Female NMRI Mice Research Articles

Effect of Intermittent Feeding on Gonadal Function in Male And Female NMRI Mice During Chronic Stress

ABSTRACT Stress can inhibit gonadal activity via Hypothalamus-Pituitary-Gonad (HPG) axis activity suppression. In the present study, effects of intermittent feeding (IF) on gonadal function under stress in male and female mice were evaluated. Twenty eight male and twenty eight female mice's were divided into four groups. The control group received adequate food and water without stress. The second group received four days of electric shock without food deprivation. The third group was deprived of food two hours/day for a week, and the fourth group was deprived of food (2 hours/day for seven consecutive days) and then electric foot shock stress was applied to them for four days. Blood samples were collected from all animals for plasma testosterone, estrogen and/or Interlukin-6 (IL-6) evaluation. The animals’ gonads were also removed and fixed for the measure of their weight. Results showed that stress reduces both testosterone and estrogen levels, whereas IF did not change the hormone levels. In addition, stress increases blood IL-6 concentration. The combination of IF and stress, increased the hormone levels in animals. Stress and IF alone had no significant effect on gonadal weight in the male mice, whereas stress decreased gonadal weight in the females. Combination of stress with IF increased gonadal weight in both male and female mice. In conclusion stress showed a negative effect on gonadal function in both animals with more effect on females. Intermittent feeding inhibits the stress effect and even promotes the gonadal function in both sexes. The effect may be due to IL-6 reduction.

The effect of Melatonin on histological changes of ovary in induced polycystic ovary syndrome model in mice

Abstract Background Antioxidants can be used as adjuvant treatment of polycystic ovary syndrome (PCOS). Melatonin (MT) is one of the antioxidant that is used nowadays. Objective In this study, the effect of MT on the histological changes of ovary in the experimental model of polycystic ovary syndrome is investigated. Methods In this study 30 immature female NMRI mice were divided into 5 groups including: (1) control group received distilled water, (2) received a dose of 10 mg/kg MT for 5 days, (3) received Dehydroepiandrostenedione (DHEA) at a dose of 6 mg/kg for 20 days to induce PCOS, (4) after induction of PCOS received MT at a dose of 10 mg/kg for 5 days, and (5) received 6 mg/kg DHEA and 10 mg/kg MT for 20 days simultaneously. Results The evaluation of ovarian tissue characteristics such as the granulosa layer, theca, number and diameter of cysts and follicles was performed. PCOS caused a significant reduction in the number of antral follicles and corpus luteum and an increase in the number of primordial, primary, pre-antral and cystic follicles in comparison with the control group (P Conclusion These findings indicate that MT have a protective effects on polycystic ovary damages induced by DHEA, although the mechanism is unclear. It is likely that this is happening by reducing oxidative damage.

Specificity of exogenous acetate and glutamate as astrocyte substrates examined in acute brain slices from female mice using methionine sulfoximine (MSO) to inhibit glutamine synthesis.

Removal of endogenously released glutamate is mediated primarily by astrocytes and exogenous 13 C-labeled glutamate has been applied to study glutamate metabolism in astrocytes. Likewise, studies have clearly established the relevance of 13 C-labeled acetate as an astrocyte specific metabolic substrate. Recent studies have, however, challenged the arguments used to anchor this astrocyte specificity of acetate and glutamate. The aim of the current study was to evaluate the specificity of acetate and glutamate as astrocyte substrates in brain slices. Acutely isolated hippocampal and cerebral cortical slices from female NMRI mice were incubated in media containing [1,2-13 C]acetate or [U-13 C]glutamate, with or without methionine sulfoximine (MSO) to inhibit glutamine synthetase (GS). Tissue extracts were analyzed by gas chromatography-mass spectrometry. Blocking GS abolished the majority of glutamine 13 C-labeling from [1,2-13 C]acetate as intended. However, 13 C-labeling of GABA was only 40-50% reduced by MSO, suggesting considerable neuronal uptake of acetate. Moreover, labeling of glutamate from [1,2-13 C]acetate in the presence of MSO exceeded the level probable from exclusive labeling of the astrocytic pool, which likewise suggests neuronal acetate metabolism. Approximately 50% of glutamate was uniformly labeled in slices incubated with [U-13 C]glutamate in the presence of MSO, suggesting that neurons exhibit substantial uptake of exogenously provided glutamate. © 2017 Wiley Periodicals, Inc.

Impact of prolonged oocyte incubation time before vitrification on oocyte survival, embryo formation, and embryo quality in mice.

Oocyte incubation time before freezing is one of the factors affecting oocyte vitrification. In the assisted reproductive technology (ART) clinics, it is sometimes decided to perform oocyte vitrification after a long period of incubation time due to various conditions, such as inability to collect semen samples, unsuccessful urological interventions (PESA, TESE, etc.), or unexpected conditions. A time factor of up to 6h has been studied in the available reports. Therefore, this study was designed to evaluate oocyte incubation time before freezing at 0, 6, 12, 18, and 24h after retrieval. Metaphase II (MII) oocytes were obtained from NMRI female mice after being randomly divided into the five groups of 0, 6, 12, 18, and 24h of freezing via hormonal stimulation following retrieval and entered into the vitrification-warming process. The thawed oocytes were evaluated according to the survival criteria and then inseminated with the sperms of male mice for in vitro fertilization. The next day, the embryo formation rate and embryo quality were assessed. Our results demonstrated that even after 24h of incubation, the survival rate of oocytes was 51.35% with the embryo formation rate of 73.21%. However, the survival and embryo formation rates significantly decreased within 12, 18, and 24h after retrieval compared to the groups vitrified at 0h. The embryo quality was significantly reduced by vitrification at 0 to 24h after retrieval. According to our data, although a prolonged incubation time before freezing reduced the survival rate, there was still a chance for oocytes to stay alive with acceptable embryo formation and quality rates after vitrification warming of oocytes.

Which Stage of Mouse Embryos Is More Appropriate for Vitrification?

Vitrification has been shown as one of the most effective methods of cryopreservation for mammalian embryos. However, there is no consensus which stage of embryonic development is the most appropriate for vitrification with subsequent maximal development after thawing. This study was carried out to explore and compare the effect(s) of vitrification on mouse 2-cell, 4-cell, 8-cell, morula and blastocyst stage embryos and subsequent blast formation and hatching after thawing. In this experimental study, 2-cell embryos were obtained from the oviducts of super ovulated female NMRI mice. Some embryos were randomly selected and vitrified through a two-step media protocol and cryotop. Other embryos were cultured to assess their development. During the ensuing days, some of these cultured embryos were vitrified at 4-cell, 8-cell, morula and blastocyst stages. After 10 to 14 days, the embryos were thawed to assess their survival and also cultured to determine the rate of blastocyst formation and hatching. The results were analyzed using one-way ANOVA and Tukey's post-hoc tests. There was no significant difference in the survival rates of vitrified embryos at 2-cell, 4-cell, 8-cell, morula and blastocyst stages after thawing (P>0.05). The blastocyst formation rate of vitrified 8-cell embryos was significantly higher than that of 2-cell embryos (P<0.05). The hatching rate of vitrified 4-cell, 8-cell and blastocysts were significantly higher than that of 2-cell embryos (P<0.05). Vitrification is suitable for cryopreservation of all stages of mouse embryonic development. However, the best tolerance for vitrification was observed at 4and 8-cell stages of development. Accordingly, the development of vitrified embryos to blastocysts, following thawing, was most efficacious for 4 and 8-cell embryos. Compared to mouse 2-cell embryos, embryos vitrified as blastocysts had the highest rate of hatching.

Antioxidant capacity and genoprotective effect of ethanol fruit extract from Detarium microcarpum Guill. and Perr. (Caesalpiniaceae)

ObjectiveTo evidence the ability of ethanol fruit extract from Detarium microcarpum (D. microcarpum) to preserve DNA integrity against oxidative genomic damage. MethodsEthanol extract from D. microcarpum fruit pulp was analyzed for its antioxidant capacity using ferric reducing antioxidant power, 2,2-diphenyl-1-picrylhydrazyl, 2,2′-azinobis-3-ethyl-ethylbenzothiazoline-6-sulphonate, superoxide anion, deoxyribose degradation and lipid peroxidation models. The genoprotective activity was assessed ex vivo by comet assay, on liver cells of NMRI female mice using cyclophosphamide (CP) as genotoxic agent. ResultsEthanol extract from D. microcarpum fruit pulp exhibited interesting antioxidant activity in 2,2-diphenyl-1-picrylhydrazyl, deoxyribose degradation and lipid peroxidation assays. The extract did not present any genotoxic effect but protected DNA against CP-induced damages with a dose-dependent manner. The genoprotective effect observed was related to the antioxidant molecules of the fruit that scavenged the hydroxyl radical (generated by the metabolism of CP) as well as the peroxyl and alkoxyl radicals issued from lipid peroxidation. Other mechanisms such as inactivation of CP metabolism to genotoxic end products, induction of the expression of antioxidant and DNA repair enzymes have been discussed. ConclusionsOur results suggest that the wild edible fruit from D. microcarpum could be beneficial on consumer's health by its antioxidant and genoprotective effects, particularly during chemotherapies exhibiting genotoxic effects like CP in cancer treatment.

Effects of Vitex agnus-castus fruit on sex hormones and antioxidant indices in a d-galactose-induced aging female mouse model

BackgroundAging is associated with the loss of endocrine function. In this study, Vitex agnus-castus (Vitex), which has antioxidant effects and high levels of phytoestrogen, was investigated with regard to the hypothalamic-pituitary-gonadal axis and antioxidant indices in natural aging and in a d-galactose induced aging model in female mice. MethodsThe mice were subcutaneously injected with d-galactose (500 mg/kg/d for 45 days). Extract of Vitex (600 mg/kg/bid for 7 days by gavage) was used to treat d-galactose-induced aging and natural aging in mice. Seventy-two female NMRI mice (48 3-month-old normal mice and 24 18-24-month-old mice), weighing 30–35 g were randomly divided into six groups: control, Vitex, d-galactose, Vitex + d-galactose, Aging, and Vitex + Aging. The antioxidant indices and sex hormone levels were subsequently measured by enzyme-linked immunosorbent assay kits. ResultsBody weight and the levels of malondialdehyde (MDA), follicle-stimulating hormone, and luteinizing hormone levels were significantly increased in the d-galactose aging and natural aging groups, whereas catalase and superoxide dismutase (SOD) activity and estrogen level were significantly decreased in these same groups. d-Galactose can also disrupt the estrous cycle and damage the uterus and ovarian tissues. Vitex could effectively attenuate these alterations. ConclusionVitex improved some aging events in the reproductive system of female mice. Therefore, because of its apparent antiaging effects, Vitex can be suitable for some aging problems such as oxidative stress, female sex hormone deficiency, and an atrophic endometrium.

Potent radioprotective effect of herbal immunomodulator drug (IMOD) on mouse bone marrow erythrocytes as assayed with the micronucleus test

Background : Although numerous natural or synthec drugs have been tested for their radioprotecve capacity, yet no suitable drug has been introduced for roune clinical use. In this study the radioprotecve effect of new herbal commercially known as IMOD, specifically made to decrease the side effects of HIV virus, was invesgated on mouse bone marrow cells. Materials and Methods : Female NMRI mice (in a group of five) were exposed to 2 Gy gamma radiaon following three days of intravenously injecon (IV) of IMOD at various doses. Mice were sacrificed 48 and 72 h a1er irradiaon. Bone marrow was flushed and slides for bone marrow smears were prepared according to standard method. A1er st aining slides in May Grunwald and Giemsa, polychromac erythrocytes (PCE) and normochromac erythrocytes (NCE) were scored for presence of micronucleus (MN). Results: The results showed that gamma irradiaon increased the frequency of micronuclei dramacally and excreted cytotoxic effect of cell proliferaon. Injecon of various dos es of IMOD before irradiaon however, led to a considerable reducon in the frequency of micronuclei in bone marrow erythrocytes as well as cellular toxicity. Conclusion: Results indicated radioprotecve capability of IMOD with a dose reducon factor (DRF) of about 2.3 at a dose of 20 mg/kg body weight. The considerably high DRF might be indicave that IMOD besides being an immunomodulator might also posses' anoxidant property.

Effect of Temporary Inactivation of Nucleus Accumbens on Chronic Stress Induced by Electric Shock to the Sole of the Foot in Female NMRI Mice

BACKGROUND AND OBJECTIVE: Activity changes in the neurons of nucleus accumbens during stress have been previously identified. However, the role of nucleus accumbens in diminishing stress-induced side-effects is not fully understood. In this study, we aimed to evaluate the effects of temporary inactivation of nucleus accumbens on stressinduced metabolic changes in female mice. METHODS: This experimental study was performed on 48 female NMRI mice with an average 27±3 g. The nucleus accumbens was unilaterally and bilaterally cannulated. After one week of recovery, 2% lidocaine or saline was administered in mice for four consecutive days (5 min per day) before inducing electric shock to the sole of the foot. Plasma corticosterone level, food and water intake, and delay in eating were assessed as stress-induced metabolic parameters. FINDINGS: Stress lonely, caused an increase in plasma corticosterone (17±0.8) compared with the control group (4.5±0.3) (p<0.001). It also, caused an increase delay in eating (%218±9.8, p<0.01) and, decrease water (%80±4.5) and food (%84±5.5) intake (p<0.05). Temporary inactivation of nucleus accumbens did not affect the stress-induced changes in plasma corticosterone, and it suppressed the effect of stress on the amount of water intake; inactivation of the left nucleus accumbens was more effective (%195±7.6, p<0.01). Temporary inactivation of nucleus accumbens neutralized the effect of stress on the amount of food intake. Temporary inactivation of the right nucleus accumbens augmented the effect of stress on delay in eating (%264±10.8, p<0.01), and inactivation of the left nucleus accumbens could suppress this effect. CONCLUSION: It seems that temporary inactivation of nucleus accumbens can be effective in diminishing stressinduced metabolic changes. However, this influence is indicative of asymmetry in the function of right and left nucleus

Effects of Portulaca oleracea ethanolic extract on reproductive system of aging female mice

Aging contains morphological and functional deterioration in biological systems. D-galactose (D-gal) generates free radicals and accelerates aging. Portulaca oleracea (Purslane) may have protective effect against oxidative stress. Purslane ethanolic extract effects were evaluated on antioxidant indices and sex hormone in D-gal aging female mice. 48 female NMRI mice (25-35 gr) were randomly divided into, 6 groups: 1- control (normal saline for 45 days), 2- Purslane (200 mg/kg for last 3 weeks), 3-D-gal (500 mg/kg for 45 days), 4-D-gal+Purslane, 5- Aging, 6-Aging+Purslane. Sex hormones, antioxidants and malondialdehyde (MDA) level of ovary and uterus were measured. Histological assessment was also done. In D-gal treated and aging animals, LH and FSH levels were significantly increased (p<0.001) while estrogen and progesterone levels were significantly reduced (p<0.001) in comparison with control group. MDA contents were significantly increased in ovaries and uterus of D-gal and aging groups (p<0.01). Superoxide dismutase (SOD) (p<0.001) and catalase (p<0.01) activities were significantly decreased in both aging and D-gal treated animals. Ovarian follicles were degenerated and atrophy on uterine wall and endometrial glands was observed in D-gal and aging groups. Alteration in hormone levels, MDA contents and antioxidant activity were significantly reversed by Purslane (p<0.05). Purslane could also improve histological changes such as atrophy of endometrium. These findings indicate that Purslane can attenuate aging alternations induced by D-gal and aging in female reproductive system.

The effect of progesterone treatment after ovarian induction on endometrial VEGF gene expression and its receptors in mice at pre-implantation time.

Progestrone is a prequisite for pre-implantation angiogenesis and induce decidual angiogenesis. It is unknown the effect of progestrone administration on the endometrium of hyperstimulated mice at pre-implantation time. Adult female NMRI mice were divided in three groups [control group, ovarian stimulated group and progestrone treated mice after ovarian stimulation]. Uterine horn samples removed at pre-implantation time in each group. Motic image Plus 2 software was used to assess the quantitative vascular parameters of endometrium. Gene expression was determined for vascular endothelial growth factor (VEGF), FMS-like tyrosine kinase (FLT) and Kinase insert domain protein receptor (FLK) genes using the real time PCR method. Data analysis was done with LinReg PCR and Rest-RG software. Comparison between progestrone treated mice after ovarian stimulation with control group showed that increase in rate of VEGF gene expression [0.775] and decrease in rate of FLK [6.072] and FLT [1.711] gene expression. Analysis of the data on quantitative vascular parameters were indicated remarkable increase in quantitative vascular parameters of progestrone treated mice compare to control group. Biological effect of progestrone on the vascular changes after ovarian stimulation resulted in an increase in VEGF receptors experession, it seems that induced angiogenesis by progesterone could result in better condition for implantation.

Evaluation of mouse embryos produced in vitro after electromagnetic waves exposure; Morphometric study.

IntroductionToday, the use of electromagnetic waves in medical diagnostic devices, such as magnetic resonance imaging (MRI), has increased, and many of its biological effects have been reported. The aim of the present study was to assess the biological effects of 1.5 Tesla (T) magnetic resonance imaging (MRI) on fertility and reproductive parameters.MethodsEighty adult male and female NMRI mice (NMRI: Naval Medical Research Institute) of age 6–8 weeks were studied and randomly divided into two study and control groups. After confirmation of pregnancy, the mice in the study group were exposed to the MRI (1.5 T) machine’s waves over the next three weeks, once a week for 36 minutes. One day and thirty-five days after the last radiation, the mice were killed in order to do the in vitro fertilization (IVF) by neck beads’ displacement and the impact on the evolution of embryos, and its quality was studied. Data were analyzed using SPSS version 20 and the significance level of less than 0.05 was considered.ResultsEmbryo morphometry showed that the total diameter and the cytoplasm diameter of the study group embryos suffered significant reduction compared to the control group, 1 day after the last irradiation (p < 0.05), but the diameter of the perivitelline space of this group’s embryos had a significant increase (p < 0.05). The qualitative results during 35 days after irradiation showed that morphologically parameters of the embryos in the study group had no significant differences from the control group.ConclusionExposure to MRI irradiation can transiently disturb the development of mouse embryos and fertility, but these effects are reversible 35 days after the last irradiation.

The Effect of Methamphetamine on Oocyte Quality, Fertilization Rate and Embryo Development in Mice

Objectives: Methamphetamine (METH) is an illicit psychoactive drug. There are different reasons of abusing METH such as for recreational use and sexual satisfaction. This study was designed to investigate the effects of short and long-term use of METH on oocyte and embryo development in mice. Materials and Methods: In this study, 75 female NMRI mice were divided into five groups of 15. The groups consisted of a control group, experimental groups A and B, which received METH (10 mg/kg/day) intraperitoneally for 2 and 14 consecutive days, respectively and two sham groups A and B, which only received saline. After the last injection, 10 international units pregnant mare serum gonadotropin (IU PMSG) and 10 IU human chorionic gonadotropin (HCG) were administered intraperitoneally for induction of ovulation. Then, all the mice were sacrificed to aspirate their oocytes for further evaluation. In vitro fertilization (IVF) was done by using mature oocytes and embryo development was investigated up to the blastocyst stage. Data was analyzed using SPSS and non-parametric Mann-Whitney test. Results: The number of fragmented oocytes in experimental group A was significantly increased in comparison with experimental group B and control group (P < 0.05). Also there were significant differences in the number of unfertilized oocytes and early cleavage stage embryos in experimental group B in comparison with other groups that indicates the reduction of fertilization rate in experimental group B (P < 0.05). Conclusion: Short-term administration of METH in mice can affect oocyte quality but had no effect on early embryo development, while the long-term administration may affect oocyte and embryo development in early cleavage stage.

Significant effects of sex, strain, and anesthesia in the intrahippocampal kainate mouse model of mesial temporal lobe epilepsy.

The intrahippocampal kainate mouse model of mesial temporal lobe epilepsy is increasingly being used for studies on epileptogenesis and antiepileptogenesis. Almost all previous studies used male mice for this purpose, and no study is available in this or other models of acquired epilepsy that directly compared epileptogenesis in female and male rodents. Epidemiological studies suggest that gender may affect susceptibility to epilepsy and its prognosis; therefore, one goal of this study was to investigate whether sex has an influence on latent period and epileptogenesis in the intrahippocampal kainate model in mice. Another aspect that was examined in the present study was whether mouse strain differences in epileptogenesis exist. Finally, we examined the effects of different types of anesthesia (chloral hydrate, isoflurane) on kainate-induced status epilepticus (SE) and epileptogenesis. Continuous (24/7) video-EEG monitoring was used during SE and the 2 weeks following SE as well as 4-6 weeks after SE. In male NMRI mice with chloral hydrate anesthesia during kainate injection, SE was followed by a seizure-free latent period of 10-14 days if hippocampal paroxysmal discharges (HPDs) recorded from the kainate focus were considered the onset of epilepsy. Anesthesia with isoflurane led to a more rapid onset and higher severity of SE, and not all male NMRI mice exhibited a seizure-free latent period. Female NMRI mice differed from male animals in the lack of any clear latent period, independently of anesthesia type. Furthermore, HPDs were only rarely observed. These problems were not resolved by decreasing the dose of kainate or using other strains (C57BL/6, FVB/N) of female mice. The present data are the first to demonstrate marked sex-related differences in the latent period following brain injury in a rodent model of acquired epilepsy. Furthermore, our data demonstrate that the choice of anesthestic agent during kainate administration affects SE severity and as a consequence, the latent period, which may explain some of the differences reported for this model in the literature.

Three dimensional in vitro culture of preantral follicles following slow-freezing and vitrification of mouse ovarian tissue

To evaluate the effects slow-freezing and vitrification on three dimensional in vitro culture of preantral follicles, ovaries of 12–14 days old female NMRI mice were isolated and randomly assigned to fresh control, slow-freezing and vitrification groups. Slow-freezing was performed using programmable freezer. Vitrification was carried out in a medium consisting of ethylene glycol (EG) and dimethyl sulphoxide (Me2SO) by needle immersion method. middle sized preantral follicles were mechanically isolated and cultured for 12 days in 0.7% sodium alginate gel. The follicles development and quantitative expression of oocyte specific genes (Bmp15, Gdf9, Fgf8) and the growth related genes (Igf1, Kit, Kit-l) were assessed after 1, 8 and 12 days of culture. Both cryopreserved groups showed reduction of follicular survival rates compared to the control group on days 8 and 12 of culture (P < 0.05). Antrum formation rates reduced in slow-freezing after 12 days of culture (P < 0.05). Evaluation of gene expression showed reduction of Bmp15, Gdf9, Fgf8, Kit and Kit-l during 12 days of culture (P < 0.05). Kit and Kit-l expression in slow-freezing group significantly reduced on day 8 of culture (p < 0.05). Igf1 expression was lower in slow-freezing group on 1st day of culture than vitrification and control groups (P < 0.05). Finally, intergroup comparison showed same expression pattern of genes after 12 days of culture. Thus, cryopreservation of mouse ovaries by both methods can preserve most developmental parameters and expression of maturation genes. However, vitrification is a better method for cryopreservation of mouse ovaries due to greater antrum formation and expression of growth related markers.

Mouse preantral follicle development in two-dimensional and three-dimensional culture systems after ovarian tissue vitrification

ObjectivesThis work on follicle culture and evaluation of expression of oocyte maturation genes helps to better understand the complicated processes of folliculogenesis and to develop new approaches for infertility treatment. Study designOvaries of 12-day-old female NMRI mice were divided into control and vitrification groups. After vitrification and warming procedures, ovarian tissue morphologies were histologically evaluated and compared to those of the control group. In the second stage, preantral follicles were mechanically isolated from non-vitrified and vitrified ovaries and cultured for 12 days in two-dimensional (2D) and three-dimensional (3D) systems. Finally, the survival and growth rate of follicles and quantitative expression of oocyte maturation genes (Gdf9, Bmp15 and Bmp6) were studied. ResultMorphological integrity of ovarian tissue in vitrification group was well preserved. Survival rates of cultured preantral follicles in control group during 2D and 3D systems were somewhat similar, but were significantly different between 2D and 3D systems in vitrification group. Although the growth rate of follicles was similar in the 3D system in both groups, substantially higher growth rate was observed for the control group in the 2D system. Expressions of oocyte maturation genes were, to some extent, similar between control and vitrification groups. There was a remarkable reduction in expression pattern of genes in 3D compared to 2D system in both experimental groups, during the 12th day of culture period. Conclusions3D in-vitro culture system could be appeared more appropriate than 2D culture system for preservation of follicles in terms of spatial morphology, growth rate and expression reduction of maturation genes.

ZINC OXIDE NANOPARTICLES ABSORPTION RATE IN THE HEART TISSUE OF FEMALE MICE

Nanotechnology researchers have identified a wide range of nanoparticle applications that may have an important role in medicine and treatment of diseases. Due to lack of detailed documentation about the toxicology of zinc oxide (ZnO) nanoparticles, this study was aimed to evaluate the absorption of ZnO nanoparticles in hearts of female NMRI mice. Overall, 20 adult NMRI female mice were studied in experimental and control groups. ZnO nanoparticles with concentration of 100 and 300 mg/kg were administered in the drinking water for 28 days and the mice were dissected after 28 days. Then, the heart tissues were isolated and dissolved in acid and the amount of ZnO deposited into the heart tissue was measured by atomic absorption spectrophotometer. ZnO nanoparticles treatment groups were significantly influenced by the nanoparticles compared with the control group. The experimental group 1 and 2 had a significant increase in ZnO NPs absorption in heart tissue compared to the control group ( P <0.01). Due to the physiological similarities between mice and humans, the results of this study can be applied in prevention of the cardiac damage during the consumption of ZnO NPs.

Comparing the growth and the development of mouse pre-antral follicle in medium with PL (Platelet Layset) and with FBS

Introduction: In recent years a great deal of effort has been made to improve the culture medium which can be used for different cell culturing and Folliculogenesis. PL (Platelet Layset) with a high percentage of growth factors and also microelements can be effective on the growth of follicle and oocyte. Material and method: Preantral follicles were collected from 12 to 14 days female NMRI mice and cultured in culture medium with a different supplement as the serum for 12 days. The different serums were, 5% and 10% FBS as the control groups and the experimental groups were enriched by 5% PL, 10% PL and a combination of 5% PL and 5% FBS. The growth and development of follicle and oocyte was monitored and also the amount of Estradiol (E2) and progesterone (P4) was detected in different mediums on days 9 and 12 of culturing. Result: Oocytes had a significant growth in all medium compared to day zero and reached to the size of a mature oocyte (p < 0.05). Although survival rate after one day had significant decrease in all medium compared to 5% FBS (p < 0.05), after 12 days of culturing, the best survival rate belonged to 5% PL. Progesterone secretion increased in experimental groups significantly (p < 0.05) while Estradiol secretion in 5% and 10% PL decreased in half of control groups. Discussion: This result demonstrates that PL can be a considerable supplement that can be added to the culture medium or a good replacement for FBS as serum used in Folliculogenesis.

Induced adverse effects of prenatal exposure to silver nanoparticles on neurobehavioral development of offspring of mice.

Steadily increased use of silver nanoparticles (Ag-NPs), has increased the amount of its exposure to humans and animals. Current scarce knowledge about the influences of prenatal exposure to Ag-NPs on postnatal outcomes, motivated us to investigate whether being exposed to it during pregnancy has any effects on neurobehavioral development of the adult offspring. Thirty virgin female NMRI mice were mated and treated subcutaneously once every three days from gestation day 3 until delivery, by 0, 0.2 and 2 mg/kg of bodyweight (BW) of Ag-NPs. Behavioral functions of adult offspring including spatial memory, passive avoidance learning, stress, anxiety-like behaviors and locomotor activities were assessed by commonly used neurobehavioral paradigms and the results were compared according to treatment and sex. Prenatal exposure to Ag-NPs significantly impaired their cognitive behavior in the Morris water maze. Although no evidence was observed indicating more anxiety-like behaviors in the treated offspring in the elevated plus maze, the number of defecations and leanings in the open field assay and number of passages in the light-dark box were greater in groups prenatally treated by Ag-NPs. Most of the impairments were more apparent in the offspring which had been prenatally exposed to high doses of Ag-NPs, particularly female ones. The present study indicated that the exposure of pregnant animals to Ag-NPs may lead to various neurobehavioral disorders in their offspring. Thus, more attention should be paid to avoid exposure to Ag-NPs, especially from pregnant females.

(p-MeC6H4Pri)2Ru2(SC6H4-p-But)3]Cl (diruthenium-1), a dinuclear arene ruthenium compound with very high anticancer activity: An in vitro and in vivo study

Dinuclear p-cymene ruthenium complexes of the type [(p-MeC6H4Pri)2Ru2(SC6H4-p-R)3]+, accessible from the reaction of p-cymene ruthenium dichloride dimer with the corresponding thiophenol and isolated in high yield as the chloride salts, were found to be highly cytotoxic towards human ovarian cancer cells, the IC50 values being in the nanomolar range. Remarkably, the in vitro anticancer activity of these complexes is at least as good for the cisplatin-resistant mutant A2780cisR as for the normal A2780 cancer cell line. The most active compound of this series, [(p-MeC6H4Pri)2Ru2(SC6H4-p-But)3]Cl (diruthenium-1) with an IC50 value of only 30 nM for both cell lines, was selected for an in vivo anticancer study using a mouse model. The effects of diruthenium-1 on the growth of a solid Ehrlich tumour, inoculated subcutaneously in female NMRI mice, and on the survival rate of the tumour-bearing mice were studied. The maximum tolerated dose of diruthenium-1 for healthy animals was 2 mg/kg. Diruthenium-1 at doses of 0.4 and 0.6 mg/kg administered i.p. twice within 8 days inhibited the tumour growth, although somewhat less than cisplatin (5 mg/kg i.p.) in the positive control group. However, it was only diruthenium-1 at a dose of 0.6 mg/kg that prolonged significantly the survival rate of the tumour-bearing mice as compared to the untreated control group.A biodistribution study by ICP-MS showed ruthenium to be present in the tumour and particularly in the excretion organs but not in the brain; the elimination half-life of diruthenium-1 was estimated to be approximately 30 h. In human breast cancer cells MCF-7 and BT-549, diruthenium-1 exhibited antiproliferative and cytotoxic effects, stimulated the expression and phosphorylation of p53, and modulated the expression of MAP kinases. The levels of ATP decreased but the lactate production rose. Supported by the observed inhibition of mitochondrial respiration, we assume that diruthenium-1 acts predominantly as an inhibitor of the aerobic metabolism. Contrary to the previous findings concerning the catalytic oxidation of glutathione by this type of complexes in solution, we did not observe any glutathione oxidation by diruthenium-1 in the cancer cells: Diruthenium-1 decreased the intracellular concentrations of both reduced and oxidised gluthathione. Altogether, diruthenium-1 turned out to be a promising compound with a potential as an anticancer drug for a future ruthenotherapy.