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Lab Attendance

Lab 1 --Z3415242 (talk) 12:45, 6 August 2014 (EST)

http://www.ncbi.nlm.nih.gov/pubmed

Pubmed

PMID2508416

<pubmed>25084016</pubmed>

Lab 2 attendance --Z3415242 (talk) 12:25, 13 August 2014 (EST)

Lab Assessment 1

Article 1

<pubmed>PMC3985938</pubmed>

The above article has suggested that vitamin D may have an impact on human reproduction. The aim of the investigation was to see if vitamin D levels can help predict rate of implantation and clinical pregnancy in infertile women after having IVF (in vitro fertilisation).

Method

The following method was used to perform the investigation. Vitamin D status as determined by serum 25(Oh) D levels were evaluated amongst a group of woman who went for IVF. 182 women were recruited and of that 172 were included for analysis. Woman who were aged 18-41 years, had FSH level 12 IU/L or lower and were able to provide informed consent were included in the study. Third party reproduction cycle, those with uterine abnormalities and language barriers were excluded. The participants underwent a normal IVF cycle. Standard agonist and antagonist protocols were used. Busrelin acetate was used in the agonist protocol and cetrolix acetate or ganirelix acetate was used in antagonist protocol. Controlled ovarian hyper-stimulation was achieved by recombinant or purified FSH with or without LH or human menopausal gonadotropin. The doses of the medication were determined on individual basis.

Ovarian response was examined by serial transvaginal ultrasonography as well as through serum lutenizing and estradiol assay. Serum25 (OH) D samples had been collected before oocytes were retrieved. Nuclear maturation was triggered with HcG.Oocytes were retrieved under ultrasound guidance transverginally 36-38 hours following injection of HCG. On day 3-5 after fertilisation, embryo was transferred and vaginal progesterone was used for luteal phase support.

Findings

Patients were classified into sufficient (greater than 75nmol/L of vitamin D) and insufficient levels(less than 75nmol/L). From results, those from sufficient group showed to be more likely to undergo embryo transfer on day 5( 71.8% as statistically analysed) compared to woman from insufficient group ( 58.9%). Higher clinical pregnancy rate per cycle was discovered amongst woman with sufficient level compare to woman with insufficient level. They also found higher clinical pregnancy per embryo transfer in the sufficient group of woman. Implantation rate was observed to be higher in sufficient group however this was not statistically significant. Multivariable analysis adjusted for age, BMI and day 5 verses day 3 embryo transfers showed that serum level was an independent predictor of clinical pregnancy. Woman in this study with sufficient vitamin D level had a higher rate of clinical pregnancy following IVF compare with woman with insufficient level. Vitamin D supplement could provide an easy and cost effective way of improving pregnancy rate in woman undergoing IVF.

Article 2

<pubmed>24834703</pubmed> Properties of Zona Pellucida are believed to be important for oocyte cytoplasmic maturation and meiotic spindle for chromosomal alignment and proper separation of maternal chromosome. This study aimed to find the possible effect of ZP birefringment properties and mitotic spindle visualisation and localisation as a predictor of IVF outcome.

Method

This was a perspective study performed over 16 months at Medical University of Bialystok Poland where 51 patients undergoing IVF-ET (In vitro fertilisation and embryo transfer), intracytoplasmic sperm injection (ICSI) were included. Of 51 couples 39 had unexplained infertility and 12 had mild male factor. Two different methods for controlled ovarian hyper stimulation were used.

1. Long protocol (n=32) – The drug used was GNRH agonist Diphereline SR. It is a recombinant form of FSH.

2. Antagonist protocol (n=19). FSH and GNRH antagonist was used.

Ovulation was induced with HCG (human chorionic gonadotropin) in all patients. Oocyte retrieved transvaginally under ultrasound guidance 36 hours after HCG injection. Oocyte denudation was performed using hyaluronidase. Before intracytoplasmic sperm injection (ICSI) non invasive measuring with Olympus X7 inverted microscope was performed. Through this 228 cumulus oophorus were selected to be examined. 31were excluded due to inadequate oocyte maturation. Examination was done in different ways. 1. ZP birefringence was evaluated in a scale of 0degree to 4degree, 2. ZP autoscoring was performed, 3. ZP numeral autoscoring was performed, 4.MS visualisation, 5.Spindle detection and localisation of MS in relation to polar body.

Detection was grouped as: 1. Oocyte with spindle forming at less than 30 degree 2. 30-45 degree 3. 45-120 degree and 4. More than 120 degree, to PB. Oocytes whose spindles were located to be in another position where rotated in such way that the injected needle did not penetrate it. After this, oocytes were able to be divided into 3 groups. Group1: Embryo selected for transfer and outcome- day 14 after ET positive pregnancy test. Group 2: Embryo selected-day 14 after ET negative pregnancy test, Group 3: Embryo was not selected for transfer.

FINDINGS

Autoscores obtained in all three groups were nearly identical and no statistically difference were found in study groups. MS localisation in relation to PB was very close ( <450) in 70.9% of oocytes. The rate of MS positive oocyte was highest in group with pregnancy but statistically not significant. Between oocyte selected and non-selected for ET, no statistically significant difference is numeral score of ZP was found. ZP manual evaluation indicated stronger bifringement when pregnancy was not achieved. The rate of MS positive oocyte was higher in group with pregnancy. None of this was statistically significant so it was concluded that polarisation microscopy imaging and rating of ZP and MS cannot be a direct predictor of IVF outcome.

Lab Online Assessment 2-IMAGE

Adriamycin treated mouse shows association of abnormal notochord branching and fore gut abnormality

A)Different angle of a control embryo showing endoderm, notochord and floor plate in white. (B-H)- Adriamycin treated embryos immunostained for HNF3β demonstrating morphological abnormalities. Abnormal notochord branching is indicated with red arrow while different fore gut abnormalities are indicated with green arrow.

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