Seyma Hascalik

Reply: Combating endometriosis by blocking proteasome and nuclear factor-κB pathways

BACKGROUND The objective of this study is to investigate the effect of pyrrolidine dithiocarbamate [PDTC; a nuclear factor-kappaB (NF-kappaB) inhibitor] and bortezomib (Velcade; a proteasome inhibitor) on the development of experimental endometriotic implants in rats. METHODS Endometriosis was surgically induced in 30 rats using the method of Vernon and Wilson. Three weeks later the viability and volume of the implants were recorded and classified. Afterwards, rats were put into three groups with equal numbers. The groups were labelled as the control, the PDTC and the bortezomib groups. Seven days after treatment, a third laparotomy was done and the volume of implants was measured again. The animals were then sacrificed, and the implants were stained with Ki67, proliferating cell nuclear antigen (PCNA), CD34, CD31 and Massons trichrome histochemical staining. RESULTS In 80% of the implanted rats, vesicles at the suture region were observed, and the rats graded according to average vesicle diameter (D) as: Grade 1 (no vesicle, 20% of rats), Grade 2 (D < 2 mm, 33.3% of rats), Grade 3 (2 mm 4.5 mm, 26.7% of rats) and Grade 4 (D > 4.5 mm, 20% of rats). After treatment with PDTC or bortezomib, these percentages were decreased for Grades 3 and 4, and increased in Grade 1. The post-treatment implant volumes were decreased in the PDTC and bortezomib groups (P < 0.002 and P < 0.001), and slightly increased in the control group (P = 0.279). In the PDTC and bortezomib groups, CD34, CD31, PCNA and Ki67 expression levels were similar but were significantly reduced compared with the control group. CONCLUSIONS PDTC and bortezomib may represent a novel therapeutic strategy for treatment of endometriosis.

Melatonin reduces torsion-detorsion injury in rat ovary: biochemical and histopathologic evaluation

Abstract:  This experimental study was designed to determine the effects of melatonin on the levels of malondialdehyde (MDA), reduced glutathione (GSH), xanthine oxidase (XO) after adnexial torsion/detorsion (ischemia/reperfusion, I/R) of the ovaries of in rats. Forty adult albino rats were divided into five groups: sham operation, torsion, I/R plus saline, I/R plus melatonin and torsion plus melatonin. Rats in the sham‐operated group underwent a surgical procedure similar to the other groups but the adnexa was not occluded. Rats in the torsion group were killed after adnexal torsion for 3 hr. Melatonin and saline were injected intraperitoneally (10 mg/kg) 30 min before detorsion to the I/R plus melatonin group and I/R plus saline group respectively. After 3 hr of ovarian detorsion, the rats were killed and ovaries were removed. Melatonin was injected intraperitoneally (10 mg/kg) 30 min before torsion to the torsion plus melatonin group. After 3 hr of ovarian torsion, the rats were killed and ovaries were harvested. The tissue levels of MDA, GSH and XO were measured. MDA and XO levels in the I/R plus saline group increased significantly when compared with torsion and sham‐operated groups (P < 0.001). MDA and XO levels in the I/R plus melatonin group were lower than I/R plus saline and differences between the two groups were statistically significant (P < 0.001). GSH levels in the I/R plus saline group decreased significantly when compared with ischemia and sham‐operated groups (P < 0.001). GSH levels in the I/R plus melatonin treated rats were significantly higher than I/R plus saline and ischemia groups (P < 0.001). The tissue levels of XO, MDA and GSH were similar between ischemia and ischemia plus melatonin groups. Morphologically, polymorphonuclear neutrophil infiltration and vascular dilatation were obvious in the I/R‐damaged ovaries, and the changes also partially reversed by melatonin. This study demonstrates that melatonin protects the ovaries against oxidative damage associated with reperfusion following an ischemic insult.

Resveratrol, a Red Wine Constituent Polyphenol, Protects from Ischemia-Reperfusion Damage of the Ovaries

Objective: The aim of this study is to investigate the effects of resveratrol on histopathological changes, antioxidant status and lipid peroxidation, in torsion-detorsion injury in rat ovaries. Method: To determine whether ischemia followed by reperfusion can induce ovarian oxidative damage, we created a model of adnexal ischemia-reperfusion by using rats. Ischemia was induced by unilateral occlusion of the tubo-ovarian vessels for 3 h. Reperfusion was achieved by releasing the occlusion and restoring the circulation for 3 h. Thirty-two adult female albino rats were divided equally into 4 groups: sham operation, torsion, saline/detorsion and resveratrol/detorsion. Rats in the torsion group were killed after 360° clockwise adnexal torsion for 3 h. Resveratrol was injected intraperitoneally 30 min before detorsion in the resveratrol/detorsion group, and saline was administered in the saline/detorsion group. After 3 h of adnexal detorsion in both of these groups, the rats were killed and adnexa were removed. The tissue levels of malondialdehyde, reduced glutathione and xanthine oxidase activity were measured. Results: Malondialdehyde and xanthine oxidase levels in the saline/detorsion group were increased significantly when compared to the torsion and sham operation groups (p < 0.001). Malondialdehyde levels in the resveratrol group were lower than in the saline/detorsion group, and differences between the two groups were statistically significant (p < 0.001). Xanthine oxidase levels in the resveratrol group were lower than in the saline/detorsion and torsion groups, and differences between these groups were statistically significant (p < 0.001). Reduced glutathione levels in the saline/detorsion group were decreased significantly when compared to the torsion and sham operation groups. Reduced glutathione levels in the resveratrol group were significantly higher than in the saline/detorsion group (p < 0.006). Histological examination showed a significant improvement in ovarian morphology in the resveratrol-treated rats compared with the ischemia and ischemia-reperfusion groups. Conclusion: Our results demonstrated that intraperitoneal resveratrol administration reduced the lipid peroxidation products of ischemic rats and ovarian damage was reduced as indicated by histological examination.

Metabolic Changes in Pelvic Lesions: Findings at Proton MR Spectroscopic Imaging

Objective: The purpose of this study is to investigate the in vivo magnetic resonance spectroscopic (MRS) features of pelvic lesions using long echo time and to characterize the spectral patterns of various pathological entities. Materials and Methods: 17patients with surgically and histopathologically confirmed pelvic lesions underwent long echo-time MRS, and the results obtained were analyzed. Before laparotomy, choline (Cho), lactate, lipid and creatine (Cr) levels of all lesions were measured by single voxel MRS (point-resolved spectroscopy technique, TE 136 ms). Voxels were placed in the center of the lesions. The MRS results of lesions were compared with the final histopathological diagnoses. Results: Spectroscopy analysis of serous, mucinous and undifferentiated carcinoma of the ovary revealed Cho, lactate and lipid signals, but granulosa-theca cell tumor showed only a lipid signal. The Cho signal was obtained from only 3 patients with mature cystic teratoma but none of the other benign ovarian tumors and pelvic abscesses. A lipid signal was detected in 3 patients diagnosed with pelvic abscess and all benign ovarian tumors. In addition to the lipid signal, a lactate signal was detected in the spectra of two pelvic abscesses. One case of endometrioma and 1 case of teratoma did not show any signal. Conclusion: MRS demonstrates significant differences in metabolite concentration between benign and malignant ovarian tumors and pelvic abscesses. MRS may therefore be helpful in the differential diagnosis of adnexal lesions.

Reversible posterior leukoencephalopathy in eclampsia

Posterior leukoencephalopathy syndrome (RPLS) is a newly recognized brain disorder that predominantly affects the cerebral white matter w1x. Its causes are diverse, but common precipitants are acute elevations of blood pressure, immunsuppressive drugs, hypertensive encephalopathy and eclampsia w1,2x. Sudden elevation in mean arterial pressure exceeds the autoregulatory capability of the brain vasculature and results in vasogenic edema w2x. T2-weighted images characteristically show symmetrical diffuse hyperintensity selectively involving the parieto-occipital lobe w1x. MR spectroscopy (MRS) is powerful and non-invasive chemical analysis tool and is widely used for brain lesions such as tumors, metabolic and neurodegenerative diseases w3x. Only isolated reports of MRS for RPLS accompanied by eclampsia have been published w4,5x. We calculated metabolite ratios in parieto-occipital lobes using single voxel MRS in a patient with RPLS initially and 4 months later. A 21-year-old, primigravid woman was seen at term with severe

Magnetic Resonance Spectroscopy Features of Uterine Leiomyomas

Objective: The purpose of this study is to investigate the in vivo magnetic resonance spectroscopy features of uterine leiomyomas using long echo time and to characterize the spectral patterns of these lesions. Methods: We calculated metabolites in 15 patients with uterine leiomyomas and myometrium of 20 healthy control subjects using single-voxel proton MR spectroscopy (point resolved spectroscopy technique, TE:136 ms). Voxels were placed at the center of the uterine leiomyomas. The peak areas of creatine, choline, lipid and lactate were determined. The MR spectroscopy results of uterine leiomyomas were compared with the spectroscopy results obtained from the myometrium of healthy control subjects. Results: The characteristically obtained signal was choline, which was detected not only in 14 of the 15 leiomyomas (93.3%) but also in 18 of the 20 myometrium of control subjects (90%). The lipid signals were determined in 9 of 15 patients with uterine leiomyomas (60%) and 8 of 20 control subjects (40%). The lactate signal was obtained from six of 15 patients with leiomyomas (40%) but only two of myometrium (10%). The creatine signal was obtained from 4 of 15 patients with leiomyomas (26.6%) and 5 of 20 myometrium (25%). Among the tested parameters only lactate peak was statistically significant (p < 0.05). Conclusion: Proton MR spectroscopic imaging may be helpful for the investigation of the underlying pathophysiology of uterine leiomyomas. The presence of lactate and lipid signals in the spectrum may be a useful indicator of metabolic pathway of uterine leiomyomas.

An appraisal of serum preptin levels in PCOS

Plasma preptin levels were higher in women with PCOS compared with the control group (mean±SD, 823.2±140.7 vs. 324.9±147.3 pg/mL). This article suggests a potential role of preptin in the pathogenesis of PCOS.

Angiotensin-converting enzyme gene polymorphism and risk of insulin resistance in PCOS

The aim of this study was to establish the frequency of angiotensin-converting enzyme (ACE) insertion (I) or deletion (D) gene polymorphism in women with polycystic ovary syndrome (PCOS) and to examine the association of this polymorphism with insulin resistance. A total of 32 women with PCOS and 31 healthy, age- and body mass index-matched controls were studied. Serum lipids, fasting glucose, insulin and other hormones concentrations were measured. Homeostasis model assessment was used to estimate insulin resistance (HOMA-IR). DNA was extracted from peripheral blood leukocytes and genotyping of ACE I/D polymorphism was carried out by polymerase chain reaction. ACE genotypes were distributed as follows: DD was present in 16 (50%), ID in 12 (37.5%) and II in four (12.5%) PCOS patients, and DD in seven (22.6%), ID in 20 (64.5%) and II in four (12.9%) of healthy subjects. The frequency of D and I alleles were found in 69% and 31% of the PCOS group and 55% and 45% in the control group, respectively. There were no significant differences regarding the genotypic distribution and allelic frequency between the groups. However the ACE DD genotype was significantly associated with serum insulin concentrations and HOMA-IR measurement (both P=0.005). ACE DD genotype is associated with an increased insulin resistance in women with PCOS.

Effects of Resveratrol, Raloxifene, Tibolone and Conjugated Equine Estrogen on Vaginal Squamous Cell Maturation of Ovariectomized Rats

Objective: The effects of estrogen replacement therapy, selective estrogen receptor modulators, or tibolone on vaginal squamous cell maturation in postmenopausal women are not well established. Resveratrol (3,5,4′-trans-trihydroxystilbene) has been shown to bind the estrogen receptor in rat uteri. The aim of this study was to cytologically evaluate vaginal smears from ovariectomized rats treated with resveratrol, raloxifene, tibolone and conjugated equine estrogen, and to compare each drug with regard to vaginal epithelial maturation. Material and Methods: Forty-two bilaterally ovariectomized Wistar albino rats were equally randomized into 6 groups: (1) control sham-operated rats; (2) ovariectomized rats administered 0.1% ethanol; (3) ovariectomized rats administered resveratrol at a dose of 5 mg/kg/day p.o.; (4) ovariectomized rats administered conjugated equine estrogen (CEE) at a dose of 0.1 mg/kg/day p.o.; (5) ovariectomized rats administered tibolone at a dose of 0.25 mg/kg/day p.o., and (6) ovariectomized rats administered raloxifen at a dose of 1 mg/kg/day p.o. Administration of drugs started 5 days after bilateral ovariectomy and continued for 35 days. After 35 days of treatment a vaginal smear was obtained from each rat. Smears were stained with the usual Papanicolaou method, and observed with a light microscope by an experienced cytopathologist. Cytological grading was made according to the extent of parabasal, intermediate, superficial and anuclear squamous cells. Results: Ovariectomized rats had lower scores for superficial and anuclear cells when compared to sham-operated rats (p < 0.05). The CEE group had higher scores for superficial and anuclear cells than those of the ovariectomized, raloxifene and tibolone groups (p < 0.05). The resveratrol-treated rats had higher scores for superficial cells but lower scores for parabasal cells than ovariectomized rats (p < 0.05). The raloxifene and tibolone groups had the same scores for intermediate, superficial and anuclear cells but lower scores for parabasal cells compared to ovariectomized rats. Conclusion: The results of our study suggest that resveratrol offsets the reduction in vaginal stratification generally observed after oophorectomy.

Effect of electromagnetic field emitted by cellular phones on fetal heart rate patterns.

The study was planned to determine the effects of electromagnetic fields produced by cellular phones on baseline fetal heart rate, acceleration and deceleration. Forty pregnant women undergoing non-stress test were admitted to the study. Non-stress test was obtained while the subjects were holding the CP on stand by mode and on dialing mode, each for 5 min. Similar recordings were taken while there were no phones around for 10 min. Electromagnetic fields produced by cellular phones do not cause any demonstrable affect in fetal heart rate, acceleration and deceleration.