Long non-coding RNAs and female infertility: What do we know?

Ten percent of people who are of reproductive age experience infertility. Sometimes the most effective therapies, including technology for assisted reproduction, may lead to unsuccessful implantation. Because of the anticipated epigenetic alterations of in vitro as well as in vitro fertilization growth of embryos, these fertility techniques have also been linked to unfavorable pregnancy outcomes linked to infertility. In this regard, a variety of non-coding RNAs such as long noncoding RNAs (lncRNAs) act as epigenetic regulators in the various physiological and pathophysiological events such as infertility. LncRNAs have been made up of cytoplasmic and nuclear nucleotides; RNA polymerase II transcribes these, which are lengthier than 200 nt. LncRNAs perform critical roles in a number of biological procedures like nuclear transport, X chromosome inactivation, apoptosis, stem cell pluripotency, as well as genomic imprinting. A significant amount of lncRNAs were linked into a variety of biological procedures as high throughput sequencing technology advances, including the development of the testes, preserving spermatogonial stem cells' capacity for differentiation along with self-renewal, and controlling spermatocyte meiosis. All of them point to possible utility of lncRNAs to be biomarkers and treatment aims for female infertility. Herein, we summarize various lncRNAs that are involved in female infertility.

Clinical Application Study of Polymeric Nanospheres Network in Methylphenidate Extraction from Urine Samples by Dispersive Solid Phase Extraction Adsorbent.

Purpose: This research introduces a polymeric nanosphere as a new dispersive solid phase extraction (DSPE) adsorbent for the extraction of methylphenidate (MPH) from urine and its high performance liquid chromatography (HPLC) analysis. Methods: Polymeric nanosphere is a kind of copolymeric network obtained by copolymerization of an ionic liquid monomer and styrene in the presence of vinyltriethoxysilane and 2-hydroxyethylmethacrylate. HPLC coupled with ultra violet detector was applied for the determination and quantification of MPH. Dominant parameters in extraction were modified by the one-parameter-at-a-time method. The results are as follow: 10 mg of polymeric nanospheres (PNS), 400 µL of acetonitrile (ACT), 5 mL of urine with the pH value of 9, and the extraction and desorption times of 2 and 5 minutes, respectively, which can be selected as the optimum extraction conditions. Results: Calibration curve was plotted through optimized conditions, and the proposed method was validated. The results demonstrated that the method presented linearity in the concentration range of 30-1200 ng/mL. Selectivity, matrix effect and metabolites interference effect were investigated and the method presented no obvious interference effect during the analysis run time. Repeatability, limit of detection (LOD) and limit of quantification (LOQ) values of the method can be reported in this section as well. The method showed satisfactory results with 98.8% relative recovery in the analysis of positive urine samples. Conclusion: The findings convinced the applicability of the introduced method for DSPE and HPLC analysis of the positive urine samples in different laboratories.

Application of neurotrophic factor-secreting cells (astrocyte - Like cells) in the in-vitro Alzheimer's disease-like pathology on the human neuroblastoma cells.

This study investigated physical proximity and paracrine activity of neurotrophic factor-secreting cells (NTF-SCs) on beta-amyloid treated cells. Mesenchymal stem cells (MSCs) - to-NTF-SCs (Astrocyte -like cells) trans-differentiation was confirmed using immunofluorescence staining of GFAP. BDNF and NGF levels were measured by ELISA. To mimic AD-like condition, SH-SY5Y cells were exposed to 10 µM Aß1-42. SH-SY5Y cells were allocated into Control; and Aß1-42-treated cells. Treated cells were further classified into three subgroups including Aß1-42 cells, Aß1-42 cells + NTF-SCs (CM) and Aß1-42 cells + NTF-SCs co-culture. Cell viability was measured by MTT assay. Anti-inflammatory and anti-tau hyperphosphorylation effects of NTF-SCs were assessed via monitoring TNF-α and hyperphosphorylated Tau protein expression level respectively. To explore the impact of NTF-SCs on synaptogenesis and synaptic functionality, real-time PCR assay was performed to measure the expression of synapsine 1, homer 1 and ZIF268. The level of synaptophysin was monitored via immunofluorescence staining. Data showed MSCs potential in trans-differentiating toward NTF-SCs indicated with enhanced GFAP expression (p < 0.05). ELISA assay confirmed the superiority of NTF-SCs in releasing NGF and BDNF compared to the MSCs (p < 0.05). Aß significantly induced SH-SY5Y cells death while juxtacrine and paracrine activity of NTF-SCs significantly blunted these conditions (p < 0.05). Trans-differentiated cells had potential to reduce Tau hyperphosphorylation and TNF-α level after treatment with Aß through juxtacrine and paracrine mechanisms (p < 0.05). Moreover, NTF-SCs significantly increased the expression rate of synapsin 1, homer 1 and zif 268 genes in Aß-treated cells compared to matched-control group coincided with induction of synaptophysin at the protein level(p < 0.05). NTF-SCs reversed AD-like neuropathological alterations in SH-SY5Y cells via paracrine and juxtacrine mechanisms.

Dispersive Micro-Solid Phase Extraction for Sensitive Determination of Methotrexate from Human Saliva Followed by Spectrophotometric Method.

For biological assessing of hospital personnel occupationally exposed to antineoplastic drugs, highly sensitive and accurate methods are required. Methotrexate (MTX) is an anticancer agent that is widely used in a variety of human cancers. For the first time, dispersive-micro solid phase extraction (D-µ-SPE) has been applied for determination of low levels of MTX in saliva samples. The method is based on rapid extraction of MTX using graphene oxide adsorbent. The sample preparation time is decreased by the fact that the adsorbent dispersed in the sample solution and extraction equilibrium can be reached very fast. This significant feature which obtained with this method is of key interest for routine trace laboratory analysis. The influence of different variables on D-µ-SPE was investigated. Under optimum conditions, the calibration graph was linear over the range of 10-1000 ng/ml. The relative standard deviations are better than 9.0%. The proposed method was successfully applied for the determination of MTX in patient samples.

Dispersive micro solid phase extraction of busulfan from plasma samples using novel mesoporous sorbent prior to determination by HPLC-MS/MS.

Determination of busulfan concentration in patients undergoing bone marrow transplantation is necessary in order to reduce toxic effects and/or graft rejection due to unadjusted dose exposure. A new extraction method namely dispersive micro solid phase extraction (DMSPE) based on mesoporous sorbent was used for cleaning-up the plasma samples. DMSPE coupling with liquid chromatography with tandem mass spectrometry (LC-MS/MS) was implemented for the determination of busulfan dosage in plasma samples. The linear range was found from 10 to 2000 ng/ml. The precision and accuracy were found better than 15% according to Food and drug Administration (FDA) guideline. This method was successfully used to determine the busulfan in patients administrated busulfan as part of the preparative regimen for bone marrow transplantation.

Pyranocoumarins from Zosima absinthifolia (Vent) Link roots.

Zosima absinthifolia (Vent) Link (Apiaceae) is a perennial herb indigenous to Iran. It has been used as a medicinal plant from ancient time in Iran, Turkey and Pakistan. In the present work, air-dried and powdered plant roots were extracted with n-hexane, dichloromethane and methanol, respectively, using Soxhlet apparatus. The dichloromethane extract was subjected to vacuum liquid chromatography (VLC) and preparative thin layer chromatography (P-TLC) to yield two pyranocoumarins, aegelinol and agasyllin. The antimicrobial assay was performed using agar dilution method. The results showed that purified compounds have modest to weak antibacterial and antifungal activity.

Comparison of treatment outcomes of infertile women by clomiphene citrate and letrozole with gonadotropins underwent intrauterine insemination.

This study was designed to compare the effect of clomiphene and letrozole in ovulatory stimulation in infertile women under intrauterine insemination who referred to Mahdiyeh infertility clinic during 2008-2009. 106 infertile women were randomly divided into two equal groups. Patients were treated with 5 mg of letrozole daily (in letrozole group) or 100 mg of clomiphene citrate daily (in clomiphene group) for five days starting on day 3 of their menses. Dose and time of FSH was similar in the two groups. Number of follicles, endometrial thickness, Pregnancy rate and prevalence of complications were compared in the two groups. Mean (±SD) of age in letrozole and clomiphene groups was 26.3 ± 3.9 and 25.2 ± 4.9 respectively (P=0.186). Average number of follicles was 2.5 ± 1.65 in letrozole group and 2.36 ± 1.4 in clomiphene group (P=0.764). ß-hCG was positive in 11 (20.8%) in letrozole and 12 (22.6%) in clomiphene groups (P=0.814). Pregnancy rate was 20.8% and 22.6% in letrozole and clomiphene group respectively (P=0.814). There was no difference in rate of abortion between groups. Endometrial thickness (ET) at the time of hCG administration in the letrozole (6.8 ± 1.5 mm) and in clomiphene (6.6 ± 1.2 mm) (P=0.615). But ET>7.4 mm was found in 2 cased (3.8%) in clomiphene group and 12 cases (%22.8) in letrozole groups (P=0.01). It appears that letrozole and clomiphene have similar outcome infertile women under intrauterine insemination and these drugs are good alternative for each others.

Intrauterine insemination with husband semen: an evaluation of pregnancy rate and factors affecting outcome.

PURPOSE: The aim was to determine pregnancy rate following intrauterine insemination (IUI) and its associated factors in a university assisted reproductive technique center in Tehran, Iran. METHODS: A retrospective analysis of 350 IUI cycles with ovarian stimulation by clomiphene citrate and/or gonadotropins was performed. RESULTS: The overall pregnancy rate was 22% (77/350). Of the 77 pregnancies, 88.3% resulted in live birth, 7.8% in spontaneous abortion, 2.6% in blighted ovum and 1.3% were ectopic. Logistic regression analysis revealed three predictive variables as regards pregnancy: number of the treatment cycle (OR:3.5 CI:1.9 - 6.4 p:0.006), duration of infertility (OR:2.1 CI:1.2 - 3.7 p = 0.001) and age (OR:2.15 CI:1.1 - 4.4 p = 0.04). Pregnancy rate did not have any independent relation to sperm count, type of infertility, number and size of follicle and side of ovulatory ovary. CONCLUSION: Our results indicate that clomiphene citrate and/or gonadotropins IUI is a convenient and useful treatment option in women with younger age ( <30 years) and fewer treatment cycles and fewer infertility duration (4 years).