Effect of Some Heavy Metals on Testosterone Hormone in Infertile Men

الخلفية: يتم تعريف العقم على أنه فشل في تحقيق الحمل بعد عام واحد وأكثر من الاتصال الجنسي غير المحمي بانتظام. ارتبط تعرض الجهاز التناسلي للمعادن الثقيلة أيضًا بالعقم عند الرجال. وهناك عدد كبير من المعادن عبارة عن مواد سامة للجهاز التناسلي ويشتبه في كونها مضطربة لنظام الغدد الصماء. المرضى والطرق: أجريت هذه الدراسة في المعهد العالي لتشخيص العقم والتقنيات المساعدة الإنجابية في جامعة النهرين ومركز استشاري التسمم / مستشفى الجراحات التخصصية في مدينة الطب في الفترة من نوفمبر 2018 إلى أبريل 2019. كان عدد الرجال 150 المسجلين في هذه الدراسة مقسمة إلى مجموعتين ، المريض ومجموعات المراقبة الطبيعية. وشملت مجموعة المرضى 100 رجل تتراوح أعمارهم بين 20-50 سنة. شملت المجموعة الثانية 50 رجلاً يتمتعون بصحة جيدة ولديهم طفلان على الأقل دون أي تاريخ سابق للأمراض الجهازية كمجموعة مراقبة. تم تقدير تركيزات المعادن الثقيلة لجميع مجموعات الدراسة. الهدف من الدراسة الحالية هو تقييم بعض تركيزات المعادن الثقيلة ، في الدم والدم الكامل للمجموعات. تقييم هرمون التستوستيرون في مصل الرجال المصابين بالعقم والضوابط الصحية ودراسة العلاقة بين هذا الهرمون والعناصر النزرة التي قدرت بين الحالات. النتائج : أظهرت تراكيز المعادن الثقيلة وهي كل من النحاس والرصاص والكادميوم زيادة معنوية عالية وتحت مستوى احتمالية اقل من 0.010.01 في مصل دم و دم الرجال العقيمين حيث سجلت (139.7+ 8.49 Mg/dl, 24.18 + 4.30 Mg/dl, 0.31 + 0.10 Mg/dl) على التوالي وعند المقارنة مع التراكيز في مصل الدم و دم الرجال السليمين ( السيطرة ) كانت النتائج  على التوالي(125.14 + 10.12 Mg/dl, 15.24 + 2.69 Mg/dl, 0.16 + 0.03 Mg/dl)  كما أظهرت متوسطات عنصر الزنك (القصدير) انخفاضا معنويا عاليا عند مستوى احتمالية اقل من 0.01      (70.82+9.48Mg/dl) في مصل دم الرجال العقيمين مقارنة مع مجموعة السيطرة حيث كانت كما أظهرت تراكيز هرمون الشحمون الخصوي انخفاضا معنويا عاليا وتحت مستوى احتمالية اقل من 0.01  (185.40 + 61.92 Ng/dl)  (450.35+131.27 Ng/dl) بالمقارنة مع مجموعة الرجال السليمين كذلك كانت علاقة الارتباط لمستويات تراكيز عنصر النحاس وهرمون الشحمون الخصوي في مصل الدم عكسية عند الرجال العقيمين. الاستنتاجات: خلصت هذه الدراسة إلى أن بعض المعادن الثقيلة أظهرت أنها مؤشرات لعقم الرجال. ارتبط مستوى النحاس في المصل سلبًا بمستوى هرمون التستوستيرون في المصل لدى الرجال المصابين بالعقم، وهذه النتيجة يمكن أن تفترض أن النحاس الزائد يعمل كمضاد هرموني. لم تظهر المعادن الثقيلة المتبقية ارتباطًا كبيرًا بالهرمونات المحددة.Background:  Infertility is defined as a failure of achieve a pregnancy after one year and more of regular unprotected sexual intercourse. The exposure of reproductive system to heavy metals has also been associated with male infertility. A large number of metals are toxicants to the reproductive system and are suspected to be endocrine system disruptors.   Patients and methods:  This study performed at the high institute of infertility diagnosis and assisted reproductive technologies in Al-Nahrain University and the Poisoning consultation center / specialized surgeries hospital in the Medical city in period from November 2018 to April 2019.  Number of 150 men were enrolled in this study divided into two groups, the patient and the normal control groups. The infertile group included 100 men aged 20-50 years who they have abnormal seminal fluid analysis parameters. The second group included 50 healthy fertile men who have at least two children without any previous history of systemic diseases as a control group. Heavy metals and testosterone concentrations were estimated for all the study groups. Aim of current study are evaluation of some heavy metals concentrations, in serum and whole blood of groups. Assessment of testosterone hormone in serum of infertile men and healthy controls and study the correlation between this hormone and trace elements which were estimated among cases. Results: The heavy metals Cu, Pb and Cd showed a highly significant increase (p<0.01) in the serum and blood of infertile men (159.77±8.49 µg/dl, 24.18±4.30µg/dl, 0.31±0.10 µg/dl) respectively when compared with their concentrations in healthy control men (125.14±10.12 µg/dl, 15.74±2.69 µg/dl, 0.16±0.03 µg/dl) respectively. The mean levels of Zn showed a highly significant decrease (p<0.01) in the serum of infertile men (70.82±9.48 µg/dl) compared to its mean level in the healthy control group (95.70±11.41 µg/dl). Serum testosterone hormone in infertile men (185.40±61.92 ng/ml) showed a highly significant decrease (p<0.01) as compared with those of healthy men (450.35±131.27 ng/ml). Serum copper level was negatively correlated with serum testosterone level among infertile men. Conclusions: this study concluded that some heavy metals were shown to be an indicators for men infertility. Serum copper level was negatively correlated with serum Testosterone level among infertile men .This result can hypothesize that excess copper acts as a hormonal disrupter. The remaining heavy metals did not show significant correlation with the selected hormones

The effect of serum and follicular fluid vitamin D on intracytoplasmic sperm injection outcome

Vitamin D is a cholesterol derived, fat soluble, steroid substance present in the body. It plays an important role in the female reproductive system and regulating its functions. It is believed that vitamin D improves the endometrial thickness. The relationship between the level of vitamin D and fertilization, embryo quality and Intracytoplasmic sperm injection outcome remains controversial till now. The aim of the study to assess the relation of vitamin D levels in serum and follicular fluid with Intracytoplasmic sperm injection outcome. Eighty-eight women were enrolled in this study. They were classified according to the cause of infertility into case group with female factor and control group with male factor. All women were undergoing Intracytoplasmic sperm injection procedure. Serum and follicular fluid vitamin D levels were measured on the day of oocyte retrieval. It was found that no significant differences in either serum or follicular fluid level of vitamin D between the case and control groups and higher serum and follicular fluid levels of vitamin D are associated with better pregnancy rates after Intracytoplasmic sperm injection. Further studies on vitamin D in male are required to prove its role in the Intracytoplasmic sperm injection. The present study concluded that no significant differences in either serum or follicular fluid level of vitamin D between the case and control groups and higher serum and follicular fluid levels of vitamin D are associated with better pregnancy rates after Intracytoplasmic sperm injection

Role of Vascular Endothelial Growth Factor (VEGF) and doppler sub-endometrial parameters as predictors of successful implantation in Intracytoplasmic Sperm Injection (ICSI) patients

Objective: to investigate the expression of vascular endothelial growth factor (VEGF) in patients having infertility due to low endometrial acceptance, and to correlate it to non-invasive ultrasound variables, endometrial thickness, and sub-endometrial Doppler parameters (PI, RI, Vs/Vd). Methods: 80 women all under the age of 40 underwent ICSI-ET; all patients were exposed to ovarian stimulation protocols. The oocytes were retrieved using an ultrasound guide, and were fertilised via injection of sperm inside the follicle (ICSI). VEGF serum level was analysed at day of embryo transfer by ELIZA test, and sub-endometrial evaluation was conducted via two-dimension power Doppler ultrasound (2D PD-US), by measuring resistance index (RI) and pulsatility index (PI) on the day of embryo transfer. Results: There was a significantly higher VEGF level and endometrial thickness in pregnant (433± 207 and 9.72± 1.35) women, compared to non-pregnant (276± 165 and 8.95 ± 1.21) respectively as p-values were (0.001 and 0.01)). Additionally, there were significantly lower RI and PI in pregnant (0.584 ± 0.124 and 0.829 ± 0.301) women compared to non-pregnant (0.651±0.132 and 1.006±0.335) women, as p-values were (0.02 and 0.02, respectively). The level of E2 was on the day of embryo transfer and Vs/Vd in pregnant women (1402± 524 and 3.14 ± 3.75) and in the non-pregnant group (1296± 611 and 3.82 ± 3.07), as p-values were 0.41 and 0.38, respectively. Conclusion: The combined analysis of endometrial receptivity was completed, and the serum level of VEGF and sub-endometrial evaluation with 2D PD-US was defined by measuring resistance index (RI) and pulsatility index (PI) on the day of embryo transfer. These can serve as useful prognostic methods for the detection of endometrial receptivity and pregnancy outcomes in infertile women undergoing ICSI protocols, and will be helpful for candidate counselling about postponing embryo transfer and cryopreservation, which may serve as a better option, to be recommended for the next cycle, when achieving better endometrial Doppler parameters

Impact of xenotransplantation of sheep ovarian cortex and follicular fluid- enriched SMART medium on the morphology of recovered of sheep oocyte

Ovarian follicles and cortical tissue can survive viable after ovariectomy and use successfully for xenotransplantation or in vitro maturation. This study designed to assess in vitro maturation (IVM) of sheep oocytes recovered from immature follicles using simple medium for assisted reproductive techniques (SMART) medium enriched with follicular fluid (FF) using xenotransplanted ovarian tissue cortex inside the body of female mice injected with or without different hormonal stimulation protocols. In this study, follicular fluid (FF) aspirated from randomly sheep ovarian follicles. Seventy-five healthy and mature female mice were used for transplantation of sheep ovarian tissue (OT) on the inner side of the peritoneum. Later on, these female mice were classified into two groups. Group A: control (without medication). Group B: hormonal programs (hormonal stimulation). The last experimental group was Group C: the direct examination of sheep ovarian cortex group. The sheep ovarian cortex was recovered from female mouse then oocytes were collected by slicing for assessment and classified into three groups. Group1: oocytes incubated within SMART medium alone (control group). Group 2: oocytes incubated within SMART medium enriched with 5% FF, and Group 3: oocytes incubated within SMART medium enriched with 10% FF. The normal oocytes morphology was assessed post-xenotransplantation and parameters were statistically analyzed. No significant (P> 0.05) difference of normal oocytes morphology was seen between treated group (GB) and control group (GA). However, significant (P= 0.043) difference was observed in the percentage of normal oocyte morphology of recovered oocytes for the group (GC) in compare to (GA). Meanwhile, there was highly significant (P< 0.001) difference between GB and GC groups. Addition 5% FF to SMART medium of GB revealed significant (P= 0.044) difference in compare to the GA. Moreover, highly significant (P< 0.001) difference was observed in GC in compare to GA and GB compared to GC. Meanwhile, addition 10% FF to SMART medium of G3 revealed significant (P< 0.05) difference in the percentages of the normal and abnormal morphology of recovered oocytes. In conclusion, this study approved that normal oocyte morphology and maturation was valuable when using ovarian tissue grafts. In addition, the combination of SMART medium with 10% FF was revealed the best oocyte morphology and maturation

Morphometric study of changes in GAD 65 expressing beta cell mass in the islets of langerhans in pregnant mice

INTRODUCTION: This study explored the adaptive changes in pancreatic islets by assessing the maternal pancreatic β-cell mass (PβCM), islets size, number, shape, distribution and vascularity using the anti-GAD65 Ab as a marker in pregnant mice. MATERIALS AND METHODS: Fourty female Swiss-Webster mice were randomly divided into two age matched groups of 20 animals each, pregnant and control (non-pregnant) groups. The pregnant mice were sacrificed at Day 19 of gestation and the control group was sacrificed 19 days after observation in the same environmental conditions. Specimens of pancreata were fixed in 10% formal-saline, and processed for light microscopy. Paraffin sections were stained with chromogen DAB. Image scope (APERIO) with Image J software was used for morphometric study. RESULTS: Islets cells showed specific reaction with GAD65 Ab. There was an increase in the cellularity and vascularity of the PβCM. A significant increase in the islets area of the pregnant group ranging between 32-723 μm2 in comparison with the control group that ranged between 5–210 μm2, the islet size and numbers also increased; there were numerous newly formed islets and coalescence of adjacent islets, positive reactivity of β-cells toward anti- GAD65 during pregnancy, the mean of positive granulation in PβCM in pregnant group was 8273.5±SE 831.35 while in control group was 34±SE 0.2. CONCLUSION: The changes for alteration in PβCM such as increase in cells number, size, positive anti-GAD65 reactivity and their vascularity during pregnancy is related to the adaptation for hormonal and nutritional demands of both foetus and mother

Comparison between the effect of GnRH agonist and HCG injection on the luteal phase support in patient undergoing IUI

Intrauterine insemination is amongst the foremost recommended procedures to reinforce the probability of pregnancy in couples with infertility. The success rate of this system depends on numerous factors, one among which is that the quality of the luteal phase which could be due to insufficient production of progesterone. To study the impact of luteal phase GnRH agonist versus HCG supplementation on pregnancy rates in females undergoing intrauterine insemination. This study was performed on random 40 infertile couples. Basal hormones measurement of FSH, LH, E2 and progesterone was done before undergoing ovarian stimulation by letrozole or clomiphene citrate. In the mid cycle transvaginal ultrasound was done to measure the dominate follicle/ s and endometrial thickness (ET). Assessment of progesterone and E2 was done on the trigger day. Triggering of ovulation was done by the HCG injection (5000IU). IUI for all females included in the study then they were divided into two groups of 20. Group A were supplemented with GnRH agonist injections (0.1 mg) on day 9,10,11,12 post IUI plus progesterone suppositories 400 mg for 14 days while group B were supplemented with HCG injections 1500IU on day 1, 4, 7 post IUI plus progesterone suppositories 400 mg for 14 days. Then Progesterone level was re measured at day 21 of the cycle. Chemical pregnancy rate was done and recorded. The hormonal levels showed no significant difference in baseline serum FSH, LH and estradiol (E2) (P > 0.05); however, there was significant difference in mean serum baseline progesterone between protocol groups (p= 0.024); additionally, there was no significant difference in mean serum estradiol (E2) and progesterone at day 12 (p> 0.05), but the difference in mean serum progesterone at day 21 was highly significant (p < 0.001); its level was highest in group B than A. The pregnancy rate of infertile women in groupA (who used GnRH as supportive to luteal phase) was higher (40.0 %).Conclusions: Luteal phase support by GnRH injection (Decapeptyl 0.1mg) significantly increased pregnancy rate in infertile females undergoing intrauterine insemination compared to HCG injection (1500IU) in spite of significant elevation in progesterone hormone at 21the day in HCG group

Histological observation related to the use of laser and ultrasound on bone fracture healing

Objective: To study the effect of both laser and ultrasound radiation on bone fracture healing process. Materials and Methods: Nd:YAG laser (1064 nm wavelength, 135 mW power, 16 joules energy) and ultrasound (1 MHz frequency, 50 mW/cm2 power intensity) were used in this work. Fifteen mature, male, albino rats, were divided into three groups and subjected to a partial fracture on the lateral aspect of femur by a sharp blade. The fi rst group of these animals served as control group. The second group was illuminated by the Nd:YAG laser for two minutes; the fi rst dose was given immediately after surgical fracture induction; the other doses were given on days two, three, six and then one dose weekly for the next three weeks while the third group were treated by the addition of the CW ultrasound perpendicular to the laser treatment in the second group. Results: The present study showed that ultrasound increases the penetration of laser power through the tissue. The histological assessments at day 28 after the fracture of fi rst group showed incomplete healing of the bone with disfi guration and disarrangement of Haversian system and the periosteum was not yet well developed. Treatment with laser showed irregularity and lack of Haversian system formation in bone healing of the second group. The laser and ultrasound treated group (third group) expressed a complete healing at the site of fracture with a complete layer of periosteum and a well arranged Haversian system. Conclusion: Combination of laser and ultrasound in therapy can enhance healing process of a fractured bone more than laser therapy alone, as ultrasound increases the depth of laser penetration in tissue

The effects of dexamethasone on Tibia development of local chick-Embryo. I: computer-assisted morphometric study

Dexamethasone is a glucocorticoid as a member of the steroidal anti-inflammatory and immunosuppressant. It has well documented effects on skeletal structures osseous and cartilaginous, commonly used to treat or control diseases. Objective To evaluate by histomorphometric study the effects of dexamethasone on the embryogenesis of long bones in chick embryos. Methods Forty-eight fertile chick eggs of Gallus gallus domesticus, were used. The eggs were divided into 2 groups; control and treated groups of 24 eggs each, these groups were subdivided into 4 subgroups (n=6 eggs). On day 10 of incubation, the control group was injected with 25 μl of distilled water while the treated group was injected with 25 μl of distilled water contained 8 μg dexamethasone. In the next days (11, 12, 13, and 14 of incubation), 12 chick embryos were sacrificed in each day. A computer-assisted morphometric/ image analysis (Motic Image Plus version 2.0ML), was used to measure length, area, perimeter of tibiae, and the area and perimeter of the perichondral osseous collar of cross section in mid-diaphyseal zone of these bones. Results These bones of chick embryos treated with dexamethasone, suffered shortening and retardation in length, weight, area and perimeter throughout the period of this study, decline area and perimeter to the perichondral osseous collar in the mid-diaphyseal zone. Conclusion Dexamethasone given at day 10 of incubation caused tibial bones growth retardation at development stages 11, 12, 13, and 14-days; this was observed in the measured parameters: bone length, area, perimeter and weight