mohammad zandi

Fetal bovine serum (FBS) is rich in nutrients, growth factors, hormones, and other essential compounds necessary for cell growth. However, its use has several disadvantages. Platelets in particular contain the majority of the potent mitogenic factors found in serum.

The present study aimed to evaluate the effect of platelet lysate (PL) on in vitro embryo production by replacing FBS, to determine if this modified Charles Rosenkrans medium (mCR2aa) can support ovine embryo development compared to the commercially available BO-IVC medium.

In the first experiment, varying concentrations of FBS (0%, 2.5%, 5%, and 10%) and PL (0%, 2.5%, 5%, and 10%) were investigated in the mCR2aa culture medium. In the second experiment, the optimized mCR2aa medium was compared to the BO-IVC medium. Ovine oocytes were matured and fertilized in vitro using BO-IVM Bioscience medium and Brackett and Oliphant medium, respectively. Subsequently, the resulting zygotes were cultured in either mCR2aa medium supplemented with amino acids or BO-IVC medium, according to the experimental design, under a humidified atmosphere of 5% CO 2 , 5% O 2 , and 90% N 2 at 38.5ºC.

The results indicated that using 5% PL in the mCR2aa culture medium reduced the serum percentage to 2.5%.

It was not possible to completely eliminate serum from the mCR2aa medium solely by substituting it with PL. Further studies are needed to explore other serum properties such as chelating agents and antioxidant activities to fully optimize the mCR2aa medium without serum.

Spermatogonial stem cells (SSCs) transmit genetic information to the next generation. For the successful transplantation of these cells, the culture medium of these cells must be optimized.

This study investigated the natural antioxidant of grape seed extract (GSE) in comparison to vitamin C in the culture medium of SSCs.

A Soxhlet extractor was used to prepare ethanolic and acetonic extracts, and the Clevenger apparatus was used to prepare aqueous extracts of grape seeds. Then, their antioxidant capacities were determined using the DPPH method. The SSCs were extracted from lamb testicular tissue by a two-step digestion method, and different concentrations of GSE and vitamin C were investigated for survival, colony formation, and expression of apoptotic-related genes.

The results showed that the acetonic extract in the concentration of 400 g/mL showed about 90% antioxidant properties based on the DPPH test; nevertheless, the aqueous and ethanolic extracts had only 50% of their antioxidant properties at this concentration. The acetonic extracts significantly decreased the viability of SSCs without any colony formation when used in a culture medium. The highest survival rate was obtained from the ethanolic extracts of grape seeds at the concentration of 50g/mL, and a significant difference was obtained with ethanolic extracts (100 to 1000 g/mL) (P < 0.05). The viability of SSCs treated with vitamin C (50 g/mL) was significantly higher (P < 0.05) than the control and 5 - 25 g/mL of vitamin C groups.

Aqueous, ethanolic, and acetonic extracts of grape seeds should not be used in the culture of SSCs. However, the use of 25 - 50 g/mL of vitamin C is recommended in the culture of SSCs. 

 More understanding of the physiological needs of the embryo during its growth from the zygote to the blastocyst stage, as well as the composition of the uterine and fallopian tubal secretions, has led to the development of single-step or sequential culture media.

 The present study aimed to investigate the utilization of mCR2aa culture medium as a sequential culture medium for embryo development, supplemented with platelet lysate (PL) to decrease the concentration of fetal bovine serum (FBS) in the embryo culture medium.

 After maturation and fertilization of sheep oocytes obtained from a slaughterhouse, potential zygotes were cultured in mCR2aa medium without FBS for two days (C1, 1 - 3 days of culture). In the second two days of culture (C2, 3 - 5 days of culture), 2.5% or 5% of FBS and 5% or 10% of PL were used. In the remaining days of culture (C3, 5 - 9 days of culture), 2.5%, 5%, or 10% of FBS, and 0%, 2.5%, 5%, or 10% of PL were used. These percentages were compared to the commercial BO-IVC™ medium and mCR2aa medium, both of which contained 10% of FBS.

 No significant difference was observed in the number of produced blastocysts between different treatments and the control. However, the number of hatched blastocysts in BO-IVC™, as a single-step culture medium, significantly differed from that of the other treatments. The number of hatched blastocysts in the sequential culture medium was higher in media supplemented with 2.5% FBS and 10% PL in the C2 mCR2aa medium, and 2.5% FBS without PL in the C3 mCR2aa medium, but no significant difference was observed in other treatments. However, increasing the concentration of FBS and PL in the C3 mCR2aa medium significantly decreased the number of hatched blastocysts.

 An increase in the percentage of PL to 10% necessitated a decrease in FBS to 2.5% in the C2 mCR2aa medium, while PL in the C3 mCR2aa medium was not required in the presence of 2.5% FBS.

Women worldwide are concerned about breast cancer, resulting in high mortality and morbidity.

In this research, to investigate the function of Taxol, Vinblastine, and Vincristine on the MCF-7 cell line, different concentration levels (10, 20, 40, 80, 160, and 320 µmol/mL) of the drugs were examined. Taxolwas also used in combination with Vinblastine and Vincristine.

The examined compounds were applied to the cells for 48 h. The MTT test was employed for assessing cytotoxicity, and the nonlinear regression method was run to estimate the values of IC50 (half maximal inhibitory concentration) values.

It was found that taxol + vincristine, taxol, and vinblastine treatments with concentrations of 41.45, 64.46, and 67.12 µmol/mL resulted in the best 50% inhibitory effect. The lowest effective concentration that had significant effects on cancer cells was vinblastine (160 µmol/mL), taxol + vincristine (80 µmol/mL), taxol (80 µmol/mL), taxol + vinblastine (80 µmol/mL), and vincristine (40 µmol/mL), respectively (P < 0.05).

From the results of the current study, a synergistic reaction was found between taxol and vincristine based on calculated CI values.

Accurate estimation of genetic variance components is essential for the correct prediction of breeding values. The discovery of SNP markers and advances in molecular genetics and the discovery of new methods for sequencing the entire genome of living organisms and the development of bioinformatics methods and computer science, etc. have provided a great deal of molecular data and created a branch of genetics called genomics. This study aimed to estimate the genomic variance components of Border sheep using SNP data.

For this study, the SNP panel, which was genotyped with a 50k marker chip from Illumina, was used. Data were collected at Falkiner Farm in Australia. The studied traits were birth and weaning weights, diameter, and length of wool yarn. To study the relationship between allelic frequency and the amount of justified genetic variance justified, SNPs were classified into five different groups of rare allelic frequency (MAF), with approximately equal numbers in each group. The analyses were performed with the approach of limited genomic maximum likelihood and the method of analysis of complex genome traits with GCTA software.

Genomic heritability estimated by SNPs with a rare allelic frequency of more than one percent for birth weights, weaning, diameter, and length of wool were 0.58, 0.47, 0.59, and 0.2, respectively. The contribution of different groups of SNPs with rare allelic frequencies in justifying genetic variance for different traits was different and in general a significant part of genetic variance was justified by SNPs with

Although the number of SNPs was different in different groups, the amount of genetic variance justified by different MAF groups was different. According to the results, a very large, ideal sample size and better coverage of low-frequency variants are needed to obtain a stronger and more reliable inference.

Annually, various types of cancer cause thousands of deaths globally, and identifying an appropriate therapeutic option for these disorders is of crucial importance. Side effects of anticancer drugs can be reduced through the promising strategy of combination therapy.

The present paper has investigated the in vitro cytotoxicity of Taxol, carboplatin, vinblastine, and vincristine alone and in combination against human malignant melanoma A375 cells and non-cancerous fibroblast HU2 cells to examine the possible side effects of the drugs.

The cells were subjected to the examined compounds for 48 h, and the MTT test was conducted to evaluate the cytotoxicity.

The results indicated that the most significant effect was related to 120 µg/mL vincristine and 7.5 µg/mL Taxol+ vincristine treatments, with the survival amounts of 24 ± 0.6 and 28 ± 0%, respectively. In addition, the best 50% inhibitory effect was found to be related to Taxol + vincristine, vinblastine, and Taxol+ vinblastine treatments at the concentrations of 0.04, 2.2, and 3.4 µg/mL, respectively.

According to the findings of in vitro toxicity, the evaluated complexes are not cytotoxic against human fibroblast HU2 cells. Also, the most significant effect on A375 cells was associated with vincristine treatment. No synergistic reaction was recorded among the different combinations of drugs based on the calculated CI values.

Prolactin is secreted from the anterior pituitary gland and plays an important role in the mammary gland development, differentiation and lactation. The aim of current study was to evaluate prolactin receptor gene polymorphisms in the Shal breed and Shal×Romanov crossbreeds. For this purpose, after blood sampling of animals and extraction of DNA using phenol chloroform method, two fragments of 176 and 215 base pairs from intron 1 of prolactin receptor gene were amplified using PCR reaction. Then PCR products were analysed by acrylamide gel electrophoresis and silver staining. Results showed that AB and AC genotypes were identified in 176 base pairs fragment of prolactin receptor gene (PRLP1) in Shal breed. The frequencies of these genotypes were 0.92 and 0.08, respectively; and the frequencies of A, B and C alleles were 0.5, 0.46 and 0.04, respectively. Also, in the position of PRLR1 of this gene in Shal×Romanov crossbreeds, the frequencies of AB and AC genotypes were 0.94 and 0.06, respectively; and the frequencies of A, B and C alleles were 0.50, 0.46 and 0.04, respectively. A polymorphism study in the 215 bp fragment of the prolactin receptor gene (PRLR2) in the Shal and Shal×Romanov crossbreeds showed only AA genotype. Results of this study showed that PRLP1 position in Shal breed and Shal×Romanov crossbreeds was polymorphic, however the PRLP2 position in the studied samples was monomorphic.

Palatal rugoscopy is a reliable method in the forensic personal identification and racial group specification. the aim of the present study is to use palatal rugae pattern in sex and ethnicity identification applications.
Four hundred individual dental casts from four different ethnic populations of Iran were randomly selected. The pattern of the palatal rugae (shape, length, and number) investigated and its reliability to classify sex and minor ethnicity for each individual cast was evaluated(P The most common rugae shapes were straight, followed by wavy and curved types. The least frequent shapes were converging and circular types. Palatal rugae patterns were unique to each person. However, they could not differentiate males and females and had low abilities to classify the racial subsets.
The palatal rugae pattern was unique to each individual and palatal rugoscopy can be considered as a reliable forensic identification tool where utilizing other methods such as DNA profiling, fingerprint, and dental record comparison is impossible or difficult. In this study, palatal rugoscopy was not a reliable method to classify the sex of an individual and to differentiate between different racial subsets.

Nowadays, it is necessary to discover new and efficient antifungal or antimicrobial drugs because of increasing drug resistance organisms. Using medicinal plants for natural treatment of diseases caused by bacterial origin has mainly been considered.
In this study, the impacts of antimicrobial medicinal plants extract were compared based on four bacteria in vitro.
In this experimental study, disc diffusion assay and the minimum inhibitory concentration (MIC) method were used to investigate the antibacterial effects of selected plant extract elicited by two different solvent on S. aureus, E. coli, P. aeruginosa and S. enteric. Data were analyzed with a statistical software program (SPSS 16).
The hydro-alcoholic extract of Myrtus communis (myrtle) and water extract of Cinnamomun zeylanicum (cinnamon) were the most active extracts screened for antimicrobial activities against different four bacteria as tested organisms. The diameter of inhibition zones ranged from 23 to 28 mm. Comparison of the antibacterial effect of plant extracts and commercial drug revealed that the size of inhibition zone of penicillin against Staphylococcus aureus bacterium was larger than the plant extracts. However, myrtle extract at the minimum inhibitory concentration (MIC) of 30 mg/mL showed more powerful antibacterial activity compared to the other extracts and even penicillin. Petroselinum crispum (parsley), Nerium oleander (Oleander) and Glycyrihiza glabra (licorice) were found to have the least effect on the tested bacteria.
In the present study, plant extracts with different compounds showed antibacterial activity (especially myrtle and cinnamon). Hence, they can be used as new source for antibacterial substances.

The goal of this paper was to study the effect of ginger extracts and vitamin E on the performance of ovine Spermatogonial Stem Cells (SSCs). For this purpose, different concentrations of hydro and hydroethanolic extracts of ginger and vitamin E on viability, colony formation and expression of inhibiting (bcl2ll and bcl2)and inducing (bax) apoptosisgenes were studied. Spermatogonial Stem Cells (SSCs) were extracted from lamb testes with slaughterhouse origin using two steps enzymatic digestion method and enrichment by differential plating method. The characterization of SSCs was carried by alkaline phosphatase staining and expression of c-kit and oct-4 genes. Results have shown that the viability of SSCs was decreased significantly by using more than 800 µg/mL of hydroethanolic extract of ginger, in comparision with control group (P

Aim and Successful in vitro embryo production extremely depends on the components of culture media during in vitro maturation. With ingredients like Anitol, Fennel has properties similar to estrogen. The aim of this study was to investigate the effect of Fennel essence on nuclear maturation of bovine oocyte.
In order to in vitro maturation off oocytes, Cumulus-oocyte complexes (No. 772) from follicles 2–8 mm in diameter were aspirated and cultured in M199 maturation media containing 10% FBS, 10% bovine follicular fluid, 5 µg/ml FSH, 1 µg/ml oestradiol- 17β, 0.81mM sodium pyruvate and 50 µg/ml gentamicin sulfate. Maturation culture media supplemented with 0, 5, 10, 20, 30 and 50 mg/ml of Fennel essence and were cultured in a CO2 incubator containing 5% CO2 and 95% humidity in air at 38.5ºC for 24 h.
Results showed that the amount of maturation of oocytes under 5, 10, 20 and 30 mg/ml of Fennel essence were not significantly different based on expansion of cumulus cells and polar body extrusion in compared with control (P>0.05). However, 50 mg/ml of Fennel essence significantly decreased expansion of cumulus cells and polar body extrusion in compared to other groups (P From the results of current study we can conclude that, Fennel essence, based on cumulus expansion and polar body extrusion, unable to improve nuclear maturation of bovine oocytes. In order to investigate the effect of Fennel essence on nucleus and cytoplasmic maturation of bovine oocytes, it is suggested to use in vitro fertilization.

Introduction A simple and efficient method for producing multi-transgenic animals is required for medical and veterinary applications. The principal technique for the production of transgenic animals is pronuclear microinjection, which has a low efficiency for the generation of transgenic farm animals expressing a single transgene. Recently, nuclear transfer has been used to clone large animals, and could allow multiple genetic manipulations to be undertaken in vitro, prior to a single nuclear transfer, rather than complex and time consuming breeding programs. However, at present the frequency of success in cloning large animals is very low and is very expensive. The production of transgenic livestock by sperm mediated gene transfer (SMGT) has a number of advantages compared to other transgenic techniques such as nuclear transfer and microinjection. Both nuclear transfer and microinjection techniques are technically demanding and labor intensive. In contrast, SMGT requires no sophisticated equipment or technical expertise. Furthermore, bovine genetics are distributed through sperm in the dairy industry consequently making it easy to distribute genetically modified sperm. SMGT is based on the ability of sperm to bind to exogenous DNA molecules and transfer it to the oocyte during fertilization. The major benefits of the SMGT technique were found to be its high efficiency, low cost and ease of use compared to other methods. SMGT was first described in a small animal model, with high efficiency reported in the mouse. Recently the technique has been successfully adapted and optimized for use in large animals. Studies have shown that spermatozoa from numerous species, including bovine, can bind and take up foreign DNA and transfer it to the embryo. In bovine studies, the efficiency of SMGT can vary widely depending on both the transgene and the gene transfer method. Liposomes have been shown to be particularly effective in transferring DNA into bovine sperm. However, not all embryos derived from transfected sperm contain the transgene, suggesting that mechanisms exist, which impede SMGT. The aim of this study was to investigate the possibility of gene transfer to bull spermatozoa by using lipofection.
Materials and Methods Ram testes collected from Meysam abattoir slaughterhouse immediately after slaughter and were brought to the laboratory in an ice chest. In the laboratory, the testes were rinsed twice with normal saline and were then trimmed to remove the extra testicular tissue and washed properly with saline containing 0.1% streptomycin sulphate. Connective tissue covering the cauda epididymis was removed by careful dissection, with care to avoid rupturing blood vessels or the epididymal duct. For detection of transfected spermatozoa, they stained with Rodamine. In order to transfection of sperm, 2 μg of Rodamine labeld DNA and 0.5 μl of TurboFect were diluted in 25 μl of transfection medium separately, and incubated for 5 min at room temperature. Then, the diluted DNA was added to diluted TurboFect (total volume=50 μl) and incubated for 20 min at room temperature. 1×106 sperm were added to 50 μl of DNA- TurboFect complexes and mixed gently by rocking the plate back and forth. To evaluation of transfected spermatozoa motility, acridine orange staining was used. Each experiment was replicated at least three times, and for each replicate, at least 50 ES cell colonies were used. Data were analysed with a statistical software program (SPSS 16). Comparisons between two treatments and multiple numeric datasets were performed using t-test and one-way ANOVA followed by Duncan multiple-range test, respectively. Results are expressed as mean±SEM and statistical significance was accepted at P0.05). The comparison of transfected and normal spermatozoa reveal that, motility of transfected spermatozoa at 60 minutes after transfection was significantly lower than normal ones (p

Many studies examine the antibacterial effects of medicinal plants; however, little research is done to evaluate their effects on different cell types, especially dermal fibroblasts.
The current study aimed to study the effect of different concentrations of Aloe Vera, henna, chamomile, myrtle, mint, licorice, cinnamon, ginger and cedar extracts and their synergistic effects on the viability of dermal fibroblasts.
To evaluate the performance of herbal extracts on dermal fibroblasts, in the first experiment different concentrations (6.25, 12.5, 25, 50, 100, 250, 500 and 1000 µg/mL) of the extracts were evaluated by the MTT cell proliferation assay. In the second experiment, the minimum effective concentrations of the plant extracts in triple combination were evaluated in the cells under study.
The minimum effective concentrations of henna, chamomile, myrtle, mint, cinnamon, ginger and cedar were 12.5, 6.25, 6.25, 6.25, 6.25, 12.5 and 12.5µg/mL, respectively. Results showed that, by comparing the minimum effective concentration of herbal extracts, the viability of dermal fibroblasts significantly increased by cedar extract (P Based on the results of the current study, it was concluded that Aloe vera, licorice and mint extracts had synergistic effects on the viability of dermal fibroblasts; in addition, the combination of Aloe vera and licorice with either henna or myrtle, and Aloe vera and mint with either cedar or ginger resulted in synergistic effects on viability of dermal fibroblasts. The third category of triple combinations of herbal extracts with synergistic effects on the cells under study was the combination of Aloe Vera and mint with either chamomile or cinnamon and also Aloe vera and licorice with either myrtle or cedar.

Herbal extracts have recently received the greatest attention in the path of finding naturally occurring chemicals with antibacterial and therapeutic value; however, each type of herbal remedy may have its own side effects..
The aim of the current experiment was to study the antibacterial effect of myrtle, parsley, mint, henna and chamomile extracts on Escherichia coli and their effects on colony formation and survival of spermatogonial stem cells (SSCs)..
Spermatogonial stem cells were isolated by two-time enzymatic digestion from slaughterhouse origin ovine testis and plant extraction by deionized water. Comparisons between different treatments were performed using analysis of variance (ANOVA) followed by Duncan’s multiple range tests..
The results showed that there was no significant difference between mint, henna and penicillin, on inhibition of Escherichia coli growth, however parsley, myrtle and chamomile were significantly different from penicillin (P The results of these experiments provide evidence that henna by antibacterial activity had no detrimental affect on SSC and Sertoli cells and is a good candidate for substitution of antibiotics..

This research studies the effects of activation and inhibition of Wnt3A signaling pathway in buffalo (Bubalus bubalis) embryonic stem (ES) cell-like cells. To carry on this experimental study, the effects of activation and inhibition of Wnt3A signaling in buffalo ES cell-like cells were examined using Bio (0.5 mM) combined with WNT3A (200 ng/ml), as an activator, and Dickkopf-1 (Dkk1, 250 ng/ml), as an inhibitor, of the pathway. ES cells were cultured up to three weeks in ES cell medium without fibroblast growth factor-2 (FGF-2) and leukemia inhibitory factor (LIF), but in the presence of Bio, WNT3A, Bio+WNT3A and Dkk1. The effects of these supplements were measured on the mean area of ES cell colonies and on the expression levels of a number of important genes related to pluripotency (Oct4, Nanog, Sox2 and c-Myc) and the Wnt pathway (β-catenin). ES cell colonies cultured in ES cell medium that contained optimized quantities of LIF and FGF-2 were used as the control. Data were collected for week-1 and week-3 treated cultures. In addition, WNT3A-transfected ES cells were compared with the respective mock-transfected colonies, either alone or in combination with Dkk1 for expression of β-catenin and the pluripotency-related genes. Data were analyzed by ANOVA, and statistical significance was accepted at P<0.05. Among various examined concentrations of Bio (0.5-5 mM), the optimum effect was observed at the 0.5 mM dose as indicated by colony area and expressions of pluripotency- related genes at both weeks-1 and -3 culture periods. At this concentration,the expressions of Nanog, Oct3/4, Sox2, c-Myc and β-catenin genes were nonsignificantly higher compared to the controls. Expressions of these genes were highest in the Bio+WNT3A treated group, followed by the WNT3A and Bio-supplemented groups, and lowest in the Dkk1-treated group. The WNT-transfected colonies showed higher expressions compared to both mock and Dkk1-treated mock transfected colonies. WNT3A functions to maintain the pluripotency of ES cell-like cells both as an exogenous growth factor as well as an endogenously expressed gene. It complements the absence of FGF-2 and LIF, otherwise propounded essential for buffalo ES cell culture. WNT3A antagonizes the inhibitory effects of Dkk1 and acts in combination with its activator, Bio, to activate the Wnt signaling pathway.

In order to retain an undifferentiated pluripotent state, embryonic stem (ES) cells have to be cultured on feeder cell layers. However, use of feeder layers limits stem cell research, since experimental data may result from a combined ES cell and feeder cell response to various stimuli. In this experimental study, a buffalo ES cell line was established from in vitro derived blastocysts and characterized by the Alkaline phosphatase (AP) and immunoflourescence staining of various pluripotency markers. We examined the effect of various factors like fibroblast growth factor 2 (FGF-2), leukemia inhibitory factor (LIF) and Y-27632 to support the growth and maintenance of bubaline ES cells on gelatin coated dishes, in order to establish feeder free culture systems. We also analyzed the effect of feeder-conditioned media on stem cell growth in gelatin based cultures both in the presence as well as in the absence of the growth factors. The results showed that Y-27632, in the presence of FGF-2 and LIF, resulted in higher colony growth and increased expression of Nanog gene. Feeder-Conditioned Medium resulted in a significant increase in growth of buffalo ES cells on gelatin coated plates, however, feeder layer based cultures produced better results than gelatin based cultures. Feeder layers from buffalo fetal fibroblast cells can support buffalo ES cells for more than two years. We developed a feeder free culture system that can maintain buffalo ES cells in the short term, as well as feeder layer based culture that can support the long term maintenance of buffalo ES cells.

Multiple choice exams are one of the most common objective exams used in medical education. So, it is important to find ways to improve the quality of these exams, especially in residency programs. Thus the aim of this study was to investigate the effect of education on quality improvement of Multiple Choice Questions (MSQ) designed in Annual Residency Exams of Dental Faculty. In this experimental study, the structure and taxonomy in all MCQs designed by dental faculty for annual residency exams in 2008 were analyzed through valid and reliable checklist. Checklist items were based on Millman’s principles and several other resources. The same process was repeated for MCQs after running a workshop for question designers. The pre- and post- workshop data were compared using Z test. From 1239 questions, 63.1% and 76.3% of the questions developed in 2008 and 2009 had no structural flaw, which revealed a significant difference between the two years (P<0.001). Regarding high taxonomy questions, a significant increase was observed in the year 2009 (P=0.039). Considering the results of this study, designing and running training workshop for question designers improved quality of MCQs and these workshops can lead to improvement of MCQ exams in other universities.