Evaluation of the expression profile of mRNAs and lncRNAs in cumulus cells associated with polycystic ovary syndrome and pregnancy.

Objectives: Polycystic ovary syndrome (PCOS), the primary cause of anovulatory infertility in women, may change the gene expression profile of cumulus cells. In human ART (assisted reproductive technology), gene expression profiling in cumulus cells, a non-invasive method, may be used to identify the most competent oocytes. We aim to identify key genes according to the network-based data and assess the suitability of these genes as markers to predict oocyte competence and PCOS diagnosis. Materials and Methods: The GSE34526 microarray dataset was obtained from the Gene Expression Omnibus (GEO) database. The function and pathway enrichment analysis for DEGs were analyzed. A protein-protein interaction (PPI) network analysis and candidate gene screening were conducted. A two-layer network consisting of mRNA and lncRNA was constructed. Expression levels of hub genes were verified using quantitative RT-PCR (qRT-PCR). Results: A total of 2721 DEGs were retained. The PPI network of selected genes associated with the biological process of "cell communication" was analyzed, and the first 10 key genes were determined by degree. Additionally, 2 hub genes and 2 hub lncRNAs, including STAT3, RHOA, GAS5, and LINC01116, were selected from the lncRNA-mRNA network. Finally, expression levels of STAT3, RHOA, GAS5, and LINC01116 were significantly increased in the cumulus cells of PCOS patients compared to the control group (P<0.05). However, there was no significant difference in expression between the pregnant and non-pregnant groups. Conclusion: STAT3, RHOA, GAS5, and LINC01116 may serve as possible diagnostic markers for PCOS. However, further studies on a larger population are needed to validate this finding.

Chondrogenic activity of two herbal products; pomegranate fruit extract and avocado/soybean unsaponifiable.

BACKGROUND AND PURPOSE: Articular cartilage defects aren't repaired by itself. Numerous studies have been conducted in the area of cartilage tissue engineering and some of them considered herbal products. An attempt was made in this study to compare the effects of pomegranate fruit extract (PFE), avocado/soybean unsaponifiable (ASU), and their equal proportional mixture on the chondrogenesis of human adipose-derived stem cells (hADSCs). EXPERIMENTAL APPROACH: PFE was prepared through the percolation method. ASU powder was dissolved in ethanol at 10 µg/mL concentration and was sterilized. The hADSCs first were isolated, expanded in monolayer culture and identified, and next seeded on fibrin scaffolds. The hADSCs/fibrin scaffolds were divided into 4 groups of control, ASU, PFE, and PFE+ ASU and subjected to in vitro induction for 2 weeks. The control group received chondrogenic medium, other groups received chondrogenic medium plus ASU, PFE, or PFE + ASU, respectively. The MTT assay was performed for cell viability evaluation, real-time polymerase chain reaction for expression of cartilage genes, and the toluidine blue, safranin-O, and immunohistochemistry for staining of the constructs. FINDINGS / RESULTS: Cell viability, cartilage genes expression, matrix staining density, and collagen II protein levels in PFE samples were significantly higher than those of the other groups (P < 0.05). Histological assessments revealed more chondrogenic centers (P < 0.05) in the PFE group compared to the other groups. CONCLUSION AND IMPLICATIONS: In this study, it was revealed that PFE can be considered as an induction factor for future chondrogenic studies.

A Study on the Presence of Osteopontin and α3ß1 Integrin in the Endometrium of Diabetic Rats at the Time of Embryo Implantation.

BACKGROUND: Embryo implantation is a critical and multifactorial phenomenon which can be affected by any alteration in molecular micro construction of endometrium. The aim of the current study was to evaluate the effects of diabetes on osteopontin (OPN) and α3ß1 integrin proteins level at the time of endometrial receptivity. METHODS: Twenty-eight female rats were divided into control, diabetic, pioglitazone-treated and metformin-treated groups. Western blot was performed to determine the OPN and α3ß1 integrin proteins in rats' endometrium at the time of implantation. Data were analyzed by analysis of variance (ANOVA) and p<0.05 was considered statistically significant. RESULTS: OPN increased significantly in the diabetic group in comparison with control (p<0.001), metformin-treated (p=0.008) and pioglitazone-treated groups (p< 0.001). Furthermore, α3ß1 integrin protein level in diabetic group had a significant difference in comparison with that of the control (p<0.001), metformin-treated (p= 0.026) and pioglitazone-treated groups (p<0.001). CONCLUSION: OPN and α3ß1 integrin proteins are involved in embryo implantation and their changes in diabetic condition can affect fertility. Treatment with pioglitazone and metformin improved the level of OPN and α3ß1 integrin proteins while pioglitazone was more effective.

Agar-based edible films for food packaging applications - A review.

Agar is a biopolymer extracted from certain red algae. The continuous and transparent film made from agar gum is becoming a common and renewable alternative for plastic-based food packaging materials. However, plain agar film suffers from brittleness, high moisture permeability, and poor thermal stability. Considerable researches have been devoted to improving the properties of agar films to extend their applications. These include reinforcements by nanomaterials, blending with other biopolymers, and incorporating plasticizers, hydrophobic components, or antimicrobial agents into their structure. This article comprehensively reviews the functional properties and defects of edible films made from agar gum. Also, it describes various strategies and components used to make an agar film with desirable properties. Moreover, the applications of agar-based edible films with improved functionality for food packaging are discussed.

A Survey on Main Semen Parameters in Natural Pregnancy and Intrauterine Insemination: Are There Any Significant Differences?

Intrauterine insemination (IUI) is a treatment of choice compared with other invasive and expensive techniques of assisted reproduction. Sperm quality is used to predict its outcome and success. Establishing threshold levels for sperm parameters is useful to avoid spending time and money to do other assisted reproductive techniques. This study was carried out to compare various semen parameters in a group of men eligible to participate in an IUI program with those of fertile men whose wives were pregnant at the time of the study. Two hundred and thirty-four semen samples were evaluated from subfertile men whose partners were candidates for IUI and 234 semen samples were evaluated from fertile men whose partners were pregnant less than 12 weeks. To assess the sensitivity and specificity of the main semen parameters, receiver operating characteristic (ROC) curves were used. Normal sperm morphology is more sensitive and specific compared with its progressive motility and concentration. No significant differences in various semen parameters of fertile men and those of the male partners of IUI candidates were observed. ROC analysis identified that sperm normal morphology using strict criteria may be a good indicator of fertility status in men. No significant difference in various semen parameters between the male partners of IUI candidates and the fertile men was seen. However, utilizing ROC curves, sperm morphology using strict criteria could be a good predictor of fertility.

The Study of Association Between Polymorphism of TNF-α Gene's Promoter Region and Recurrent Pregnancy Loss.

BACKGROUND: According to the literature review, polymorphisms of tumor necrosis factor alpha's (TNF-α) promoter region are probably the genetic risk factors of recurrent pregnancy loss. This study has investigated five single nucleotide polymorphisms in the TNF-α gene's promoter region to evaluate their relationship with recurrent pregnancy loss disorder. METHODS: Blood samples were taken from 65 women with recurrent pregnancy loss (Case group) and 65 healthy women with a history of successful pregnancy (Control group). Polymerase chain reaction with high resolution melting (PCR-HRM) analysis was done to determine the promoter region of -308G/A, -850T/C, -238G/A, -1031T/C and -863A/C TNF-α polymorphisms. The data were assessed using logistic regression models. P values less than 0.05 were considered statistically significant. RESULTS: Significant associations were found between recurrent pregnancy loss and -863C/A (p=0.000), -308G/A (p=0.045), and -238G/A (p=0.034) polymorphisms. TNF-α polymorphisms of -863C and -238G may be susceptible factors of recurrent pregnancy loss cases. The -308G polymorphism has an important role in maintaining pregnancy. CONCLUSION: The -863C/A and -238G/A TNF-α polymorphisms are possible genetic risk factors of recurrent pregnancy loss and might be its predictive markers.

PROTAMINE1 and PROTAMINE2 genes expression in the sperms of oligoasthenospermic individuals and intrauterine insemination candidates couples: Is there any significant differences?

BACKGROUND: Male infertility refers to a male's inability to cause pregnancy in a fertile female. It seems the large portion of this category of infertility, has roots in genetic factors. PROTAMINE family is one of the most important genes which are involved in male factor infertility. Hence, the aim of this study is to evaluate PROTAMINE1 and PROTAMINE2 (P1 and P2) genes expression in oligoasthenospermic individuals and intrauterine insemination (IUI) candidate couples' sperms. MATERIALS AND METHODS: Samples were gathered from the patients referred to the Isfahan Infertility Center of Shahid Beheshti, 80 semen samples were in IUI candidates groups and 16 semen samples were in oligoasthenospermia group was collected. The outcome of IUI procedure was followed up after 14 days. Through these samples, 16 couples achieved pregnancy (IUI+) and from the top of the list, 16 semen samples with negative ß-HCG were obtained (IUI-). After RNA extraction from sperms, PROTAMINE genes family expression was evaluated in our three groups by real time-reverse transcription polymerase chain reaction. RESULTS: Our study revealed that P1 gene expression has no significant differences between IUI-, IUI+, and oligoasthenospermia groups, whereas P2 gene expression showed significant differences between oligoasthenospermia with two IUI groups. Main sperm parameters have no significant differences between IUI groups. CONCLUSION: This study reveals P1 and P2 genes expression value have no significant differences between IUI- and IUI+. On the other hand, P2 gene expression value has significant differences between oligoasthenospermia with two IUI groups.

Preparation and characterization of tragacanth-locust bean gum edible blend films.

The present work introduces the structure and physicomechanical properties of a novel blend film made from binary solutions of gum tragacanth (GT) and locust bean gum (LBG) at different mixing ratios. Apparent viscosities and surface tensions of individual and blend gum solutions were also investigated. The viscosity data indicated that there was a distinct synergism between the two gums at all mixing ratios. FTIR spectra showed the existence of noncovalent intermolecular interactions between gums. The surface tensions of binary solutions were significantly lower than those of individual gums which is advantageous for coating applications. All films had homogenous and smooth surface morphology and their transparency, water vapour barrier and mechanical properties were improved by incorporating LBG in blend. The results of this study suggest that GT-LBG blend film, owing to its desirable properties, has the potential to be used as a new degradable food packaging material.

Effect of T3 hormone on neural differentiation of human adipose derived stem cells.

Human adult stem cells, which are capable of self-renewal and differentiation into other cell types, can be isolated from various tissues. There are no ethical and rejection problems as in the case of embryonic stem cells, so they are a promising source for cell therapy. The human body contains a great amount of adipose tissue that contains high numbers of mesenchymal stem cells. Human adipose-derived stem cells (hADSCs) could be easily induced to form neuron-like cells, and because of its availability and abundance, we can use it for clinical cell therapy. On the other hand, T3 hormone as a known neurotropic factor has important impressions on the nervous system. The aim of this study was to explore the effects of T3 treatment on neural differentiation of hADSCs. ADSCs were harvested from human adipose tissue, after neurosphere formation, and during final differentiation, treatment with T3 was performed. Immunocytochemistry, real-time RT-PCR, Western blotting techniques were used for detection of nestin, MAP2, and GFAP markers in order to confirm the effects of T3 on neural differentiation of hADSCs. Our results showed an increase in the number of glial cells but reduction in neuronal cells number fallowing T3 treatment.

Comparative Study of Microtubule-associated Protein-2 and Glial Fibrillary Acidic Proteins during Neural Induction of Human Bone Marrow Mesenchymal Stem Cells and Adipose-Derived Stem Cells.

BACKGROUND: In recent years, adipose tissue, due to the stem cells contained within, has found a new special place in laboratory and clinical applications. These adipose-derived stem cells (ADSCs) have the same characteristics of bone marrow mesenchymal stem cells (BMSCs). Although bone marrow (BM) is not easily accessible and its procurements may be painful, most patients possess excess fat which can be obtained by less invasive methods; this makes adipose tissue ubiquitous, available and an ideal large-scale source for research on clinical applications. METHODS: BMSCs and ADSCs were harvested from three healthy human and were characterized using flow-cytometry. After they were treated for neurosphere formation using basic fibroblast growth factor, epidermal growth factor, B 27; terminal differentiation was performed. In this study, we used immunocytochemistry, real time-polymerase chain reaction and western blotting techniques for detection and comparison of Nestin, microtubule-associated protein-2 (MAP-2) and glial fibrillary acidic protein (GFAP) markers in human ADSCs and BMSCs. RESULTS: Under appropriate conditions ADSCs can differentiate into neuron-like cells and express neural markers the same as BMSCs, also the expression of GFAP marker in differentiated cells derived from ADSCs was significantly lower than the cells derived from BMSCs (P < 0.05). While the expression of MAP-2 marker in both groups was the same. CONCLUSIONS: However, due to its advantages and according to our results based on the expression levels of GFAP and MAP-2, adipose tissue rather than BM could represent a more appropriate stem cell source for investigating the application of these cells in understanding the pathophysiology and in treatment of neurodegenerative disorders.

Co-culture with neurotrophic factor secreting cells induced from adipose-derived stem cells: promotes neurogenic differentiation.

Adipose-derived stem cells (ADSCs) and bone marrow stem cells (BMSCs) can be equally proper in the treatment of neurodegenerative diseases. However, ADSCs have practical benefits. In this study, we attempted to induce the secretion of neurotrophic factors (NTF) in human ADSCs. We then evaluated the effects of co-culture with NTF secreting cells in neural differentiation of human ADSCs. Isolated human ADSCs were induced to neurotrophic factors secreting cells. To evaluate the in vitro effects of NTF-secreting ADSCs on neurogenic differentiation of ADSCs, we used neurogenic induction medium (control group), the combination of neurogenic medium and conditioned medium, or co-cultured NTF-secreting ADSCs which were encapsulated in alginate beads (co-culture) for 7 days. ELISA showed increased (by about 5 times) release of brain-derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in NTF-secreting ADSCs compared to human ADSCs. Real time RT-PCR analysis revealed that NTF-secreting ADSCs highly expressed NGF and BDNF. In addition, co-culture with NTF-secreting ADSCs could also promote neuronal differentiation relative to gliogenesis. Overall, NTF-secreting ADSCs secrete a range of growth factors whose levels in culture could promote neuronal differentiation and could support the survival and regeneration in a variety of neurodegenerative diseases.

Comparing brain-derived neurotrophic factor and ciliary neurotrophic factor secretion of induced neurotrophic factor secreting cells from human adipose and bone marrow-derived stem cells.

Adipose derived stem cells (ADSCs) and bone marrow stem cells (BMSCs) may be equally beneficial in treating neurodegenerative diseases. However, ADSCs have practical advantages. In this study, we aimed to induce neurotrophic factors secreting cells in human ADSCs. Then, we compared the level of brain-derived neurotrophic factor (BDNF) and ciliary neurotrophic factor (CNTF) secretion in neurotrophic factors secreting cells from human adipose and bone marrow-derived stem cells. Isolated human ADSCs and BMSCs were induced to neurotrophic factor (NTF)-secreting cells. The levels of expression and secretion of BDNF and CTNF of induced cells were assessed using immunocytochemical, Real-Time polymerase chain reaction, and enzyme linked immunosorbent assay (ELISA). The level of BDNF significantly increased in both the induced mesenchymal stem cells (MSCs) relative to ADSCs and the BMSCs (P < 0.01). Moreover, ELISA analysis showed that the release of BDNF in the induced BMSCs was almost twofold more than the induced ADSCs. Overall, NTF-secreting factor cells derived BMSCs and ADSCs could secret a range of different growth factors. Therefore, the variation in neurotrophic factors of different induced MSC populations suggest the possible beneficial effect of each specific kind of neurotrophic factor secreting cells for the treatment of a particular neurodegenerative disease.