Assessment of bone metabolism and biomechanical properties of the femur, following treatment with anastrozole and letrozole in an experimental model of menopause.

The aim of the present study was to investigate the impact of anastrozole and letrozole supplementation following surgically induced menopause on bone metabolism and biomechanical properties. A total of 45 Wistar rats underwent ovariectomy and were then randomly allocated to receive no treatment, anastrozole or letrozole. At 2 and 4 months following the initiation of the present study, the serum levels of osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) were determined, and the animals were sacrificed at the end of the 4-month period to assess the biomechanical properties of the femoral bones. The applied force and the deflection of the central section were recorded during the test. Taking advantage of these quantities, the fracture force, the stiffness of the bone and the energy absorbed until fracture were determined. At 2 months following the initiation of the experimental protocol, the mean OPG levels were significantly increased in the control group compared with the anastrozole-treated group (P<0.01). Similarly, RANKL levels were significantly increased in the control rats compared with the anastrozole-treated animals (P<0.001) and animals that received letrozole (P<0.05). Notably, these trends were not observed at the end of the experiment (4 months). A biomechanical study of the femoral bones revealed significantly decreased stiffness among animals that received anastrozole (P<0.05) and letrozole (P<0.01) compared with their control counterparts. The results of the present study indicate that treatment with anastrozole and letrozole significantly increases the levels of OPG and RANKL in bone, an effect that appears to be directly associated with the biomechanical properties of bones.

The impact of Anastrazole and Letrozole on the metabolic profile in an experimental animal model.

Anastrazole and Letrozole are used as endocrine therapy for breast cancer patients. Previous studies suggested a possible association with metabolic and liver adverse effects. Their results are conflicting. Fifty-five 4-week-old female Wistar rats were allocated in 4 groups 1) ovariectomy control (OC), 2) ovariectomy-Anastrazole (OA) 3) ovariectomy -Letrozole (OL), 4) control. Serum glucose, cholesterol, triglycerides, HDL-c and LDL-c were measured at baseline, 2 and 4 months. At the end, the animals' liver were dissected for pathology. At 4 months, total cholesterol differed among the OC and OL groups (p = 0.15) and the control and OL groups (p = 0.12). LDL-C differed between the control and OC groups (p = 0.015) as well as between the control and OA (p =0 .015) and OL groups (p = 0.002). OC group triglycerides, differed from those of the OL group (p =0 .002) and the control group (p = 0.007). The OA also significantly differed from the OL (p = 0.50). Liver pathology analysis revealed differences among groups with favored mild steatosis and ballooning. Anastrazole and Letrozole seem to negatively influence the lipid profile in our experimental model. This information should be taken in caution by medical oncologists when addressing patients with altered lipid metabolism.

Can anti-Mullerian hormone (AMH) predict the outcome of intrauterine insemination with controlled ovarian stimulation?

OBJECTIVE: To assess whether the levels of anti-Mullerian hormone (AMH) are related to outcome of intrauterine insemination (IUI) in patients treated with gonadotropins. INTERVENTION(S): A total of 195 patients underwent controlled ovarian stimulation (COS) with recombinant follicle stimulating hormone (rFSH) (50-150 IU/d). All patients were submitted upto three cycles of IUI. OUTCOME: Primary outcome was the ability of AMH levels to predict clinical pregnancy at first attempt and the cumulative clinical pregnancy probability of upto three IUI cycles. Secondary outcomes were the relation of AMH, LH, FSH, BMI, age, parity and basic estradiol levels with each other and the outcome of IUI. RESULTS: The area under the receiver operating characteristic (ROC) curve in predicting clinical pregnancy for AMH at first attempt was 0.53 and for cumulative clinical pregnancy was 0.76. AMH levels were positively correlated with clinical pregnancy rate at first attempt and with cumulative clinical pregnancy rate, but negatively correlated with patient's age and FSH levels. Patient's FSH, LH levels were negatively correlated with cumulative clinical pregnancy rate. CONCLUSIONS: AMH levels seem to have a positive correlation and patient's age and LH levels had a negative correlation with the outcome of IUI and COS with gonadotropins. AMH concentration was significantly higher and LH was significantly lower in patients with a clinical pregnancy after three cycles of IUI treatment compared with those who did not achieve pregnancy.

Role of hysteroscopy prior to assisted reproduction techniques.

STUDY OBJECTIVE: To determine whether diagnostic hysteroscopy before assisted reproduction techniques (ΑRT) in women without known disease of the uterine cavity is necessary. DESIGN: Prospective cohort clinical study. SETTING: Reproductive medicine clinic. PATIENTS: The study group consisted of 217 infertile women attending the Reproductive Clinic for examination before undergoing ART, either in vitro fertilization or intracytoplasmic sperm injection. INTERVENTIONS: Patients underwent transvaginal sonography (TVS) and hysterosalpingography (HSG) for initial evaluation. If there were no abnormal intrauterine findings, diagnostic hysteroscopy was additionally performed. MEASUREMENTS AND MAIN RESULTS: The safety and diagnostic value of hysteroscopy before ART was examined. Diagnostic hysteroscopy was performed successfully, without complications, in all 217 women. Ninety-five (43.7%) had a history of ART failures (group 1), and 122 (56.3%) had undergone no previous ART attempts (group 2). In 148 women (68.2%), findings at hysteroscopy were normal, whereas in 69 (31.8%), hysteroscopy revealed intrauterine lesions (polyps, septa, submucosal leiomyomas, or synechiae) that led to operative hysteroscopy. The most common intrauterine abnormality was the presence of endometrial polyps in 26 patients (12%). The total percentage of abnormal intrauterine findings was higher in women with a history of repeated ART failures in comparison with those with no history of ART attempts. No statistically significant difference in the outcome of in vitro fertilization or intracytoplasmic sperm injection was observed between women with normal hysteroscopic findings and patients with hysteroscopically corrected endometrial disease. CONCLUSION: Sensitivity of diagnostic hysteroscopy is significantly higher than TVS and HSG in the diagnosis of intrauterine lesions. Diagnostic hysteroscopy should be performed before ART in all patients, including women with normal TVS and/or HSG findings, because a significant percentage of them have undiagnosed uterine disease that may impair the success of fertility treatment.

Changes in hormonal profile and seminal parameters with use of aromatase inhibitors in management of infertile men with low testosterone to estradiol ratios.

OBJECTIVE: To compare the effects of 2.5 mg letrozole with those of 1 mg anastrazole daily on the hormonal and semen profiles of a subset of infertile men with low T/E(2) ratios. DESIGN: Prospective, nonrandomized study. SETTING: Reproductive medicine clinic. PATIENT(S): The study group consisted of 29 infertile men with a low serum T/E(2) ratio (<10). INTERVENTION(S): Patients were divided into two groups. Group A included 15 patients treated with 2.5 mg letrozole orally once daily for 6 months, and Group B consisted of 14 patients treated with 1 mg anastrazole orally every day for 6 months. MAIN OUTCOME MEASURE(S): Hormonal evaluation included measurement of serum FSH, LH, PRL, T, and E(2). In all sperm analyses pretreatment and posttreatment total motile sperm counts (ejaculate volume × concentration × motile fraction) were evaluated. RESULT(S): The use of aromatase inhibitors (either letrozole or anastrazole) in cases of infertile men with low T/E(2) ratios improved both hormonal and semen parameters. CONCLUSION(S): This study suggests that some men with severe oligospermia, low T levels, and normal gonadotropin concentration may have a treatable endocrinopathy.

Maternal and umbilical resistin levels do not correlate with infant birth weight either in normal pregnancies and or in pregnancies complicated with gestational diabetes.

OBJECTIVE: To evaluate the role of resistin in the pathophysiology of insulin resistance during pregnancy and on the birth weight of infants born from women with gestational diabetes (GDM). MATERIAL AND METHODS: Thirty women diagnosed with GDM were compared to 30 normal pregnant controls. Maternal serum resistin and insulin levels were measured at the time of the oral glucose tolerance test screening. In addition, umbilical levels of resistin and insulin were measured at the time of delivery. RESULTS: There was no difference in maternal serum resistin levels in women with GDM as compared to normal controls at 24-26 weeks. There was no difference in umbilical resistin levels between the infants born in the two groups. There was no correlation between infant weight and either maternal resistin at 24-26 week or umbilical resistin levels. CONCLUSION: There were no significant differences in umbilical resistin levels between infants born of women with GDM as compared to normal pregnant women. In addition, there was no correlation between resistin levels during pregnancy, as well as between umbilical resistin levels and neonatal birth weight. In conclusion, resistin seems to play a rather minor role in the pathophysiology of GDM and the energy metabolism during fetal life.

Effect of pentoxifylline on vascular endothelial growth factor C and flk-1 expression on endometrial implants in the rat endometriosis model.

OBJECTIVE: To investigate the effects of pentoxifylline, on vascular endothelial growth factor (VEGF)-C and flk-1 expression in the rat endometriosis model. DESIGN: Prospective, randomized, placebo-controlled study. SETTING: Academic institution. ANIMAL(S): Twenty Wistar rats with surgically induced endometriosis. INTERVENTION(S): Animals were evaluated after surgical induction of endometriosis and random allocation to a group that received pentoxifylline and a control group that received NaCl 0.9%, for 3 weeks. At the end of the treatment period the animals were killed and the implants evaluated macroscopically as well as by immunohistochemistry. MAIN OUTCOME MEASURE(S): Morphologic changes of the endometriotic implants; and evaluation of VEGF-C and flk-1 expression by a semiquantitative analysis (HSCORE) for the intensity of immunohistochemical reactivity. RESULT(S): A significant reduction was observed in the mean volume of the endometriotic implants per animal in the treatment group as compared with the control group. There was a significant reduction not only in the mean volume of implants per animal but also in the mean number of implants per animal after treatment. By immunohistochemical evaluation (HSCORE), there was a significant reduction in VEGF-C expression after treatment in all areas examined. A significant reduction of flk-1 expression was also noted in the glandular compartment after treatment but not in the epithelial surface or stroma. CONCLUSION(S): Pentoxifylline may cause suppression of endometriotic lesions by suppressing angiogenesis through VEGF-C and flk-1 expression.

Randomized controlled trial comparing superovulation with letrozole versus recombinant follicle-stimulating hormone combined with intrauterine insemination for couples with unexplained infertility who had failed clomiphene citrate stimulation and intrauterine insemination.

OBJECTIVE: To compare the efficacy of letrozole to recombinant FSH for ovarian stimulation combined with IUI in a group of patients that had failed to conceive after clomiphene citrate (CC) and IUI. DESIGN: Prospective randomized trial with human subjects. SETTING: University-based fertility center. PATIENT(S): Fifty couples with unexplained infertility that failed to conceive after three cycles of CC combined to IUI. INTERVENTION(S): Couples were randomized to undergo superovulation either with letrozole or with recombinant FSH combined to IUI. MAIN OUTCOME MEASURE(S): Clinical pregnancy per cycle of treatment and clinical pregnancy per couple. RESULT(S): Pregnancy rate (PR) per cycle was 8.9% in the letrozole group as compared with 14% in the gonadotropin IUI group. This resulted in a cumulative PR per couple of 24% versus 36% and a take home baby rate of 20% versus 28%. Endometrial thickness was significantly lower in the letrozole group (7.1 +/- 2.3 vs 8.6 +/- 1.8). CONCLUSION(S): Ovarian stimulation with letrozole is associated with acceptable PRs compared with gonadotropin with significant less cost, risks, and patient inconvenience.

Prevention and management of ovarian hyperstimulation syndrome.

The Ovarian Hyperstimulation Syndrome (OHSS) represents one of the biggest nightmares of all physicians involved in Assisted Reproductive Technologies (ART). Every year, several hundreds of women are hospitalized and to date several deaths have been reported. The pivotal event in the development of OHSS is the disruption of capillary integrity that results in leakage of intravascular fluid and proteins into third space. On the molecular level, human chorionic godadotropin (HCG) either exogenous or endogenous, functions as the triggering point for the production of vascular endothelial growth factor (VEGF) that is the main mediator to increase permeability on the vascular bed. Spontaneous OHSS has also been reported, either due to inappropriate activation of a mutant FSH receptor or due to very high levels of HCG during pregnancy. The available evidence on the several preventive and therapeutic approaches with special attention to level 1 evidence when available is also presented. OHSS is a self-resolving condition and the main role of the physician is to correct and maintain the intravascular volume, to support renal function and respiration and prevent thrombotic events. An algorithm on the management of OHSS on an outpatient basis and in the hospital is based on the previous mentioned principles.

Effect of oral contraceptive treatment on bone mass acquisition in skeletally immature young female rats.

The objective of the present study was to investigate the effect of oral contraceptive (OC) treatment on bone mass accrual in skeletally immature young female rats. Animals in the baseline group were killed at the beginning of the experiment and were subjected to bone density assessment by peripheral quantitative computerized tomography (pQCT). The control group was fed a base diet free of phytoestrogens, while animals in the contraceptive group received the same base diet mixed with 2.67 microg desogestrel/100 g body weight and 0.0533 microg ethinyl estradiol/100 g body weight. The duration of the treatment period was 16 weeks. Densitometric measurements by dual energy x-ray absorptiometry and serum bone markers assessment were carried out at baseline, at 8 weeks and at 16 weeks, while pQCT densitometry took place after sacrifice. All bone mineral density and bone mineral content indices measured by dual energy x-ray absorptiometry increased significantly throughout the study period in both the OC and control group. Concerning pQCT measurements, animals in both the OC and the control group had significantly higher cortical density compared with baseline (midtibia: p=.0003 and .0003, respectively). Total area and periosteal circumference were significantly higher in OC group, both in proximal (p=.003 and .003, respectively) and midtibia (p=.048 and .042, respectively) compared with baseline. Osteoprotegerin serum levels increased in both groups, and at the end of the experiment, circulating osteoprotegerin was significantly higher in the OC group compared with controls (p=.032). At the end of the experiment, carboxyl-terminal telopeptides of collagen type I levels were significantly lower in the OC-treated animals compared with controls (p=.046). Our results suggest that OC administration to skeletally immature female rats allows normal bone accrual and may even improve bone geometry. This effect may be mediated through enhanced inhibition of bone resorption.

Thyroid hormone resistance and enlargement of the sella turcica during pregnancy.

CASE REPORT: A 26-year-old pregnant woman was admitted to our institution running her 30 weeks of gestation. The patient had a past history of total thyroidectomy cause of a thyroid papillary carcinoma and presented with increased supraphysiological TSH levels under 250 microg T4, while slightly hyperthyroid, from the clinical point of view. Partial resistance to thyroid replacement therapy or TSH-secreting tumour was evoked. Pituitary MRI revealed a pituitary enlargement without excluding a pituitary adenoma. To avoid further stress on pituitary a caesarean section was performed at 38 weeks of gestation. MRI 7 months later was normal, while the patient remained under high doses of T4 replacement therapy and TSH was found at the upper limits of normalcy, while T3, T4 and FTI were above normalcy. CONCLUSION: We conclude that, in the absence of thyroid gland, high TSH levels due to thyroid hormone resistance could be erroneously attributed to a pituitary TSH secreting tumour, when associated with a pregnancy-related pituitary enlargement.

Continuous combined replacement therapy in postmenopausal women: effect on plasma lipoproteins, bone mass and the endometrium.

OBJECTIVE: The purpose of this study was to examine the effects of Hormone Replacement Therapy (HRT) on plasma lipoprotein levels, bone mass and the endometrium. METHODS: Thirty healthy women were enrolled in the study for climacteric symptoms and received a combination of 17beta-estradiol and norethisterone acetate continuously for 2 years. An untreated group, consisting of 25 healthy postmenopausal women presenting the same symptoms and not willing to receive HRT, were also studied and served as controls. Plasma lipoprotein levels, bone mineral density as well as thickness of the endometrium measured by ultrasound and biopsy of the endometrium were evaluated at baseline and after 1 and 2 years. There were no differences between the two groups regarding clinical and demographic baseline data. RESULTS: After 2 years of HRT, total cholesterol and LDL-cholesterol levels were significantly reduced (p<0.05). Changes in serum triglyceride and HDL-cholesterol levels were not statistically significant. Levels of lipoprotein (a) showed a statistically significant decrease at 1(st) and 2(nd) year (p<0.05) in both groups while levels of ApoA1 and ApoB were significantly lower only in the HRT group compared to the baseline values. After 2 years of HRT, biomarkers of bone metabolism showed a significant decline, while endometrium was atrophic in 93.3% of the cases. CONCLUSIONS: Hormone replacement therapy (17beta-estradiol combined with norethisterone acetate) was associated with favorable changes in the lipid status and bone metabolism. Endometrium was atrophic in most of the treated subjects.