Effects of Panax ginseng, zearalenol, and estradiol on sperm function.

BACKGROUND: Estrogen signaling pathways are modulated by exogenous factors. Panax ginseng exerts multiple activities in biological systems and is classified as an adaptogen. Zearalenol is a potent mycoestrogen that may be present in herbs and crops arising from contamination or endophytic association. The goal of this study was to investigate the impact of P. ginseng, zearalenol and estradiol in tests on spermatozoal function. METHODS: The affinity of these compounds for estrogen receptor (ER)-alpha and beta (ERα and ERß)-was assessed in receptor binding assays. Functional tests on boar spermatozoa motility, movement and kinematic parameters were conducted using a computer-assisted sperm analyzer. Tests for capacitation, acrosome reaction (AR), and chromatin decondensation in spermatozoa were performed using microscopic analysis. RESULTS: Zearalenol-but not estradiol (E2)- or ginseng-treated spermatozoa-decreased the percentage of overall, progressive, and rapid motile cells. Zearalenol also decreased spontaneous AR and increased chromatin decondensation. Ginseng decreased chromatin decondensation in response to calcium ionophore and decreased AR in response to progesterone (P4) and ionophore. CONCLUSION: Zearalenol has adverse effects on sperm motility and function by targeting multiple signaling cascades, including P4, E2, and calcium pathways. Ginseng protects against chromatin damage and thus may be beneficial to reproductive fitness.

Impact of kudzu and puerarin on sperm function.

The goal of this study was to investigate the impact of kudzu (Pueraria mirifica) and the isoflavone puerarin in functional toxicological tests on spermatozoa and to assess the affinity of extracts and pure isoflavones for estrogen receptor (ER)-alpha and -beta (ERα, ERß) in receptor binding assays. Capacitation, acrosome reaction and chromatin decondensation in spermatozoa were analyzed using microscopic analysis. Kudzu, but not puerarin, reduced motility of sperm. Puerarin reduced the percent spontaneous acrosome reaction in spermatozoa. The pathways used by kudzu that affect sperm function are not fully mirrored by puerarin. Puerarin, kudzu and its other phytoestrogenic components displayed preferential affinity for ERß, however the diverse effects of kudzu and puerarin on sperm function implicate the involvement of multiple signaling systems.

Accu-Beads as a quality control measure for manual and automated methods of measuring sperm concentration-an observational study.

PURPOSE: To determine the precision and accuracy of Accu-Beads and their utility as a quality control product for manual and automated measurements of sperm concentration. METHODS: This observational study was performed at an Assisted Reproductive Technology laboratory in a tertiary-care, university hospital. To simulate sperm concentration, bead concentrations were measured with the use of a manual and an automated method. RESULTS: The manual counts did not vary significantly from the automated counts regardless of the concentration. However, the counts did vary between lots of low concentration of Accu-Beads and between the two different types of fixed counting chambers. The two bead concentrations used in this study were below the 95 % confidence interval for the values listed by the manufacturer. CONCLUSION(S): In our laboratory, Accu-Beads met enough of the requirements of a good control material to be acceptable for daily quality control use, especially if we set our own ranges of acceptability for each vial of Accu-Beads. It is necessary to evaluate each new lot of Accu-Beads when they are received and again if they are used with a different counting chamber.

Temperature variations within and between incubators-a prospective, observational study.

PURPOSE: To determine if there is a temperature variation within and between incubators. METHODS: This prospective, experimental trial with external controls was performed at an Assisted Reproductive Technology laboratory in a tertiary-care, university hospital. Temperature values were taken at various locations within and between incubators. RESULTS: Even though they were both set to 37.0 °C, the same make and model incubators had significantly different internal temperatures. Temperatures differed significantly among top, middle and bottom shelves and between fronts and backs of shelves. CONCLUSION(S): We found temperatures differed within and between our front-loading incubators. Thus, laboratory personnel should evaluate their incubators for temperature variations within and between incubators and, if temperatures differ significantly, develop a plan to deal with discrepancies.

The heat is on: room temperature affects laboratory equipment--an observational study.

OBJECTIVE: To evaluate the effect of ambient room temperature on equipment typically used in in vitro fertilization (IVF). DESIGN: We set the control temperature of the room to 20 °C (+/-0.3) and used CIMScan probes to record temperatures of the following equipment: six microscope heating stages, four incubators, five slide warmers and three heating blocks. We then increased the room temperature to 26 °C (+/-0.3) or decreased it to 17 °C (+/-0.3) and monitored the same equipment again. We wanted to determine what role, if any, changing room temperature has on equipment temperature fluctuation. RESULTS: There was a direct relationship between room temperature and equipment temperature stability. When room temperature increased or decreased, equipment temperature reacted in a corresponding manner. Statistical differences between equipment were found when the room temperature changed. What is also noteworthy is that temperature of equipment responded within 5 min to a change in room temperature. CONCLUSIONS: Clearly, it is necessary to be aware of the affect of room temperature on equipment when performing assisted reproductive procedures. Room and equipment temperatures should be monitored faithfully and adjusted as frequently as needed, so that consistent culture conditions can be maintained. If more stringent temperature control can be achieved, human assisted reproduction success rates may improve.

Changes in long luteal protocol affects the number of days of stimulation: evolution of an assisted reproductive technology practice.

OBJECTIVE: To determine if the types of drugs used in controlled ovarian hyperstimulation influence the number of days of stimulation. STUDY DESIGN: This retrospective study of couples presenting at an assisted reproductive technology (ART) facility January 1997 through December 2007 included patients who were < 40 years of age, had a body mass index (BMI) of 20-35, used fresh sperm or oocytes, had > or = 3 embryos available on day 3, never had previous ART, and did not use a gestational carrier. Data analysis was limited to patients treated with long luteal stimulation. Data were separated into two groups: patients stimulated with urinary drugs and those stimulated with recombinant drugs. RESULTS: The groups were not different in age and day 3 follicle-stimulating hormone values, but were different in BMI, diagnoses, and fertilization methods. There were a significantly greater number of days of stimulation, total number of follicles produced, and peak estradiol level in the recombinant group. The recombinant group had less drug given per day, less endometrial thickness, and fewer embryos transferred. CONCLUSION: A change in our long luteal stimulation protocol appears to have lengthened the days of stimulation for our patients without altering pregnancy rate.

The Implementation of Virtual Instruction in Relation to X-ray Anatomy and Positioning in a Chiropractic Degree Program: A Descriptive Paper.

This article provides information regarding the introduction of virtual education into classroom instruction, wherein a method of classroom instruction was developed with the use of a computer, digital camera, and various software programs. This approach simplified testing procedures, thus reducing institutional costs substantially by easing the demand for manpower, and seemed to improve average grade performance. Organized files with hundreds of digital pictures have created a range of instructor resources. Much of the new course materials were organized onto compact disks to complement course notes. Customizing presentations with digital technology holds potential benefits for students, instructors and the institution.

Blastocyst rate and live births from vitrification and slow-cooled two-cell mouse embryos.

OBJECTIVE: The purpose of this study was to develop a closed vitrification system, compare vitrification to a slow-cooled cryopreservation method, and compare the pup rate between both methods using two-cell mouse embryos. DESIGN: Randomized, prospective animal study. SETTING: Hospital-based IVF practice. ANIMAL(S): B6C3F1 mouse embryos. INTERVENTION(S): Two-cell mouse embryos were cryopreserved using a slow-cooled or vitrification method and then thawed at a later date. The embryos were cultured and transferred to recipient females. MAIN OUTCOME MEASURE(S): Embryos were observed for blastocyst rate and pups were observed for phenotypic anomalies and weighed at 30, 60, and 90 days after birth. RESULT(S): Neither the blastocyst rate, pup rate, nor pup weights were significantly different when the two cryopreservation methods were compared. CONCLUSION(S): Because there were no differences in blastocyst rates, pup rates, or pup weights, we plan to further investigate the potential effects of vitrification on genotypic damage via the Comet Assay.

Effect of human granulosa cell co-culture using standard culture media on the maturation and fertilization potential of immature human oocytes.

OBJECTIVE: To determine whether culture of immature human oocytes with and without autologous cumulus cells (CC) in standard culture medium would provide additional oocytes for use in IVF procedures. DESIGN: Prospective study. SETTING: Hospital-based IVF practice. PATIENT(S): Sixty-one women undergoing 62 IVF-embryo transfer procedures between August 2004 and June 2005. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Oocyte maturation after culture with and without CC, oocyte fertilization, and zygote cleavage. RESULT(S): Culture with CC did not alter the maturation rate for metaphase I (M I) oocytes to the metaphase II (M II) stage. Germinal vesicle (GV) stage oocytes cultured with CC matured at a significantly higher rate than did GV oocytes cultured without CC. Although acceptable fertilization rates were obtained (62.1%), "usable" embryo yield was very low (13.0%). CONCLUSION(S): This study demonstrated good maturation of M I oocytes but poor maturation of GV oocytes in standard culture medium. Of the immature oocytes that did mature, were fertilized, and cleaved to embryos, only a very small number were actually suitable for patient use. Therefore, these extended culturing techniques were inefficient in maturing and providing additional oocytes/embryos for patient use. The use of specialized in vitro maturation medium may improve the utility of this process.

Quantitative and qualitative analysis of microorganisms in an assisted reproductive technology facility.

OBJECTIVE: To perform a quantitative and qualitative analysis of the environmental microorganisms associated with two assisted reproductive technology (ART) laboratories. DESIGN: Prospective descriptive study. SETTING: Two ART laboratories at Greenville Hospital System (GHS) University Medical Center in Greenville, South Carolina. PATIENT(S): None. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Quantitative and qualitative analysis of colony-forming units (CFU; bacteria and fungi) and particles from an embryology clean room and an andrology laboratory. RESULT(S): Within these laboratories, three of the sites sampled differed significantly in the mean number of CFU per plate. These sites included external laboratory surfaces, laboratory room air, and air within cell-culture incubators. The mean number of viable CFU per cubic meter of laboratory room air differed significantly between the two laboratories. A total of 23 unique microbial species were isolated from the andrology laboratory, whereas 10 unique species were isolated from the embryology clean room. CONCLUSION(S): The data herein suggest that the increased positive control measures used in the embryology clean room are effective in minimizing environmental biologic contamination.

Incubator management in an assisted reproductive technology laboratory.

OBJECTIVE: To study the effect of incubator management on assisted reproductive technology (ART) outcomes. DESIGN: Series of retrospective and controlled, randomized studies. SETTING: Tertiary care infertility practice. PATIENT(S): Mammalian gametes/embryos. INTERVENTION(S): Evaluation of human and bovine oocytes/embryos cultured in various environmental conditions. MAIN OUTCOME MEASURE(S): Fertilization and embryo development rate as well as clinical pregnancy rate (PR). CONCLUSION(S): Here we review the general topic of incubator management as it pertains to ART. Discussed within the context of this article will be our experiences as they relate to incubator management. Details as they apply to incubator environment also will include gamete/embryo positions within incubator, air quality, and quality control.

Assisted Reproductive Technology (ART) in the upstate: effectiveness of surgical sperm removal techniques in conjunction with ICSI to promote the success of pregnancy.

With aspiration of sperm directly from the epididymis (MESA) or extracted from the testis (TESA) and intracytoplasmic sperm injection, it is possible to provide men who are incapable of producing viable sperm in an ejaculate the ability to overcome this severe sperm defect and produce a successful pregnancy. In our practice, 10 of 26 ART cycles have had successful pregnancies with a total of 17 children being delivered. Thus, MESA/TESE techniques coupled with ICSI provide this population of patients with a viable option for producing offspring.

Economics of flexible vs. rigid catheters for intrauterine insemination.

A decision tree model was created to investigate pregnancy rates, as well as cost factors associated with rigid vs. flexible catheters. Although the flexible catheter costs more than the rigid catheter, it is associated with a higher pregnancy rate, which decreases costs of IUI treatment because fewer cycles are needed.

Comparison of pregnancy rates for two embryo-transfer catheters.

OBJECTIVE: To compare two flexible catheters and determine whether pregnancy rates (PRs) differed. DESIGN: Prospective, randomized pilot study. SETTING: Tertiary-care infertility practice. PATIENT(S): The subjects for this study were 100 women undergoing assisted reproductive technology (ART) cycles between September 2003 and October 2005. INTERVENTION(S): None. MAIN OUTCOME MEASURE: Clinical PR for Edwards-Wallace (Cooper Surgical, Shelton, CT) and Cook World (Cook Urological, Spencer, IN) embryo-transfer (ET) catheters. RESULT(S): The outcomes of pregnancy between the Edwards-Wallace and Cook World ET catheters were not significantly different. Use of the flare stimulation protocol resulted in a significantly lower PR compared to long luteal and antagonist protocols. No other variable affected PR. CONCLUSION(S): A 5% difference in PR was demonstrated between ET catheters; however, this study does not have the power to detect a significant difference in clinical PR between the two catheter groups. The pilot study demonstrates good design due to the lack of significant differences between the variables when stratifed by cather as well as pregnancy status. In a larger study, the flare protocol should be excluded.

Factors affecting assisted reproductive technology (ART) pregnancy rates: a multivariate analysis.

PURPOSE: To determine factors instrumental in achieving a clinical pregnancy in assisted reproductive technology (ART) patients. METHODS: This study included 205 women undergoing their first ART cycle. Univariate and multivariate analyses were performed for patient demographics, in vitro production data, and factors associated with embryo transfer. Odds ratios (OR) were performed where appropriate. RESULTS: Our analyses indicated that age (OR: 0.879), specific year in which the cycle was performed (OR: 2.959), and use of intracytoplasmic sperm injection (OR: 2.867) altered potential pregnancy rate. In addition, percent fertilization (OR: 1.028), number of embryos transferred (OR: 1.842), type of catheter used to transfer the embryos (OR: 0.377), presence of blood on the catheter (OR .414), and embryologist (OR: 2.338) also altered pregnancy rate. CONCLUSIONS: Our data indicate patients' age, use of the Cook catheter, and presence of blood on the catheter reduce pregnancy rates. Performing ART in 1999, using ICSI, increasing fertilization rate, increasing number of embryos transferred (albeit less desirable when the chance of multiple gestation may occur), and transferring embryos via a particular embryologist, improve pregnancy rates.

Defining the typical work environment for assisted reproductive technology laboratories in the United States.

OBJECTIVE: To determine time and staffing requirements as they relate to laboratory personnel in the field of assisted reproductive technology (ART). DESIGN: Descriptive study. SETTING: Assisted reproductive technology clinics in the United States. PERSONNEL: All personnel working in ART laboratories, which included laboratory directors and technicians (those laboratory personnel trained in specific areas of embryology, andrology, and endocrinology). INTERVENTION(S): None. MAIN OUTCOME MEASURES(S): Frequencies were determined for time and staffing requirements as they pertained to individuals and procedures (e.g., oocyte retrieval, embryo transfer, semen analysis, hormone assay, etc.) performed in embryology, andrology, and endocrine laboratories. RESULT(S): Two different surveys of ART clinics in the United States were combined in this report. The average technician worked 40 hours a week, whereas the average laboratory director worked 49 hours per week. The average embryologist performed 181 procedures (not cases) per year. The average andrologist performed 648 procedures (not cases) per year, while the average endocrinologist completed 2,673 procedures (not cases) per year. CONCLUSION(S): This is the first report to describe time and staffing requirements as they relate to the laboratory personnel in an ART facility.

Developing techniques for determining sperm morphology in today's andrology laboratory.

PURPOSE: To investigate three areas: the staining of spermatozoa; the Computer Assisted Semen Analysis (CASA); and the variability of technicians. METHODS: Staining experiment: sperm from 15 beef bulls were randomized to one of three staining protocols. CASA experiment: slides were evaluated using the Integrated Visual Optical System and compared to technician results. Variability of technicians: five laboratorians analyzed the same set of 20 slides six different times. RESULTS: Staining experiment: the size of the sperm increased in proportion to increased time and heat associated with each successive protocol. CASA experiment: coefficient of variation ranged from 18.3 to 101.7% (12 slides). Variability of technicians: the mean sperm morphology results ranged from 7.3 to 15% normal forms. CONCLUSIONS: Until laboratories adhere to the universal standard set by the World Health Organization to evaluate sperm morphology, a laboratory must rely on its own quality control to insure repeatable results.

Flexible versus rigid intrauterine insemination catheters: a prospective, randomized, controlled study.

OBJECTIVE: To prospectively compare pregnancy rates for couples undergoing IUI by a single healthcare provider with random assignment to either rigid (Tomcat; Kendall Sovereign, Mansfield, MA) or flexible (Soft Pass; Cook, Spencer, IN) catheters. DESIGN: Randomized, controlled study. SETTING: Tertiary care infertility practice. PATIENT(S): One hundred infertile women enrolled from a single infertility practice who met the inclusion and exclusion criteria for IUI were enrolled. Of the 100 patients, 95 underwent ovarian stimulation before IUI: 35 with clomiphene citrate alone, 60 with clomiphene citrate combined with a single dose of gonadotropins. INTERVENTION(S): Timed IUI was performed with either rigid (Tomcat) or flexible (Cook) insemination catheters. MAIN OUTCOME MEASURE(S): Pregnancy rate per cycle. RESULT(S): No differences were noted between groups for the following: age, length of infertility, day-3 FSH level, number of prior IUI cycles, total motile sperm count, days of abstinence, infertility diagnoses, insemination ratings, and stimulation protocols. No difference in pregnancy rates was observed between rigid and flexible catheter groups. CONCLUSION(S): There is no statistically significant difference between flexible and rigid catheters for IUI.

Changes in the freezing protocol for human zygotes alter embryonic development and pregnancy rates.

OBJECTIVE: To determine the effect of various freezing protocols on postthaw development and pregnancy rates resulting from transfer of human zygotes. DESIGN: Prospective study. SETTING: Tertiary care center. PATIENT(S): Couples undergoing assisted reproductive technology (ART) procedures who wished to have their excess zygotes cryopreserved. INTERVENTION(S): We cryopreserved zygotes with one of three protocols. MAIN OUTCOME MEASURE(S): Post-thaw survival and development of the zygotes as well as pregnancy rate after transfer of these zygotes. RESULT(S): A 3-minute hold time after seeding, followed by a final preplunging temperature of -180 degrees C, resulted in a clinical pregnancy rate of 28.6%. In contrast, a 15-minute postseed hold time and a -30 degrees C final chamber temperature resulted in a 37.3% clinical pregnancy rate. When we combined the protocols to provide a 15-minute postseed holding time and a -180 degrees C before plunging into liquid nitrogen, we achieved a 69.6% clinical pregnancy rate. CONCLUSION(S): By increasing the postseeding hold time and decreasing the temperature of the freezing chamber before plunging the zygotes into liquid nitrogen, significant improvements can be made in postthaw development and pregnancy rates.