In vivo MRI visualization of growth and morphology in the orthotopic xenotrasplantation U87 glioblastoma mouse SCID model

Glioblastoma multiforme (GBM) is the most common and lethal type of brain cancer with the average lifespan of patients about 9–12 months. The study of tumor formation and the evaluation of new therapies for GBM require accurate and reproducible experimental brain tumor animal models. In this study we used MRI for investigation of tumor morphology and growth dynamic in an orthopic xenotransplantation immunodeficient mouse model (SCID mouse line). Comparison of T1- and T2-weighed MRI scans preformed with a high-field MRI scanner (Bruker, BioSpec, 11,7 T) revealed insufficient tumor/normal tissue T1-contrast because of high longitudinal magnetization of the magnetic field in our scanner. Intravenous injection of paramagnetic manganese oxide (MnO) nanoparticles dramatically increased the tumor/normal tissue contrast in T1-weigthed MRI scans. The study of glioblastoma growth with T2-weighed images showed that a significant tumor development began not earlier than 3 weeks after cell culture intracranial injection and then the tumor grew exponentially. Thus, we developed a protocol of the characterization of glioblastoma U87 growth and morphology by T1- and T2-weighed and MnO-enhanced MRI in the orthopic xenotransplantation mouse model. The results demonstrate that this SCID model may be used as an in vivo preclinical model to test the efficacy and putative side effects of novel anticancer therapies

GC-based chemoprofile of lipophilic compounds in Altaian Ganoderma lucidum sample

The presented data contains information about component composition of lipophilic compounds in Ganoderma lucidum fungal body sample obtained using gas chromatography and subsequent mass spectrometry

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 <em>db/db</em> 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

Metabolic and motor activity effects of microalgae (Chlorella vulgaris) in laboratory mice

In recent years, the microalgae (Chlorella vulgaris) have increasingly attracted great interest as a potential source of pharmacologically active compounds. Showing anticoagulation, antioxidant and antitumor activities of Chlorella revealed its hypotensive properties. The aim of this study was to evaluate the effects of Chlorella suspension on the weight of the animals, their moving activity, and erythropoiesis. The study was performed on males and females of ICR mice. The animals from the experimental group drank only the Chlorella suspension during 3 weeks and were given standard food. Control animals drank during this period only water and had the same food. The body weight of males in the control and the experimental group with Chlorella did not change, while females in the experimental group showed an increase of body weight in a week. A similar pattern was obtained for estimation of animal body weight changes relative to food consumption. The number of red blood cells in females and males from group with Chlorella increased only after 3 weeks after the start of the experiment. Hemoglobin also increased only after 3 weeks after the start of Chlorella consumption, but only for females. All groups of animals had the same motor activity during experiment. Blood sampling resulted in a reduction of activity in control males and females as well as in males with Chlorella. The motor activity of females with Chlorella after blood sampling did not change. So, consumption of the Chlorella suspension by females leads to more effective digestion and resulted in increased body weight, improved erythropoiesis resulted in increased red blood cells and hemoglobin and also increased their resistance to acute stress. The males in the same situation increased only the erythropoiesis

Zika virus has an oncolytic activity against human glioblastoma U87 cells

Glioblastoma is a highly lethal brain cancer. Virotherapy with the use of oncolytic viruses has since recently been regarded as a promising approach for the clinic treatment of human glioblastomas. The purpose of this work was to perform a primary evaluation of the Zika virus as a potential oncolytic agent against glioblastomas. In vitro experiments showed that the Zika virus strain MR 766 is able to selectively infect and lyse neoplastic cells of the human glioblastoma cell line U87 MG. The selectivity index (SI, the ratio of infectious titer for tumor cells to titer on normal untransformed cells) was 2·102. The selectivity of the replicative activity of Zika virus in relation to U87 MG glioblastoma cells was additionally confrmed by indirect immunofluorescence. Using the model of immunodefcient SCID mice with subcutaneous xenografts of human glioblastoma U87 MG, a strong antitumor activity of the Zika virus under a course (daily for 4 days) of intratumoral administration of 5·105 TCID50 of Zika virus was shown. Treatment with Zika virus resulted in more than a 10­fold reduction in mean volumes of tumors. The tumor growth inhibition index was 92.63 %. Recurrences (metastases) of tumor re­growth were not registered within 64 days of observation. This result demonstrated the prospect of further in­depth studies of the Zika virus as a potential oncolytic agent against human glioblastomas

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

ЭФФЕКТ ЭКСТРАКТА МИЦЕЛИЯ DUDDINGTONIA FLAGRANS НА ПОДКОЖНЫЕ КСЕНОГРАФТЫ КЛЕТОК C33a КАРЦИНОМЫ ШЕЙКИ МАТКИ ЧЕЛОВЕКА

The purpose of the study was to analyze the antitumor effects of the extract of mycelium from Duddingtonia flagrans (strain F-882) on xenografts of human C33a cervical cancer cells.Material and Methods. To evaluate the antitumor effect, we used the absolute values of xenograft volumes and calculated the tumor growth inhibition and the index of tumor growth.Results. At the first stage of the experiment, a 4-week subcutaneous injection of the water extract of F-882 resulted in an almost twofold slowdown in xenograft growth, with the tumor growth inhibition value of 50.6 %. In the second stage of the experiment, a 2.5-week subcutaneous injection followed by a 1.5-week intratumoral injection of F-882 also caused the tumor growth inhibition. After completing F-882 injections, the effect of tumor growth inhibition continued for 2.5 weeks and the tumor growth inhibition value was 58.7 %.Conclusion. The mycelium extract F-882 was shown to have an antitumor effect on subcutaneous xenografts of human C33a cervical carcinoma cells.Цель исследования – изучение противоопухолевого действия экстракта мицелия Duddingtonia flagrans (штамм F-882) на ксенографты карциномы шейки матки человека C33a.Материал и методы. Для оценки противоопухолевого действия использовали абсолютные значения объёмов ксенографтов и рассчитывали показатели: торможение роста опухоли и индекс прироста опухоли.Результаты. На первом этапе эксперимента после 4 нед подкожных введений F-882 значение торможения роста опухоли составило 50,6 %, т. е. инъекции привели к почти двукратному замедлению роста опухолей в экспериментальной группе. На следующем этапе комбинированное введение, подкожное в течение 2,5 нед, а затем внутриопухолевое в течение 1,5 нед, также показало ингибирование роста  ксенографтов, после окончания инъекций F-882 эффект замедления роста опухолей продолжался в течение 2,5 нед и показатель торможения роста опухоли составлял 58,7 %.Заключение. Впервые было выявлено, что экстракт мицелия F-882 способен оказывать интенсивное противоопухолевое действие на подкожные ксенографты карциномы шейки матки человека клеток C33a

QUEST-DMC superfluid ³He detector for sub-GeV dark matter

The focus of dark matter searches to date has been on Weakly Interacting Massive Particles (WIMPs) in the GeV/c2-TeV/c2 mass range. The direct, indirect and collider searches in this mass range have been extensive but ultimately unsuccessful, providing a strong motivation for widening the search outside this range. Here we describe a new concept for a dark matter experiment, employing superfluid 3He as a detector for dark matter that is close to the mass of the proton, of order 1 GeV/c2. The QUEST-DMC detector concept is based on quasiparticle detection in a bolometer cell by a nanomechanical resonator. In this paper we develop the energy measurement methodology and detector response model, simulate candidate dark matter signals and expected background interactions, and calculate the sensitivity of such a detector. We project that such a detector can reach sub-eV nuclear recoil energy threshold, opening up new windows on the parameter space of both spin-dependent and spin-independent interactions of light dark matter candidates

Possibility of using a mouse SCID as a model animal to variola virus for evaluating anti-smallpox drug efficacy

At present, there is no animal model for smallpox that reflects the weakened immune system in people and can therefore help assess the prophylactic (highly preventive) efficiency of antiviral drugs. To fill in the gap, we have explored the possibility of using outbred immunodeficient SCID mice as a model animal for smallpox with the aid of virolo­gical, histological and electron microscopic and sta­tistical methods. There was no clinical evidence of disease by intranasal infection of mice at a dose of 5.2 log10 PFU (plaque forming units). At the same time, the 50 % infective dose (ID50) of VARV estimated for animals by registering the presence of the virus in their lungs after 4 days post i.n. infection was 3.5 log10 PFU and was relatively similar to that in humans, theoretically determined by identification of the clinical picture of the disease. Virus replication was detected only in the respiratory organs of mice challenged i.n. with VARV at a dose of 5.2 log10 PFU (50 ID50). The values for its concentrations in the lungs and nose resembled those for affected people and well-known animal models (Macaca cynomolgus and ICR mice), respiratorily infected with VARV at similar doses. The existing model animals were not significantly different from SCID mice in the duration of viral presence in the lungs. Moreover, in SCID mice, as in humans and other animal models, similar pathomor- phological changes of inflammatory necrotic nature in the respiratory organs have been reported. Using SCID mice in assessing the prophylactic efficacy of the antiviral drugs NIOCH-14 and ST-246 demonstrated the adequacy of the results obtained to those described in the literature. This opens up the prospect of using SCID mice as an animal model for smallpox to develop antiviral drugs intended for people with severe immuno­suppressive states