Comparison of different types of liposomal nano structures for microRNA transfection to human mesenchymal stem cell line S1939.

Background: Liposomes are utilized as a drug delivery carrier in various fields of biomedicine. They are synthesized in the nanometer-size range and are becoming a viable drug delivery carrier for the treatment of different diseases. MicroRNAs as regulatory elements could be transferred to cells for changing their morphology or physiology. The study's major aim is to find the optimized formula of liposomes for transfection of microRNA to human mesenchymal stem cell line S1939 (HMSCs). Materials and Methods: Various ratios of soybean phosphatidylcholine (SPC), cholesterol, 1, 2 dioleoyloxy-3- (trimethylammonium) propane (DOTAP), and polyethylene glycol (PEG) were combined. The mean diameter of all formulations and their surface properties were determined by a zeta sizer device and scanning electron microscope, respectively. The cytotoxicity of formulations was assessed using MTT (3,4,5-dimethyl thiazol-2-yl) (2,5-diphenyltetrazolium bromide) assay. The transfection effectiveness of liposomal miRNA vs empty liposomes was determined using agarose gel electrophoresis. Results: The optimized liposome vesicles were prepared using 45:30:27.5:5 molar ratios of SPC:DOTAP:cholesterol: DSPE-PEG. The liposome formulations F10 and F18 were the best in terms of biocompatibility because of the higher viabilities of treated cells. The best formulation (F18, containing 0.7 µg of miRNA and 10 µg of liposome) was nearly 100% efficient in sequestering and fixing miRNA. Phase-contrast and fluorescent microscopic examinations showed intra-nuclear as well as intracytoplasmic localization of the particles. Conclusion: Some easily prepared liposomal formulation vehicles are quite efficient in the transfection of miRNA into the HMSCs and could be used for in vitro applications in regenerative medicine.

The Viability of Human Testis-Derived Cells on Human Serum Albumin-Based Scaffold as An Artificial Male Germ Cell Niche.

Azoospermia is one of the challenging disorders affecting couples who are afflicted with infertility. Human testisderived cells (hTCs) are suitable candidates for the initiation of in-vitro spermatogenesis for these types of patients. The current study aimed to assess the proliferation of hTCs through the cell culture on the three-dimensional (3D) porous scaffolds. Cells harvested from the testicular sperm extraction (TESE) samples of the azoospermic patients were cultured on the 3D porous scaffolds containing human serum albumin (HSA)/tri calcium phosphate nanoparticles (TCP NPs) for two weeks. The proliferation/viability of the cells was assessed using the MTT assay, along with H and E histological staining method. The MTT assay showed that hTCs could stay alive on this scaffold with 50 and 66.66% viability after 7 and 14 days, respectively. Such viability was not significantly different when compared with cells grown on monolayer flask culture (P>0.05). Therefore, 3D HSA/TCP NPs scaffolds could be used for the reconstitution of the artificial human somatic testicular niche for future applications in regenerative medicine for male infertility.

The impacts of pill contraceptive low-dose on plasma levels of nitric oxide, homocysteine, and lipid profiles in the exposed vs. non exposed women: as the risk factor for cardiovascular diseases.

BACKGROUND: Consuming oral contraceptive pills is one of the methods for preventing pregnancy worldwide. As using the pills has always caused the greatest concern for the likelihood of developing cardiovascular diseases and also given the limited conducted studies in this regard, this study was carried out to determine the impacts of low dose birth control pills on plasma levels of nitric oxide, homocysteine, and lipid profiles in the exposed vs. non exposed women as the risk factors for cardiovascular diseases. METHODS: This was a combined cohort study conducted on 100 women, having the age range 20-35 years and normal menstrual cycles, referred to the health care centers in Yazd, Iran. The demographic data were obtained through face to face interviews performed by the researcher. Anthropometric indexes were measured and biochemical factors were determined by testing blood samples. Then, using SPSS 16 and statistical independent t-test and Chi- square, the data were analyzed. RESULTS: The mean ± standard deviations of plasma levels of homocysteine, nitric oxide, cholesterol, triglyceride, Low Density Lipoprotein, and High Density Lipoprotein levels in the group consumed low dose contraceptive pills were 3.84 ± 2.35 µmol/l, 181.36 ± 90.44 µM, 180.7 ± 38.28 mg/dl, 129.82 ± 47.92 mg/ dl, 101.42 ± 30.66 mg/dl, and 56.46 ± 8.42 mg/dl, There were significant statistical differences between those consuming the pills and those not consuming the pills regarding cholesterol (P < 0.05). CONCLUSION: Consuming Low Dose contraceptive pills can increase the plasma levels of cholesterol, triglyceride, and Low Density Lipoprotein levels; i.e. this condition is called dyslipidemia. On the other hand, there were no changes in the levels of homocysteine and nitric oxide in the healthy women consuming the pills; therefore, the pills may not develop cardiovascular diseases in healthy women. Accordingly, it is recommended that the health care providers prescribe the pills for the women with cautions.

Molecular Identification of Hemoprotozoan Parasites in Camels (Camelus dromedarius) of Iran.

BACKGROUND: Although camels represent a valuable source of food, wool and hide in many countries, in-depth information about their vector-borne pathogens is scarce compared to other animals. The aim of the current study was to characterize vector-borne protozoa in the blood of dromedaries from Iran by molecular tools. METHODS: From June to July 2014, 200 peripheral blood samples were collected from asymptomatic one-humped camels in two provinces of Kerman and Sistan- va-Baloochestan in central and southeastern Iran. Microscopic examination was performed on Giemsa-stained blood smears, and drops of blood were spotted on Whatman FTA® cards for further analyses. Genomic DNA was extracted from the cards, and PCR was carried out for the detection of piroplasms and trypanosomes, followed by sequence analysis of positive samples. RESULTS: One sample was positive Trypanosoma spp. trypomastigotes in light microscopy. PCR results revealed one positive sample each with Theileria annulata and Trypanosoma evansi. CONCLUSION: Camels were identified as hosts for bovine Mediterranean theileriosis in the investigated area. The presence of Tr. evansi, the causative agent of surra disease, was also confirmed in camels of Iran. Further studies are recommended in order to investigate their impact on the health and productivity of camels and other livestock in this region.

The viability of mouse spermatogonial germ cells on a novel scaffold, containing human serum albumin and calcium phosphate nanoparticles.

BACKGROUND: In spermatogenesis, spermatogonial cells differentiate to the haploid gametes. It has been shown that spermatogenesis can be done at in vitro condition. In vitro spermatogenesis may provide an open window to treat male infertility. OBJECTIVE: The aim of this study was to evaluate the effects of a novel scaffold containing human serum albumin (HSA)/tri calcium phosphate nanoparticles (TCP NPs) on the mouse spermatogonial cell line (SCL). MATERIALS AND METHODS: First, TCP NPs were synthesized by reaction of calcium nitrate and diammonium phosphate at pH 13. Then, serial concentrations of TCP NPs were separately added to 500 mg/mL HSA, and incubated in the 100(o)C water for 30 min. In the next step, each scaffold was cut (2×2mm), placed into sterile well of microplate, and then incubated for 1, 2, and 3 days at 37(o)C with mouse SCL. After incubation, the cytotoxicity of the scaffolds was evaluated by different tests including 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) assay, vital staining, and cell counting. On the other hand, the release of TCP NPs and HSA from the scaffolds was measured. RESULTS: Based on microscopic observation, the size of cavities for all scaffolds was near 200-500 µm, and the size of TCP NPs was near 50-100 nm. All toxicity tests showed that the increase of TCP concentration in the scaffold did not affect mouse SCL. It means that the percentage of cell viability, LDH release, vital cells, and cell quantity was 85%, 105%, 90%, and 110%, respectively. But, the increase of incubation time led to increase of LDH release (up to 115%) and cell count (up to 115%). Also, little decrease of cell viability and vital cells was seen when incubation time was increased. Here, no release of TCP NPs and HSA was seen after increase of TCP concentration and incubation time. CONCLUSION: It can be concluded that the increase of TCP concentration in HSA/ TCP NPs scaffold does not lead to cytotoxicity. On the other hand, the increase of incubation time leads to increase of mouse SCL cell death. In this study, it was found that TCP NPs and HSA could not release from the scaffolds. In future, both proliferation and differentiation of mouse SCL on HSA/TCP NPs scaffold must be checked over more wide incubation times.

Diagnostic Value of the Risk of Malignancy Index (RMI) for Detection of Pelvic Malignancies Compared with Pathology.

INTRODUCTION: Pelvic masses are among most the common causes of patient admission into gynecology clinics and one of the most common reasons for referral to gynecologic oncology departments due to the risk of uterine or ovarian malignancies. The aim of this study is to compare the four indices of the risk of malignancy index (RMI 1-4), as a combination of menstrual status, radiological findings, and serum CA125 concentration, for discrimination of benign from malignant pelvic masses. METHODS: This retrospective descriptive and analytic study was conducted on 200 patients with pelvic mass, post-surgery, and who were referred to the oncology department in Shahid Sadoughi hospital of Yazd (Iran) between June 2007 and September 2011. Data regarding demographics, pathology reports, paraclinical and clinical tests were analyzed. The four RMI indices were separately used for determination of benign vs. malignant masses using the optimized cutoff points, ROC curve, sensitivity, specificity, predictive value of positive and negative, and accuracy. Finally, p value for each index was calculated, and a final discrimination power was measured by using SPSS version 17 software. RESULTS: The calculated p values in the four RMI indices in ultrasound findings indicated statistical significance, and the RMI 2 showed the highest level of accuracy or diagnostic performance. RMI 2 had a cutoff point of 90, an under-chart area 86.7, 79.36% sensitivity, 78.95% specificity, 58.44%, positive predictive value, 90.08% negative predictive value, and 78.93% accuracy, and a p value of 0.004. However, this relationship was found not to be meaningful using CT scan images. CONCLUSIONS: Using RMI 2 for differentiation of malignant from benign pelvic masses is a reliable method with ultrasound findings.

Incidence of sexual dysfunction in men after cardiac surgery in Afshar hospital, Yazd.

BACKGROUND: Successful rehabilitation of cardiac surgery patients should include consideration of their sexual activity, but there is paucity of data regarding this matter. OBJECTIVE: This study determined the incidence and type of sexual dysfunction in our patients. MATERIALS AND METHODS: Two hundred-seventy nine men with age under 70 years old who had coronary artery bypass graft (CABG), valvular, or other types of cardiac surgery from Dec. 2006 until Dec. 2007 were enrolled in this descriptive-analytical study. They were interviewed before and 12 weeks after the operation in regard to the impact of surgery on their sexuality. The statistical methods used included analysis of variance, Kappa test, and chi-square analysis. RESULTS: The mean age of the patients was 55.7 ± 10.66 (25-69) years. The incidence of sexual dysfunction was 20.1% before, and76.4%, 12 weeks after the operation. P-valueas tested by Kappa test was 0.0001, which means that cardiac surgery had adverse effect on sexual activity of the patients. Types of sexual dysfunction were impotence, premature ejaculation, and decreased or loss of libido in 6.5%, 4.3% and 9.3%, respectively before operation, and 34.8%, 21.5% and 20.1%, respectively 12 weeks after the operation. Concurrence of more than one dysfunction was not reported. CONCLUSION: Sexual dysfunction is common after cardiac surgery, and sexual counseling is still not being addressed adequately. The role and responsibility of the physician and the rehabilitation nurse becomes evident, together with the need for the patient's partner to participate in counseling.