. In was shown that MnO NP in these doses significantly suppressed the growth of U-87MG glioblastoma xenografts: on day 21 from the start of the treatment, the tumor growth inhibition index was 61.1 and 99.22%, respectively. These results indicate the necessity of the further studies of MnO NP as a potential oncolytic agent for the therapy of human glioblastomas.

Treatment of Intervertebral Disc Degeneration in Wistar Rats with Mesenchymal Stem Cells.

We studied the effect of erythropoietin on functional properties of mesenchymal stem cells under conditions of oxidative stress and their therapeutic potential in the treatment of intervertebral disc degeneration in Wistar rats. It was shown that erythropoietin stimulates proliferation under conditions of oxidative stress. Injection of bone marrow mesenchymal stem cells into the damaged intervertebral disc was followed by an increase in the height of the intervertebral disc and activation of repair processes in the nucleus pulposus. The combination of mesenchymal stem cells with erythropoietin provides the best effect of cell therapy in case of intervertebral disc damage.

Effect of Silicon Dioxide Nanoparticles on Syrian Hamsters Infected by Opisthorchis felineus: 1H MRS Study of the Brain.

In recent years, silicon dioxide nanoparticles have been widely used in medicine and the pharmaceutical industry, however, their effect on the brain has hardly been studied. We assessed the effects of long-term consumption of 5-nm amorphous silicon dioxide nanoparticles (SiO2-NPs) by Syrian hamsters infected with the trematodes Opisthorchis felineus on the hippocampus and frontal cortex. Spectroscopic determination of brain neurometabolites, performed using a horizontal Magnetic Resonance Imaging system at 11.7 Tesla magnetic field, has shown that the ratio of the excitatory neurotransmitters (glutamate + glutamine + aspartate) to the inhibitory ones (GABA + glycine) was higher in the animals infected with O. felineus. However, pre-consumption of the SiO2-NPs solution prevented this imbalance. In addition, the protective effect of SiO2-NPs on the level of myo-inositol and glycine was found. It is concluded that the use of SiO2-NPs can neutralize the negative effects of infectious factors on the brain.

Effects of Melatonin on the Body Composition, Physical Performance, and Blood Erythrocyte Indexes of C57Bl/6J Mice Exposed to Continuous Illumination.

Male C57Bl/6J mice were exposed to daily 24-h illumination over 14 days and daily intragastrically received melatonin (1 mg/kg) or water (placebo). Controls were kept under standard day/night (14/10 h) conditions. Melatonin prevented the development of anemia in mice exposed to continuous illumination, which was proven by higher blood hemoglobin levels by the end of the experiment in melatonin-treated animals in comparison with the placebo group. Studies by the low-field NMR spectrometry detected lower lean body mass, total body water, and especially, fat content (by ~13%) in animals receiving placebo. Melatonin treatment led to an increase in the lean body mass and total body water on day 7 (in comparison with the placebo group) without affecting fat mass. On day 14 of continuous illumination, lean body mass increased in comparison with the corresponding parameter in the control and placebo groups. Melatonin had no effect on the physical endurance of mice exposed to continuous illumination (assessed by the grid hanging test).

Virotherapy of the Malignant U87 Human Glioblastoma in the Orthotopic Xenotransplantation Mouse SCID Model.

The possibility of glioblastoma virotherapy at intravenous injection of the LIVP-GFP recombinant virus was studied in experimental model of orthotopic xenotransplantation of human glioblastoma cell line U87 to SCID laboratory mice. The LIVP-GFP recombinant virus deficient for thymidine kinase exhibited a significantly greater oncolytic capacity than the original LIVP virus, and an intravenous injection of LIVP-GFP at the early stages of tumorigenesis in mouse brain in most cases resulted in the lysis of the tumor.

Specificity of hexarhenium cluster anions for synthesis of Mn2+-based nanoparticles with lamellar shape and pH-induced leaching for specific organ selectivity in MRI contrasting.

The present work demonstrates the structure variation of hexarhenium anionic cluster units [{Re6S8}(CN)(6-n)(OH)n]4- (n = 0, 2, 4) as the strategy to develop Mn2+-containing nanoparticles (NPs) exhibiting pH-dependent leaching. The dicyanotetrahydroxo complex [{Re6S8}(CN)2(OH)4]4- is the optimal for the synthesis of the Mn2+-based NPs with a lamellar shape exhibiting the pH-dependent aggregation and magnetic relaxation behavior. The pH-dependent behavior of the NPs derives from the easy protonation of the apical hydroxo ligands of [{Re6S8}(CN)2(OH)4]4- cluster, which triggers partial leaching of Mn2+ ions and aggregation of the NPs driven by the surface neutralization. The in vivo MRI scanning of the mice intravenously injected with the NPs indicates the preferable accumulation of the lamellar NPs within mouse intestine over liver and kidneys. This differs from the spherical NPs constructed from [{Re6Se8}(CN)6]4- units, which provide the preferable brightening of mouse liver over kidneys and intestine.

In vivo MRS study of long-term effects of traumatic intracranial injection of a culture medium in mice.

Orthotopic transplantation of glioblastoma cells in the brain of laboratory mice is a common animal model for studying brain tumors. It was shown that 1H magnetic resonance spectroscopy (MRS) enables monitoring of the tumor's occurrence and its development during therapy based on the ratio of several metabolites. However, in studying new approaches to the therapy of glioblastoma in the model of orthotopic xenotransplantation of glioma cells into the brain of mice, it is necessary to understand which metabolites are produced by a growing tumor and which are the result of tumor cells injection along the modeling of the pathology. Currently, there are no data on the dynamic metabolic processes in the brain that occur after the introduction of glioblastoma cells into the brain of mice. In addition, there is a lack of data on the delayed effects of invasive brain damage. Therefore, this study investigates the long-term dynamics of the neurometabolic profile, assessed using 1H MRS, after intracranial injection of a culture medium used in orthotopic modeling of glioma in mice. Levels of N-acetylaspartate, N-acetylaspartylglutamic acid, myoinositol, taurine, glutathione, the sum of glycerophosphocholine and phosphocholine, glutamic acid (Glu), glutamine (Gln), and gamma aminobutyric acid (GABA) indicate patterns of neurometabolites in the early stage after intracranial injection similar to brain trauma ones. Most of the metabolites, with the exception of Gln, Glu and GABA, returned to their original values on day 28 after injection. A progressive increase in the Glu/Gln and Glu/GABA ratio up to 28 days after surgery potentially indicates an impaired turnover of these metabolites or increased neurotransmission. Thus, the data indicate that the recovery processes are largely completed on day 28 after the traumatic event in the brain tissue, leaving open the question of the neurotransmitter system impairment. Consequently, when using animal models of human glioma, researchers should clearly distinguish between which changes in neurometabolites are a response to the injection of cancer cells into the brain, and which processes may indicate the early development of a brain tumor. It is important to keep this in mind when modeling human glioblastoma in mice and monitoring new treatments. In addition, these results may be important in the development of approaches for non-invasive diagnostics of traumatic brain injury as well as recovery and rehabilitation processes of patients after certain brain surgeries.

CD-1 mice females recognize male reproductive success via volatile organic compounds in urine.

Sexual selection is considered as one of the leading factors of evolutionary development. In the conditions of incessant competition, specialized methods of attracting individuals of the opposite sex as well as criteria for assessing the quality of a sexual partner have been formed. In order for animals to rely on signaling from sexual partners, the signal must reflect the morpho-physiological status of animals. A high reproductive efficiency of male mice is a good advantage for mate selection and thus must be somehow demonstrated to potential mates. The aim of our study was to find out if male mice could demonstrate their reproductive efficiency through urine volatile organic compounds. The experiment implies cohabiting one male with two mature females for 6 days. The reproductive success of the male was assessed by the presence or absence of pregnant females. At the same time, naive females, who did not participate in reproduction, assessed the urine of the successful males as more attractive, which was expressed in shorter Latency time of sniffs in the Olfactory test. Using a rapid headspace GC/MS analysis, we have found volatile organic compounds (VOCs) in male urine that correlated with female behavior. It turned out that these substances are derivatives of mouse pheromone 6-hydroxy-6-methyl-3-heptanone. The amplitude of peaks corresponding to this pheromone correlated with the testosterone level in blood and the weight of preputial glands. The amplitude of peaks increased in males after mating with whom the females turned out to be pregnant. It is important to note that body weight, weight of testes, weight of seminal vesicles, weight of preputial glands, and plasma testosterone level alone are not reliable indicators of male reproductive success. Thus, the content of the pheromone 6-hydroxy-6-methyl-3-heptanone in the urine of males can serve as a good predictor of the quality of the male as a sexual partner for female CD-1 mice.

The expression of apoptosis-regulating proteins Bcl-2 and Bad in liver cells of C57Bl/6 mice under light-induced functional pinealectomy and after correction with melatonin.

The presence of humans and animals under long-term continuous lighting leads to a suppression of melatonin synthesis, that is, to light-induced functional pinealectomy (LIFP), and the development of desynchronosis. To create LIFP, C57Bl/6 mice were kept under 24-hour lighting (24hL) for 14 days. The animals in the control group were kept under standard lighting conditions. In the next series of experiments, mice with LIFP received daily intragastrically either melatonin (1 mg/kg body weight in 200 µl of distilled water) or 200 µl of water as a placebo. The comparison group consisted of intact animals that received placebo under standard lighting conditions. Immunohistochemical analysis (using an indirect avidin-biotin peroxidase method) revealed the expression of the antiapoptotic protein Bcl-2 and the proapoptotic protein Bad in sinusoid liver cells (a heterogeneous population consisting of the endotheliocytes, Kupffer cells, Ito cells, and Pit cells) and in individual hepatocytes. The Bad expression area in the liver of LIFP mice increased 4 times against a background of the unchanged Bcl-2 expression area. Changes in the brightness (a parameter inversely proportional to the marker concentration) of Bad and Bcl-2 areas did not reach significance. Our results indicate a weakening of the antiapoptotic protection of liver cells of LIFP animals, which creates conditions for activation of the "mitochondrial branch" of apoptosis. Melatonin treatment of LIFP mice resulted in a 3.3-fold increase in Bcl-2 expression area and a 2.7 % decrease in Bcl-2 region brightness compared with the experimental untreated group. Bad protein parameters were unreliable. Thus, melatonin treatment of animals cancels the effect of LIFP, restoring the Bcl-2 expression area and increasing this protein concentration, which indicates an increase in antiapoptotic protection and creates conditions for blocking the development of the "mitochondrial branch" of apoptosis in liver cells.

Apoptosis in the liver of male db/db mice during the development of obesity and type 2 diabetes.

Obesity and diabetes mellitus are known to lead to the development of metabolic syndrome and non-alcoholic fatty liver disease (NAFLD). The mechanisms of programmed cell death are actively involved in maintaining cellular homeostasis along development of NAFLD. Proteins of the BCL-2 family are key regulators of physiological and pathological apoptosis. Homozygous males of BKS.Cg-Dock7mLeprdb/+/+/J mice (db/db mice) are characterized by progressive obesity and the development of type 2 diabetes mellitus (DM2) with severe hyperglycemia at 4-8 weeks and organ lesions at 8-10 weeks of age. The aim of this research was to study the expression of molecular cell regulators of apoptosis in liver cells of db/db mice males at different stages of obesity and diabetes development (at the age of 10 and 18 weeks). Immunohistochemical analysis (using the indirect avidin-biotin peroxidase method) and morphometric evaluation of the expression of the antiapoptotic protein Bcl-2 and the proapoptotic protein Bad in liver cells of studied animals at different stages of obesity and DM2 were carried out. An excess of the value of the Bcl-2 protein staining area over the Bad protein staining area was revealed in the liver of 10-week-old animals. The Bcl-2/Bad expression area ratio in 10-week-old animals was twice as high as in 18-week-old animals, which indicates the presence of conditions for blocking apoptosis in the liver of younger db/ db mice. At the 18th week of life, db/db mice displayed an almost threefold increase in the expression area of the Bad protein against the background of an unchanged expression of the Bcl-2 protein. The decrease in the Bcl-2/Bad staining area ratio in 18-week-old animals was due to the increase in the Bad expression area, which indicates the absence of antiapoptotic cell protection and creates conditions for activation of the mitochondrial pathway of apoptosis in the liver of male db/db mice with pronounced signs of obesity and DM2.

Erratum to: "Anxiety and neurometabolite levels in the hippocampus and amygdala after prolonged exposure to predator-scent stress".

Vavilovskii Zhurnal Genetiki i Selektsii = Vavilov Journal of Genetics and Breeding. 2019;23(5):582-587 (in Russian) Page 587, in Acknowledgements instead of The animals and behavioral testing are supported by the budget project (No. 0324-2019-0041). The MRI study is supported by the budget project (No. 0259-2019-0004). All studies are implemented using the equipment of Center for Genetic Resources of Laboratory Animals at ICG SB RAS, supported by the Ministry of Education and Science of Russia (Unique ID# of the project: RFMEFI62117X0015). should read The animals and behavioral testing are supported by the budget project (No. 0324-2019-0041). The MRI study is supported by the budget project (No. 0259-2019-0004). All studies are implemented using the equipment of Center for Genetic Resources of Laboratory Animals at ICG SB RAS, supported by the Ministry of Education and Science of Russia (Unique ID# of the project: RFMEFI62117X0015). The study was conducted within the basic part of the state task of the Ministry of Science and Higher Education of the Russian Federation (No. 17.7255.2017/8.9). The original article can be found under DOI 10.18699/VJ19.528.