Advanced strategies for single embryo selection in assisted human reproduction: A review of clinical practice and research methods.

Among the primary objectives of contemporary assisted reproductive technology research are achieving the births of healthy singletons and improving overall fertility outcomes. Substantial advances have been made in refining the selection of single embryos for transfer, with the aim of maximizing the likelihood of successful implantation. The principal criterion for this selection is embryo morphology. Morphological evaluation systems are based on traditional parameters, including cell count and fragmentation, pronuclear morphology, cleavage rate, blastocyst formation, and various sequential embryonic assessments. To reduce the incidence of multiple pregnancies and to identify the single embryo with the highest potential for growth, invasive techniques such as preimplantation genetic screening are employed in in vitro fertilization clinics. However, new approaches have been suggested for clinical application that do not harm the embryo and that provide consistent, accurate results. Noninvasive technologies, such as time-lapse imaging and omics, leverage morphokinetic parameters and the byproducts of embryo metabolism, respectively, to identify noninvasive prognostic markers for competent single embryo selection. While these technologies have garnered considerable interest in the research community, they are not incorporated into routine clinical practice and still have substantial room for improvement. Currently, the most promising strategies involve integrating multiple methodologies, which together are anticipated to increase the likelihood of successful pregnancy.

Cerumenolytic Effects of Carbamide Peroxide in Patients with Ear Wax Obstruction.

Introduction: Accumulated and compacted ear wax or cerumen can cause conductive hearing loss, discomfort and vertigo, and infection. This study investigates the effect of Carbamide peroxide (CP) compared with Phenol glycerin (PG) ear drops on cerumen. Materials and Methods: This experimental study investigated the effect of PG and CP ear drops on cerumen in ex vivo and in vivo phases. In the ex vivo phase cerumen degredation was scored following PG and CP treatments. In the in vivo phase, 29 patients with bilateral cerumen impaction were randomly entered the study. PG and CP were applied 3 times a day (each time 5 drops) for 4 days by patients. After treatments, the time of cerumen removal was measured. Results: Instant changes showing degredation of cerumen (grade 1) was evident when it was exposed to CP, on the other hand degredation changes (grade 1) in cerumen treated with PG was only evident after 20 min incubation at 37 oC, while grade 3 degredation was evident in cerumen treated with CP after the same time incubation. Although the time needed for removal of cerumen was lower in CP treatment (54.10±31.77) compared to PG treatment (67.10±35.54), the difference was not statistically significant. Conclusion: Based on the literature and our results, carbamide peroxide is suggested as a proper treatment for patients with EAC obstruction caused by cerumen compaction, because not only it is significantly effective in cerumen degredation, but also no side effects have been reported.

In search of subcortical and cortical morphologic alterations of a normal brain through aging: an investigation by computed tomography scan.

Morphologic changes in the brain through aging, as a physiologic process, may involve a wide range of variables including ventricular dilation, and sulcus widening. This study reports normal ranges of these changes as standard criteria. Normal brain computed tomography scans of 400 patients (200 males, 200 females) in every decade of life (20 groups each containing 20 participants) were investigated for subcortical/cortical atrophy (bicaudate width [BCW], third ventricle width [ThVW], maximum length of lateral ventricle at cella media [MLCM], bicaudate index [BCI], third ventricle index [ThVI], and cella media index 3 [CMI3], interhemispheric sulcus width [IHSW], right hemisphere sulci diameter [RHSD], and left hemisphere sulci diameter [LHSD]), ventricular symmetry. Distribution and correlation of all the variables were demonstrated with age and a multiple linear regression model was reported for age prediction. Among the various parameters of subcortical atrophy, BCW, ThVW, MLCM, and the corresponding indices of BCI, ThVI, and CMI3 demonstrated a significant correlation with age (R2≥0.62). All the cortical atrophy parameters including IHSW, RHSD, and LHSD demonstrated a significant correlation with age (R2≥0.63). This study is a thorough investigation of variables in a normal brain which can be affected by aging disclosing normal ranges of variables including major ventricular variables, derived ventricular indices, lateral ventricles asymmetry, cortical atrophy, in every decade of life introducing BW, ThVW, MLCM, BCI, ThVI, CMI3 as most significant ventricular parameters, and IHSW, RHSD, LHSD as significant cortical parameters associated with age.

Upregulation of apoptotic genes and downregulation of target genes of Sonic Hedgehog signaling pathway in DAOY medulloblastoma cell line treated with arsenic trioxide.

Sonic hedgehog (SHH) medulloblastoma etiology is associated with the SHH molecular pathway activation at different levels. We investigated the effect of arsenic trioxide as a downstream-level inhibitor of the SHH signaling pathway on morphology, cytotoxicity, migration, and SHH-related and apoptotic gene expression of DAOY cells. Cells were treated at various arsenic trioxide (ATO)concentrations (1, 2, 3, 5, and 10 µM) for different times (24 and 48 hr). Following treatments, the morphology of the cells was investigated at ×20 and ×40 magnification by an inverted microscope. Then, cytotoxicity was investigated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and trypan blue assays. Cell migration was analyzed through the wound-healing assay. Furthermore, the expression of SHH-related (GLI1, GLI2, SMO, and MYCN) and apoptotic genes (BAX, BCL2, and TP53) was assessed by real-time quantitative polymerase chain reaction (qPCR). Finally, GLI1, SMO, and MYCN markers were analyzed through immunocytochemistry. Data were analyzed by SPSS (version 16) and P≤0.05 was considered significant. Morphological changes were seen at 3 and 2 µM in 24 and 48 hr of treatment, respectively. The MTT assay showed a dose-dependent cytotoxicity indicating an IC50 value of 3.39±0.35 and 2.05±0.64 µM in 24 and 48hr treatment, respectively. In addition, the trypan blue assay showed higher IC50 values of 4.29±0.25 and 3.92±0.22 µM in 24 and 48 hr treatment, respectively. The wound-healing assay indicated a dose-dependent reduction of cell migration speed showing a 50% reduction at 2.89±0.26 µM. Significant downregulation of GLI1 and GLI2, as well as the upregulation of BAX, BAX/BCL2 ratio, and TP53 were evident. Significant increases in GLI1 and MYCN markers were also evident in immunocytochemistry. ATO, as a downstream effective inhibitor of the SHH pathway, substantially leads to cell death, cell migration inhibition, apoptosis upregulation, and downregulation of SHH target genes in DAOY medulloblastoma. Since ATO is a toxic chemotherapeutic agent, it must be used at low concentrations (2 µM) in order not to damage healthy cells.

Evaluation of the prevalence of bovine leukemia virus DNA in peripheral blood mononuclear cells of multiple sclerosis patients.

Multiple sclerosis (MS) is an immune system-mediated neurodegenerative disease. Recent studies suggest that viral agents, especially the Epstein Barr virus (EBV), are etiological agents for MS. The roles of other viruses in MS have been investigated. Studies have shown an increase in the level of antibodies against bovine leukemia virus (BLV) in patients with MS. In this regard, our study aimed to examine the presence of BLV DNA in peripheral blood mononuclear cells (PBMCs) of MS patients in Iran. In this cross-sectional study, the presence of BLV in 109 Iranian MS patients and 60 healthy controls was evaluated. The isolated PBMCs were used for DNA extraction and PCR, using specific primers for two distinct genes. The mean age of the participants was 39 ± 9.5 years, and 27 (24.77%) of them were male. Clinical evaluation of these patients showed the most frequent MS type to be relapsing-remitting MS (RRMS) (71; 65.14%). BLV evaluation did not show any BLV DNA presence in the PBMCs of individuals in either the MS or healthy control groups. Therefore, our study showed no evidence of BLV infection in Iranian MS patients.

Effects of dietary supplementation of different oils and conjugated linoleic acid on the reproductive and metabolic aspects of male mice.

The present study was carried out to examine first, if diets enriched with 320 g of the base diet with common dietary oils including fish oil, olive oil, hydrogenated sunflower seed (H-SFS) oil, flaxseed oil and sunflower seed oil (SFS) could induce weight gain and alter reproductive and metabolic characteristics of male mice. Second, whether the addition of conjugated linoleic acid (CLA, 10% of the diet) could ameliorate any negative effects. In this cross-sectional study, 90 four-week-old male NMRI mice were used in two consecutive experiments. A high level of dietary oils negatively affected some reproductive and metabolic characteristics of male mice (p < 0.05), specifically, sunflower seed oil enrichment resulted in higher HDL levels and apoptosis of germinal epithelial cells. An olive oil-enriched diet caused an increase in plasma triglyceride concentrations and germinal cell apoptosis, as well as a decrease in sperm concentration and perturbed spermatogenesis. When CLA was fed in conjunction with dietary oils it successfully mitigated some of the negative reproductive and metabolic characteristics. We conclude that male reproductive processes are affected by high dietary oils, even before signs of obesity are evident. Inclusion of dietary CLA may provide some benefit to offset negative effects, although further studies are required.

Frequency, shape, and estimated volume of intracranial physiologic calcification in different age groups investigated by brain computed tomography scan: a retrospective study.

Intracranial calcification is referred to calcification of parenchyma and vascular structures in brain which can be physiologic or pathologic. This study was conducted with the purpose of investigating the frequency, location, pattern, dimensions and estimated volume of intracranial physiologic calcification (IPC) by computer tomography in different age groups. In this cross-sectional retrospective study, brain computed tomography scans of 216 patients were analyzed in 9 age groups each containing 24 patients from 2 to 89 years old. Data were analyzed by SPSS software using one way analysis of variance (ANOVA, post hoc Tukey), chi square, and linear regression tests (P≤0.05 was considered significant). Rate of calcification in different areas were as follows: pineal gland (75.0%), habenula (36.4%), pineohabenula (15.0%), right lateral ventricle choroid plexus (RCP) (67.7%), left lateral ventricle choroid plexus (LCP) (62.7%), falx cerebri (26.8%), petroclinoid ligament (13.2%), tentorium cerebelli (6.8%), third ventricle choroid plexus (0.9%), fourth ventricle choroid plexus (2.7%), basal ganglia (0.9%). A significant correlation exists between the presence of calcification in pineal, habenula, RCP, and LCP (P≤0.001). Nodular shape of calcification was dominant (47.9%). Estimated volume of pineal calcification showed increased levels in group 8 (70-79 years old) compared to group 2 (10-19 years old) (P≤0.05). Since the accurate description of radiologic appearance of IPCs (location, shape, and size) accompanied with age and clinical manifestation is of great importance in diagnosis and distinguishing from pathologic calcification-for example in patients with melatonin dysregulation or schizophrenic patients-this study was required.

Possible ameliorating effects of Glycyrrhiza Glabra (Licorice) on the sperm parameters in rats under high fat diet.

OBJECTIVES: Adverse effects of obesity, which is caused by an imbalance between the energy intake and expenditure, on the male reproductive system have been reported. Considering the anti-obesity effect of Glycyrrhiza Glabra (GC), we conducted this study to elucidate whether it can ameliorate the sperm parameters. METHODS: In this experimental study, male Wistar rats of 6-8 weeks old were divided into four groups: control, high fat diet (HFD), GC50 (HFD plus 50 mg/kg GC extract), and GC100 (HFD plus 100 mg/kg GC extract). During the 16 weeks of the study course, the rats consumed the extract through gavage, daily. Body mass index (BMI), body weight gain, serum lipid profile, leptin concentration, and sperm parameters were investigated. Data were analyzed by one-way analysis of variance (ANOVA) (post hoc Tukey) to express the significance of mean differences of variables between groups, and linear regression test was used to express the correlation model of variables. Both tests were performed by SPSS software; p≤0.05 was considered significant. RESULTS: BMI was significantly decreased by the GC50 and GC100 groups compared to HFD group. GC50 group considerably decreased leptin level compared to HFD group. A significant positive correlation between leptin and triglyceride levels was evident. GC50 and GC100 extensively increased the total sperm motility and ameliorated the sperm abnormal morphology and count compared to HFD group. CONCLUSION: Glycyrrhiza Glabra extract may exert its ameliorating effects on the sperm parameters through its anti-obesity impact. Both doses of the extract were effective, however, the GC100 was more effective in improving the sperm parameters.

Differentiation of bone marrow derived mesenchymal stem cells into male germ-like cells in co-culture with testicular cells.

OBJECTIVE: Stem cell therapy, specifically, pre-induction of mesenchymal stem cells toward male germ-like cells may be useful in patients with azoospermia. The aim of this study was to evaluate in vitro differentiation of mouse bone marrow-derived mesenchymal stem cells (BMSCs) into male germ-like cells by indirect co-culture with testicular cells in the presence of bone morphogenetic protein 4 (BMP4). METHODS: Experimental groups included: control (mouse BMSCs), treatment group-1 (BMSCs treated with BMP4), treatment group-2 (indirect co-culture of BMSCs with mouse testicular cells in the presence of BMP4) and treatment group-3 (indirect co-culture of BMSCs with testicular cells). BMSCs-derived male germ-like cells were evaluated by the expression of Dazl, and Stra8 using RT-qPCR. RESULTS: Stra8 gene expression was significantly increased in the treatment group-2 and Dazl gene was significantly increased in the treatment group-1 compared to other groups. In conclusion, indirect co-culturing of BMSCs with testicular cells and BMP4 leads to the differentiation of BMSCs into male germ-like cells which express specific male germ-like genes. Testicular cells released factors that contributed to the differentiation of BMSCs into male germ progenitor cells. CONCLUSION: This study suggests that mesenchymal stem cells may be differentiated into male germ-like cells and therefore, may be a novel treatment option for men with azoospermia.

The effect of hydroethanolic extract of Matricaria chamomilla on the reproductive system of male rats exposed to formaldehyde.

Formaldehyde is a common agent in our surrounding environment and can adversely affect the male reproductive system. In this study, the effectiveness of Matricaria chamomilla (MC) extract as an antioxidant was investigated in rats treated with formaldehyde. Thirty-two male Wistar rats were randomly divided into six groups: F (10 mg/kg formaldehyde), M200 (200 mg/kg MC extract), M500 (500 mg/kg MC extract), FM200 (10 mg/kg formaldehyde and 200 mg/kg MC extract), FM500 (10 mg/kg formaldehyde and 500 mg/kg MC extract) and control group (0.9% normal saline). Formaldehyde and MC extract were administered daily for 30 consecutive days via intraperitoneal injection. Hormonal status, sperm parameters, testis tissue histology, germinal cells apoptosis and stereological analyses of testis tissue were investigated. Testosterone and LH levels were significantly increased in FM200, FM500, F200 and F500 groups compared to F group (p ≤ 0.05). Sperm count, motility and viability were significantly enhanced in FM200, FM500, F200 and F500 groups compared to F group (p ≤ 0.05). A decrease in the number of apoptotic germ cells in FM200, FM500, M200 and M500 groups (p ≤ 0.05) was evident. In particular, the MC extract in dose 500 mg/kg is seen to reduce the adverse effects of formaldehyde on the reproductive system of male rats.

The effect of lithium chloride on BDNF, NT3, and their receptor mRNA levels in the spinal contusion rat models.

OBJECTIVE: Nowadays, there seems to be no decisive way for treatment of spinal cord injury (SCI).Extensive cell death (apoptosis and necrosis) occurring in SCI can cause considerable progressive sensorimotor disabilities. Preventing cell death by improving endogenous regenerative capability could an effective strategy for the treatment of SCI. This study was designed to evaluate the effects of lithium chloride (LiCl) on the cell survival through overexpression of BDNF and NT3 mRNA level and their receptors in the contusion rat models. METHODS: Rats were randomly divided into four experimental groups (eight rats/group) including: contused animals (the non-treatment group); contused animals (the control group) which received laminectomy; contused animals received normal saline (vehicle)and contused animals received intraperitoneal injection of 20 mg/kg LiCl three days after surgery. Injection continued for 14 days as treatment. Basso, Beattie, Bresnahan (BBB) rating scale was used to assess the motor function of the rats. To evaluate the histopathological and gene expression analysis, rats were sacrificed 28 days after surgery. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) was performed to obtain the relative levels of mRNA for BDNF, NT3 and their receptors. RESULTS: The results showed LiCl ameliorates BBB scores via up-regulation of BDNF and TrkB receptors. Also, histological analysis showed that the numerical density per area of TUNEL- positive cells and the percentage of cavity significantly decreased in the LiCl-treated group. CONCLUSION: Our findings suggest that LiCl protects neural cells and effectively enhances locomotor function, which was done through up-regulation of endogenous BDNF expression in rats with SCI. ABBREVIATIONS: SCI: spinal cord injury; LiCl: lithium chloride; BDNF: Brain-derived neurotrophic factor; NT3: Neurotrophin-3; BBB: Basso, Beattie, Bresnahan; TrkB: Tropomyosin receptor kinase B; TUNEL: Terminal deoxynucleotidyl transferase dUTP nick end labeling.

Indirect Co-Culture of Testicular Cells with Bone Marrow Mesenchymal Stem Cells Leads to Male Germ Cell-Specific Gene Expressions.

OBJECTIVE: Non-obstructive azoospermia is mostly irreversible. Efforts to cure this type of infertility have led to the application of stem cells in the reproduction field. In the present study, testicular cell-mediated differentiation of male germ-like cells from bone marrow-derived mesenchymal stem cells (BM-MSCs) in an in vitro indirect co-culture system is investigated. MATERIALS AND METHODS: In this experimental study, mouse BM-MSCs were isolated and cultured up to passage three. Identification of the cells was evaluated using specific surface markers by flow-cytometry technique. Four experimental groups were investigated: control, treatment with retinoic acid (RA), indirect co-culture with testicular cells, and combination of RA and indirect co-culture with testicular cells. Finally, following differentiation, the quantitative expression of germ cell-specific markers including Dazl, Piwil2 and Stra8 were evaluated by real-time polymerase chain reaction (PCR). RESULTS: Molecular analysis revealed a significant increase in Dazl expression in the indirect co-culture with testicular cells group in comparison to the control group. Quantitative expression level of Piwil2 was not significantly changed in comparison to the control group. Stra8 expression was significantly higher in RA group in comparison to other groups. CONCLUSION: Indirect co-culture of BM-MSCs in the presence of testicular cells leads to expression of male germ cell-specific gene, Dazl, in the induced cells. Combination of co-culture with testicular cells and RA did not show any positive effect on the specific gene expressions.

Neuroprotective effects of selegiline on rat neural stem cells treated with hydrogen peroxide.

Oxidative stress and reactive oxygen species generation have been implicated in the pathogenesis of several neurological disorders including Parkinson's disease, Alzheimer's disease, amyotrophic lateral sclerosis and multiple sclerosis. In the present study, the neuroprotective effects of selegiline against hydrogen peroxide-induced oxidative stress in hippocampus-derived neural stem cells (NSCs) were evaluated. NSCs isolated from neonatal Wistar rats were pretreated with different doses of selegiline for 48 h and then exposed to 125 µM H2O2 for 30 min. Using MTT and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays, acridine orange/ethidium bromide staining and reverse transcription-quantitative polymerase chain reaction, the effects of selegiline on cell survival, apoptosis and the expression of B-cell lymphoma 2 (Bcl-2) and heat shock protein 4 (Hspa4) in pretreated stem cells were assessed compared with a control group lacking pretreatment. The results indicated that the viability of cells pretreated with 20 µM selegiline was significantly increased compared with the control group (P<0.05). Additionally, 20 µM selegiline increased the mRNA expression of Bcl-2 and Hspa4 (P<0.05 vs. control) and suppressed oxidative stress-induced cell death (apoptosis and necrosis; P<0.05 vs. control and 10 µM groups). From these findings, it was concluded that selegiline may be a therapeutic candidate for the treatment of neurological diseases mediated by oxidative stress.

Applied anatomy, today's requirement for clinical medicine courses.

Anatomy as an indispensable part of the medical curricula, offering impeccable knowledge, prepares the students to enter the practical atmosphere. The aim of this study was to evaluate the clinical application of anatomy courses of the medical students in Zanjan University of Medical Sciences. This cross-sectional study was conducted in 2015 with census sampling on all clinical students (trainees and interns). To collect feedback from students, the questionnaire designed by researchers was used. The Likert rating scale of very high, high, medium, low, and very low was considered and scores of 5 (very high) to 1 (very low) were applied. Data were analyzed by SPSS software. Among the courses of anatomy, trunk anatomy has the greatest impact on clinical courses of medical students (P<0.001). Subjects of muscular system, lymphatic system, vascular system, and nervous system were of significant clinical application during clinical periods; however, no significant clinical application observed for skeletal system (P<0.05). Teaching clinical tips by professors can help improve the performance of medical students in clinical education. In addition, using three-dimensional anatomical software is suggested as well.