Functional and structural assessment of the possible protective effect of platelet-rich plasma against ischemia/reperfusion-induced ovarian injury in adult rats.

This study aimed to evaluate the possible protective effect of platelet-rich plasma (PRP) on ischemia reperfusion (I/R)-induced ovarian injury in a rat model. Forty adult female albino rats were randomly assigned to four groups: control, ischemia, I/R, and I/R + intraperitoneal PRP. Induction of ischemia was done by bilateral ovarian torsion for 3 h, while reperfusion was done by subsequent detorsion for another 3 h. PRP was injected 30 min before detorsion. Histological assessment and measurement of ovarian anti-Mullerian hormone (AMH) were done to assess the degree of tissue damage and the remaining ovarian reserve. Ovarian malondialdehyde (MDA) and total antioxidant capacity (TAC) levels were measured to evaluate the oxidant-antioxidant balance. Tumor necrosis factor-α (TNF-α) was measured to assess degree of inflammation. Immunohistochemical assessment of ovarian vascular endothelial growth factor-A (VEGF-A) was also done. PRP treated I/R group revealed a significant decrease in MDA (P = 0.007), TNF-α (P = 0.001), and a significant increase in TAC (P = 0.001) and VEGF-A (P = 0.003) in comparison to the untreated I/R group. Furthermore, limited vascular congestion and inflammatory infiltration were observed after PRP treatment. However, no significant difference was detected in AMH after PRP treatment. Our results denoted that PRP may help in preservation of ovarian function and structure during surgical conservative detorsion of the torsioned ovary. These protective effects could be attributed to its ability to reduce oxidative stress, inflammation and also to its high content of growth factors especially VEGF.

An assessment of the protective effect of gonadotropin-releasing hormone agonist and antagonist on bleomycin-induced ovarian toxicity in rats.

The aim of this study is to evaluate the effect of GnRH agonist or GnRH antagonist therapy on bleomycin-administered rats by examining ovarian follicle counts and AMH levels. A total of 30 female Wistar albino rats aged 4-6 months were randomly divided into 4 groups. First, an intramuscular injection of bleomycin (30 mg/m2) was administered to all except the control group on the 1st, 8th and 15th days. The control group (Group I) was administered 0.1 mL intramuscular saline on those days. The bleomycin group (Group II) was followed up without any further treatment. The bleomycin + GnRH agonist group (Group III) was administered subcutaneous GnRH agonist triptorelin (1 mg/kg) at the same time as the bleomycin injections. The bleomycin + GnRH antagonist group (Group IV) was administered 1 mg/kg cetrorelix acetate subcutaneously, concurrently with the bleomycin. Although AMH levels were lower in the bleomycin group than in all the other groups, there was no statistically significant difference between the groups in terms of AMH levels (p > .05). In the bleomycin + cetrorelix acetate and bleomycin + triptorelin groups, significantly higher primordial, secondary and tertiary follicle counts were determined compared to the bleomycin group (p < .001). In conclusion the harmful effects of bleomycin on ovarian reserve can be reduced by the simultaneous administration of GnRH agonist or GnRH antagonist.

Ethanol Sclerotherapy versus Laparoscopic Surgery for Endometrioma Treatment: A Prospective, Multicenter, Cohort Pilot Study.

STUDY OBJECTIVE: To compare the cost-effectiveness of ultrasound (US)-guided aspiration and ethanol sclerotherapy versus laparoscopic surgery for benign-appearing ovarian endometrioma. DESIGN: Prospective, cohort pilot study. SETTING: Multiple centers, Spain. PATIENTS: Forty patients with suspected ovarian endometrioma identified by US, with a maximum diameter of 35 to 100 mm, of whom 33 met inclusion criteria. INTERVENTIONS: The study group (n = 17) underwent US-guided aspiration plus sclerotherapy with ethanol, and the control group (n = 14) underwent laparoscopic cystectomy. MEASUREMENTS AND MAIN RESULTS: Recurrence, complications, and direct costs were compared. One of 17 sclerotherapy patients recurred (5.9%) compared with 4 of 14 laparoscopic surgery patients (28.6%) (odds ratio 0.18, 0.01-1.53). No serious adverse effects (Clavien-Dindo ≥ III) were observed in the sclerotherapy group; 1 patient in the surgery group had a Clavien-Dindo IIIb complication. Median hospital direct costs were significantly lower in the sclerotherapy group than those in the surgery group-266 euros versus 2189 euros. CONCLUSION: Ethanol sclerotherapy seems to be cost-effective for endometrioma and also appears to reduce complications. In this pilot study, recurrence was not higher than with conventional surgery.

Pathological assessment of follicular loss in laparoscopic endometrioma excision: effects of cyst size and surgeon's experience.

AIM: We planned to histologically demonstrate unintentional ovarian harm caused by excision of endometriomas. METHODS: One hundred and seven patients with 135 endometriomas had laparoscopic excision of endometriomas at a tertiary health-care clinic over a 6-year period. Records were obtained retrospectively and pathological specimens were reevaluated to demonstrate follicular loss. Tissue damage was scored morphologically and compared according to surgeons' experience and endometrioma size. RESULTS: Ovarian follicles were detected in 80% of endometrioma specimens near the cyst wall. Morphological scores of ovarian follicles were not affected by the surgeon's experience or endometrioma size. CONCLUSION: Surgical excision of endometriomas causes inevitable ovarian follicular loss. As observed in our study, this loss rate could not be traced to surgical experience or ovarian size.

Multi-modal magnetic resonance elastography for noninvasive assessment of ovarian tissue rigidity in vivo.

For centuries, physicians have relied on touch to palpate tissue and detect abnormalities throughout the body. While this time-tested method has provided a simple diagnostic examination for large, superficial abnormalities, it does not permit quantifiable measurements of stiffness in deeper, small organs. Advances in noninvasive imaging to measure tissue rigidity represent important extensions of manual palpation techniques. Tissue fibrosis occurs with age in many organs; in the ovary, it is thought to be a marker of polycystic ovary syndrome and age-related idiopathic infertility, although quantitative assessment of fibrosis in this deep, abdominal tissue has not been possible. We used noninvasive methods to quantify ovarian tissue rigidity and clarify the role of tissue stiffness in reproductive health. With proper validation against accepted standards, noninvasive imaging techniques may become the quantitative counterpart to interior probing palpation methods and invasive (surgical) diagnoses, with applications across many clinical settings, including evaluation of adolescent and young adult ovarian function.

Subjective assessment of ovarian masses using pattern recognition: the impact of experience on diagnostic performance and interobserver variability.

PURPOSE: To compare diagnostic performance and interobserver variability in a group of 36 examiners, with four different levels of experience. METHODS: Nine junior trainees, eight level I senior trainees, 11 level II senior gynecologists, and eight level III expert sonologists classified 105 ultrasound images of adnexal masses into three subgroups of ovarian lesions (malignancies, functional cysts, and dermoid cysts). RESULTS: The level III sonologists obtained the best diagnostic results together with the lowest interobserver variability (κ = 0.70, SD = 0.04). They achieved significantly better results in comparison with the junior trainees and also the senior trainees (κ = 0.51, SD = 0.12, p < 0.001; and κ = 0.51, SD = 0.09, p < 0.001). Differences between level III sonologists and the group of level II observers did not reach statistical significance (κ = 0.65, SD = 0.09, p = 0.70). There were no significant differences between senior and junior trainees (p = 1.0) and both groups achieved a significantly poorer diagnostic performance in comparison with the level II observers (p < 0.01 and p < 0.01). For all observers, the largest differences were seen for classifying malignancies, the best results for classifying functional cysts, and the poorest for evaluating dermoid cysts. CONCLUSIONS: Diagnostic performance of pattern recognition significantly improves with an increasing level of experience, emphasizing the importance of standardized ultrasound training programs with supervision by experts.

Assessment of cyst content using mean gray value for discriminating endometrioma from other unilocular cysts in premenopausal women.

OBJECTIVE: To assess whether the analysis of cyst content using mean gray value (MGV) can discriminate ovarian endometriomas from other unilocular ovarian cysts in premenopausal women. METHODS: Stored three-dimensional (3D) volumes from 54 unilocular ovarian cysts diagnosed in 50 premenopausal women (mean age, 37 (range, 22-50) years) were analyzed to calculate the MGV from cyst content. Cysts with solid components or septations were excluded. MGV was calculated in all cases with the Virtual Organ Computer-aided AnaLysis technique. The B-mode presumptive diagnosis based on the examiner's subjective impression was also recorded. RESULTS: Sixteen of the cysts resolved spontaneously and were given a final clinical diagnosis of hemorrhagic functional cyst, while 38 cysts were removed surgically (diagnosed histologically as seven simple cysts, three hemorrhagic cysts, 20 endometriomas, five mucinous cysts and three paraovarian cysts). B-mode diagnoses were as follows: seven simple cysts, 18 hemorrhagic cysts, 24 endometriomas, three mucinous cysts and two paraovarian cysts. MGV was significantly higher in ovarian endometrioma when compared with all other kinds of cyst. The receiver-operating characteristics curve showed that using an MGV cut-off >or= 15.560 had a sensitivity of 85% and a specificity of 76.5% for diagnosing ovarian endometrioma (area under the curve, 0.831; 95% CI, 0.718-0.944). These figures were similar to those for B-mode diagnosis (sensitivity, 90%; specificity, 82%) (McNemar test, P = 1.000). Combining B-mode and MGV gave a sensitivity of 80% and a specificity of 91%. CONCLUSION: Cyst content MGV is higher in ovarian endometrioma than it is in other unilocular ovarian cysts. The diagnostic performance of MGV is similar to that of the examiner's subjective impression. The combination of both criteria achieves the highest specificity.

Preoperative serum tetranectin, CA125 and menopausal status used as single markers in screening and in a risk assessment index (RAI) in discriminating between benign and malignant ovarian tumors.

OBJECTIVES: To improve the poor survival in ovarian cancer (OC) patients, the research has been focused on new OC markers. One aim is to find markers to identify the cancers in early preclinical stages by screening. Another aim is to find new diagnostic markers, which may select patients at high risk for OC for quick referral to highly specialized centers in gynecologic oncology. These aims were addressed in the present study by evaluating serum tetranectin (TN) and serum CA125 on a large number of pre- and postmenopausal women with ovarian tumors and controls. METHODS: The potential ability of the markers to discriminate between the four groups (208 benign ovarian tumor, 153 borderline ovarian tumor (BOT), 445 OC and 1333 age matched controls) in OC screening was examined. We also constructed a risk assessment index (RAI) for discrimination between tumor groups based on these variables and menopausal status. RESULTS: Highly significant differences in both TN and CA125 levels were found between all the four groups as well as between the different FIGO stages of OC patients. A very high probability of having OC or a benign tumor, respectively, was predicted by the RAI. CONCLUSIONS: In the case-control part of the study, we found that TN and CA125 deserve to be validated on pre-clinical samples by inclusion in future marker panels. The RAI is also a potential new candidate for a diagnostic tool for selecting patients at high risk for having OC; hence it deserves further evaluation in a prospective clinical study.

Clinical assessment, gross examination, frozen section of ovarian masses: do patients benefit?

OBJECTIVE: The overall risk of malignancy in ovarian neoplasm is 13% in premenopausal women and 45% in postmenopausal women. Differentiating benign and malignant disease with frozen section is possible during operation; however, information on patients' history, physical examination, paraclinical criteria (tumour markers, imaging) and gross examination of tumour can also be helpful in planing the surgery. METHODS: This study was conducted on 150 women who underwent laparotomy due to adnexal mass between April 2003 and October 2005 at Vali-e-Asr Hospital, Tehran, Iran. Sensitivity and specificity of clinical assessment (history, tumour marker and imaging), gross examination and frozen section were calculated. RESULTS: Based on our findings frozen section had the highest sensitivity for diagnosing malignant tumour comparing with other methods of diagnosis (88.9%). Sensitivity was 71.3% for preoperative clinical examination, 83% for ultrasonography, 89.8% for CT scan, 70% for CA125 and 84.1% for gross examination, likewise the highest specificity was seen for frozen section (93.5%). CONCLUSION: This data confirm that frozen section diagnosis is a reliable method for the surgical management of patients with an ovarian mass, but history of disease, Para clinical criteria and gross examination can help to surgeon to perform on appropriate operation in the areas where frozen section is not possible.

Power Doppler in the assessment of pelvic masses in a low risk group.

Discrimination between benign and malignant adnexal masses aids in optimising the management of women with pelvic tumours. A grading system incorporating grey-scale and power Doppler measurements might increase the accuracy of pre-operative assessment. The present study aimed to assess the sensitivity and specificity of a grading system including grey-scale and power Doppler findings. A total of 179 women who had undergone power Doppler Ultrasound for a pelvic mass between April 2001 and March 2003 were identified. Masses were surveyed by grey-scale and power Doppler and graded in relation to the estimated risk of malignancy. Clinicians were provided with the findings. The results were compared with the findings at surgery. A total of 74.3% were premenopausal. Some 94.4% (n = 169) had benign; 1.7% borderline and 3.9% malignant histology. A total of 69% underwent surgery. Specificity was 72.2 - 95.9%; sensitivity 55.6 - 88.9% and negative predictive value 97.6 - 99.2%. Power Doppler as an additional diagnostic aid may help the clinician in the management of pelvic masses. Further research is needed to clarify sensitivity and specificity.

Assessment of cadmium impregnation in women suffering from endometriosis: a preliminary study.

Environmental chemicals with oestrogen-like activity are suspected aetiological factors of endometriosis. In animal experiments, cadmium was recently shown to possess oestrogen-like properties. In the frame of a case-control study designed to investigate environmental risk factors for endometriosis, we compared cadmium urinary excretion (CdU) and blood concentration in 59 patients with peritoneal endometriosis, deep endometriotic (adenomyotic) nodules of the recto-vaginal septum and controls. After standardisation for age (30 years) and smoking status, the mean levels of cadmium in urine were (geometric mean [geometric S.D.]) 0.25 [1.50], 0.29 [1.76] and 0.26 [1.46] microg/g creatinine, respectively. Cadmium concentrations in blood did not differ among the three groups. These data, therefore, do not support a role for cadmium in the onset or the growth of endometriosis or deep endometriotic (adenomyotic) nodules of the recto-vaginal septum.

Mucosal epithelial proliferation of the fallopian tube: prevalence, clinical associations, and optimal strategy for histopathologic assessment.

The prevalence and clinical significance of mucosal epithelial proliferation or hyperplasia of the fallopian tube are controversial in the few studies reported. Some authors have retrospectively examined "routine" sections (one or two submitted from each tube), whereas others have prospectively blocked the entire tubes. In the current study, we prospectively studied a total of 168 tubes from 98 women who had various indications for salpingectomy and compared the diagnosis in an initial single section (to simulate the usual practice) with that in the remainder of the entirely sectioned and submitted tube (mean total number of sections, 9.0). Some degree of mucosal epithelial proliferation was found in 83% of all tubes examined, with no difference between the tubes removed for routine tubal ligation and those in women who had benign ovarian lesions, malignant gynecologic tumors, uterine leiomyomata, or benign tubal lesions (salpingitis or ectopic pregnancy). Mucosal epithelial proliferation graded as more than mild, however, was seen in only 4.5% of the otherwise normal ligated tubes versus 35 to 46% of tubes associated with the other lesions. When the initial sections were compared with the subsequent ones, the diagnosis was identical in 96 tubes (57%). In the other 72 tubes (43%), the difference in diagnosis was never greater than one grade (no, mild, moderate, severe mucosal epithelial proliferation), with the diagnosis more often upgraded (50 tubes) than downgraded (22 tubes) in the additional sections. It is concluded that there is no reason to submit an entire tube for histologic examination to detect clinically significant lesions, and the usual practice of submission of one or two sections is clinically appropriate.

Morphometric assessment of the murine ovarian toxicity of 7,12-dimethylbenz(a)anthracene.

7,12-Dimethylbenz(a)anthracene (DMBA) is a polycyclic aromatic hydrocarbon and a component of cigarette smoke that has been identified as a murine reproductive toxicant. The morphometric parameters of total ovarian volume, individual corpus luteum volumes, and total corpora lutea volume were measured in C57BL/6N mice treated with DMBA. Each group received single intraperitoneal injections of 0, 0.1, 1.0, or 10 mg/kg and were sacrificed at 1, 2, 3, or 4 weeks after treatment. DMBA produced a dose-dependent decrease in ovarian volume and number of corpora lutea in each ovary. The observed reduction in total corpora lutea volume did not fully account for the loss in total ovarian volume. This is consistent with previous descriptions of a toxic effect on all ovarian components including growing and resting follicles. Growing follicles that escaped the toxic effects of DMBA and achieved ovulation resulted in a corpus luteum that appeared histologically normal. Morphometric analysis of this animal model further defines the dynamic changes in the mouse ovary in response to DMBA.

Comparison of random and serial sections in assessment of ovarian toxicity.

Assessment of ovarian toxicity by follicle quantitation and morphometric analysis of serial sections is time consuming and expensive. This report compares the estimation of follicle number obtained from counting oocytes in serial sections or 5 random sections of mouse ovaries. Ovaries were obtained from C57BL/6N and B6C3F1 mice treated with ovarian toxicants. C57BL/6N mice were treated with cyclophosphamide (0, 75, 200, and 500 mg/kg, ip) and killed at 24, 72, and 168 h. B6C3F1 mice were treated daily, ip, with 4-vinylcyclohexene (0, 100, 400, and 800 mg/kg/day for 30 days), vinylcyclohexene diepoxide (0, 10, 40, and 80 mg/kg/day for 30 days), or benzo(a)pyrene (100 mg/kg, single dose) and killed on day 31. Ovarian serial sections were prepared and oocytes counted in every tenth section. When serial sections were evaluated, 30 to 60 sections were counted. Random section counting involved randomly selecting 5 sections from the 30 to 60 sections previously counted by the serial method. Chemically-induced follicle loss was evident by the reduction in follicle counts relative to control animals when estimating follicle number using serial or random section counting. Furthermore, a linear regression analysis of follicle counts over all treatment groups showed that the highest correlation between random and serial section counting and was for primordial follicles. Correlation coefficients (R2) for each follicle type were: primordial--C57BL/6N 0.82, B6C3F1 0.86; growing--C57BL/6N 0.16, B6C3F1 0.46; antral--C57BL/6N 0.07, B6C3F1 0.11. Although other screens may need to be developed for growing and antral follicles, primordial follicle toxicity is adequately defined by counting 5 random sections.

Morphometric assessment of ovarian stromal proliferation--a clinicopathological study.

Ovarian stromal hyperplasia is commonly observed in peri- and post-menopausal ovaries. Assessment has, hitherto, been largely subjective or dependent on arbitrary criteria, which has made comparison of studies difficult. In this study, the stroma of a large number of ovaries was assessed morphometrically. Data were related to clinicopathological and cytomorphological information. A significant association was noted between the degree of stromal proliferation and post-menopausal endometrial adenocarcinoma (P = 0.004). There were no significant correlations between the degree of stromal proliferation and average cytomorphological dimensions.