Fatty Acid Content and Tumor Growth Changes in Mice After Exposure to Extremely High-Frequency Electromagnetic Radiation and Consumption of N-3 Fatty Acids.

The topical problem is to find new, more effective and safe treatments for cancer. The purpose of the present work was to study the combined effects of low-intensity extremely high-frequency electromagnetic radiation (EHF EMR) and consumption of n-3 polyunsaturated fatty acids (PUFAs) on tumor growth and the content of FAs in the thymus and tumor tissue in mice. Fatty acid composition was determined using gas chromatography. Exposure of tumor-bearing mice with solid Ehrlich carcinoma to EHF EMR with effective parameters (42.2 GHz, 0.1 mW/cm2, 20 min daily for 5 consecutive days beginning on the first day after the tumor inoculation) led to delaying the tumor growth and restored the content of almost all FAs in thymic tissue to the level of intact animals. Animal intake of the preparation enriched with n-3 PUFAs increased the content of n-3 PUFAs in thymic tissue significantly, but did not affect the tumor growth, even in combination with EHF EMR exposure. Combined action of EHF EMR exposure and n-3 preparation promoted recovery of thymus weight in tumor-bearing animals. The data obtained assume a complex interaction between the immune system and the tumor, and the important role of FAs in the regulation of this interaction.

Minimal uranium accumulation in lymphoid tissues following an oral 60-day uranyl acetate exposure in male and female C57BL/6J mice.

High levels of uranium (U) exist in soil, water, and air in the Southwestern United States due, in part, to waste generated from more than 160,000 abandoned hard rock mines located in this region. As a result, many people living in this region are chronically exposed to U at levels that have been linked to detrimental health outcomes. In an effort to establish a relevant in vivo mouse model for future U immunotoxicity studies, we evaluated the tissue distribution of U in immune organs; blood, bone marrow, spleen, and thymus, as well as femur bones, kidneys, and liver, following a 60-d drinking water exposure to uranyl acetate (UA) in male and female C57BL/6J mice. Following the 60-d exposure, there was low overall tissue retention of U (<0.01%) at both the 5 and the 50 ppm (mg/L) oral concentrations. In both male and female mice, there was limited U accumulation in immune organs. U only accumulated at low concentrations in the blood and bone marrow of male mice (0.6 and 16.8 ng/g, respectively). Consistent with previous reports, the predominant sites of U accumulation were the femur bones (350.1 and 399.0 ng/g, respectively) and kidneys (134.0 and 361.3 ng/g, respectively) of male and female mice. Findings from this study provide critical insights into the distribution and retention of U in lymphoid tissues following chronic drinking water exposure to U. This information will serve as a foundation for immunotoxicological assessments of U, alone and in combination with other metals.

Aging of lymphoid organs: Can photobiomodulation reverse age-associated thymic involution via stimulation of extrapineal melatonin synthesis and bone marrow stem cells?

Thymic atrophy and the subsequent reduction in T-cell production are the most noticeable age-related changes affecting lymphoid organs in the immune system. In fact, thymic involution has been described as "programmed aging." New therapeutic approaches, such as photobiomodulation (PBM), may reduce or reverse these changes. PBM (also known as low-level laser therapy) involves the delivery of non-thermal levels of red or near-infrared light that are absorbed by mitochondrial chromophores, in order to prevent tissue death and stimulate healing and regeneration. PBM may reverse or prevent thymic involution due to its ability to induce extrapineal melatonin biosynthesis via cyclic adenosine monophosphate (AMP) or NF-kB activation, or alternatively by stimulating bone marrow stem cells that can regenerate the thymus. This perspective puts forward a hypothesis that PBM can alter thymic involution, improve immune functioning in aged people and even extend lifespan.

EMSA Eritin Drives Expansion of Regulatory T Cells and Promotes T Cells Differentiation in Irradiated Mice.

Sublethal irradiation therapy in cancer treatment causes generalized immunosuppression, which results in a range of DNA damage. We examined the significance of a polyherbal medicine called "EMSA Eritin" on immunological responses in sublethally irradiated mice focusing on the involvement of Treg, naïve T cell, and also the development and differentiation of T cells in thymus. Normal BALB/c mice were sublethally irradiated with dose of 600 rad. The irradiated mice were then orally administered by EMSA Eritin once a day at different doses: 1.04, 3.12, 9.37 mg/g body weight. The treatment was performed for 14 days. On day 15, immunological responses were observed by analyzing the status of Treg and differentiation of T cells in thymus. The administration of EMSA Eritin to irradiated mice resulted in a significant increase of pre T cells, Treg cells, and naïve T cells, which in general could maintain and normalize healthy condition in mice.

Carbon Ion Irradiated Neural Injury Induced the Peripheral Immune Effects in Vitro or in Vivo.

Carbon ion radiation is a promising treatment for brain cancer; however, the immune system involved long-term systemic effects evoke a concern of complementary and alternative therapies in clinical treatment. To clarify radiotherapy caused fundamental changes in peripheral immune system, examinations were performed based on established models in vitro and in vivo. We found that brain-localized carbon ion radiation of neural cells induced complex changes in the peripheral blood, thymus, and spleen at one, two, and three months after its application. Atrophy, apoptosis, and abnormal T-cell distributions were observed in rats receiving a single high dose of radiation. Radiation downregulated the expression of proteins involved in T-cell development at the transcriptional level and increased the proportion of CD3⁺CD4(-)CD8⁺ T-cells in the thymus and the proportion of CD3⁺CD4⁺CD8(-) T-cells in the spleen. These data show that brain irradiation severely affects the peripheral immune system, even at relatively long times after irradiation. In addition, they provide valuable information that will implement the design of biological-based strategies that will aid brain cancer patients suffering from the long-term side effects of radiation.

Polysaccharide extracted from Rheum tanguticum prevents irradiation-induced immune damage in mice.

AIM: To investigate the protective effect of purified fraction 1 polysaccharide extracted from Rheum tanguticum RTP1 on irradiation-induced immune damage in mice. METHODS: Kunming mice were randomly divided into five groups: normal group (NC), irradiation control group (IC), RTP1 low dose (200 mg/kg), middle dose (400 mg/kg) and high dose (800 mg/kg) groups. RTP1 was administered by the gastric route for 14 d, mice in the NC and IC groups being given by 0.9% sodium chloride solution in the same way. The mice in all groups except NC group were irradiated with 2.0 Gy6°Co γ-ray on the fourteenth day. Immune indives of non-specific immune function, cellular immunity and humoral immunity were assessed at the 24th hour after radiation. RESULTS: Compared with the IC group, the spleen index, thymus index, rate of carbon clearance, phagocytic function of macrophages, lymphocyte proliferation, hemolysin value of blood serum and NK activity were increased markedly (P < 0.05 or P < 0.05). CONCLUSION: RTP1 has an obvious protective effects on damage in γ-ray radiated mice.

[Muscle regeneration and the state of the thymus in adult rats under laser irradiation and alloplasty of the gastrocnemius muscles and diaphragm of newborn rats].

The regeneration of gastrocnemius muscles of adult rats under implantation conditions in areas of muscle tissue damage in newborn rats has been studied. Alloplasty was performed using minced gastrocnemius and diaphragm muscles, which differs at birth in animals by degree of differentiation. The rat-recipient area of alloplasty was subjected to He-Ne laser radiation before operation, with the aim of reducing the immune response to allogenic muscle tissue. It has been shown that the number of regenerating myofibers produced in implanted gastrocnemius muscles is more than in alloplants from diaphragms. However, the formation of cartilage, bone, and adipose tissue foci were observed in the alloplastic region throughout the whole regeneration period. After implantation of minced diaphragm muscles, cartilage nodes were observed only in 7-day regenerates. At the end of observation, in the first instance, the area of muscle trauma in adult rat muscles was replaced by adipose tissue, even in the case of initial laser irradiation. During the implantation of diaphragm muscles, the area of trauma was filled with regenerating muscle tissue.

[Modern approaches to correction and prophylaxis of premature aging using adaptogenes of plant origin].

Last years, the big attention is given studying of the methods promoting increase of the immune status of an organism, got under influence of extreme factors, in particular, ionas radiations. In this connection perspectivity of application of laboratory diagnostics for testing a degree of defeat of immune system and application phytoadaptogenes is proved, namely: tinctures of a Panacis ginseng, Schizandrae chinensis, an extract Elaetherococcus for correction of such conditions.

[The use of drinking sulfate mineral water in combination with laser and magnetic-laser irradiation for primary prophylaxis of post-radiation disorders(experimental study)].

Experiments on laboratory rats have demonstrated that prophylactic use of drinkable sulfate mineral water in combination with laser and magnetolaser irradiation of adrenal glands produces a radioprotecive effect that was especially well apparent in the liver and less so in the testes. Simultaneously, the functional activity of the thymus decreased. Protection of the liver by the combination of the above factors was more pronounced than the effect of each of them applied separately.

Radioprotective effects of black seed (Nigella sativa) oil against hemopoietic damage and immunosuppression in gamma-irradiated rats.

BACKGROUND AND AIM: Sixty male Wistar rats, divided into 4 groups, 15 each, were designed as I-control rats, II-rats orally intubated with Nigella sativa oil (1 ml/kg b.wt./day) for 5 days/week, III-whole body gamma irradiated rats with the estimated LD50/30 (4 Gray) and IV-rats daily intubated with Nigella sativa oil then subjected to whole body gamma irradiation, to investigate the radioprotective potential of Nigella crude oil against hemopoietic adverse effects of gamma irradiation. RESULTS: Irradiation resulted in significant reduction in hemolysin antibodies titers and delayed type hypersensitivity reaction of irradiated rats, in addition to significant leukopenia and significant decrease in plasma total protein and globulin concentrations and depletion of lymphoid follicles of spleen and thymus gland. Furthermore, there was a significant increase in malondialdehyde concentration with a significant decrease in plasma glutathione peroxidase, catalase and erythrocyte superoxide dismutase activities were recorded. Oral administration of Nigella sativa oil before irradiation considerably normalized all the above-mentioned criteria; and produced significant regeneration in spleen and thymus lymphoid follicles. CONCLUSION: Our results strongly recommend Nigella sativa oil as a promising natural radioprotective agent against immunosuppressive and oxidative effects of ionizing radiation.

[Induced thymus aging: radiation model and application perspective for low intensive laser radiation].

The influence of gamma-radiation on morphofunctional state of thymus is rather like as natural thymus aging. However gamma-radiation model of thymus aging widely used to investigate geroprotectors has many shortcomings and limitations. Gamma-radiation can induce irreversible changes in thymus very often. These changes are more intensive in comparison with changes, which can be observed at natural thymus aging. Low intensive laser radiation can not destroy structure of thymus and its effects are rather like as natural thymus aging in comparison with gamma-radiation effects. There are many parameters of low intensive laser radiation, which can be changed to improve morphofunctional thymus characteristics in aging model. Using low intensive laser radiation in thymus aging model can be very perspective for investigations of aging immune system.

Scorpion venom peptides accelerate hematopoietic recovery of myelosuppression in irradiated mice.

Sublethally irradiated mice were administered with scorpion venom peptides (SVP) or with PBS in the saline control group, 3 days before and 7 consecutive days after irradiation. Hematopoietic recovery was assessed by bone marrow (BM) cell proliferation index (PI) and colony forming unit-granulocyte/macrophage (CFU-GM), spleen weight index (SI) and thymus weight index (TI), colony-forming unit-spleen (CFU-S) and peripheral leukocyte counts. In addition, IL-1alpha and SCF levels in BM, IL-6 and GM-CSF levels in serum were determined. In SVP treated groups, PI was improved dramatically versus control mice on day 22 after irradiation. The number of CFU-GM colonies in all SVP treated groups was higher than the control groups. The difference of the number of CFU-GM colonies between SVPV group (0.2 mg/kg) and the control was significant on day 5 and 10 after irradiation (p < 0.05). SVPIV (0.2 mg/kg) could activate the CFU-S formation on day 10 after irradiation. SI was in peak value on day 15 after irradiation in all groups and the SI value of SVPV treated group was higher than control group (p < 0.05). Our results suggest that SVP may be valuable natural peptides that relieve myelosuppression caused by radiation. The effect of SVP accelerating the hematopoietic recovery was potentially through a mechanism of stimulating the release of cytokines.

Indigowood root extract protects hematopoietic cells, reduces tissue damage and modulates inflammatory cytokines after total-body irradiation: does Indirubin play a role in radioprotection?

Radix of Isatis indigotica (indigowood root, IR) has been used in traditional medicine for its potential anti-inflammatory effect. The purpose of this study is to investigate the radioprotective effects of radiation caused damages in hematopoietic system and normal tissues in mice. A total of 57 BALB/c mice were randomized into six treatment groups: control, IR treatment (0.195, 0.585 and 1.170 g/kg, p.o. daily), L-glutamine (0.520 g/kg) and sham group. All mice except the sham group were irradiated and then administered for one week. The radioprotective effect on hematopoietic system, serum cytokines, and intestinal toxicity was studied. Protective effects on spleen and thymus are found in IR-treated groups. IR assisted in restoration of leukocytopenia after whole mice irradiation with significant reduction of serum TNF-alpha, IL-1beta, and IL-6. These enhancements of hematopoietic effects are due to an increase in the serum G-CSF concentration in IR treated groups. In histopathological assessment, significant improvement of intestine toxicity is observed in high-dose IR and L-glutamine group. Evidences show that IR has potentials to be a radioprotector, especially in recovery of hematopoietic system, reduction of inflammatory cytokines and intestinal toxicity. Indirubin may play a crucial role, but the underlying mechanism is not very clear and warrants further studies.

Radioprotective activity of betalains from red beets in mice exposed to gamma irradiation.

We investigated the radioprotective activity of betalains from red beets in mice irradiated by a (60)Co gamma (gamma) ray (6.0 Gy, at a dose of 1.5 Gy min(-1)). Mice were randomly divided into five groups, namely the control group and four experimental groups which were given one of four concentrations of betalains from red beets (0, 5, 20 and 80 mg/kg, equivalent to betanin) for 30 days. The four experimental groups of mice were then exposed to the (60)Co gamma-rays and were given betalains from red beets for a further 3 days. The number of white blood cells, karyota of the femur and the number of micronuclei in polychromatophilic erythrocytes of bone marrow in mice were determined. The activity of superoxide dismutase, catalase, glutathione peroxidase, malondialdehyde, spleen index and thymus index were also determined. The results indicate that the administration of betalains from red beets is radioprotective in mice irradiated by (60)Co in vivo. The underlying mechanism remains unclear but appears to be mediated by the antioxidant activity of the betalains from red beets and modulation of the immune system.

Polypeptide from Chlamys farreri attenuates murine thymocytes damage induced by ultraviolet B.

AIM: Polypeptide from Chlamys farreri (PCF, molecular mass is 879) is a new marine polypeptide compound isolated from Chlamys farreri. This study investigates the possible protective roles and the mechanism of PCF against ultraviolet B (UVB)-induced apoptosis in murine thymocytes. METHODS: The rate of apoptosis and caspase-3 activation was measured by flow cytometry. The expression of stress-response genes c-fos and c-jun was observed by RT-PCR. Western blot analysis was performed to determine the release of cytochrome c. RESULTS: It was found that UVB induced murine thymocyte death. The cells treated with UVB showed an increase in cytochrome c release, caspase-3 activity, as well as in the expression of c-fos and c-jun. In addition, all were involved in UVB-induced cell apoptosis. CONCLUSION: Our present observations pointed to the ability of PCF to avert UVB-induced apoptosis in thymocytes by modulating c-fos and c-jun expression, cytochrome c release, and the consequent activation of caspase-3, which were essential components of the UV-induced cell apoptotic pathway. The results suggested that PCF is a promising protective substance against UV radiation.

Radiation-induced incidence of thymic lymphoma in mice and its prevention by antioxidants.

Previous reports from our laboratory have shown that in Swiss female mice exposed to an acute dose (3 Gy) of whole body irradiation (WBI), induced thymic lymphoma (TL) resulted after three to four weeks of exposure. The present study was aimed to further evaluate dependency on gender and effect of age of mice at the time of irradiation on TL incidence. A significant decrease in body weight gain was observed in female mice exposed to WBI, which was found to be correlated with the increase in weight and size of thymus, compared to their respective controls. An increase in TL incidence was observed with the increased postirradiation time, which was 47, 80, and 93% after 90, 120, and 150 days of WBI, respectively, in female mice. In irradiated female mice, the TL incidence was significantly higher and the growth of tumor in terms of weight and size was more aggressive than in males of the same age. Moreover, mice with higher age groups at the time of irradiation showed substantial decrease in TL incidence and its aggressiveness; and these effects were more conspicuous in males than in females. In mice irradiated at the age group of three to four weeks, the TL incidence was 83 and 72% in female and male, respectively, which was decreased to 74% in female and 14% in male in the age group of 12-13 weeks. It was further observed that the postirradiation feeding of animals with antioxidants resulted in a significant decrease in TL incidence, and the prevention in TL incidence was more in animals fed with curcumin (55%) than with ascorbic acid and eugenol (20%). These results have provided significant new findings on the phenomenon of radiation-induced TL incidence related to gender and age at the time of irradiation and its prevention by postirradiation antioxidant feeding to mice.

[Structure peculiarities of muscle regenerates and state of thymus under He-Ne laser therapy in different periods after muscle trauma].

We studied the gastrocnemius muscle regeneration and the reactive changes in thymus of rats under different regimens of He-Ne laser therapy of both operated legs (632.8 nm; 2.5-3.0 mW/cm(2) ). Laser radiation (10 exposures by 3 min within 1-15 days after muscle trauma, 4.5-5.4 J/cm(2) total dose per each leg) stimulated inflammatory reaction, muscle healing and favored preservation of muscle tissue in regenerates. The changes in thymus mass, its histological structure, size of cortex and thymocite mitotic index pointed to the increase of the functional load on thymus and delay of its recovery. The same dose of laser therapy of muscles within 16-30 days after trauma led to the increase of muscle tissue sclerotization in regenerates. The reactive changes in thymus were less pronounced. Threefold decrease of laser dose (10 exposures by 1 min for 1-15 days, 1.5-1.8 J/cm(2)) suppressed inflammatory reaction, impaired the muscle regeneration. The increase of functional activity in thymus was not observed.

Effects of four Si-Wu-Tang's constituents and their combination on irradiated mice.

Effects of four Si-Wu-Tang (SWT)'s constituents, fructose (Fru), paeoniflorin (Pae), ferulic acid (FA), tetramethyl pyrazine (TP), and their combination on irradiated mice as model of anaemia were investigated, with the purpose of further understanding the relationship between SWT's constituents and activities. Similarly to SWT, oral administration of Fru, Pae, FA, TP and their combination, to some extent, all showed effects of increasing the number of peripheral leukocyte and increasing four types of progenitor cells in bone marrow, including colony-forming unit-granulocyte-macrophage (CFU-GM), colony-forming unit-mature erythroid (CFU-E), colony-forming unit-immature erythroid (BFU-E) and colony-forming unit-multipotential (CFU-mix). Pae and FA showed significant body weight reducing effect, which were largely abolished when they were combined with Fru and TP. The SWT, Fru and combination significantly increased the thymus index while Pae significantly decreased it. Both SWT and TP significantly increased the spleen index but the combination did not. The results suggested that multiple constituents contribute to the promoting effect of SWT on hematopoiesis. Although being a very common compound in plants, the Fru has a special contribution to SWT's effect, which cannot be neglected. It may be an important active constituent that is responsible for SWT's promoting effect on hematopoiesis and immunity. Another suggestion is that when being combined, some effect of one constituent, sometimes is unexpected side effect, may be abolished by other. This may reflect the advantage of multiple constituent characteristics possessed by most TCMs.

[Effects of exposure of different skin areas to low-power laser light].

The effect of helium-neon laser light of extremely low power of 0.2 mW/cm2 and wavelength 632.8 nm on the immune status of mice bearing solid tumors was studied. The evaluation of the status of tumor-bearing animals was provided by taking into account the number of immune cells, cytokine concentration (tumor necrosis factor, interleukin 2, production of nitric oxide, expression of heat shock proteins (Hsp70 and Hsp90), and activity of natural killers. The model of a solid tumor was formed by subcutaneous injection of Ehrlich carcinoma cells, and average life span of tumor-bearing mice achieved about 55 days. Different areas of the skin of tumor-bearing mice were subjected either to a single (1 min, dose 0.012 J/cm3) or repeated exposure to laser light (1 min, 48-h intervals, 30 days). Two different areas were irradiated: the thymus projection area or a hind limb with solid tumors. The results showed that chronic exposure of tumor-bearing mice in the thymus projection area, and especially, hind limb, reduced the resistance, which manifested itself in the acceleration of tumor growth and a tendency of mouse life span to decrease. On the contrary, a single exposure stimulated the antitumor immunity for several days after the exposure. The results show the expediency of further investigation of the immunomodulative effects of low-power laser light and the necessity of monitoring the immune system during laser therapy.

Effects of low-power laser radiation on mice immunity.

BACKGROUND/PURPOSE: Because of large interest in biological effects of laser radiation used in laser therapy, the effect of extremely low-level red laser light intensity on the immune cell activity has been studied in the animal model with well-characterized macrophage and T cell populations as responder cells producing cytokines, protective proteins, active oxygen, and nitric compounds. To study of the possible side effects of laser immunotherapy we monitored the productions of cytokines, nitric oxide (NO), and heat shock protein 70 (Hsp70) in mice subjected to a periodic laser exposure for 1 month. METHODS: Helium-neon laser radiation with the power of 0.2 mW/cm2 and wavelength of 632.8 nm was applied on two different mouse skin surfaces, i.e. a thymus projection area or a hind limb. Healthy NMRI male mice were irradiated repeatedly with laser light for 1 min with 48-h intervals for 30 days. The animals were divided into three groups of 25 mice. The first and the second groups were exposed to laser light, on the thymus and hind limb area, respectively. The third, sham-irradiated group served as a control. Early and prolonged effects of laser radiation on the levels of NO (by Griess assay), Hsp70 (by Western blot assay), tumor necrosis factors (TNF-alpha and TNF-beta) (by cytotoxic assay using L929 cells as targets), and interleukin-2 (IL-2) (by ELISA assay) were determined. RESULTS: The dynamics of immune responses to low-power laser light intensity was shown to be dependent on two factors, i.e. the cumulative dose and the localization of the irradiated surface. Besides, various populations of cells demonstrated different sensitivity to laser radiation, with T cells being more responsive among examined populations of the cells. Low intensity laser light induced an immune cell activity when the exposure duration did not exceed 10 days, while a more prolonged period of treatment generated more severe changes in the immune system, up to immunosuppression. The treatment of the thymus zone resulted in more pronounced changes in the cytokine production as well as in NO and Hsp70 synthesis. CONCLUSION: Low-power laser irradiation showed more effective immunomodulatory effects when applied on the thymus projection area. The rise in IL-2 and Hsp70 production related to a short-term effect of laser application may be reversed after repeating laser treatment. We suggest that for the support of immune system stability, the prolonged laser therapy should be accompanied by supplementary methods.