The spermatozoa were first seen in ejaculates in the 17^th century. However, the basic mechanisms of human fertilization have been only fully understood after the discovery of ovum in 1827. As a result, the interest in developing technologies for semen analysis arose from the early 1900s. Indeed, standard methodologies for semen analysis were designed mostly along the first half of the 20^th century. Before the 1930s, semen analysis was nearly unavailable clinically, since there were still no robust methodologies for assessing sperm characteristics, as well as to set up standard references that could be able to assess the reproductive capacity of men. However, joining some methodologies reported from 1910 up to 1930, standardization was attained and thereby semen analysis increasingly assumed its role in laboratory practice for investigating men in barren marriage. This article aims in reviewing historical backgrounds on the semen analysis, up to its insertion in laboratory practice. Emphasis is given to the major studies that contributed either directly or indirectly in developing the earliest routine for the semen analysis.

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The azoospermia factor (AZF) region on the Y chromosome consists of genes required for spermatogenesis. Among the three subregions, the AZFc subregion located at the distal portion of AZF is the driver for genetic variation in Y chromosome. The candidate gene of AZFc is known as deleted in azoospermia gene, which is studied with interest because it is involved in germ cell development and most frequently deleted genes leading to oligozoospermia and azoospermia. Recently, two partial deletions in AZFc gr/gr and b2/b3 are characterized at the molecular level which showed homologous recombination between amplicons, affecting spermatogenesis process. There are novel methods and commercially available kits for accurate screening and characterization of microdeletions. It is important to detect the AZFc microdeletions through genetic screening and counseling those infertile men who planned to avail assisted reproduction techniques such as undergoing intracytoplasmic sperm injection or in vitro fertilization.

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Background: Premature progesterone rise (PPR) has long been implicated as contributing to implantation failure. Despite the use of gonadotropin-releasing hormone (GnRH) analogues, subtle increases in serum progesterone (P₄) levels beyond a threshold progesterone concentration were observed on the day of trigger in controlled ovarian hyperstimulation cycles. Aims: The purpose of the study was to evaluate the incidence of PPR on the day of trigger in conventional IVF/ICSI cycles and its impact on clinical pregnancy rate. Settings and Design: A total of 235 patients undergoing conventional IVF/IVF–ICSI by fresh embryo transfer cycles from January 2016 to December 2016 at the infertility unit of a tertiary care hospital were prospectively analyzed. Material and Methods: Patients included in the study were subjected to GnRH agonist long/antagonist protocol. Ovulation induction was given with rFSH and/or HMG in both the protocols. The cutoff for defining PPR was P₄≥ 1.5 ng/ml, and an analysis of the role of P₄on clinical pregnancy rate was performed. Statistical analysis was performed with the Statistical Package for the Social Sciences trial version 23.0 software for Windows and Primer software. Results and Conclusion: The overall clinical pregnancy rate per embryo transfer was 30.6%. The clinical pregnancy rate in the patients with P₄ <1.5 ng/ml was significantly higher than those with elevated levels, P₄≥ 1.5 ng/ml (33.3% vs. 12.9%; P = 0.037). Premature progesterone elevation in ART cycles is possibly associated with lower clinical pregnancy rates.

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Objective: Is a single dose of gonadotropin-releasing hormone agonist (GnRHa) trigger to induce final oocyte maturation in polycystic ovarian syndrome (PCOS) undergoing in vitro fertilization (IVF) cycles with GnRH antagonist protocol sufficient to provide optimal oocyte maturity? Design: This is a prospective, randomized, double-blind, proof-of-concept study. Setting: This study was carried out at a tertiary care center. Material and Methods: A total of 125 patients diagnosed with PCOS defined as per the ESHRE/ASRM Rotterdam criteria (2003) undergoing IVF in antagonist protocol were randomized into two groups. Group A: single dose of GnRHa 0.2 mg, 35 h prior to oocyte retrieval, and Group B: 0.2 mg GnRHa 35 h prior to oocyte retrieval + repeat dose of 0.1 mg 12 h following the 1^st dose. 12 h post-trigger, luteinizing hormone (LH), progesterone (P4), and follicle-stimulating hormone (FSH) values were estimated. Statistical Analysis: Continuous variables were expressed as mean ± standard deviation and categorical variables as proportions where applicable. Independent sample t-test was used for continuous variables which were normally distributed and Mann–Whitney U-test for data not normally distributed. Chi-square test or Fisher's exact test was used for categorical variables where appropriate. Odds ratio (OR) with 95% confidence intervals (CIs) was calculated. In addition, receiver operating characteristic curve was used to evaluate the post-trigger LH, P4, and FSH values at 12 h as predictors of oocyte maturity. Main Outcome Measures: Primary outcome: maturity rate of the oocytes. Secondary outcomes: oocyte yield, fertilization rate, availability of good quality embryos on day 3, blastocyst conversion, OHSS rates, post-trigger serum LH (IU/L), FSH (IU/L), and P4 (ng/mL) levels implantation rate and clinical pregnancy rate. Results: A higher number of mature (metaphase II) oocytes were obtained in Group B compared to Group A (OR of 0.47; CI: 0.38–0.57; P < 0.01). Significantly a higher number of blastocysts were obtained in Group B than Group A (4.00 vs. 3.04; P = 0.023). The odds of clinical pregnancy per patient were higher in Group B (OR = 0.56; CI [0.27–1.24]), with a trend towards better clinical pregnancy in Group B than in Group A. Conclusions: A repeat dose of GnRHa trigger 12 h following the first dose probably by maintaining a sustained level of gonadotropins yielded a better maturity of oocytes, higher number of blastocysts, and a trend towards higher clinical pregnancy than a single dose in PCOS patients undergoing IVF in antagonist cycles.

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Objective: This study aims to study the difference in etiology and outcome in terms of implantation rate and abortion rate in fresh (self-stimulated) versus frozen (oocyte donation cycle) in vitro fertilization (IVF) and in transient versus persistent fluid. Material and Methods: This retrospective study was conducted in the Department of Reproductive Medicine of tertiary care center from January 2012 to November 2015. Data were collected retrospectively from the departmental files. Twenty-four patients from fresh IVF-stimulated cycles and 24 from frozen oocyte donation cycle with their endometrium prepared by hormone replacement treatment were included in the study. All patients selected in the study had grade-A embryo transfer of day 3–4 with maximum three embryo transferred. Pregnancy was defined by rising serum beta-human chorionic gonadotrophin levels performed after 14 days of embryo transfer and further confirmed by ultrasonographic visualization of gestational sac at 6 weeks. All biochemical pregnancies were included in implantation failure. All pregnant patients were followed till the termination of pregnancy and further noted as live birth or abortion. Results: Clinical pregnancy rate was seen more in self-stimulated cycle (62.5%) with live birth rate of 50% than hormone replacement treatment cycle, in which clinical pregnancy rate was 45.83% with live birth rate of 33.33%. Clinical pregnancy rate was highest in group with very less fluid in cavity (1–2 mm) 63% and with live birth of 52.63%. Clinical pregnancy was seen only in two patients of group B with anterior and posterior (AP) diameter of fluid in cavity of 2–3 mm with live birth of only one, whereas in group C, with AP diameter of 3–5 mm, none of the patient conceived. This difference was statistically significant. Clinical pregnancy rate was 65.62% in transient fluid accumulation with live birth rate of 53.25%, which was significantly higher than persistent fluid accumulation (P value − 0.0337 for pregnancy rate and 0.0312 for live birth rate). Conclusion: Fluid accumulation seen in fresh cycles are generally associated with better outcome because it may be associated with good prognostic factors – small AP diameter of fluid, with transient fluid accumulation and more with poly cystic ovarian syndrome as an etiological factor; however, in frozen cycle, it can be associated with poor outcome.

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Unconsummated marriage are mainly due to vaginismus or erectile dysfunction. They contribute to about 5% of couples in an infertility clinic. Their incidence is increasing in the metropolitan cities because of stressful lifestyles. Many of couples are advised intrauterine insemination as fertility treatment. However, home insemination is a simple, inexpensive and effective way of achieving pregnancy in such couples. We present the first study to document pregnancy rates of artificial home intravaginal insemination in couples with unconsummated marriage. Aim: To assess the pregnancy rates (PR) with intravaginal insemination (IVI) by couples with nonconsummation of marriage. Setting: Infertility outpatient clinic, New Delhi. Design: Retrospective analysis. Material and Methods: 55 couples of unconsummated marriage were evaluated in a period of two years. Group 1 contains 29 patients aged between 20 to 33 years; group 2 includes 14 patients aged between 33 to 36 years and group 3 includes 12 patients aged more than 36 years. Result(s): Unconsummated marriage was caused by vaginismus in 67% of the cases, erectile dysfunction in 22% and premature ejaculation in 11%. Clinical pregnancy rate was 69% in group 1; 43% in group 2 and 25% in group 3 after 6 cycles of AI. Conclusion(s): Intravaginal insemination is a simple, short, economical, effective and painless procedure for conception in nonconsummation of marriage.

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Background: Turner syndrome (TS) is a chromosomal disorder associated with dysmorphic features and comorbidities, with recent trends focusing on early diagnosis for adequate management. Aim: The aim is to study the age and mode of presentation of TS, associated comorbidities and look for any correlation with the genotype. Material and Methods: This was a retrospective analysis of girls with TS attending the endocrinology clinic of a tertiary care center. Their age, mode of presentation, and clinical features were noted. All participants underwent ear examination, echocardiography, and ultrasonography of the abdomen. Laboratory investigations included serum T4, thyroid-stimulating hormone, thyroid peroxidase antibodies, follicle-stimulating hormone, fasting, and 2-h plasma glucose after 75 g glucose load and a karyotype. Simple descriptive statistical methods were used. Results: Seventeen cases of TS were seen with a median age of presentation of 18 years (range 14–42 years). Primary amenorrhea was the most common reason for seeking medical attention (76.4%) followed by short stature and diabetes mellitus (11.8% each). The mean height at presentation was 137.5 ± 5.4 cm. Monosomy of X chromosome (45,X) was the most common karyotype obtained in 58.8% of the patients, followed by 45,X/46, XX in 17.6%, 45,X/46X,i(X)(q10) in 11.8%, and 45,X/47,XXX and 46X,delXp11.2 in 5.9% patients each. Bicuspid aortic valve was seen in two patients having a 45,X/46,XX karyotype. Conclusion: Primary amenorrhea is the most common presenting feature in girls with TS leading to a delayed age of presentation. Short stature and dysmorphic features are often overlooked in infancy and childhood due to socioeconomic factors. This late age of presentation is a cause of concern as early detection and management is important for height outcomes, bone health, and psychosocial support. Assessment of comorbidities becomes important in this setting.

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Swyer syndrome with complete gonadal dysgenesis is associated with an absence of testicular differentiation in a phenotypic female with a 46, XY karyotype. A 14-year-old unmarried girl was referred with complaints of primary amenorrhea and nondevelopment of breast. Her built was normal. Examination of her secondary sexual characteristics revealed no breast development, absent axillary hairs, and sparse pubic hairs. External genitalia was of female type. Karyotype showed genotype of 46, XY. Magnetic resonance imaging revealed hypoplastic uterus with absent fallopian tubes and ovaries. A diagnosis of Swyer syndrome was made. Laparoscopy showed infantile uterus, normal fallopian tubes, and streak gonads. Laparoscopic removal of streak gonads was done as there is a risk of gonadoblastoma in such cases. The patient was started on hormonal replacement therapy. Swyer syndrome results mainly due to mutation in certain genes such as SRY gene, which leads to failure of development of testis.

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Background: About 15%–20% of couples get affected by recurrent miscarriages (RM) and chromosomal abnormality in one partner affects 3%–6% of RM couples. Aims: The present study aimed to determine the prevalence of cytogenetic anomalies in couples with RM. Settings and Design: A case–control study was undertaken, in which 243 couples who had experienced 2 or >2 miscarriages were investigated for chromosomal abnormalities and compared with 208 healthy, age-matched control couples who had at least one healthy live born and no history of miscarriages. Material and Methods: Peripheral blood (PB) lymphocytes were cultured using PB-Max Karyotyping medium (GIBCO) for chromosomal analysis and 20 metaphases were analyzed for each individual. Statistical Analysis: Student's t-test was used for statistical evaluation and P < 0.05 was considered statistically significant for all instances. Results: The current study revealed 3.1% RM cases showing structural chromosomal aberrations, of which balanced translocations and Robertsonian translocations constituted 66.7% and 26.7% cases, respectively, while inversions constituted 6.7% abnormal RM cases. Polymorphic variations were observed in 1.9% RM patients and 1.2% controls as well. However, the number of abortions were significantly more (P = 0.027) in male carriers of balanced translocations as compared to female carriers in the RM group. There was no significant difference for age (P = 0.539) between RM women and control women. Conclusions: Although similar studies exist in literature, our study is the first of its kind from our region that has compared the chromosomal anomalies between the RM group and the control group. We observed 3.1% of balanced translocations and an increased number (though nonsignificant) of polymorphic variations and satellite associations in the RM group as compared to the control group.

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Background: The vascularization status of ovarian follicles affects reproductive competence of oocytes and in turn embryo quality by regulating its oxygen supply. Transvaginal power Doppler ultrasound can noninvasively map this vascularity of the ovarian follicles. Thus, we aimed to study the association of perifollicular vascularity and pregnancy outcome in women while on treatment for an in vitro fertilization cycle. Material and Methods: A prospective study on 200 participants evaluated the vascularity of 1008 follicles on the day of oocyte retrieval to outline a map depicting perfusion of each follicle. The vascularity was graded based on percentage of the perifollicular outline in the map depicting vascularity which was Grade 1 ≤25%, Grade 2 26-50%, Grade 3 51-75%, Grade 4 76-100%. Results: Of 1008 follicles aspirated, only 733 follicles were analyzed as per the exclusion criteria. Grades III and IV follicles were high vascular grade follicle whereas Grades I and II were low perfused follicles. Six hundred and twenty-seven oocytes were retrieved from 733 follicles with majority from Grade III and IV vascularity (75.8%: Grade III and IV vs. 24.2%: Grade I and II). The number of oocytes exhibiting maturity and their fertilization rates were significantly higher in high vascularity follicles. Three hundred and forty-one Grade I embryos formed and 89.1% were from better-perfused follicles versus 10.9% from lower ones. Conclusions: The association between perifollicular perfusion and follicular oxygenation and oocyte maturation does exist which ultimately gets translated to quality of embryos. If other confounding factors such as endometrial receptivity and transfer technique are controlled, it influences the implantation potential too.

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Objective: To determine the efficacy of the stair-step protocol (SSP) using clomiphene citrate (CC) in patients with polycystic ovary syndrome (PCOS) and compare it with traditional regimen. Design: This was randomized control trial. Setting: Infertility Clinic. Patient(s): Sixty infertile PCOS women. Intervention(s): Patients were randomized into the study (SSP – 30 patients) and control group (traditional protocol – 30 patients). In the SSP, patients were treated with CC 50 mg/day for 5 days and in nonresponsive patients, the dosage was increased to 100 mg/day for 5 days in the same cycle. Maximum dose of 150 mg was given until the dominant follicle was generated. In control group, the dose increment in nonovulatory cases was done in subsequent cycle. Ultrasonography follow-up was done to detect ovulation. Main Outcome Measure(s): Ovulation rate and duration of treatment. Results: Ovulation (66.7% vs. 50% respectively) and pregnancy rates (26.7% vs. 15.7%) were similar between the stair step and the control group. The duration of treatment was significantly shorter in stair step compared to traditional protocol (17.23 vs. 53 days). CC 100 mg was the most effective dose for ovulation in either group. There were no significant differences in the systemic side effect. Conclusions: By using SSP, effective treatment is provided in significantly shorter time period without any detrimental effect on the ovulation and pregnancy rates.

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