Estimation Methods for Infertility Treatment Success: Comparison of Four Methods.

Objective: To analyze and compare four methods for estimating the chance of treatment success in infertile couples. Materials and methods: In a retrospective cohort study, information on demographic and clinical features, including age, body mass index (BMI), duration of infertility, semen analysis, previous history of treatment and clinical examination of infertile couples were analyzed. Treatment success (childbearing) was calculated with four methods as live birth ratio, conditional probability and survival analysis (life table and Kaplan-Meyer method) and results are compared. Results: The fertility ratio for the first treatment cycle was 29.72% which decreased to 23.13% by total treatment cycles. The success rate was 75.4%. With conditional probability calculation at the end of the five treatment cycles. With the life table method in a five-year period, the probability for live birth was 78% and by Kaplan-Meyer method 73.1% and the median of treatment time was 562 days. Conclusion: Calculation of infertility treatment success rate by only simple live birth ratio of childbearing couples is associated with underestimation. Using the conditional probability method reduces that underestimation, but it is not considered the censored cases in the treatments. It seems life table (as a proxy of survival analysis) presents the closest estimation to clinical facts with considering the repetition of the treatment cycle and the duration of treatment.

The Success Rate and Factors Affecting the Outcome of Assisted Reproductive Treatment in Subfertile Men.

BACKGROUND: This study was conducted to evaluate the success rate of male infertility treatment and the factors affecting its outcome. METHODS: In a historical cohort study, from Mar 2013 to Mar 2014, 323 couples with male factor were investigated. Couples had treated with IUI or/and ICSI were included randomly. Assisted reproduction technology (ART) outcome (treatment success) was defined as a live birth. Age, duration of infertility, type of infertility, treatment history and clinical examination results were investigated. The logistic regression and survival analysis were applied. RESULTS: The average of men age, duration of infertility and BMI were 33.5, 4.7 (yr) and 26.6 (kg/m2) respectively. 87.9% of men have primary infertility and average duration of treatment was 14.1(month). Previous treatment, type of infertility, treatment method, man's BMI, normality of sperm and sperm head were important variable that affecting outcome. The rate of live birth in the first attempt was 29.7%, and 44.9% of the couples succeeded to give live birth after several treatment cycles. Couples who had no previous history of treatment were 8.5 times more successful in live birth. The Cox analysis showed that "BMI of man" and percentage of "Sperm with normal head" are predictors that had a significant effect on live birth. CONCLUSION: Live birth in the first treatment cycles was influenced by four variables but two other variable were affecting several treatment cycles outcome. The chances of successful treatment were higher with taking into account the length of time and having live birth was determined as 78% for five years of continuous treatment.

Preimplantation High-Resolution HLA Sequencing Using Next Generation Sequencing.

Hematopoietic stem cell transplantation (SCT) is the only therapeutic option in a number of heritable hematologic disorders and hematologic cancers. Many parents and families fail to find an HLA-identical donor for their affected family member. In such cases, conceiving for a "savior baby" remains the only option, especially in countries without access to national registries. By means of next generation sequencing (NGS) techniques, in a single experiment on single-cell products of in vitro fertilization, a healthy HLA-identical embryo can be implanted in the uterus of a concerned mother. The patient can therefore benefit from cord blood SCT along with confirming that the fetuses are not suffering from the heritable disorder. This study is an attempt to study the feasibility of preimplantation HLA sequencing on single blastomeres using NGS. Two couples who had previously undergone preimplantation genetic diagnosis of ß-thalassemia and their overall 10 embryos were studied and their 5 HLA loci were typed in high resolution through multiple displacement amplification and NGS of single cells. For 88.9% of the 90 HLA alleles, conclusive HLA typing in 4 digit sets was made. HLA alleles were typed; 1 ambiguity in the allelic group and 4 ambiguities in the protein level were observed that were then unraveled by haplotype analysis. Amplification efficiency was 93.3% with an allele drop-out (ADO) rate of 22.2% (6 alleles dropped from a maximum of 27 possible ADOs). In this study the feasibility of a new method of preimplantation HLA sequencing via combining the state-of-the-art techniques used in single-cell whole genome amplification, preimplantation genetic diagnosis, and high-resolution HLA typing by NGS has been shown. This method can make preimplantation HLA sequencing a practicable technique in families desperate for an HLA-matched donor.

HIV/STI RISK-TAKING SEXUAL BEHAVIOURS AND RISK PERCEPTION AMONG MALE UNIVERSITY STUDENTS IN TEHRAN: IMPLICATIONS FOR HIV PREVENTION AMONG YOUTH.

Recent evidence indicates a rising trend in premarital sexual activity among young people in Iran. However, little is known about the extent to which young people's sexual behaviours expose them to HIV and STI risks. This study aimed to assess HIV/STI-related sexual risk-taking behaviours (correlates and determinants) and HIV/STI risk perception among male university students in Tehran. A representative sample of male university students (N=1322) studying in government and private Tehran universities completed an anonymous questionnaire survey in 2013-14. Respondents were selected using two-stage stratified cluster sampling. About 35% of respondents had ever had premarital sex (n=462). The majority (about 85%) of the sexually experienced students reported having multiple sexual partners in their lifetime. More than half (54%) reported inconsistent condom use over the previous month. Despite this exposure to HIV/STI risk, the respondents had a very low level of HIV/STI risk perception. Only 6.5% were highly concerned about contracting HIV over the previous year, and an even lower percentage (3.4%) were concerned about contracting STIs in the near future. Early sexual debut (<18 years), studying in a private university, ever watching pornography and work experience were found to be significant predictors of having multiple sexual partners. Younger age at sexual debut, having one lifetime sexual partner and poor HIV knowledge were significant predictors of inconsistent condom use over the preceding month. HIV prevention programmes among Iranian youth need to focus on the postponement of first sex and enhancement of HIV/STI knowledge in the light of increasing access of young people to pornography.

Comparison of three methods for mitochondria isolation from the human liver cell line (HepG2).

AIM: The aim of this study was to evaluate and compare three available methods for mitochondrial isolation from a human cell line to predict the best method for each probable application. BACKGROUND: Organelle isolation is gaining importance in experimental laboratory settings. Mitochondrial dysfunction may affect tumorgenesis process. There are some evidences that transplantation of healthy, intact and active mitochondria into cells containing defective mitochondria may reduce the proliferation. Therefore, isolated mitochondria could be considered as an effective tool for assessment and management of mitochondrial related disorders. PATIENTS AND METHODS: Mitochondrial isolation from the human liver cell line (HepG2) was performed using two commercially available kits, including Qproteome (Qiagen) and MITOISO2 (Sigma-Aldrich), as well as a manual method. Integrity of inner membrane of mitochondria was assessed by JC-1 staining. Activity of isolated mitochondria was evaluated by DCFH-DA staining, and total yield of isolated mitochondria determined by micro-Lowry method. Finally, relative quantification using Real-time PCR was conducted to compare the mtDNA copy number of mitochondria isolated by three different methods. RESULTS: Compared to other methods, manual kit resulted in higher yields of total amount of mitochondrial protein and mtDNA copy numbers. Isolated mitochondria by Qproteome kit, showed a higher activity. Finally, the integrity of inner-membrane of isolated mitochondria was significantly higher in Qproteome when compared with the other two methods. CONCLUSION: Due to differences in quality, quantity and activity of isolated mitochondria using three techniques discussed here, the method in which best-suited to each research project should be selected according to the distinct features of isolated mitochondria.