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Biomedical subjects

W C Patton

Publications and source records attributed to W C Patton.

At least 37 records · Page 2Linked to original sources
PubMed1998

Enhanced fertility after heat-induced hyperactivation.

OBJECTIVE: To determine sperm hyperactivation, kinematic parameters, and fertilizing capacity after pretreating sperm at 40 degrees C for 4 hours. DESIGN: Prospective study involving pooled donor sperm that were colloid washed and incubated at either 23 degrees C (control) or 40 degrees C (heat-treated) for 4 hours as pretreatment. After incubation, analyses were performed with a computer-assisted sperm analyzer, whereas separate portions of sperm were evaluated with the sperm penetration assay at 37 degrees C. SETTING: Clinical and academic research environment. PATIENT(S): Cryopreserved-thawed sperm from different donors (n = 5). MAIN OUTCOME MEASURE(S): Sperm kinematic and fertilizing parameters. RESULT(S): Heat pretreatment of sperm resulted in over 22 times higher hyperactive motility at hour 4 compared with the control. The other kinematic parameters were also different. The heat-pretreated sperm group had a significantly higher percent penetration of zona-free oocytes with more swollen sperm heads per oocyte and enhanced sperm binding. CONCLUSION(S): The results showed that hyperactivation was induced by pretreatment of sperm with 40 degrees C heat, suggesting the involvement of heat factors in hyperactivation. The fertilizing capacity of sperm may be improved by the mild heat pretreatment when marked by the presence of heat-induced hyperactivation.

Female↗
PubMed1998

Ofloxacin: the next generation of antibiotic in sperm and embryo cultures for assisted reproductive technologies.

OBJECTIVE: To analyze the effect of different concentrations of ofloxacin on sperm kinematic parameters and to determine the embryotoxicity of ofloxacin at physiologic and at 100x concentrations. DESIGN: Prospective comparative study. SETTING: Clinical and academic research environment. PATIENT(S): Pooled cryopreserved donor sperm (n = 7). INTERVENTION(S): Human sperm were processed through two-layer discontinuous Percoll gradients, and the resultant pellet was resuspended in either HEPES-buffered human tubal fluid medium containing different concentrations of ofloxacin or the control medium. After measuring the kinematic parameters, the percentages of apoptosis and viability were obtained. Next, the sperm DNA was extracted and polymerase chain reaction of beta-globin gene was performed followed by denaturing gradient gel electrophoresis. Mouse embryos recovered at the one-cell pronuclear or zygote stages were cultured in the presence or absence of ofloxacin up to the hatched blastocyst stage and differences in development were recorded. MAIN OUTCOME MEASURE(S): Sperm kinematic parameters, sperm beta-globin gene, and number of embryos reaching the hatched blastocyst stage. RESULT(S): The number of embryos exposed to control and physiologic ofloxacin concentrations showed comparable excellent growth. However, the 100x concentration significantly arrested development. Rates of sperm viability and apoptosis measured 48 hours after exposure to the above concentrations were not different from controls. No differences were noted in the sperm kinematic parameters of sperm exposed to ofloxacin concentrations (1x, 10x, and 100x) or control medium after 0, 4, and 48 hours of incubation. Denaturing gradient gel electrophoresis of beta-globin genes from DNA exposed to varying ofloxacin concentrations failed to show any point mutations. CONCLUSION(S): Ofloxacin was embryotoxic at pharmacologic concentrations (100x). At physiologic or higher concentrations, ofloxacin appears to be safe and does not affect sperm kinematic parameters when compared with controls. This may indicate that sperm motility parameters alone cannot be relied on to evaluate the effects of drugs on fertility and that in vitro embryologic studies are essential. Ofloxacin at any concentration did not alter the rates of sperm apoptosis or viability. Ofloxacin does not appear to be mutagenic as evidenced by the beta-globin gene analysis.

Analysis of Variance↗
PubMed1998

A convenient mixed immunobeads screen for antisperm antibodies during routine semen analysis.

OBJECTIVE: To determine if leftover unused IgG, IgA, and IgM immunobeads could be combined as a convenient screen for antisperm antibodies in unwashed sperm. For comparisons, the mixed antiglobulin reaction (SpermMar) and immunobead test were performed. DESIGN: Prospective comparative study. SETTING: Clinical and academic research environment. SUBJECT(S): Donor human sperm negative for antisperm antibodies. INTERVENTION(S): Sperm specimens (n = 9) were equally divided and sera with antisperm antibodies added to one portion (positive control). MAIN OUTCOME MEASURE(S): Assessment of antisperm antibodies in sperm. RESULT(S): The mixed immunobeads screen qualitative results were the same for both the SpermMar and immunobead tests. The mixed immunobeads screen (positive when > or =10% motile sperm was bound) had a sensitivity and specificity of 100%. The percentages of bound sperm for the mixed immunobeads screen correlated with the SpermMar but not the immunobead test. The mixed immunobeads screen intraassay and interassay coefficients of variation were 28.9% and 45.6%, respectively. CONCLUSION(S): Matching results between the mixed immunobeads screen and the SpermMar or immunobead test suggested that the screen could be used to detect antisperm antibodies in unwashed sperm. There was no advantage to washing the sperm, although the screen was flexible and applicable for both unwashed and washed sperm.

Autoantibodies↗
PubMed1998

Heat-induced hyperactivation.

PURPOSE: The objectives of this study were (1) to determine the sperm hyperactivation and related kinematic parameters at 40 degrees C after using four sperm wash procedures and (2) to correlate the heat-induced hyperactivation data with cases of clinical pregnancies from either artificial insemination or standard in vitro fertilization (IVF). METHODS: Semen samples (n = 51) were collected by ejaculation, and semen analyses were carried out to determine the pretreatment data. Sperm kinematic measurements were performed using the Hamilton Thorn HTM-C computer-aided sperm analyzer. Hyperactivation was determined using the sort module on the HTM-C. Membrane integrity was assessed using the hypoosmotic sperm swelling procedure. Sperm morphology and acrosomal status were also determined using the Spermac stain. Each semen specimen was divided and processed through either the swim-up wash, the 1-h test-yolk buffer (TYB) wash, the 1 mg/ml pentoxifylline stimulant procedure, or the two-layer 90:47% gradient colloidal solution procedure. The washed sperm were incubated at 25 or at 40 degrees C for 4 hr. After incubation, kinematic parameters were assessed for the posttreatment data. Semen specimens were obtained on different occasions for artificial insemination or standard IVF. Data from intracytoplasmic sperm injection cases were not included to avoid confounding factors. Live births and/or pregnancies with fetal heart-beat examined by ultrasound were considered clinical pregnancies. RESULTS: Heat-induced hyperactive motility was significantly higher in sperm of the male partner of pregnant (n = 7) patients compared with nonpregnant (n = 44) patients (mean +/- SE, 10.0 +/- 3.3 versus 5.5 +/- 0.8%) after TYB processing followed by 4 hr of incubation at 40 degrees C. This was also observed after colloid (Percoll) processing (11.6 +/- 4.6 versus 5.8 +/- 0.8%). There were no differences in hyperactivation after 4 hr at 23 degrees C between pregnant and nonpregnant cases. Parameters such as count, volume, motility, viability, and acrosomal status were not different for the groups. However, the percentage of sperm with normal morphology (WHO classification) was twice as high in the pregnant group versus the nonpregnant group. CONCLUSIONS: Heat-induced hyperactivation was associated with fertile sperm and was predictive of pregnancy obtained after artificial insemination or IVF. The association was evident only after TYB or Percoll sperm processing. The study could not confirm the finding of significant decreases in motility after heat treatment of sperm derived from infertile males. The mechanism for heat-induced hyperactivation did not involve membrane integrity or the sperm acrosome, although an involvement of heat shock proteins was postulated. Interestingly, there were no pregnancies when sperm did not exhibit heat-induced hyperactivation.

Acrosome↗
PubMed1997

Silane-coated silica particle colloid processing of human sperm.

PURPOSE: The purpose of this study was to determine differences in the quality of human sperm processed through different lots of silane-coated silica particle colloid solutions. The objectives were to compare (a) sperm kinematic parameters, (b) the sperm acrosome status, (c) the membrane integrity of the head and tail regions, (d) the DNA normality, and (e) the heat-inducible hyperactivation motility after processing sperm through either a Silane-coated silica particle colloid solution, a Percoll solution, or a simple centrifuge sperm wash (control). METHODS: Sperm cells were derived from pooled cryopreserved-thawed specimens of several donors (n = 10). The pooled sperm were divided and processed through either the centrifuge wash, the 90:47% two-layer Percoll, or one of three lots of silane-coated silica particle colloidal solutions from three vendors. Aliquots of sperm cells were analyzed using the Hamilton-Thorn HTM-C motility analyzer for differences in kinematics and hyperactivation. Sperm were also analyzed for membrane integrity at both head and tail regions, normal morphology, acrosome status, and viability. Sperm undergoing apoptosis were determined using the acridine orange stain. Processed sperm were also incubated at 40 degrees C for 4 hr and the quality of the sperm was assessed using the heat-induced hyperactivation and motility parameter. RESULTS: The data showed that after sperm processing, the number of sperm recovered was higher for the three lots of colloids (silane-coated silica particle colloid solutions) compared with Percoll processing. Total sperm motility was higher in the colloidal washes compared with the control. There were no differences in motility between Percoll- and colloid-processed sperm. In contrast, the percentages of sperm exhibiting progressive motility or hyperactivation varied among the different lots of colloid solutions. The Percoll wash solution yielded the highest percentage of sperm with intact tail membranes, whereas some lots of colloid solutions disrupted sperm head membranes. The percentages of sperm undergoing apoptosis varied for the different lots of colloid solutions. There was a marked increase in hyperactivation associated with one colloid solution after heat induction. CONCLUSIONS: The results demonstrated variability in the different lots of silane-coated silica particle colloid solutions for processing sperm. Each lot of colloid solution excelled at improving different sperm parameters. The silane-coated silica particle colloid solutions were shown to be effective in recovering motile sperm compared with Percoll but the types of motility and sperm quality varied for the different lots of colloid solutions. Due to the variability in lots of silane-coated silica colloid solutions, reported studies based on only one lot or one source of colloid solution may be difficult to interpret. Furthermore, it may be advantageous to select the best lot of silane-coated silica particle colloid solution to produce the highest number of sperm exhibiting the ideal parameters for use in assisted reproduction technologies.

Acridine Orange↗
PubMed1997

Antibiotics: effect on cryopreserved-thawed human sperm motility in vitro.

OBJECTIVE: To analyze the motility and fertilizing capacity of sperm treated with different antibiotics. DESIGN: Prospective comparative study. SETTING: Clinical and academic research environment. PATIENT(S): Pooled cryopreserved donor sperm (n = 14). INTERVENTION(S): Sperm were washed with Percoll and resuspended in HEPES-buffered human tubal fluid medium containing either amoxicillin, ofloxacin, ciprofloxacin hydrochloride, nitrofurantoin monohydrate, doxycycline hyclate, cefuroxime axetil, or control medium. MAIN OUTCOME MEASURE(S): Sperm kinematic and fertilizing parameters. RESULT(S): Sperm hyperactivation was decreased in physiologic concentrations of ciprofloxacin hydrochloride and doxycycline hyclate over the course of 48 hours. At pharmacologic concentrations, ciprofloxacin hydrochloride, cefuroxime axetil, and nitrofurantoin monohydrate adversely affected motility with decreased rapid progression. Cessation of motility occurred in cefuroxime axetil and nitrofurantoin monohydrate. Sperm hyperactivation was also absent. Cefuroxime axetil decreased the percentage of intact acrosomes. In contrast, physiologic doses of ciprofloxacin hydrochloride or ofloxacin enhanced sperm fertilizing capacity. CONCLUSION(S): Ciprofloxacin affected hyperactivation by altering membrane properties, whereas doxycycline inhibited the capacitation process. Cessation of motility in cefuroxime axetil was linked to disrupted sperm head membranes. Sperm motility and fertilizing capacity were decreased in nitrofurantoin because of decreased metabolism. The positive effect of ofloxacin on fertilizing capacity did not involve changes in acrosome.

Analysis of Variance↗
PubMed1996

Analysis of the flow cytometer stain Hoechst 33342 on human spermatozoa.

Several procedures exist for processing sperm cells for sex preselection. Flow cytometric separation using the fluorochrome stain Hoechst 33342, chemically known as bisbenzimide, is the most promising. The objective of this study was to determine the effect of bisbenzimide on spermatozoa assessed by means of the sperm survival test and to analyse the beta-globin gene in sperm DNA after exposure to increasing concentrations of bisbenzimide. Donor (n = 16) sperm specimens were pooled and washed in a discontinuous Percoll gradient 95:47%, divided and incubated in tubes containing bisbenzimide at concentrations 0 (control), 0.9, 9, 90, 900 and 9000 microM at 25 degrees C and scanned on a computer-aided sperm motility analyser at 0, 1, 4 and 24 h. Spermatozoa were also incubated in a known mutagen, ethidium bromide, as positive control. After 24 h of incubation, the treated sperm cells were processed through DNA extraction and polymerase chain reaction (PCR) performed with primers targeting the beta-globin gene. The amplified DNA products were analysed for evidence of mutation in 5% polyacrylamide gel electrophoresis and 20:80 denaturing gradient gel electrophoresis (DGGE) and further confirmed in 30:40 DGGE. The results showed complete cessation of motility in sperm incubated in the presence of 900 microM or higher concentrations of bisbenzimide. The beat cross frequency sperm parameter was significantly different at the 90 microM or higher concentration of bisbenzimide compared with the control. At concentrations < 900 microM bisbenzimide, there were no differences in the remaining sperm kinematic parameters (percentage rapid progressive, percentage total progressive, sperm velocities, linearity, straightness, amplitude of lateral head displacement and percentage hyperactive motility). PCR and DGGE analyses of spermatozoa treated with bisbenzimide showed no evidence of mutation in the representative region of the beta-globin gene at concentrations < 900 microM. The data suggest an inhibitory effect of bisbenzimide on human sperm motility at 900 microM or higher concentrations of bisbenzimide. The decrease in sperm motility and rapid progression were not due to changes in pH. Point mutation in the representative region of the beta-globin gene in human spermatozoa was detected only at high concentrations (> or = 900 microM) of bisbenzimide. The data suggest that incubating sperm in low concentrations of bisbenzimide (< 90 microM) for up to 24 h does not significantly affect all the sperm kinematic parameters including the beat cross frequency parameter when compared with the control.

Benzimidazoles↗
PubMed1978

On the etiologic role of ureaplasma urealyticum (T-mycoplasma) infection in infertility.

Two hundred consecutive infertility patients and sixty-seven controls subjects were studied for the incidence of infection with Ureaplasma urealyticum (T-mycoplasma). On the basis of a complete infertility investigation, the infertility patients were subdivided into those with explained infertility and those with unexplained infertility. Of the patients with unexplained infertility, 55% had a positive culture for T-mycoplasma as compared with a 32% incidence of positive cultures in the control population. The differences were statistically significant. The 6-month pregnancy rate following successful antibiotic treatment in patients with unexplained infertility was 42%. The 6-month pregnancy rate in a comparable group of patients with unexplained infertility, seen during a 3-year period prior to mycoplasma culture and treatment, was 32%. The difference in pregnancy rates between the two groups was not statistically significant. No correlation was found between a poor postcoital test and the presence of T-mycoplasma infection nor between T-mycoplasma infection and poor cervical mucus. The role of T-mycoplasma infection in infertility was neither proven nor disproven by this study.

Cervix Uteri↗
PubMed1976

Modulation of pituitary responses to synthetic LH-RH by gonadal steroids in women with secondary amenorrhoea.

LH-RH stimulation tests were performed on two successive days in 25 women with secondary amenorrhoea of probable hypothalamic origin by intravenous injection of 150 mug of synthetic LH-RH. The patients were selected by exclusion of definite pituitary or ovarian disease and in the absence of clinical or laboratory evidence of androgen excess. Five women received successive LH-RH stimulation tests only without administration of steroid hormone. Twenty women were treated with gonadal steroids in addition to the double LH-RH stimulation test by intramuscular injection of the steroid 4 h after the first LH-RH injection. Of these 20, 9 received oestradiol (Oe2), 5 received progesterone (P) and 6 received Oe2 + P. Gonadotrophin responses to LH-RH in successive tests without steroid were not significantly different. Oe2 or P alone each produced suppression of LH and FSH responses to LH-RH in the second test as compared to the first. Combined Oe2 + P produced augmentation or suppression of the second gonadotrophin responses depending on the dose of Oe2 administered. The results demonstrate a direct effect of oestrogen or progesterone alone and in combination on the pituitary responsiveness to LH-RH. The effect of the combined steroids can be modified by variation in the oestrogen: progesterone ratio.

Amenorrhea↗
PubMed1975

Pituitary gonadotropin responses to synthetic luteinizing hormone-releasing hormone in patients with typical and atypical polycystic ovary disease.

Synthetic luteinizing hormone-releasing hormone (LH-RH) was administered intravenously to 17 women with polycystic ovary disease (PPCO), 16 women with hypothalamic amenorrhea (HA), and nine women with normal cycles. The serum levels of follicle-stimulating hormone (FHS) and luteinizing hormone (LH) were measured at frequent intervals before and after LH-RH injection. The PCO patients were arbitrarily divided into two groups on the basis of ovarian morphology. The eight patients with large, "typical" PCO's (Type I) showed a greater LH response and a lower FSH response to LH-RH than did the nine patients with smaller, "atypical" PCO's (Type II). The LH response in Type I co was also greater than that in HA, but the FSH response in both types of PCO was significantly less than that in HA. The gonadotropin responses to LH-RH in Type I PCO were quite similar to those occurring in normal women during the two to three days prededing ovulation. The results suggest that excessive LH secretion and/or impaired FSH secretion may be etiologically significant in PCO but do not clearly differentiate whether the stimulus for this pattern of gonadortopin secretion occurs at the hypothatamic or pituitary level.

Amenorrhea↗
PubMed1975

Gonadotropin levels and secretory patterns in patients with typical and atypical polycystic ovarian disease.

Gonadotropin levels and secretory patterns were studied in 28 oligomenorrheic patients with various types of polycystic ovary disease (PCO). On the basis of ovarian morphology and histology, the patients PCOuld be separated into two distinct categories arbitarily designated "typical" (type I) and "atypical" (type II) PCO. Although no differences were noted in symptomatology or 17-ketosteroid, testosterone, or follicle-stimulating hormone levels, the 12 type I patients demonstrated widely fluctuating, but markedly elevated, luteinizing hormone (LH) levels, while the 16 type II patients demonstrated lower and less fluctuating LH levels which were comparable to those found during the normal follicular phase. It is likely that type I PCO is a distinct entity similar to that described by Stein and Leventhal, while type II co represents a heterogenous spectrum of disorders, many of which remain obscure.

17-Hydroxycorticosteroids↗
PubMedDate not supplied

Sperm velocity in seminal plasma and the association with gender of offspring.

The gender of the offspring is determined by the fertilizing sperm. Previous gender studies were based on washed sperm, but not on sperm in seminal plasma. The objective was to correlate motility parameters assessed during semen analyses with the offspring gender. For comparison, fixed sperm head DNA quantitated by Hoechst 33342 fluorescence microscopy was also analyzed. Forty-six patients undergoing assisted reproduction procedures resulted in livebirth deliveries with either male or female-predominant offsprings. Sperm head fluorescence was weakly correlated to the gender in 61% of the cases. Sperm of patients with male offsprings had slower curvilinear (44.2 +/- 1.8 mean +/- SEM, versus, 49.9 +/- 2.7 micro /sec) and slower average path velocities (32.4 +/- 1.2 versus 36.3 +/- 1.7 micro /sec). Using cut-off values for the curvilinear (< 49 micro /sec) and average path (< 36 micro /sec) velocities of sperm swimming in seminal plasma, the two parameters predicted 75 and 68% of the male offspring births, respectively. The data suggest that sperm movement in seminal plasma is a marker for factors that skew the ratio of the X- to Y-sperm populations.

Benzimidazoles↗
PubMedDate not supplied

Sperm apoptosis in nonpregnant luteal phase sera after in vitro fertilization as assessed by comparative genomic hybridization.

Toxicity in serum has been reported in cases of recurrent spontaneous abortions and endometriosis. The null hypothesis was that serum toxicity was not involved in failed pregnancies after in vitro fertilization procedures. The objective was to expose donor sperm to pregnant versus nonpregnant patient sera and analyze for sperm DNA damaging effects using a novel comparative genomic hybridization method. Luteal phase sera (N = 21 cases) were drawn one week after embryo transfer. Colloid-washed donor sperm were incubated (48 h, 37 degrees C, 5% CO2 in air) in 0% or 50% sera. Single-stranded DNA (ssDNA) of control sperm were stained in Hoechst 33342 and hybridized to Sybr Gold-stained ssDNA of sera-treated sperm. Image analyses were performed and fluorescent intensities analyzed. Nonpregnant patient sera (57% of cases) were associated with DNA fragmentation (64.4 +/- 8.8 pixels; mean +/- S.E.M.) when compared with pregnant patient sera (106.3 +/- 8.4 pixels). There were no differences in the sera of biochemical (108.2 +/- 15.3) versus clinical pregnancy cases (105.3 +/- 11.4). The results suggest that nonpregnant patient sera contained factor(s) that cause DNA fragmentation leading to pregnancy losses.

Blood Proteins↗
PubMedDate not supplied

Correlation of sperm parameters with apoptosis assessed by dual fluorescence DNA integrity assay.

Failed fertilization after intracytoplasmic sperm injection or miscarriages occurs in cases involving apoptotic and necrotic sperm. Identifying normal sperm is important for successful assisted reproductive technologies (ART) procedures. The study was conducted to correlate sperm parameters with intact sperm with normal DNA assessed by the dual stain assay in 118 separate individuals. The results showed differences in percent DNA intact sperm in individuals with normal W.H.O. sperm features (62 +/- 1.1; mean +/- S.E.M.) compared with oligoasthenoteratozoospermia patients (38 +/- 5.3). Individuals whose sperm had fertilizing capacity had higher percentages of intact DNA (60 +/- 1.3 versus 47 +/- 2.4). The percentages of intact DNA sperm were significantly correlated to total motility in semen (R = 0.7), post-wash motility (R = 0.6), rapid progression (R = 0.6), intact acrosome (R = 0.5), and strict morphology (R = 0.5). There were no correlations with the remaining parameters. The dual stain assay identified sperm with normal physiology and fertilizing capacity. The dual stain assay measures DNA integrity and is a promising method to select normal sperm for ART.

DNA↗
PubMedDate not supplied

A follow-up expanded study of the correlation of sperm velocity in seminal plasma and offspring gender.

A preliminary study reported finding higher sperm velocity in seminal plasma in males of partners that conceived female offsprings. The null hypothesis was that sperm velocity was not related to the offspring gender. The objectives were: (a) to expand the previous study, and (b) to correlate offspring gender results with motility parameters determined through the computer-aided sperm analyzer (CASA) system. In combined fresh and frozen cycles (N = 187), sperm from cases with all female offsprings displayed higher curvilinear (48 +/- 1.0 mu/sec versus male 46 +/- 1.0, P < 0.05) and average path velocities (36 +/- 0.7 mu/sec versus male 34 +/- 0.7, P < 0.01). A criteria of less than 30 mu/sec or over 41 mu/sec average path velocity predicted 73 or 72% of the male or female offspring cases, respectively. A curvilinear velocity of less than 49 mu/sec or over 55 mu/sec predicted 58 or 59 % of the male or female offspring cases, respectively. Semen viscosity reflected in sperm velocity was linked to predominantly male or female sperm populations. Paracrine signals from the gender-skewed sperm precursor populations controlling viscosity merit further exploration.

Female↗