Biomedical subjects

M E Hammadeh

Publications and source records attributed to M E Hammadeh.

At least 19 recordsLinked to original sources

PubMed2006

Comparison of reactive oxygen species concentration in seminal plasma and semen parameters in partners of pregnant and non-pregnant patients after IVF/ICSI.

The aims of this study were (i) to determine and compare the concentration of reactive oxygen species (ROS) and total antioxidant status (TAS) in seminal plasma and sperm parameters of the male partners of patients undergoing IVF or intracytoplasmic sperm injection (ICSI) treatment and (ii) to establish the relationship between ROS and TAS concentrations and sperm quality and their effect on fertilization and pregnancy rate of patients who achieved a pregnancy and those who were unsuccessful. Twenty-six IVF and 22 ICSI patients were included in this study. The ROS concentration in seminal plasma and sperm concentration, vitality (eosin test), motility, morphology, membrane integrity (HOS test), maturity (chromomycin, CMA3) and DNA fragmentation (TUNEL) results and their relationship to fertilization and pregnancy were analysed. ROS concentrations were similar regarding the seminal plasma of male partners of patients who achieved a pregnancy and those who were unsuccessful. The other semen parameters, concentration, motility, vitality, membrane and DNA integrity, were comparable in both groups. However, both groups demonstrated a negative correlation between ROS concentration and sperm vitality, membrane integrity and morphology. Moreover, an inverse correlation was found between TUNEL, vitality, and membrane integrity. In conclusion, ROS concentration in seminal plasma affects the quality of spermatozoa. A negative correlation between the ROS concentration in seminal plasma and fertilization rate in both IVF/ICSI programmes was shown.

Adult↗

PubMed2005

[Body mass index, protein metabolism profiles and impact on IVF/ICSI procedure and outcome].

OBJECTIVE: Higher risks of infertility have been found in overweight women. The purpose of the present study was to explore whether protein metabolism profiles related to body mass index (BMI) and to find out whether these parameters should affect IVF/ICSI outcome. PATIENTS AND METHODS: 52 patients were enrolled in this study. All patients underwent an ovarian stimulation either with recombinant follicle stimulating hormone (Gonal-F) or human menopausal gonadotropin (Menogon) after pituitary down-regulation with Goserelin (Zoladex) or Triptorelin (Decapeptyl Gyn). Five blood samples were taken: before treatment, at the beginning of ovarian stimulation, on the day of HCG injection for the ovulation induction, on the day of follicle aspiration and 14 days after embryo transfer. The blood samples were analysed with regard to the serum concentrations of total protein, albumin, total bilirubin and urea. According to the BMI values the patients were divided into two groups: BMI < 25 kg/m (2) (GI, n = 28) and BMI > 25 kg/m (2) (GII, n = 24). The results of IVF/ICSI outcome were compared in both groups. RESULTS: In both groups, the serum concentrations of total protein, albumin, total bilirubin and urea decreased during ovarian stimulation. In GII, albumin concentration decreased significantly on the day of follicle aspiration (46.0 +/- 2.3 g/l versus 43.5 +/- 2.5 g/l, p < 0.001) and 14 days after embryo transfer (46.8 +/- 2.5 g/l versus 44.7 +/- 2.3 g/l, p < 0.002), whereas the concentration of total bilirubin was not significantly decreased on the day of HCG injection (0.57 +/- 0.29 mg/dl versus 0.49 +/- 0.26 mg/dl, p = 0.11). Furthermore, pregnancy rate in women with BMI < 25 kg/m (2) was 46.4 % and in women with BMI > 25 kg/m (2) 33.1 % (p = 0.34). CONCLUSIONS: Serum concentrations of albumin and total bilirubin are influenced by BMI. Excess weight defined as BMI > 25 kg/m (2) has a negative impact on IVF outcome leading to decreased chances of pregnancy.

Bilirubin↗

PubMed2004

[Effects of ovarian stimulation on serum concentrations of lipids and ketone bodies in patients undergoing IVF/ICSI treatment].

OBJECTIVE: The ovarian stimulation increase the estradiol concentration in comparison to the one observed in the menstrual cycle. The aim of the study was to determine the influence of ovarian stimulation on serum concentrations of lipids and ketone bodies in patients undergoing IVF/ICSI treatment. PATIENTS AND METHODS: 50 patients were included in this study. All patients underwent an ovarian stimulation either with recombinant follicle stimulating Hormone (Gonal-F, n = 28) or human menopausal gonadotropin (Menogon, n = 22) after pituitary down-regulation with Goserelin or Triptorelin. Five blood samples were taken: before treatment, at the beginning of ovarian stimulation, on the day of HCG injection for the ovulation induction, on the day of follicle aspiration and 14 days after embryo transfer. Afterwards, the samples were analysed with regard to the concentrations of triglyceride, total-cholesterol, HDL-/LDL-cholesterol, acetoacetate, beta-hydroxybutyrate and alpha-ketoglutarate. RESULTS: During the ovarian stimulation, total-cholesterol andA'LDL-cholesterol decreased significantly (p(Chol) < or = 0.001; p(LDL) < or = 0.001), whereas the concentration of HDL-cholesterol increased significantly (p(HDL) < or = 0.001). Furthermore, a significant increase of beta-hydroxybutyrate and alpha-ketoglutarate has been detected (p(HBS) = 0.039; p (KG) = 0.023). CONCLUSIONS: The observed changes in serum concentrations of lipids and ketone bodies began with the application of gonadotrophins for stimulation and persisted until the end of the treatment. Therefore, the increase of estradiol, stimulated by gonadotrophins, seems to be responsible for the changes in the serum concentrations of lipids and ketone bodies. All women undergoing ovarian stimulation should be monitored for long-term thrombogenic and atherogenic risks.

3-Hydroxybutyric Acid↗

PubMed2003

Fibroblast growth factor (FGF), intracellular adhesion molecule (sICAM-1) level in serum and follicular fluid of infertile women with polycystic ovarian syndrome, endometriosis and tubal damage, and their effect on ICSI outcome.

PROBLEM: The objective of this study was to determine the concentration of fibroblast growth factor (FGF) and soluble intracellular adhesions molecule (sICAM-1) in serum and follicular fluid (FF) of polycystic ovary (PCO), endometriosis and tubal factor infertility and male factor infertility patients, and to investigate the relationship between these parameters and the outcome of intracytoplasmic sperm injection (ICSI). METHOD OF STUDY: The concentration of FGF and sICAM-1 in serum and FF were determined in patients undergoing controlled ovarian hyperstimulation (COH) for ICSI therapy for various etiology of infertility and the results of cytokines concentration and ICSI outcome were compared between the groups. Twenty patients with PCO (G.I), 17 with endometriosis (G.II), 19 with tubal damage (G.III) and 19 with male factor infertility (G.IV) were enrolled in this study. Quantitative determination of levels of FGF and sICAM-1 was performed using enzyme-linked immunosorbent assays (ELISAs). RESULTS: The FGF level in serum of PCO patients (G.I) were 4.8 +/- 2.3 and in FF were 104.0 +/- 39.0 pg/mL. The corresponding values in the endometriosis patients group (G.II) were 5.9 +/- 3.1 and 125.4 +/- 74.9 pg/mL. The concentration of FGF in tubal factor infertility group (G.III) in serum was significantly higher (P = 0.009) than those observed in the PCO group (G.I) 7.4 +/- 4.5 pg/mL, whereas the concentration in FF was at the same level like the other groups investigated, 128.7 +/- 75.9 pg/mL. Besides, the sICAM-1 (pg/ml) concentration in FF showed a significant difference between the groups investigated (G.I, 175.3 +/- 52.8; G.II 194.4 +/- 32.2; G.III 233.1 +/- 54.3; and G.IV 215.1 +/- 54.4 ng/mL; P = 0.003). The sICAM-1 levels in serum were not significantly different between the groups (217.0 +/- 42.9; 216.3 +/- 73.6; 254.8 +/- 79.6; 237.56 +/- 78.4 ng/ml; P = 0.267). The fertilization rate was significantly higher in G.III (66.0 +/- 23.89%) in comparison to G.II (38.8 +/- 33.9%; P = 0.014) or G.IV (38.7 +/- 22.7%; P = 0.012). The pregnancy rates were similar in all groups (30, 35.3 and 35.0, 38.6%, respectively). CONCLUSION: Both, FGF and sICAM-1 are present in serum and FF of patients undergoing controlled ovarian hyperstimulation for ICSI therapy. The FGF concentration in serum differs significantly between the groups investigated, whereas, no significant difference could be observed in the FF concentration of FGF. On the other hand, the sICAM in serum showed no significant difference between the groups, whereas, sICAM in FF demonstrated a significant difference between the patient groups investigated. On the whole, the ICSI outcome was not related to serum or FF concentrations of FGF or sICAM-1. Therefore, the mean concentration of FGF and sICAM-1 in serum and in FF could not be used to predict the fertilization rate in an ICSI program.

Biomarkers↗

PubMed2003

Comparison between cytokine concentration in follicular fluid of poor and high responder patients and their influence of ICSI-outcome.

OBJECTIVE: The aim of this study was to compare the cytokine concentration in follicular fluid (FF) of low and high responder intracytoplasmic sperm injection (ICSI) patients and to find out the impact of these cytokines in FF on ICSI outcome. DESIGN: The levels of insulin-like growth factor (IGF)-I, IL-6, IL-8, epidermal growth factor (EGF), platelet-derived growth factor (PDGF), granulocyte-macrophage-colony stimulating factor (GM-CSF) were measured from low and high responder ICSI patients, the results were compared between the two groups and their influence on ICSI outcome was analysed. MATERIAL AND METHODS: A total of 49 low (G.I) and 34 high (G.II) responder patients were enrolled in this study. FF was collected at the time of oocyte retrieval and measured either by enzyme-linked immunosorbent assay (IL-6, IL8, EGF, PDGF, GM-CSF) or radio immuno assay (IGF-I). RESULTS: The concentration of IL-6 (pg/mL), IL-8 (pg/mL), IGF-I (ng/mL), PDGF (pg/mL), EGF (pg/mL), GM-CSF (pg/mL) in G.I was 6.0 +/- 4.3, 288.1 +/- 139.2, 0.416 +/- 0.089, 249.8 +/- 150.1, 9.12 +/- 5.5 and 1.45 +/- 2.10 and the corresponding value in G.II was 7.4 +/- 4.8, 208.6 +/- 64.0, 0.431 +/- 0.094, 387.6 +/- 36.0, 8.9 +/- 5.4 and 1.8 +/- 3.3, respectively. Only the PDGF concentration showed a significant (P = 0.007) difference between the two groups. Besides, negative correlations were found between PDGF and fertilization rate (r = -0.287; P = 0.046) of G.I. The mean number of retrieved (6.4 +/- 2.3 versus. 15.7 +/- 5.4) and fertilized (3.6 +/- 1.6 versus 7.0 +/- 4.5) oocytes differ significantly (P = 0.001) between the two groups. The fertilization rate was significantly higher in G.I than in G.II (60.9 +/- 25.1 versus 43.4 +/- 20.7%). CONCLUSION: There was no significant difference between IGF-I, IL-6, IL-8, EGF and GM-CSF concentrations of low and high responder patients. Besides, PDGF was significantly (P = 0.007) higher in high responder compared with low responder patients. Moreover, in poor responder patients, a negative correlation was found between PDGF and fertilization rate. However, the cytokine levels in FF of the patients undergoing controlled ovarian hyperstimulation for ICSI could not be used as a marker of oocyte fertilization and implantation potential.

Adult↗

PubMed2003

[Association between ovarian stimulation regime and protein metabolism of patients undergoing IVF/ICSI therapy].

The aim of this study was to determine the relationship between the ovarian stimulation protocol (with HMG or FSH) after down-regulation with GnRH anologa and protein (total protein and albumin) as well as bilirubin and urea in serum. Furthermore, it was intended to find out the effect of these parameters on IVF/ICSI outcome. 50 patients were included in this study. All patients underwent controlled ovarian hyperstimulation for assisted reproduction therapy either with FSH (Gonal-F) or HMG (Menogon). Ovulation induction was induced by human chorionic gonadotrophine (HCG, Predalon) 10 000 IU i. m. The protein concentration (total protein, albumin) as well as bilirubin and urea concentrations were determined before down-regulation with GnRHa, at the beginning of ovarian stimulation with FSH or HMG, on the day of ovulation induction with HCG, during oocyte retrieval and fourteen days after embryo transfer. The age, body mass index and etiology of infertility showed no significant difference between patients stimulated with HMG or FSH. Total protein and albumin concentration decreased significantly (p=0.001) from 77.45 +/- 5.90 g/L and 47.02 +/- 3.41 g/L to 74.60 +/- 4.6 g/L and 45.04 +/- 2.39 g/L, respectively at the time of oocyte retrieval. Whereas, no significant change with bilirubin and urea concentration was observed. However, the mean concentration of total protein, albumin, bilirubin of patients who become pregnant was higher of those who did not. In conclusion, this study shows that total protein and albumin concentration in plasma decreased during the follicular phase significantly with the application of exogenous gonadotrophins and steroid hormones in comparison to the value before down-regulation. The mean value of total protein, albumin, bilirubin of patients who become pregnant was higher (but not significant) of those who did not. However, the high individual variation in the present results shows that these parameters are not useful as a predictor of IVF/ICSI outcome.

Blood Proteins↗

PubMed2002

Relationship between ovarian stimulation regimen and interleukin level in pre-ovulatory follicular fluid and their effect on ICSI outcome.

PROBLEM: The aims of this study were (i) to identify the presence and concentration of interleukin-6 (IL-6), IL-8 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in pre-ovulatory follicular fluid (FF) in patients undergoing controlled ovarian hyperstimulation regimen for intracytoplasmic sperm injection (ICSI) with recombinant human follicle stimulating hormone (rFSH) (G.1), hMG (G.2) or rFSH/ hMG (G.3) treatment and (ii) to determine whether controlled ovarian hyperstimulation stimulation regimen could affect the concentration of these investigated interleukins and as a consequences the ICSI outcome. (iii) Moreover, it was intended to clarify the association between these investigated parameters and the peripheral blood level of estradiol 17-beta, progesterone, leuteinizing hormone (LH) at the day of oocyte retrieval and (iv) finally to find out whether these interleukin concentrations in FF could be considered as an additional parameter for predicting fertilization and pregnancy outcome in ICSI patients. MATERIAL AND METHODS: Interleukin-6, IL-8 and GM-CSF concentrations were determined in the FF of 85 patients. The levels of these interleukins were measured by the enzyme-linked immunosorbent assay (ELISA) technique, using commercially available kits. RESULTS: Interleukin-6 (6.5 +/- 4.6 pg/mL), IL-8 (222.9 +/- 163.4 pg/mL) and GM-CSF (1.6 +/- 2.7 pg/mL) were present in pre-ovulatory FF in ICSI patients. No significant difference in the mean concentration of IL-6, IL-8 and GM-CSF was found between the groups. Moreover, there was no correlation between these interleukin levels in FF and steroid hormone concentration in the peripheral blood. CONCLUSIONS: Interleukin-6, IL-8 and GM-CSF were found in FF of patients undergoing ovarian hyperstimulation for ICSI treatment. The stimulation protocols showed no influence on these interleukin levels in FF. Moreover, no correlation was found between these investigated interleukins and steroid hormone concentrations in peripheral blood or ICSI outcome. Therefore, the determination of interleukin level in FF of patients undergoing ovarian hyperstimulation for ICSI therapy could not be recommended as a prognostic factor for ICSI outcome.

Adult↗

PubMed2001

Evaluation of cryoinjury of spermatozoa after slow (programmed biological freezer) or rapid (liquid nitrogen vapour) freeze-thawing techniques.

PURPOSE: This study was initiated to determine the negative effect (cryodamage) on human spermatozoa after freeze-thawing and to find out whether freezing of spermatozoa with a computerized biological freezer is more advantageous than freezing above static liquid nitrogen vapour with regard to spermatozoa vitality, chromatin normality, morphology, and membrane integrity. METHODS: Forty-four semen samples were obtained from patients attending andrology laboratory, and each sample was divided into two aliquots. One aliquot was frozen using static liquid nitrogen vapour (G.II) and the second with a computerized biological freezer (G.III). Acridine orange was used for assessment of chromatin cryoinjury, whereas the morphology was evaluated according to WHO criteria. Hypoosmotic swelling test was used to identify membrane integrity and eosin-nigrosin staining was used to determine the vitality of spermatozoa. RESULTS: The mean percentage of normally condensed chromatin in the native semen sample (G.I) decreased significantly (p < .001) after freeze-thawing by using either liquid nitrogen vapour (G.II), or a biological freezer (G.III), which was significantly higher (p < .001) after freezing with liquid nitrogen vapour than after freezing with the biological programmed freezer. Morphologically normal spermatozoa decreased significantly (p < .001) in both freezing methods in comparison to the native semen samples. In addition, membrane integrity of spermatozoa (HOS-test positive) was significantly lower (p < .001) after the freeze-thawing procedure in G.II and G.III compared to G.I. In both these parameters the deterioration was similar among the two freezing procedures. Finally the mean percentage of live spermatozoa decreased significantly (p < .001) in both freezing techniques in relation to the mean value in the neat semen samples. CONCLUSIONS: Freeze-thawing procedure has a detrimental effect on chromatin, morphology, membrane integrity, and vitality of human spermatozoa not only by freezing above static liquid nitrogen vapour but even by using a computerized biological freezer. However, the chromatin deterioration rates are significantly higher by freezing above static liquid nitrogen vapour in comparison to freezing with a programmed biological freezer. Therefore, we recommend the use of this technique for freezing semen especially when ICSI technique is considered as the main therapeutic procedure.

Cell Survival↗

PubMed2001

Comparison between computerized slow-stage and static liquid nitrogen vapour freezing methods with respect to the deleterious effect on chromatin and morphology of spermatozoa from fertile and subfertile men.

The purpose of this study was to determine the negative effects (cryodamage) on human spermatozoa after freeze-thawing and to determine whether freeze-thawing of spermatozoa with a programmed slow freezer is better than freezing with liquid nitrogen vapour (rapid freezing) with regard to alterations in sperm chromatin and morphology in semen from fertile (donor) and subfertile, IVF/ICSI, patients. Ninety-five semen samples were obtained either from patients attending our IVF unit for treatment (n=34) or from donors (n=25) with proven fertility and normal sperm quality according to WHO guidelines. Each semen sample was divided into two parts after liquefaction and addition of the cryoprotectant. The first part was frozen using a programmed biological freezer and the second part was frozen by means of liquid nitrogen vapour. Smears were made before the freezing and after the thawing procedure to assess morphology (strict criteria) and chromatin condensation (Acridine Orange test). The mean percentage of chromatin condensed spermatozoa in the samples from donors (control group) was 92.4 +/- 8.4% before freezing and this decreased significantly (p < 0.0001) to 88.7 +/- 11.2% after freeze-thawing with the computerized slow-stage freezer and to 87.2 +/- 12.3% after using static liquid nitrogen vapour (p < 0.001). The corresponding values for semen obtained from patients was 78.9 +/- 10.3% before freezing which decreased to 70.7 +/- 10.8 and 68.5 +/- 14.8%, respectively (p < 0.001). On the other hand, the mean percentage of normal sperm morphology in the control group decreased from 26.3 +/- 7.5% before freezing to 22.1 +/- 6.4% (p < 0.0001) after thawing with the computerized slow-stage freezer and to 22.2 +/- 6.6% (p < 0.0001) after the use of static liquid nitrogen vapour. In the patient group, the mean percentage of normal morphology decreased from 11.7 +/- 6.1% after freezing with the biological freezer to 9.3 +/- 5.6% and to 8.0 +/- 4.9% after freezing with static liquid nitrogen vapour. This study demonstrates that chromatin packaging and morphology of human spermatozoa decrease significantly after the freeze-thawing procedure, not only after the use of static liquid nitrogen vapour but also after the use of a computerized slow-stage freezer. However, the chromatin of semen samples with normal semen parameters (donor sperm) withstand the freeze-thaw injury better than those with low quality semen samples. Therefore, the computerized slow stage freezer could be recommended for freezing of human spermatozoa, especially for subnormal semen samples, for example, ICSI and ICSI/TESE candidates and from patients with testicular tumours or Hodgkin's disease, in order to avoid further damage to the sperm chromatin structure.

Chromatin↗

PubMed2001

Predictive value of chromatin decondensation in vitro on fertilization rate after intracytoplasmic sperm injection (ICSI).

This study was undertaken to identify the relationship between sperm chromatin decondensation after incubation with sodium dodecyl sulphate (SDS) and ethylene diamine tetra acetic acid (EDTA), or heparin at various points of time. Likewise, this study will determine chromatin stability within definite time intervals, chromatin decondensation after intracytoplasmic sperm injection (ICSI), and whether chromatin decondensation in vitro could be used as a predictive test for fertilization capability after ICSI. Sixty-five infertile couples undergoing ICSI therapy were included in this prospective study. Male factor infertility was the main indication for inclusion. One millilitre from each semen sample after washing was mixed with SDS-EDTA (group 1) or SDS/heparin (group 2) and incubated for 120 min. Many smears were made within 10 min of mixing the spermatozoa with detergent and the reducing agents and at the following points of time 30, 60 and 120 min and after 24 h. Chromatin decondensation was evaluated after staining with acridine orange (AO). The mean percentage of uncondensed chromatin of spermatozoa in the semen sample in the first group before addition of SDS/EDTA was 26.1 +/- 19.0 and 22.3 +/- 18.9% in the second one. After incubation of spermatozoa for 30, 60 and 120 min and 24 h, the chromatin decondensation increased in the first group to 64.0 +/- 28.6, 83.0 +/- 21.1, 87.9 +/- 14.6, 92.1 +/- 16.2 and 98.0 +/- 6.75%, respectively. The corresponding values in the second group were 69.5 +/- 29.9, 78.6 +/- 22.4, 86.9 +/- 17.1, 95.13 +/- 6.5 and 98.3 +/- 5.6%. On the other hand, no correlations were found between the chromatin decondensation or chromatin decondensation rate in vitro and the fertilization rates in all investigated groups. In conclusion, neither the chromatin decondensation ability in vitro nor the rate of chromatin decondensation between various points of time after using SDS/EDTA, SDS/heparin could predict the chromatin decondensation of spermatozoa (fertilization capability) after ICSI.

Adult↗

PubMed2001

Comparison of sperm preparation methods: effect on chromatin and morphology recovery rates and their consequences on the clinical outcome after in vitro fertilization embryo transfer.

The aim of this study was to evaluate the efficacy of swim-up, PureSperm gradient centrifugation and glass-wool filtration methods for semen preparation and to assess the possible enhancement of the quality of the subpopulation of spermatozoa in terms of sperm concentration, morphology and chromatin condensation. Moreover, to determine the effect of this semen processing technique on the clinical outcome after in vitro fertilization embryo transfer (IVF-ET). A total of 180 semen samples of patients' husbands who were undergoing IVF therapy were prepared by swim-up (G1, n = 60), PureSperm gradient centrifugation (G2, n=60) or glass-wool (G3, n=60) methods. Chromatin condensation was assessed by Chromomycin (CMA3), whereas sperm morphology was evaluated according to strict criteria. In all three semen processing methods, the percentage of chromatin condensed and morphologically normal spermatozoa was higher after semen processing in comparison with native semen samples. The proportion of normal chromatin condensed spermatozoa prepared in glass-wool filtration was significantly higher than that in swim-up (G.I, p=0.02) or PureSperm (G.II, p=0.001). In addition semen processing with PureSperm yields significantly a higher percentage of morphologically normal spermatozoa than swim-up (p < 0.001) or glass-wool method (p < 0.002). However, the fertilization, implantation and pregnancy rates, in turn were similar in all semen preparation methods. In conclusion, PureSperm gradient centrifugation yields a higher percentage of morphologically normal spermatozoa than shown in traditional swim-up or glass-wool filtration. However, the percentage of chromatin condensed spermatozoa was significantly higher after semen processing via glass-wool in comparison with the other two methods. Nevertheless, there were no significant difference in the fertilization, implantation and pregnancy rates of sperm prepared by means of swim-up, PureSperm or glass-wool filtration. Therefore, glass-wool filtration should be recommended as the first choice for semen preparation for Intracytoplasmic sperm injection (ICSI) technique as the natural selection is bypassed. Whereas, swim-up and PureSperm should be used for semen processing in IVF programme.

Chromatin↗

PubMed2001

Association between freezing agent and acrosome damage of human spermatozoa from subnormal and normal semen.

This experimental study compares the effects of human sperm preservation medium (HSPM) with TEST-yolk buffer (TYB) as cryoprotectants of human spermatozoa with respect to the integrity of the acrosome after the freeze-thawing procedure. Fifty-six semen samples were included in this study; 18 were subnormal (G1) and 38 were normal (G2) based on World Health Organization criteria, except for morphology, which was evaluated according to strict criteria. Each semen sample was divided into two parts: the first part was prepared for cryopreservation by the addition of HSPM (1:1) and the second by addition of TYB (1:1). Freezing was performed in liquid nitrogen vapour. Smears were made before freezing and after the thawing process for evaluation of acrosome integrity using fluorescent-lectin labelling. The mean percentage of spermatozoa with intact acrosomes in the subnormal group was 77.0 +/- 7.2% before freezing and decreased significantly (P < 0.001) after thawing: to 63.7 +/- 8.2% with the use of HSPM and 66.8 +/- 8.7% with the use of TYB. The corresponding values in the normal semen samples were 83.4 +/- 9.2%, 76.0 +/- 8.8% and 77.9 +/- 9.2%, respectively. It is obvious that the decrease in the mean percentage of spermatozoa with intact acrosome was significantly higher when using HSPM in comparison with TYB, not only for G1 (-14.9 +/- 1.9% versus -11.8 +/- 1.4%) but also for G2 samples (-13.8 +/- 1.5% versus -11.9 +/- 1.3%). In conclusion, TYB should be recommended for freeze-thawing of human spermatozoa as the first-choice cryoprotectant, for normal as well as subnormal semen samples, in order to protect the sperm acrosome from the deleterious effects of the freeze-thawing procedure.

Acrosome↗

PubMed2001

Association between interleukin concentration in follicular fluid and intracytoplasmic sperm injection (ICSI) outcome.

PROBLEM: The aim of this study was to determine the presence and concentration of interleukin IL-6, IL-8, and granulocyte-macrophage-colony-stimulating factor (GM-CSF) in pre-ovulatory ovarian follicular fluid (FF) of patients undergoing controlled ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI) therapy on one hand, and to find out whether these cytotokine concentrations could be used as a predictive parameter for ICSI outcome. DESIGN: The levels of IL-6, IL-8, and GM-CSF were measured from women that underwent ICSI therapy and the results were compared between the patients who became pregnant after IC

Adult↗

PubMed2001

Comparison between the quality and function of sperm after semen processing with two different methods.

AIM: To compare the recovery rate of morphologically normal and chromatin condensed spermatozoa from native semen samples using the SpermPrep filtration columns and Percoll gradient centrifugation and to determine the influence of the two processing techniques on fertilization and pregnancy rates in an IVF-ET program. METHODS: Sixteen semen samples obtained from patient's husband were included in this study. Each was divided into two aliquots. The first aliquot was processed with SpermPrep filtration columns and the second, Percoll gradient centrifugation. Smears were made before and after semen processing with both methods for the evaluation of chromatin condensation (chromomycine CMA3) as well as morphology (strict criteria) of spermatozoa. One hundred and seventy oocytes were retrieved from the patients and the oocytes from each patient were subdivided into two sets: one set was inseminated using spermatozoa processed with SpermPrep and the other inseminated after semen processing with Percoll gradient centrifugation. RESULTS: The Percoll method yielded a significantly higher percentage of chromatin condensed (90.8 +/- 6.5% vs 82.3 +/- 8.8%, P = 0.017) and morphologically normal spermatozoa (12.9 +/- 7.4% vs 6.9 +/- 4.8%, P = 0.001) in comparison to SpermPrep. Whereas, sperm count recovery rate was significantly higher after the use of SpermPrep than after the Percoll gradient centrifugation. The fertilization rate was similar between the two methods. CONCLUSION: Semen processing with Percoll should be recommended for intracytoplasmic sperm injection as the natural selection is bypassed and the SpermPrep technique could be recommended for IVF and IUI programs as the sperm concentration plays a more significant role in these procedures.

Centrifugation, Density Gradient↗

PubMed2000

Uterine rupture in pregnancy subsequent to previous laparoscopic electromyolysis. Case report and review of the literature.

Reports about uterine rupture in pregnancy subsequent to previous laparoscopic surgery are not frequent. This may be due to the lack of long term follow up of patients who had undergone this surgery rather than the rarity of this complication. A case of uterine rupture subsequent to laparoscopic myomectomy is reported. An increasing rate of the occurrence of this complication is reviewed in current literature, thus reiterating the need for more stringent selection criteria for patients who benefit from this surgical technique.

Adult↗

PubMed2000

Relationship between ovarian stimulation regimen and cytokine concentration in follicular fluid and their effect on fertilization and pregnancy outcome of patients undergoing ICSI program.

PROBLEM: The aims of this study were to evaluate the presence of insulin-like growth factor (IGF)-I, platelet-derived growth factor (PDGF), and epidermal growth factor (EGF) in pre-ovulatory follicular fluid (FF) in patients undergoing ovarian hyperstimulation for intra-cytoplasmic sperm injection (ICSI) treatment, to determine the differences between the concentrations of these cytokines in relation to ovarian stimulation regimens, and to find the relationship between these parameters and estradiol 17-beta, progesterone, and luteinizing hormone (LH) concentration in serum, as well as ICSI outcome. METHOD: IGF-I and PDGF were measured in the FF of 85 patients. The IGF-I levels were measured by radioimmunoassay, whereas the concentrations of PDGF and EGF were measured by enzyme-linked immunosorbent assay technique, using commercially available kits. RESULTS: IGF-I (0.42 +/- 0.09 ng/mL), PDGF (307.3 +/- 274.5 pg/mL), and EDF (8.88 +/- 6.4 pg/mL) were present in pre-ovulatory FF in patients undergoing ovarian hyperstimulation for ICSI treatment. The mean concentration of IGF-I in the follicle-stimulating hormone (FSH) group was significantly higher (P = 0.036) than that found in the human menopausal gonadotrophin (hMG)/FSH group, whereas no significant difference in the mean concentrations of PDGF (P = 0.58) and EGF was shown between all investigated groups. CONCLUSION: Controlled ovarian stimulation regimens affect only IGF-I levels in FF and the cytokine concentrations of all investigated groups, in turn, showed no correlation either with steroid hormones in serum or ICSI outcome.

Adult↗

PubMed2000

Complications of triple pregnancy following intracytoplasmic sperm injection: a case report.

A case is presented of pregnancy and delivery of triplets following intracytoplasmic sperm injection (ICSI) therapy. Although the outcome was satisfactory, with the birth of normal children free from any malformation, most of the obstetric and particularly the neonatal complications that can be associated with this therapy are illustrated in this case. In addition, from point of view of medical costs, concerns are raised about the current policy of multiple embryo transfer which is directly responsible for the high rate of multiple gestations observed in the IVF/ICSI programme. The authors consequently recommend a policy of transferring not more than two embryos per treatment cycle.

Adult↗

PubMed1999

[Remifentanil for oocyte retrieval: A new single-agent monitored anaesthesia care technique].

INTRODUCTION: Transvaginal puncture for oocyte retrieval is a short-lasting but painful procedure. We hypothesized that a sole infusion of the ultra-short acting mu-agonist remifentanil may be a suitable and well-controllable single-agent analgesic technique that can dose-dependently be applied to spontaneously breathing patients. METHODS: Fifty consenting adult women were enrolled in this prospective trial. A sedative premedication was omitted, all patients received 3 L/min of inhaled oxygen, and a sole remifentanil infusion was started with 0.25 microg/kg/min. Remifentanil was adjusted as needed for pain relief (in steps of 0.05 microg/kg/min) and finished after the last puncture. Dosage requirements, vital functions, oxygen saturation (as achieved by pulse oximetry, psO(2)), adverse drug effects and the level of sedation (LOS 1-5; 1 = asleep/unarousable, 4 = calm/awake) were recorded. Remifentanil plasma concentrations were achieved by STANPUMP pharmacokinetic simulation. Data are presented as mean +/- SD. RESULTS: A total of 50 women (31.8 +/- 5.1 yr, 67.3 +/- 14. ASA I or II ) were investigated. Follicular aspiration lasted 10.8+/-5.2 min, and remifentanil was infused for 19.7+/-8.3 min. Dosage requirements were 0.25 microg/kg/min in 70% of all patients, 0.3 microg/kg/min in 22%, 0.2 microg/kg/min in 6%, and 0.4 microg/kg/min in 2% of all cases. Vital signs (baseline, after 1(st) puncture, end of surgery) nearly remained unchanged: heart frequency = 85 +/- 15, 87 +/- 17, 90 +/- 17 bpm, systolic blood pressure = 129 +/- 12, 132 +/- 13, 131 +/- 14 mmHg; respiratory rate = 116 +/- 4, 15 +/- 4 breaths/min; psO(2) = 99 +/- 1, 99 +/- 1, 99 +/- 2%. LOS was 4.0 (all), 3.9 +/- 0.3, 3.9 +/- 0.3. Remifentanil plasma concentrations were 5.0 +/- 1.3 ng/mL at the start, 6.6 +/- 1.3 at the end of surgery and 1.2 +/- 0.5 at PACU arrival. Adverse drug effects: 54% itching, no muscle rigidity. 94% of all women would choose this technique again. CONCLUSIONS: The sole infusion of remifentanil is a suitable and satisfying single-agent monitored anaesthesia care technique for oocyte retrieval. However, close anaesthetic observation - especially of the respiratory function - is mandatory.

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