Cytotoxic effect of magnetic iron oxide nanoparticles coated with seaweed aqueous extract against ovarian cancer cells

Iron oxide nanoparticles are one of the most efficient metal nanoparticles, which have prominent properties in biomedical applications. In previous studies, these nanoparticles were prepared using a green synthesis method with the aqueous extract of brown seaweed [Sargassum muticuni]. This study aimed to examine the cytotoxic effect of these synthesized nanoparticles on A2780cp ovarian cancer cells which indicate resistance to chemotherapy regimen. The cytotoxic effect of synthesized nanoparticles on A2780cp ovarian cancer cells were examined using the methylthiazol tetrazolium assay, acridine orange/ propodium iodide and caspase assay. Results indicated that Fe[3]O[4] nanoparticles encapsulated in seaweed water extract possess a cytotoxic effect on ovarian cancer cells resistant to chemotherapy [IC[50] values of 250 microg/ml, 125 microg/ml and 62.5microg/ml for 24, 48 and 72 h, respectively]. The nanoparticles through the induction of caspase-3 and -9 dependent apoptosis [intrinsic pathway] significantly exerted a cytotoxic effect against A2780cp ovarian cancer cells. The aqueous extract of brown seaweed is an appropriate candidate for stability and reduction of colloidal solution of iron oxide nanoparticles. Due to the apoptotic effect of this extract on A2780cp ovarian cancer cells, it can be a good choice for ovarian cancer treatment

[ effect of shoots of allium sativum L., ferula assa-foetida aqueous extract and low frequency electromagnetic field on angiogenesis in chick chorioallantoic membrane [in vivo]]

Background: Inhibition of angiogenesis is one of the goals of cancer treatment. In traditional medicine, Allium sativum L. and Ferula assa-foetida are consider as the important medicinal plants. The effect of electromagnetic field [EMF] on different aspects of cell growth has been confirmed

Objective: The aim of this study was to investigate the effect of shoots of Allium sativum L., Ferula assa-foetida extract and low frequency EMF on angiogenesis

Methods: 100 fertilized eggs were randomly divided into 10 groups: control [treated with distilled water], sham one [normal saline], sham two [EMF off], experimental one [shoot of Allium sativum L. extract], experimental two [shoot of Ferula assa-foetida], experimental three [combination of Allium sativum L. and Ferula assa-foetida], experimental four [EMF with intensity of 100 gauss], experimental five [Allium sativum L. + EMF], experimental six [Ferula assafoetida + EMF], experimental seven [combination of Allium sativum L. and Ferula assa-foetida + EMF]. Extract was injected on the 8th day of incubation and samples were exposed to EMF on 10th day. On 12th day, Chorioalantoic membrane was photographed and the length and numbers of vessels were measured

Results: There was a significant decrease in mean number and length of vessels in experimental groups one, two, four, five and six compared with the control group. The most significant decrease was created in group three, especially in group seven [P<0.05]. In addition, extract of Ferula assa-foetida has anti-angiogenesis effect more than Allium sativum L

Conclusion: Allium sativum L. and Ferula assa-foetida have inhibitory effects on angiogenesis and these effects amplifying by EMF

[ effects of different doses of atorvastatin on angiogenesis of chorioallantoic membrane of chick embryo]

Statins promote the proliferation, migration and survival of endothelial cells and bone marrow -derived endothelial progenitor cells [Angioblasts]. Angiogenesis, the formation of new blood vessels, is a dynamic and complex activity which is needed for embryogenesis and other physiological processes. However, in many pathological conditions such as solid tumor progression, the disease appears to be associated with persistent upregulated angiogenesis. In this research we used atorvastatin [0.1microm] and [10 microm] on angiogenesis of chick embryo. In this experimental study 42 Ross fertilized eggs were randomly divided into 3groups as follows: 1] control group, 2] group treated with atorvastatin [0.1microm], 3] group treated with atorvastatin [10microm]. In day 2 a window was opened on eggs in sterile condition and In day 8 gelatin sponge was placed on chorioallantic membrane [CAM] and was soaked with 10 microliters atorvastatin [0.1microm] and [10 microm] in group 2 and group 3. In day12 CAMs were examined and photographed by research photo-stereomicroscope in all cases. Data were analyzed statistically by ANOVA and Tukey tests. The average number and length of vessels in control and the group treated with atorvastatin [0.1microm] showed a significant increase [P<0.05] and the average number and length of vessels in the group treated with atorvastatin [10 microm] showed a significant decrease compare to control group [P<0.05]. The results of this study showed that atorvastatin has a stimulatory effect on angiogenesis in CAM and atorvastatin [10 microm] has an inhibitory effect on angiogenesis in CAM. It is suggested that astatine can be used as a new medicine for angiogenesis balancing in treat ment of diseases related to angiogenesis

[Comparison of the percentages of peripheral blood CD4+ CD25+ T lymphocytes in recurrent abortion and normal pregnancy]

Background and Aims: Recent evidences indicate that parts of the immunoregulation system such as CD4+CD25+Tcells [Treg] and Th2 cells and Th1 cells, play very important roles in the maintenance of pregnancy. The deficiency in proper recognition of fetal alloantigen by the maternal immune system is associated with recurrent pregnancy failure. Here, we investigate the proportional changes of CD4+CD25+Tcells in peripheral blood of women with unexplained recurrent spontaneous abortion in comparison to women with normal pregnancy by using flowcytometry

Methods: The case group was comprised of 24 women who had at least three successive miscarriages with unexplained etiology. They had normal karyotypes, anticardiolipin and prolactin and their husbands had normal spermograms. The percentages of TCD4+CD25+cells in peripheral blood of these patients were compared with those of 21 women who had normal pregnancy with no history of pregnancy loss. Anti-CD4, anti-CD25 and anti-CD3 antibodies were added to lymphocytes isolated from peripheral blood. Then samples were incubated, centrifuged and washed. Finally cells were analyzed using FACS Caliber system and data of the two groups were compared

Results: Mean percentage of CD4+CD25+bright T cells in peripheral blood in case group was significantly lower compared to the control group [P=0.000]. Mean percentage of CD4-CD25 bright cells in the CD4+Tcell peripheral blood was significantly higher in case group campared to the control group [P=0.021]

Conclusion: Decrease of CD4+CD25 bright T cells plays a major role in tolerating conceptus antigens and cytokine and might contribute to the maintenance of pregnancy. Inadequate CD4+CD25+Tcells or their functional deficiency may link with miscarriage. Therefore, alteration of CD4+CD25+T cells can be used as an immunologic marker for monitoring of patients with unexplained recurrent spontaneous abortion

[Synergistic effects of vitamin A and extremely low frequency electromagnetic field [50HZ] on limb bud development in balb/c mouse]

Vitamin A is involved in vertebrate morphogenesis, growth, cellular differentiation, and tissue homeostasis. Vitamin A and its metabolites are essential for adequate embryo development. Electromagnetic fields [EMFs] have been used effectively to treat some diseases, such as certain musculoskeletal and chondrogenic disorders. In the present study, we aimed to investigate the synergistic effects of EMFs and Vitamin A on limb bud development in Balb/C mice in vivo. The Balb/C mice were used as experimental model. After mating, the pregnant mice were divided randomly into 3 groups as control, sham and experimental groups. Control and experimental groups received a single dose injection of Vitamin A [15000IU/Kg] on 10.5[th] gestational day, interaperitonealy. In addition, animals in experimental group were exposed to EMFs [50 Hz /100 gauss, for 4[th] during three days]. The animals were killed on the 15.5[th] day of gestation and were submitted to caesarian section. The obtained fetuses were examined externally with an analyzed under a stereoscopic microscope and were photographed. Data were analyzed statistically by t-test and ANOVA and Ktuskal-Wallis. Morphological and histological examinations showed significant changes in limb buds as compared with sham exposed and control groups. In both fore and hind limb buds significant decrease obtained in proximo-distal [P-D] and anterior posterior [A-P] axes [P<0.05]. Chondrocytes counts revealed a significant delay in the development of chondrocytes in experimental groups as compared with sham exposed and control groups [P<0.05]. Synergistic effects of vitamin A and extremely low frequency electromagnetic field [50Hz] cause delay in development

synergic effects of rapamycin and extremely low frequency electromagnetic field on angiogenesis

Angiogenesis, the formation of new blood vessels, is a dynamic and complex activity which is needed for embryogenesis and other physiological processes. However, in many pathological conditions such as solid tumor progression, the disease appears to be associated with persistent up-regulated angiogenesis. In this research we used 0.04 T [tesla] electromagnetic field as a synergic treatment with rapamycin on angiogenesis. In this experimental study, 70 Ross fertilized eggs were randomly divided into 5 groups as following: 1] control, 2] sham-exposed, 3] a group treated with rapamycin, 4] a group treated with electromagnetic field and 5] a group treated with both rapamycin and electromagnetic field. In day 2, a window was opened on eggs in sterile condition. In day 8 a gelatin sponge was placed on chorioalantoic membrane [CAM] and was soaked with 5 micro l rapamycin in group 3 and group 5. Groups 4 and group 5 were placed in 400 Gauss magnetic field for 4 hours in day 10. In day 12, CAMs were examined and photographed by research photo-stereomicroscope in all cases. Data were analyzed using ANOVA and Mann-Whitney tests. Comparison between average number and length of vessels in controls and sham-exposed didn't show any significant differences. In group 3 and 4, a significant decrease was shown in the average number and length compared with controls. Finally, comparison between group 3 and group 5 showed a significant decrease in the average number and length of vessels in group 5 which had been treated with both rapamycin and 400 [Gauss] G electromagnetic field. The results of this study showed that 0.04 T magnetic field has an inhibitory effect on angiogenesis in CAM and can enhance the effect of rapamycin as an anti-angiogenesis drug

[Effect of cell phone radiation [940 MHz] on the learning and memory of Balb/c mice]

The possible risks of radio-frequency electromagnetic fields for human body are a growing concern for our society. In the modern world, the increase of using cell phones is remarkable in all generations especially young people. In addition many users hold phone close to their head. Therefore, we have studied the effect of cell phone radiation [940 MHz] on memory and learning of Balb/C mice which were exposed after delivery. A purpose-designed exposure at a specific absorption for 3h/day from day 1 to 20 after delivery was used. Any stress response in the brain was detected in hippocampus. Behavioral performance began 2.5 month after delivery and testing was conducted in the water maze and the radial maze distinguished working and reference memory performance. After behavioral studies, fetal heads were collected, fixed in 10% paraformaldehyde and paraffine embedded. Microwave-exposed mice were slower than sham-exposed, and cage control in swim speed [WM]. Error Analyses rates reveal significant exposure effect in RAM and MWM. However, in this study the exposed group had not significantly lost their hippocampal CA3 neurons comparing to controls or sham exposed group. Increased time to locate a submerged in water maze, reference memory [entries into unbaited arms], working memory [repeated entries into baited arms], show that acute exposure to pulsed microwaves caused a deficit in spatial reference memory in the mouse

[ effect of HCG on in vitro maturation of human immature oocytes and rate of 8 cell embryos mediated by ICSI]

The infertility treatment by using in vitro matured oocyte has many potential applications. It minimizes the risk of ovarian hyper stimulation and is an alternative treatment for women with polycystic ovarian syndrome who may have problems regarding stimulation for IVF. This method may prove important for subjects in need of fertility preservation and also provides information about the final stage of oocyte maturation. The aim of this study was to determine the effect of HCG on the rate of in vitro matured oocyte and 8 cell embryos mediated by ICSI. This experimental study was done in Montaserieh IVF center and Islamic Azad University at Mashhad. One hundred sixty eight immature oocytes aspirated from women undergoing IVF cycles were divided into 3 equal groups and allocated for maturation in 3 medium cultures. All of 3 culture groups contained G1 medium supplemented with HSA 10% [Human Serum Albumin]. Control group had no gonadotropin, but the first experimental group contained HCG 10 ILJ/ml and the second experimental group contained HCG 5 ILJ/ml. Matured oocytes were fertilized by ICSI and numbers of 8-cell embryos were investigated 72 hours after fertilization by invert microscope. The results showed that both of the HCG concentrations significantly increased the rate of matured oocytes and also the rate of 8-cell embryos with respect to control group [p<0.05]. No significant differences were found in rate of matured oocytes between the two experimental groups [p>0.05] while the number of 8-cell embryos was significantly higher in HCG 5IU/ml with respect to HCG 10 IU/ml and control group [p<0.05]. The results suggests that both of the HCG concentrations increase the rate of maturation of immature oocytes and the rate of 8-cell embryos but HCG [5IU/ml] has more effect on the number of 8-cell embryos mediated by ICSI

[ effect of extremely low frequency electromagnetic field[50Hz] on induction of chromosomal damages on bone marrow erythrocytes of male Balb/C mouse]

The increasing use of the electromagnetic field producer sets in daily living causes concerns about these waves on human health. The effects of extremely low frequency electromagnetic field [50 Hz] on induction of chromosomal damages on bone marrow erythrocytes of male Balb/C mouse has been investigated in this research. This is an experimental study in which the laboratory system of producing electromagnetic field with low frequency [50 Hz] was used. Five week old male Balb/C mice were divided into three controls, Sham-exposed and experimental groups. The experimental mice exposed were exposed to electromagnetic field [50 gauss] for 4 days [12 hours/day]. After treatment, the chromosomal damages were assessed using micronucleus test in polychromatic erythrocytes and resultant quantity data were analyzed using t and Mann-Whitney test. Results showed that frequency of micronucleated polychromatic erythrocytes of experimental group [14/35 +/- 1/589] was more than Sham-exposed [8/958 +/- 1/049] and control group [7/65 +/- 0/768] significantly [p<0/05]. Extremely low frequency electromagnetic field [50 Hz] causes chromosomal damages induction in polychromatic erythrocytes of bone marrow male Balb/C mouse

Identification and localization of certain glycoconjugates during the development of thymus in the mice embryos Balb/C

Glycoconjugates molecules showed a programmed changes during the development. These components in the cell-surface and extracellular matrix plays important roles for different developmental processes. In this study lectin histochemistry technique was used for identification and localization of some glycoconjugates during the development of thymus. Balb/C mice embryos from days 10 to 15 and also day 18 fetuses iixed in formalin and provided 5m serial sections of these samples for histochemical study. Slices were incubated with three types of HRP-conjugated lectins [HRP] include: SBA specified for alpha, [beta-D-N-acetylgalactoseamin [alpha, beta-D-GalNAc], PNA specified for beta-D-Galactose -[l-3]-D-N-acetylgalactoseamin [beta-D-Gal-[l-3]-D-GalNAc], MPA specified for D-Galactose [D- Gal]. SBA lectin was presented with decreasing intensity in the Golgi zone [GZ] and cell surfaces of mesenchymal cells, epithelial cells and T-cells microenvironment in studying days. In first day [12 and 13] T-cells had high reaction with SBA in membranes and GZ. PNA lectin was revealed with several intensity in basal membranes and membranes of mesenchymal cells, epithelial and microenvironment of T-cells in studying days. MPA lectin weakly presented in the endodermal and mesenchymal cells and microenvironment of T-cells in all days. Results indicated that glycoconjugates molecules with terminal carbohydrate of GalNAc might have a role in the development of thymus gland and maturation of the T-cells. Glycoconjugates with terminal disaccharide of Gal/GalNAc probably have significant effect on the total developmental process of thymus gland. Further, Glycoconjugates with terminal carbohydrate of Gal probably had not a key role in the development of thymus gland and T-cells or with low quantity effect

[ effects of mobile phone waves [940MHz] on embryonic development of hematopoiesis system in Balb/C mouse]

Today, increasing usage of microwaves generator sets such as cell phones, have been caused much worry related to their waves effects on human health. In this research, simulated waves effects of cell phones with frequency of 940MHz have been studied on hematopoiesis system of Balb/C mouse embryo. In an experimental study pregnant mice [Balb/C] were used as a laboratory model. The mice were divided into control, sham-exposed and experimental groups. The experimental groups were exposed to the waves on the 8[th] day of embryonic development for 4 or 10 days [everyday for 4 hours]. At the end of treatment period, the experimental samples, sham-exposed and control related to morphology and histology studies of hematopoiesis organs were done by light microscope. Using t-test and Mann-Whitney test, the quantitative data were analyzed. Morphologic study of embryos which were 18 days old didn't show any abnormality but the weight of experimental embryos [1.263 +/- 0.017 gr] compared with sham-exposed [1.268 +/- 0.006 gr] showed significant increase [P<0.05]. The study of tissue sections in embryos which were 12 days old didn't show significant change in tissue and cellular position of yolk sac of experimental samples as compared with control group. Also, tissue studies of embryos which were 18 days old didn't show any changes in general structure of liver, spleen, marrow bone but the number of Megakaryocytes of spleen and erythrocytes which had not nucleus in bone marrow of experimental samples showed significant increase as compared with the sham-exposed. The number of erythrocytes in liver which had not nucleus also showed significant increase as compared with the sham-exposed [P<0.05]. Waves of cell phones with frequency of 940MHz don't affect on general structure of liver, spleen, bone marrow of Balb/C embryos but it causes significant changes in the number of cells of these tissues

[Genotoxic effect of hydrogen peroxide on bone marrow erythrocytes of female Balb/C mouse]

Hydrogen peroxide is a strong oxidant which is being widely used in hairdresser places. Genotoxic effect of such substances, referred to as "interaction between gene and environment, is one of the most worrying problems of international health society. On the basis of the wide use of hydrogen peroxide, its capability on induction of chromosomal damages on polychromatic erythrocytes of bone marrow of exposed female Balb/C mice was investigated. The female Balb/C mice were used as experimental model. The mice were divided to three different groups including: control, Sham-exposed and experimental groups. The experimental group was exposed to inhalation of 9% oxidant vapor, containing hydrogen peroxide, for 2 and 3.5 hours per day for 7 days. Bone marrow cells of exposed as well as control and sham-exposed mice were collected using FCS, and micronucleus assay was performed. The chromosomal aberration was investigated by scoring the micronucleus containing polychromatic erythrocytes. The data was statistically analyzed by t-test. The results showed that mean No. of polychromatic erythrocytes in sham-exposed 1 and treated 1 [2 hours exposure to H2O2] were 8.87 +/- 1.01 and 5.75 +/- 3.05 respectively [P<0.05], and sham-exposed 2 and treated 2 [3.5 hours exposed to H2O2] were 8 +/- 2.36 and 42.37 +/- 4.8 respectively [P<0.01]. Also increasing the exposure treatment time with H2O2 could increase the chromosomal abnormality [P<0.05]. Results of this study revealed the genotoxic effects of inhalation of 9% H2O2 on bone marrow cells of female Balb/C mouse

[ effects of exposure to simulation cell phone waves on gonads of male mouse]

The increasing users of microwave appliances such as cell phones caused an increasing interest on investigation of its adverse effects on human health and development of animals. This study is done to investigate the effects of simulating cell phone waves on gonads and sex cells of male Balb/C mouse. This is an experimental study in which with the use of generating apparatus of simulating cell phone waves, adult male mice were exposed to cell phone waves for 10 days [4 hours per day]. Then structure and ultrastructure of testes and number of sex cells were examined by light and transmission electron microscope. Data was analyzed using t and Mann Whitney testes. The results did not show any significant differences in the size and weight of testes in mice exposed to the simulating cell phone waves. Results also showed that the number of spermatogonia cells and primary spermatocytes and spermatids and sperms were significantly increased in exposed mice [p<0.05], but the number of sertoli cells were significantly decreased [p<0.05]. The study of micrographs showed changes in ultrastructure of sexual cells, such as cleft and hetrochromatined nucleus and decrease of cell organelles and vacuolization of cytoplasm. Results indicate the effect of simulating cell phone waves on number and ultrastructure of sex cell in male Balb/C mouse