Anatomical Sciences Journal
Volume:13 Issue: 3, Summer 2016

In the present study, we assessed the effects of β-carotene on Titanium oxide Nanoparticle (TNP) induced mouse Spermatozoon Stem Cells (SSCs) apoptosis, at molecular level.
After isolation from cryptorchid mouse testis and characterization, spermatogonial stem cells were divided into four groups. In the control group, spermatogonial cells were cultured in α-MEM supplemented with 2% BSA (Bovine Serum Albumin). β-Carotene (BC) group was composed of control culture condition supplemented with 1 µg/ml β-carotene. TNP group comprised control culture condition supplemented with 1 µg/ml titanium oxide (TiO2). Ultimately the last group contained control culture condition supplemented with both 1 µg/ml BC and 1 µg/ml TiO2for three days. After that, spermatozoon viability was evaluated by MTT (3-[4,5-dimethylthiazolyl-2]-2,5-diphenyltetrazolium bromide) assay, apoptotic and necrotic indices with Annexin V/PI kit and gene expression of CASP3 and MAPK14 using qRT-PCR method.
TiO2 could significantly decrease viability of the cultured spermatozoon in TNP group compared to the control group. In BC group, we determined increased frequency of live spermatozoon compared to TNP or control group. Expression of apoptotic related genes significantly increased in TNP group. Spermatozoon induced by titanium oxide might be useful in clinical procedures. Measurement of apoptosis index using Annexin V/PI method also showed significant increase in apoptotic index of germ cells in TiO2 treated spermatozoon (P Expression of apoptotic related genes in cultured spermatozoon could efficiently be decreased by β-carotene treatment. Application of BC had a potential protective effect in preventing apoptosis in germ cells and might be useful in clinic.

In adult mammalian brain, neural stem cells are isolated from both the dentate gyrus and subventricular zone. This study aimed to isolate neural stem cells from adult rat subventricular zone and differentiate them into neurons and astrocytes.
In this study, the whole brain was removed after full anesthesia and creating cervical dislocation. Under a microscope, subventricular zone was dissected by a coronal incision in optic chiasm zone. Enzymatic digestion was performed using trypsin-EDTA. The isolated cells were cultured in serum free DMEM/F12 medium, containing bFGF (basic Fibroblast Growth Factor) and EGF (Epidermal Growth Factor) growth factors.
Neurospheres were observed five days after culturing. Immunocytochemistry was used to investigate nestin gene expression and identify neural stem cells. Neural stem cells were differentiated in poly-L-lysine coated plates in the absence of growth factors. The expression of GFAP, β tubulin III, and nestin genes were analyzed by RT-PCR. The results of immunocytochemistry confirmed nestin gene expression in the neural stem cells. Phenotype of neurons and astrocytes were observed 5 days after cell culture in differentiation medium. RT-PCR analysis revealed the expression of GFAP and β tubulin III genes.
The results of this research show that only one rat brain is needed for neural stem cells isolation and differentiation to neurons and astrocytes.

Despite technological advances and numerous published investigations, sexual dimorphism of Corpus Callosum (CC) remains a matter of ongoing controversy. In the present study on neurologically healthy Iranian adults, we investigated the possible gender- and age-related variations in anthropometric callosal measurements.
Our sample comprised 35 male and 35 female subjects with the mean (SD) age of 42.8 (14.7) and 44.7 (15) years, respectively, who referred to Partow Magnetic Resonance Imaging (MRI) center in North of Iran for headache work-up. Individuals with known neurologic disorders, history of head trauma, left handed subjects, and those younger than 20 and older than 80 years old were excluded. We measured callosal and brain dimensions on the midsagittal section and analyzed the data using Independent sample t test, analysis of variance, analysis of covariance, Pearson correlation coefficient, and linear regression.
The unadjusted dimensions were larger in male participants compared to female ones. Corpus callosum area on the midsagittal plane, the longitudinal brain and callosal measurements and dimensions related to the width of CC were significantly larger in males than females (P We found apparently larger callosal dimensions in the male participants, which could be an artifact caused by the significantly larger male brain dimensions. Our investigations on the less studied racial groups also provide further evidence regarding the confounding effect of brain volume on the observed sexual dimorphism of CC.

Contrary to a common belief, most mammalian females lose the ability of Germ Cell (GC) renewal and oogenesis during fetal life. Although, it has been claimed that germ line stem cells preserve oogenesis in postnatal mouse ovaries, that postnatal oogenesis keeps producing functional and sufficient GCs in the case of infertility (caused by different reasons) is doubtful. On the other hand, there are many studies showing derivation of primordial GCs and late GCs from Embryonic Stem Cells (ESCs) in vitro. This study aimed to clarify the role of ESC-derived GCs in oogenesis.
Mouse ESCs via Embryoid Body (EB) formation were differentiated into GC lineage by adding Bone Morphogenetic Protein 4 (BMP4) and Retinoic Acid (RA) to the culture medium. Expression of GC markers was characterized by using Reverse Transcription Polymerase Chain Reaction (RT-PCR) and immunohistochemistry. Several 6- to 10-week-old female mice, sterilized using chemical agents, were injected with ESCs-derived GCs thorough their tail veins. To track the transplanted cells, their ovaries were immunohistochemically stained after two months.
Expression of GC specific markers such as mouse vasa homologue (Mvh) and Deleted in Azoospermia-Like (DAZL) indicated that GCs were successfully developed from ESCs. Interestingly, there was no evidence of homing of GCs in the transplanted ovaries after transplantation of ESCs-derived GCs.
Our findings do not suggest any contribution of ESC-derived GCs within the sterilized mice ovaries.

We aimed to study the effects of different CO2 pressures on expression of P33 gene and apoptosis in liver and spleen cells during CO2 pneumoperitoneum.
This study was performed on 30 male Sprague-Dawley rats, weighing between 280 and 340 g (procured from Tehran Pasteur Institute’s animal house). They were randomly divided into 3 equal groups. Groups 1 and 2 received 10 and 20 mm Hg CO2 pressures during pneumoperitoneum, respectively, and group 3 was the control group. CO2 gas was insufflated through a cannula into abdominal cavity of rats in groups 1 and 2 for one hour; then perfusion was performed for half an hour. In group 3, cannula was put into the rats’ abdominal cavities without releasing any gas. Then the rats were killed, and their livers and spleens were removed after laparotomy to study expression of gene P33 and apoptosis using RT-PCR and TUNEL techniques.
The TUNEL technique revealed a significant rise in apoptosis in liver cells of rats that received 20 mm Hg pressure of gas compared to rats that received 10 mm Hg pressure of gas and the control group (P Pressure level and duration of CO2 gas administration affect viability of liver and spleen cells. Too high a pressure or too long a duration may release cytokines and free radicals from cells of these organs, which can lead to transient or serious dysfunction.

Mesenchymal stem cells (MSCs) are suitable candidates for the treatment of liver diseases. However, their low survival rate limits their efficacy following transplantation. This study aimed to evaluate the therapeutic potentials of H2O2-preconditioned umbilical cord-derived MSCs (UCMSCs) on acute liver failure (ALF) in mice.
UCMSCs were pre-conditioned with different concentrations of H2O2. Cell viability was evaluated by WST-1 (water soluble tetrazolium) assay followed by exposure to lethal doses of H2O2. ALF was induced in NMRI mice using CCl4 and the cells therapy was performed using H2O2-preconditioned and normal UCMSCs. After 24, 48, and 72 hours, regenerative potentials of different UCMSCs groups were evaluated compared to sham group (that receive no MSCs) using biochemical and histological methods.
Lower liver enzymes was significantly evident in mice transplanted with H2O2-preconditioned UCMSCs compared with the other groups. Interestingly, histological results revealed a significant improvement in liver regeneration in these mice.
Preconditioning of UCMSCs with H2O2 not only enhances their survival but also increases the efficacy of MSCs-based cell therapy in acute liver failure.

Cisplatin is a platinum based antineoplastic drug, which is widely used for treatment of solid tumors. The present investigation was carried out to study the nephrotoxic effects of double dose injection of cisplatin in rats, as an experimental model.
In this experimental study, 45 adult male Sprague Dawley rats with average weight of 200±30 g were randomly divided into two experimental (n=30) and one control (n=15) groups. Rats of experimental groups received two repeated doses of cisplatin intraperitoneally (2.5 mg/kg, experimental group E1 & 5 mg/kg, experimental group E2) in the beginning of first and fifth week of the experiment. Eight weeks after injection, rats of all groups were given deep anesthesia and killed. Blood samples were collected directly from their hearts for biochemical evaluation. Tissue samples were removed and prepared sections were stained with H&E, PAS, Masson trichrome, and PNA methods. Prepared microscopic slides were utilized for both histopathological and morphometrical studies. Collected data were analyzed by ANOVA and Tukey post hoc test using SPSS.
Cisplatin administration induced a significant decrease in urinary space diameter of renal corpuscles in the experimental groups compared to the control group. This ultimately led to the urinary space obstruction in up to 95% of nephrons in experimental groups (P Cisplatin induces acute tubular necrosis and urinary space obstruction and some other morphological changes in rat kidney, in a dose dependent manner.

Understanding anatomical variations can help physicians achieve better results in clinical practice. During routine dissection, we found variations in the branches of Internal Iliac Artery (IIA) of a 65-year-old Iranian female cadaver. The IIA ended by dividing to an unusual smaller anterior and a greater posterior trunk which was branched to the inferior gluteal arteries. Both pelvis and the uterine artery were separated from the umbilical artery in the left side. In addition, the left iliolumbar artery arose from the main trunk of IIA. Although this cadaver had some uncommon variations, the findings are significant from clinical perspective.