Successful intravaginal culture of human embryos for the first time in Pakistan--an experience at the Sindh Institute of Reproductive Medicine, Karachi.

Approximately 10-15% of the married couples remain childless. Conventional in vitro fertilisation (IVF) has been the treatment option for most of these cases. However, it is expensive and only available to a small fraction of the infertile population. The Sindh Institute of Reproductive Medicine (SIRM), Karachi, Pakistan, has introduced a new method of IVF for the first time in Pakistan. Intravaginal culture (IVC) is a simple, reliable and cost-effective alternative to IVF. The procedure can be performed in an office set-up with minor capital equipment. Here, we report a case of a subfertile woman treated by IVC, which resulted in successful fertilisation of the eggs (fertilisation rate 71.42%) and embryo development, with an intrauterine pregnancy. Since this is the first successful case of IVC in Pakistan, comparative success rate and take-home baby rate have yet to be established.

Cryopreservation of embryos by vitrification at a private sector reproductive medicine facility in Karachi.

OBJECTIVE: To assess the survival of freezing cleaved human embryos through vitrification. METHODS: The prospective study was conducted at the Karachi-based Sindh Institute of Reproductive Medicine between June 2008 and June 2009. The cryopreservation of embryos being a new technology in Pakistan, only 19 couples, picked through convenience sampling, comprised the study population. The couples were treated for infertility by in virto fertilisation (IVF) or intracytoplasmic sperm injection (ICSI); 125 surplus embryos were vitrified. Subsequently, 15 embryos were thawed, and transferred in a controlled cycle. SPSS version 11 was used for statistical analysis. RESULT: After the surplus embryos were vitrified and subsequently thawed and transferred, the survival of the embryos was assessed by the number of blastomeres that were intact. The overall embryo survival rate was (14/15) 93.33%. CONCLUSION: Vitrification is a simple procedure that requires less time and is likely to become safer and more cost-effective with time. Survival rate after thawing and preserving is high, but comparative success rates in terms of pregnancy and taking-home-baby rates are yet to be established in Pakistan.

Clinical results of the intra-cytoplasmic sperm injection with surgically retrieved sperms in azoospermic men at Sindh Institute of Reproductive Medicine, Karachi.

OBJECTIVE: To analyse the result of intra-cytoplasmic sperm injection (ICSI) by using surgically retrieved sperms in the azoospermic male. METHODS: Eight (9 cycles) azoospermic men were given the intra-cytoplasmic sperum injection after sperms were collected through surgical retrieval either by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) at the Sindh Institute of Reproductive Medicine (SIRM), Karachi, which is an assisted reproductive technology centre. Fertilisation, cleavage and pregnancy rates were calculated in their spouses with surgical sperm collection (SSC) and intra-cytoplasmic sperm injection. The study related to the period between September 2007 and February 2009. Statistical analysis were done using SPSS version 11.0. RESULT: After the intra-cytoplasmic sperm injection procedure, fertilisation rate of 72.72% and cleavage rate of 78.12% were achieved in the eight patients. A total of 3 (33.33%) clinical pregnancies were obtained through the transfer of embryo from surgical sperm collection in the azoospermic men. CONCLUSION: Intra-cytoplasmic sperm injection with surgical sperm collection proved to be the only successful treatment for men with azoospermia. It gave 33.33% chances of fatherhood in men who were previously labelled infertile. Results were found to be promising and are expected to improve with time and experience.

Two novel mutations in exon 2 of bone morphogenetic protein (BMP) 15 gene in Pakistani infertile females.

OBJECTIVE: To determine the proportion of fertility in Pakistani infertile females and discover if there are considerable connection among BMP15 gene polymorphism, follicle maturation and hormonal regulation in Pakistani infertile females. METHODS: All selected participants were initially examined through follicle-stimulating hormones (FSH), luteinizing hormone (LH), thyroid-stimulating hormone (TSH), Prolactin, and Trans-vaginal scan (TVS). BMP15 gene polymorphism among infertile and fertile females was done by extracted Genomic DNA from whole blood. Sanger sequencing was performed for the identification of mutation in exons-intron boundaries of the BMP15 gene. Bioinformatics tools were used to assess the protein structure. RESULTS: The total five mutations including two novel missense variants of BMP15 in exon 2, whereas three previously reported i.e. two cosmic mutations (c.615delC), (c.584InsG) and one frame shift mutations (c.635delA) were also observed. The first novel mutation was found at (c.1038InsGG) (p.346Gln < Gly) in which the insertion of GG at DNA position 1038 of exon 2 resulting in a substitution of glutamine into glycine at 346th amino acid of BMP15 protein. The second novel variant (c.1049delT) (p. Ser334Pro) was also observed in exon 2 of the BMP15 gene, which substituted serine into proline at 334th amino acid of the BMP15 protein. CONCLUSION: It is concluded that there are various missense mutations present in exon 2 of the BMP15 gene of Pakistani infertile females, consequently expected function of protein changes due to change in codons of amino acids. Provean and SIFT suggest the two novel variants as potentially deleterious. Although three other variants were also found in Pakistani infertile females which were previously reported. These mutations may result in early blockage of folliculogenesis and ovaries become streaky. Further research is required to resolve the actual allusion of these variations in the BMP15 gene.

Post dural puncture headache after spinal anaesthesia for caesarean section: a comparison of 25 g Quincke, 27 g Quincke and 27 g Whitacre spinal needles.

OBJECTIVE: To compare the frequency and severity of post dural puncture headache in obstetric patients using 25G Quincke, 27G Quincke and 27G Whitacre spinal needles. DESIGN: Comparative, randomized, double-blind, interventional study. PLACE AND DURATION OF STUDY: Liaquat University Hospital Hyderabad from October 2005 to December 2006. PATIENTS AND METHODS: 480 ASA I-II full term pregnant women, 18 to 45 years of age, scheduled for elective Caesarean section, under spinal anaesthesia, were randomized into three groups: Group I (25G Quincke spinal needle: n=168), Group II (27G Quincke spinal needle: n=160) and Group III (27G Whitacre spinal needle: n=152). Spinal anaesthesia was performed with 1.5-2.0 ml 0.75% hyperbaric bupivacaine using 25G Quincke spinal needle (Group I), 27G Quincke spinal needle (Group II) and 27G Whitacre spinal needle (Group III) at L3-4 inter-vertebral space. Each patient was assessed daily for four consecutive days following Caesarean section. Frequency and severity and of postdural puncture headache (PDPH) were recorded. Data were analyzed using SPSS-11. RESULTS: Frequency of PDPH following the use of 25G Quincke (Group I), 27G Quincke (Group II) and 27G Whitacre (Group III) spinal needles was 8.3% (14/168), 3.8% (6/160) and 2.0% (3/152) respectively. In Group I, PDPH was mild in 5 patients, moderate in 7 patients and severe in 2 patients. In Group II, it was mild in 2, moderate in 3 and severe in 1 patient. In group III, it was mild in 2 and moderate in 1 patient. Severe PDPH did not occur in Group III. Most of the patients with PDPH developed it on 1st and 2nd postoperative day. CONCLUSION: When using a 27G Whitacre spinal needle, the frequency and severity of PDPH was significantly lower than when a 25G Quincke or 27G Quincke needle was used.