Downregulation of Stemness Genes and Induction of Necrosis in Rat LA7 Cancer Stem Cells Induced Tumors Treated with Starved Fibroblasts Culture Supernatant.

BACKGROUND: Stem cell differentiation therapy is a promising strategy in cancer treatment. we show that protein cocktail prepared from serum starved fibroblasts has therapeutic potential based on this strategy. METHODS: The condition medium was prepared from foreskin isolated fibroblasts and analyzed by Liquid chromatography electrospray ionization mass spectrometry-mass spectrometry (LC-ESI-MS/MS). LA7 mammary gland cancer stem cells originated tumors were induced in Sprague Dawley rats. The rats treated subcutaneously with DMEM (group A), condition medium (group B), or normal saline (group C) once daily for 7 days. Then the tumors were removed and divided into the two parts, one part was used to quantify gene expression by stem-loop RT-qPCR assay and the other part was used for Hematoxylin & Eosin (H & E), Giemsa, and immunohistochemistry (IHC) staining. RESULTS: All induced tumors appeared as sarcomatoid carcinoma (SC). Immunohistochemistry staining confirmed this conclusion by recognizing the tumor as Ki67+, cytokeratin+, vimentine+, and estrogen receptor negative SC. RT-qPCR analysis revealed that Oct4-, Sox-2, Nanog- gene expression was much reduced in the condition medium treated tumors versus proper controls (p< 0.05). Tissue necrosis was more prevalent in this group while tumors volume was diminished almost by 40%. The LC-ESI-MS/MS analysis unrevealed the stemness reducing and the cell death inducing proteins such as, pigment epithelium-derived factor (PEDF), insulin like growth factor binding protein-5 (IGFBP-5) and -7 (IGFBP-7) in the condition medium. CONCLUSION: This study showed that the substances released from starved human fibroblasts were able to down-regulate the stemness-related genes and induce necrosis in LA7 derived tumors.

SCF Improves In Vitro Differentiation of SSCs Through Transcriptionally Up-regulating PRTM1, STRA8, c-KIT, PIWIL2, and OCT4 Genes.

Several lines of evidence strongly suggest that retinoic acid (RA) and stem cell factor (SCF)/c-Kit signal transduction pathways are involved in the differentiation of spermatogonial stem cells (SSCs). This study was aimed to investigate the effect of RA and SCF on in vitro differentiation of SSCs via evaluation of the mRNA expression of meiosis-specific genes in cultured testicular tissues. Testicular tissue samples were obtained from bilaterally vasectomized rats and also healthy adult rats and then were cultured for 25, 30, and 35 days on different conditions. The cultured testicular pieces were sectioned and stained with PAS to histological analysis. The total RNA was extracted from cultured testicular samples, and the expression of ACR, PRTM1, SYCP3, STRA8, c-KIT, PIWIL2, and OCT4 genes at mRNA level was quantified using real-time polymerase chain reaction (qPCR) procedure. After 1-month surgery, bilateral testicular weight showed a significant decrease in vasectomized adult rats compared with healthy adult rats (P < 0.05). Reduction in the diameter of the seminiferous tubules and depletion of advanced germinal elements in vasectomized rats compared with healthy adult rats were also observed. Our findings also demonstrated that the mRNA expression level of PRTM1, STRA8, c-KIT, PIWIL2, and OCT4 genes in cultured testicular tissues significantly up-regulated in experimental group II compared with the control group (P < 0.001). Our findings lead us to conclude that SCF improves in vitro differentiation of SSCs in the OA rats, at least partially, by transcriptionally upregulating PRTM1, STRA8, c-KIT, PIWIL2, and OCT4 genes.

Targeting LA7 breast cancer stem cells of rat through repressing the genes of stemness-related transcription factors using three different biological fluids.

Down-regulation of stemness genes expression is important in differentiation therapy against cancer stem cells (CSCs). The aim of this study was to evaluate the Oct4 , Sox2, Nanog, and C-myc expression in rat breast cancer stem cells (LA7) which treated with human ovarian follicular fluid (FF), replicative senescent fibroblast culture supernatant (P14), and 16 h serum starved fibroblast supernatant (16 h-SFS). The cells were exposed to these biological fluids for 24 h, 72 h, and 7 days. Stem-loop RT-qPCR assay was used to quantify the expression of above mentioned genes. Results showed that FF had the least cytotoxic effect on the LA7 cells. Except for Nanog gene, exposure of LA7 cell line to 16 h-SFS and P14 decreased significantly expression of the three other genes after 24 h (P < 0.05). Nanog and Sox2 genes expression was also decreased in LA7 cells which have been already treated with FF for 24 h. Moreover, compared to the control solution, the expression of Oct4 increased significantly after 7 days exposure to FF (P < 0.05). Annexin V-PE /7-AAD-, acridine orange/ethidium bromide staining and doubling time assays revealed apoptosis and necrosis induction by these biological fluids in LA7 cells. Moreover, in an in vitro model of metastasis assay, i.e., scratch test, these fluids exhibited anti-LA7 migration activity which culminated in 16 h-SFS treated cells. Generally, this study showed that FF, 16 h-SFS, and P14 have positive effects on down-regulation of Nanog, Oct4, Sox2 and C-myc expression, and consequently can increase the differentiation of breast cancer stem cells. For the first time, this study provided some evidence indicating that some biological fluids have potential to differentiate the CSCs, show anti- survival, growth-, and cell migration activity.

Comparison of Germ Cell Gene Expressions in Spontaneous Monolayer versus Embryoid Body Differentiation of Mouse Embryonic Stem Cells toward Germ Cells.

BACKGROUND: Genetic and morphologic similarities between mouse embryonic stem cells (ESCs) and primordial germ cells (PGCs) make it difficult to distinguish differentiation of these two cell types in vitro. Using specific GC markers expressed in low level or even not expressed in ESCs- can help recognize differentiated cells in vitro. We attempted to differentiate the mouse ESCs into GC-like cells spontaneously in monolayer and EB culture method. MATERIALS AND METHODS: In this experimental study, we attempted to differentiate ESCs, Oct4-GFP OG2, into GC-like cells (GCLCs) spontaneously in two different ways, including: i. Spontaneous differentiation of ESCs in monolayer culture as (SP) and ii. Spontaneous differentiation of ESCs using embryoid body (EB) culture method as (EB+SP). During culture, expression level of four GC specific genes (Fkbp6, Mov10l1, Riken and Tex13) and Mvh, Scp3, Stra8, Oct4 were evaluated. RESULTS: In both groups, Mov10l1 was down-regulated (P=0.3), while Tex13 and Riken were up-regulated (P=0.3 and P=0.04, respectively). Fkbp6 and Stra8 were decreased in EB+SP and they were increased in SP group, while no significant difference was determined between them (P=0.1, P=0.07). Additionally, in SP group, gene expression of Mvh and Scp3 were up-regulated and they had significant differences compared to EB+SP group (P=0.00 and P=0.01, respectively). Oct4 was down-regulated in the both groups. Flow-cytometry analysis showed that mean number of Mvh-positive cells in the SP group was significantly greater compared to ESCs, EB+SP and EB7 groups (P=0.00, P=0.01, and P=0.3, respectively). CONCLUSION: These findings showed that ESCs were differentiated into GCLCs in both group. But spontaneous differentiation of ESCs into GCLCs in SP group (monolayer culture) compared to EB+SP (EB culture methods) has more ability to express GCs markers.

Protective Effect of N-Acetyl Cysteine on Chlorpyrifos-Induced Testicular Toxicity in Mice.

BACKGROUND: Chlorpyrifos (CPF), an organophosphate pesticide, is widely used in farms in order to preserve crops and fruits. Previous studies have shown that CPF exposure might cause chronic toxicity in male genital system. The present study investigated the protective effect of N-Acetyl Cysteine (NAC), a potent antioxidant against testicular toxicity of CPF in male mice. MATERIALS AND METHODS: In this experimental study, 42 adult male mice were divided into seven groups, CPF low (0.5 mg/kg.b.w) and high (5 mg/kg.b.w) doses groups, NAC group (35 mg/kg.b.w), NAC+CPF 0/5 mg/kg.b.w, NAC+CPF 5 mg/kg.b.w, dimethyl sulfoxide (DMSO, 0.75% solution mg/kg.b.w) and control group. All treatment were done intraperitoneally. Treatment was conducted for four consecutive weeks (five days each week). However NAC was injected to NAC+CPF groups five days before initiation of the treatment procedure. One week after the last injection, mice were sacrificed using anesthetic gas to evaluate alterations in testicular histology and sperm parameters. RESULTS: Seminiferous tubules area and diameter were significantly diminished in the group treated with 5 mg/kg CPF (P<0.05). CPF also statistically reduced sperm parameters (count and motility) and damaged sperm morphology) at both doses (P<0.05). However, NAC significantly improved spermatogonia, spermatocytes, spermatid cell counts as well as sperm parameters in mice treated with both CPF concentrations (P<0.05). CONCLUSION: According to our results, NAC may significantly ameliorate CPF-induced damages to spermatogonia, spermatocytes, spermatids cell counts and sperm parameters.

Effect of Alpinia officinarum Hance rhizome extract on spermatogram factors in men with idiopathic infertility: A prospective double-blinded randomised clinical trial.

Despite scientific advances, many of the treatments in male infertility remained indeterminate. In recent years, the attention to herbal remedies as an effective treatment for male infertility is considerable. We designed this study to determine the effects of Alpinia officinarum on the results of semen analysis in men with idiopathic infertility. In this clinical trial, seventy-six participants with idiopathic infertility were included in the intervention (plant treatment: n = 31; placebo: n = 29). Participants were randomised to take capsules containing dried extract of A. officinarum rhizome or placebo on a daily (total daily dosage of 300 mg) basis for 3 months. After 12 weeks of intervention, the sperm count and total number of spermatozoa with normal morphology were increased in participants treated with A. officinarum extract compared with the placebo group. The mean sperm count was initially 52 × 106  ± 24 × 106 /ml which changed to 71 × 106  ± 23 × 106 /ml, after intervention (p = 0.043). Also, the mean percentage of spermatozoa with normal morphology was 14.34% ± 9.16% before the treatment which significantly increased to 19% ± 14.89% (p < 0.001). Alpinia officinarum, a traditional medicine remedy, can be effective in the improvement of sperm morphology and sperm count in idiopathic infertility without causing adverse effects.

Evaluation of specific germ cell genes expression in mouse embryonic stem cell-derived germ cell like cells treated with bone morphogenetic protein 4 in vitro.

BACKGROUND: Bone morphogenetic protein 4 (BMP4) is a significant signaling molecule that involves in initiating of differentiation and performs multifunctional effects on embryonic stem cells (ESCs) and embryos. OBJECTIVE: The goal of the present study was to evaluate an in vitro differentiation model of mouse embryonic stem cells into germ cells, using BMP4. MATERIALS AND METHODS: in this experimental study, we used Oct4-GFP mouse ESCs to form embryoid body (EB) aggregations for two days. Then, single cells from EB were cultured for four days with BMP4. Using MTT assay and gene expression levels for evaluation of Mvh and Riken by real-time RT-PCR of six concentrations, 12.5 ng/ ml BMP4 was determined as an optimized dose. Then, the expression level of Fkbp6, Mov10l1, 4930432K21Rik, Tex13, Mvh, Scp3, Stra8, Oct4 were evaluated. Flow cytometry and immunostaning were used to confirm the findings of the real-time RT-PCR. RESULTS: In the +BMP4 group, the genes encoding Riken (p≤0.001) and Mvh (p≤0.001) were found to be increased with significant differences compared with the control group. Mov10l1 (p=0.22), Tex13 (p=0.10), Fkbp6 (p=0.90), Scp3 (p=0.61) and Stra8 (p=0.08) were up-regulated without significance differences compared with control group. Flow cytometry analysis showed that the mean number of Mvh-positive cells in the +BMP4 group was greater when compared with ESCs, -BMP4 and EB groups (p=0.03, p≤0.001, p=0.02, respectively). CONCLUSION: Down-regulation of Oct4, expression of germ cells genes and meiosis markers expression raise this hypothesis that ESCs were differentiated by BMP4, and may be introduced into the first meiosis as germ cell-like cells.

Evaluation of Novel Mouse-Specific Germ Cell Gene Expression in Embryonic Stem Cell-Derived Germ Cell-Like Cells In Vitro with Retinoic Acid Treatment.

We designed a study to induce differentiation of Oct4-GFP (expression of Green Fluorescent Protein of oct4) embryonic stem cells (ESCs) by embryoid body (EB) culture system into germ cells (GCs) using retinoic acid (RA) and evaluated the expression level of (Fkbp6, Mov10l1, 4930432K21Rik, and Tex13) in differentiated cells. The expression levels of four GC-related genes, Oct4, Mvh, Scp3, and Stra8, was determined by quantitative real-time polymerase chain reaction (q-RT-PCR). Immunostaining and flow cytometry were used as additional tests to confirm q-RT-PCR findings. A significant increase occurred in the expression of meiotic markers and specific genes, Fkbp6 (p = 0.00), Mov10l1 (p = 0.01), and Tex13 (p = 0.00) in ESCs treated with RA (+RA) compared with the controls (-RA). Oct4 expression was decreased in all studied groups. The expression levels of 4930432K21Rik, Mvh, Stra8, and Scp3 in the +RA group was higher than that of the -RA group. Flow cytometry analysis showed that mean number of Mvh-positive cells in the +RA group was greater as compared with ESCs, -RA and EB7 groups (p = 0.00). Downregulation of Oct4 as a pluripotency factor as well as the expression of meiosis markers, this hypothesis is raised that ESCs are differentiated by RA, and have been introduced into the zygote/pachytene of first meiosis as GC-like cells.

Auto-fluorescence of a silk fibroin-based scaffold and its interference with fluorophores in labeled cells.

Silk fibroin is increasingly emerging as an important biomaterial for tissue engineering applications. The ability to fluorescently image silk matrices under a microscope would be helpful in differentiating embedded labeled cells from background signal, critical for the study of silk-based engineered tissues. In this study, we fabricated a scaffold using freeze drying and confirmed its structure by X-ray diffraction and Fourier transform infrared spectroscopy. We then examined the fluorescence of the silk fibroin scaffold using confocal microscopy, both before and after cell seeding and fluorescent labeling. We subsequently investigated the fluorescent signature of the silk fibroin scaffold chemically. Fluorophore-labeled cells seeded into the scaffold showed the same fluorescent color as the scaffold itself when excited by the same wavelength of light. UV-Vis and fluorescence spectroscopy of a silk fibroin solution indicated absorption and emission maxima at 277 and 345 nm, respectively, which is a typical protein-derived signal. HPLC and GC-MS were used to detect quercetin and quercetin derivatives, without success. We therefore conclude that unlike silk cocoons, the fluorescent behavior of silk fibroin scaffolds does not derive from quercetin and its derivatives but from the intrinsic fluorescence of fibroin protein. We also find that the fluorescent signals deriving from a scaffold and from labeled cells embedded in it can be distinguished when the different optical channels are merged.

Impact of pre-incubation time of silk fibroin scaffolds in culture medium on cell proliferation and attachment.

Cell behaviours such as proliferation and attachment can be affected by the length of pre-incubation period of the scaffolds in the culture medium for long term. The aim of this study was to investigate the long term pre-incubation of 3D silk fibroin scaffolds in complete culture medium on cell attachment and proliferation. After the preparation of silk fibroin scaffolds by the technique of freeze drying, the scaffolds were pre-incubated in complete culture medium for 2 d, 6 d or 10 d before apical papilla stem cells (SCAP) seeding. Modifications of the scaffold surface and wettability were examined by FE-SEM and water contact angle, respectively. Results showed a decrease both in roughness and water contact angle as pre-incubation time increases. DNA measurement after 18h and 10 d cell seeding showed a significant increase of DNA concentration which represents better attachment and proliferation with pre-incubation time increase. Qualitative examination, live&dead assay or H&E staining method after 30h and 10 d cell seeding respectively, indicated that pre-incubation of scaffolds has time dependent effect on cell proliferation and attachment. This suggests that improvement of cell attachment and proliferation may be mediated by differences in the amount of wettability (decreased water contact angle) after exposure of scaffold to culture medium for long term which, in turn, causes more protein adsorption in the surface of silk fibroin scaffold (decreased roughness).

Effect of Fetal Mouse Lung Tissue Co-Culture on In Vitro Maturation of Mouse Immature Oocytes.

OBJECTIVES: The aim of this study was to evaluate the fetal mouse lung tissue co-culture on in vitro maturation (IVM) of mouse immature oocytes. MATERIALS AND METHODS: In this experimental study, germinal vesicle (GV) oocytes from ovaries of a group of 25 female mice, 6-8 weeks of age, were dissected after being stimulated by 7.5 IU pregnant mare serum gonadotropin (PMSG) through an intraperitoneal (IP) injection. The fetal lung tissues were then prepared and cultured individually. A total number of 300 oocytes were cultured in the following three groups for 24 hours: control group (n=100) containing only base medium, group I (n=100) containing base medium co-cultured with 11.5- to 12.5-day old fetal mouse lung tissues, and group II (n=100) containing base medium co-cultured with 12.5- to 13.5-day old fetal mouse lung tissues. The proportion of GV and metaphase І (MI) oocytes matured into MІІ oocytes were compared among the three groups using analysis of variance (ANOVA). Correlation test were also used to evaluate the successful rate of IVM oocytes. RESULTS: The proportions of GV oocytes reaching MІІ stage were 46, 65, and 56%, in control, I and II groups, respectively (P<0.05). The percentage of the oocytes remaining at the GV stage were higher in control group as compared with two treatment groups (P<0.05). CONCLUSIONS: This study indicated that fetal mouse lung tissue co-culture method increased the percentage of GV oocytes reaching MII stage.

Cytochrome P-450 1B1 Leu432Val Polymorphism Does Not Show Association With Breast Cancer in Northern Iranian Women With a History of Infertility.

The Cytochrome P-4501B1 (CYP1B1) Leu432Val polymorphism has been previously shown to be associated with some types of cancer and affects CYP1B1-mediated metabolism of various infertility drugs. To establish the frequency of CYP1B1 Leu432Val polymorphism among women with a history of infertility drug use, we studied the genotypes of 147 patients with breast cancer with a history of infertility and 150 cancer-free, infertile women (control group) in Northern Iran. A polymerase chain reaction-based restriction fragment length polymorphism assay was used to detect GG (Val/Val), CG (Leu/Val), and CC (Leu/Leu) genotype frequencies, which did not vary significantly between the 2 patient groups (P = .847). We established for the first time that the incidence of CYP1B1 Leu432Val polymorphism is 46.6% among women with infertility history and breast cancer in Northern Iran. Finally, our results do not show any significant association between CYP1B1 Leu432Val polymorphism and breast cancer in infertile women in this region, who have also received infertility treatment.

Does different BMI influence oocyte and embryo quality by inducing fatty acid in follicular fluid?

OBJECTIVE: We aimed to assess the effects of obesity-related follicular fluid (FF) fatty acids (FAs) on the number and quality of oocytes, good embryo quality rate, and pregnancy rate. MATERIALS AND METHODS: This prospective cohort study was conducted on 105 infertile women under the age of 38, who underwent intracytoplasmic sperm injection (ICSI) from March 2015 to October 2015. They were grouped into three body mass index (BMI) categories. The fatty acids composition of the FF was analyzed by GC-MS head space method. We studied the FAs correlation with BMI and ICSI outcomes. RESULTS: The distribution of fatty acids did not differ significantly in each BMI group, with the exception for stearic that was marginally significant (p=0.05). The mean number of mature oocytes did not differ significantly between the BMI groups, the percent of Metaphase II (MII) oocytes was inversely associated with the BMI (rs=-0.21, p=0.03). Kruskal-Wallis test showed that the distribution of good quality embryos' percentages were different in at least two categories of studied BMI groups (p=0.009, p=0.02). The mean concentration of palmitic acid was higher in nonpregnant patients for all of the studied BMI classes (p=0.02, p=0.03, p=0.05), however, stearic (p<0.001) and linolenic acids (p=0.01) were higher in nonpregnant normal weight patients. CONCLUSION: Differences in BMI are not associated with the fatty acid composition of the FF. The FF fatty acid possibly affects the outcome of ICSI through the achievement of clinical pregnancy. Therefore, it is essential to provide patients with nutritional counseling before they use assisted reproductive techniques.

Comparison of pregnancy rate between fresh embryo transfers and frozen-thawed embryo transfers following ICSI treatment.

BACKGROUND: The use of assisted reproductive technology (ART) is increasing in the world. The rate, efficacy and safety of ART are very different among countries. There is an increase in the use of intra cytoplasmic sperm injection (ICSI), single fresh embryo transfer (ET) and frozen-thawed embryo transfer (FET). OBJECTIVE: The objective of this study was to compare pregnancy rate in fresh ET and FET. MATERIALS AND METHODS: In this retrospective cross-sectional study 1014 ICSI-ET cycles (426 fresh ET and 588 FET) from 753 women undergoing ICSI treatment referred to Fatemezahra Infertility and Reproductive Health Research Center in Babol, Iran from 2008 to 2013 were reviewed. RESULTS: There were no significant differences between biochemical pregnancy rate (23% versus 18.8%, OR 1.301; 95% CI .95-1.774), gestational sac (95.6% versus 100% in FET, OR 0.60; 95% CI 0.54-0.67), and fetal heart activity (87.2% versus 93.6% OR .46; 95% CI .16-1.32) in fresh ET and FET cycles, respectively. P< 0.05 was considered statistically significant for all measures. CONCLUSION: Although, the result showed no significantly difference between the fresh ET and the FET cycles, however the embryos are able to be stored for subsequent ART. Therefore, we recommend FET cycles as an option alongside the fresh ET.

Histopathological, histomorphometrical, and radiographical evaluation of injectable glass-ceramic-chitosan nanocomposite in bone reconstruction of rat.

Background. Bone defects following tumor resection and osteolysis due to bone lesions, periodontal tissue disorders, and bone reconstruction are challenges that surgeons face. Gass-ceramic-chitosan nanocomposite contains chitosan, a derivative of crustaceans' exoskeleton. Methods. Thirty-two 6-8-week-old male Wistar rats were chosen. One hole on each right and left tibia was made. The right tibia holes were filled with injectable glass-ceramic-chitosan nanocomposite, and the left tibia holes were left empty. After 7, 14, 28, and 60 days, histopathological, histomorphometrical, and radiographical assessments were performed. Results. Radiographic density on days 7 and 14 was significantly higher in the right tibias than in the left tibias. Trabecular bone thickness, which was higher in the right tibias, increased from day 7 to day 60 in both right and left tibias, although not significantly. Conclusions. Glass-ceramic-chitosan nanocomposite is suggested for use in bone repair in cases of bone loss. More histopathological, histomorphometrical, and radiographical assessments are also recommended.

Total antioxidant capacity and lipid peroxidation in semen of patient with hyperviscosity.

Semen hyperviscosity (SHV) is one of the factors involved in deficiency in sperm function. This research aimed to evaluate seminal plasma total antioxidant capacity (TAC) and malondialdehyde (MDA) levels in infertile patients with hyperviscous and non-hyperviscous semen samples to understand whether hyperviscous semen is associated with oxidative damage in infertile subjects. In this cross sectional study, 59 semen samples were provided by fertile (n=12) individuals as control, infertile patients with normal viscosity (n=25) and infertile patients with hyperviscosity (n=22). After semen parameters examination, semen viscosity was studied by glass pipettes. Seminal plasma TAC and MDA levels were measured by ferric reducing of antioxidant power (FRAP) and thiobarbituric acid reaction (TBAR) methods, respectively. A probability less than 0.05 was considered statistically significant throughout the article. The mean of sperm parameters including: counts, motility and normal morphology in patients with hyperviscosity were significantly lower than those in non-hyperviscosity patients (p<0.05, p<0.01 and p<0.001, respectively). The mean of seminal plasma TAC value in seminal plasma of non-hyperviscosity patients (1710.31 ± 458.67 µmol/l) was significantly (p<0.01) higher than that of hyperviscosity group (1230.25 ± 352 µmol/l). A trend toward a higher mean of seminal plasma MDA value was estimated for hyperviscous group compared with non-hyperviscous (1.01 ± 0.41 nmol/ml vs. 0.94 ± 0.28 nmol/l); however, it was nonsignificant. Hyperviscous semen impairs seminal plasma TAC which is eventually associated with sperm membrane lipid peroxidation.

Immunotoxicity effects of carbaryl in vivo and in vitro.

Carbaryl is a pesticide for controlling pests in agricultural industry. To determine of immunotoxicity effects of carbaryl, rats were exposure with carbaryl for 28 days. The lymphoid organ weight, lymphocyte proliferation, IL-2, IFN-γ, IL-4, IL-10, IL-1ß and TNF-α cytokines level were measured, respectively. Exposure with carbaryl significantly reduced both thymus and spleen weight and also suppressed lymphocyte proliferation. In addition, carbaryl significantly decreased IL-2, IFN-γ, IL-1ß and TNF-α and also increased IL-4, IL-10 cytokines. These findings suggest that exposure to carbaryl can induce immunotoxicity effects on lymphoid organ weight, suppresses the functions of lymphocyte and macrophage, Th2 polarization in Th1/Th2 balance by reducing of IFN-γ and increasing of IL-4 and IL-10 cytokines. Therefore, carbaryl can contribute to the development of allergic, autoimmune, cancer or infection diseases through immunotoxicity effects and unbalancing of Th1/Th2 immune response however, further study is necessary.

The effect of camphor on sex hormones levels in rats.

In some traditional therapies, it has been claimed that camphor (a crystalline ketone obtained from cinnamomum camphora) would be a suppressor of sexual behaviors and sex hormones. This study evaluated the effects of camphor on sex hormones, like luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone. In this experimental study, 56 male rats were divided into 5 groups, including control (n=12), sham (n=11) and three treatment groups (n=11) in three different doses. The sham groups received daily intra peritoneal (IP) injections of the vehicle (ethanol 10%) for 30 days. Three treatment groups received different daily IP injections of the camphor (1, 2 and 5 mg/Kg) for 30 days and the control groups didn't received anything. Serums were used for assaying LH, FSH and testosterone. The level of LH significantly increased in all doses of camphor among the treatment groups as compared to the control (p<0.05), but camphor in doses 2 and 5 mg/Kg significantly reduced the FSH level as compared to control group (p<0.05). No significant changes were seen in testosterone levels. Camphor increased level of LH, decreased level of FSH, whereas it failed to change level of testosterone. The claim of inhibitory effect of camphor on sexual activity could not be confirmed by this study. More investigations in this field are suggested.

The effect of 8-methoxypsoralen on pituitary-gonad axis and ovarian function in mice.

OBJECTIVE: 8-Methoxypsoralen (8-MOP) is a photoactive compound widely used in the treatment of proliferate disorders. The present study investigates the effects of 8-MOP on ovary function and pituitary-gonad axis in mice. MATERIALS AND METHODS: : In this experimental analytical study, 45 female Balb/C mice were divided into three groups (n=15), control, sham (olive oil injection) and experimental. The experimental group were received an intraperitoneal (i.p.) injection of the LD50 dose of 60 mg/kg 8-MOP. At 30 days after injection, the animals were sacrificed while in the proestrus stage and examined for morphological and histological changes their ovaries. Blood samples were collected and estrogen, luteinizing hormone (LH) and follicle stimulating hormone (FSH) levels were assessed by radioimmunoassay. Data were analyzed using one-way ANOVA and the t test. RESULTS: The mean levels of estrogen and progesterone in the experimental group significantly decreased (p< 0.001). However, there was a significant increase in LH and FSH levels in this group compared to the control groups (p< 0.001). The mean number and diameter of the corpus luteum (CL) and the number of growing follicles in the experimental group significantly reduced compared to the control and sham groups (p< 0.001). The mean granulosa thickness in the experimental group also significantly decreased compared to the control and sham groups (p< 0.001). CONCLUSION: Our data indicated that 8-MOP can affect the levels of LH, FSH, estrogen and progesterone. Our findings further suggest that consecutive doses of 8-MOP may impair the female reproductive tract (or development).