Enhancement of the luteinising hormone surge by male olfactory signals is associated with anteroventral periventricular Kiss1 cell activation in female rats.

Olfactory stimuli play an important role in regulating reproductive functions in mammals. The present study investigated the effect of olfactory signals derived from male rats on kisspeptin neuronal activity and luteinising hormone (LH) secretion in female rats. Wistar-Imamichi strain female rats were ovariectomised (OVX) and implanted with preovulatory levels of 17ß-oestradiol (E2 ). OVX+E2 rats were killed 1 hour after exposure to either: clean bedding, female-soiled bedding or male-soiled bedding. Dual staining for Kiss1 mRNA in situ hybridisation and c-Fos immunohistochemistry revealed that the numbers of Kiss1-expressing cells and c-Fos-immunopositive Kiss1-expressing cells in the anteroventral periventricular nucleus (AVPV) were significantly higher in OVX+E2 rats exposed to male-soiled bedding than those of the other groups. No significant difference was found with respect to the number of c-Fos-immunopositive Kiss1-expressing cells in the arcuate nucleus and c-Fos-immunopositive Gnrh1-expressing cells between the groups. The number of c-Fos-immunopositive cells was also significantly higher in the limbic system consisting of several nuclei, such as the bed nucleus of the stria terminalis, the cortical amygdala and the medial amygdala, in OVX+E2 rats exposed to male-soiled bedding than the other groups. OVX+E2 rats exposed to male-soiled bedding showed apparent LH surges, and the peak of the LH surge and area under the curve of LH concentrations in the OVX+E2 group were significantly higher than those of the other two groups. These results suggest that olfactory signals derived from male rats activate AVPV kisspeptin neurones, likely via the limbic system, resulting in enhancement of the peak of the LH surge in female rats. Taken together, the results of the present study suggests that AVPV kisspeptin neurones are a target of olfactory signals to modulate LH release in female rats.

Evidence of involvement of neurone-glia/neurone-neurone communications via gap junctions in synchronised activity of KNDy neurones.

Pulsatile secretion of gonadotrophin-releasing hormone (GnRH)/luteinising hormone is indispensable for the onset of puberty and reproductive activities at adulthood in mammalian species. A cohort of neurones expressing three neuropeptides, namely kisspeptin, encoded by the Kiss1 gene, neurokinin B (NKB) and dynorphin A, localised in the hypothalamic arcuate nucleus (ARC), so-called KNDy neurones, comprises a putative intrinsic source of the GnRH pulse generator. Synchronous activity among KNDy neurones is considered to be required for pulsatile GnRH secretion. It has been reported that gap junctions play a key role in synchronising electrical activity in the central nervous system. Thus, we hypothesised that gap junctions are involved in the synchronised activities of KNDy neurones, which is induced by NKB-NK3R signalling. We determined the role of NKB-NK3R signalling in Ca2+ oscillation (an indicator of neuronal activities) of KNDy neurones and its synchronisation mechanism among KNDy neurones. Senktide, a selective agonist for NK3R, increased the frequency of Ca2+ oscillations in cultured Kiss1-GFP cells collected from the mediobasal hypothalamus of the foetal Kiss1-green fluorescent protein (GFP) mice. The senktide-induced Ca2+ oscillations were synchronised in the Kiss1-GFP and neighbouring glial cells. Confocal microscopy analysis of these cells, which have shown synchronised Ca2+ oscillations, revealed close contacts between Kiss1-GFP cells, as well as between Kiss1-GFP cells and glial cells. Dye coupling experiments suggest cell-to-cell communication through gap junctions between Kiss1-GFP cells and neighbouring glial cells. Connexin-26 and -37 mRNA were found in isolated ARC Kiss1 cells taken from adult female Kiss1-GFP transgenic mice. Furthermore, 18ß-glycyrrhetinic acids and mefloquine, which are gap junction inhibitors, attenuated senktide-induced Ca2+ oscillations in Kiss1-GFP cells. Taken together, these results suggest that NKB-NK3R signalling enhances synchronised activities among neighbouring KNDy neurones, and that both neurone-neurone and neurone-glia communications via gap junctions possibly contribute to synchronised activities among KNDy neurones.

Lack of pulse and surge modes and glutamatergic stimulation of luteinising hormone release in Kiss1 knockout rats.

Kisspeptin, encoded by the Kiss1 gene, has attracted attention as a key candidate neuropeptide in controlling puberty and reproduction via regulation of gonadotrophin-releasing hormone (GnRH) secretion in mammals. Pioneer studies with Kiss1 or its cognate receptor Gpr54 knockout (KO) mice showed the indispensable role of kisspeptin-GPR54 signalling in the control of animal reproduction, although detailed analyses of gonadotrophin secretion, especially pulsatile and surge-mode of luteinising hormone (LH) secretion, were limited. Thus, in the present study, we have generated Kiss1 KO rats aiming to evaluate a key role of kisspeptin in governing reproduction via pulse and surge modes of GnRH/LH secretion. Kiss1 KO male and female rats showed a complete suppression of pulsatile LH secretion, which is responsible for folliculogenesis and spermatogenesis, and an absence of puberty and atrophic gonads. Kiss1 KO female rats showed no spontaneous LH/follicle-stimulating hormone surge and an oestrogen-induced LH surge, suggesting that the GnRH surge generation system, which is responsible for ovulation, does not function without kisspeptin. Furthermore, challenge of major stimulatory neurotransmitters, such as monosodium glutamate, NMDA and norepinephrine, failed to stimulate LH secretion in Kiss1 KO rats, albeit they stimulated LH release in wild-type controls. Taken together, the results of the present study confirm that kisspeptin plays an indispensable role in generating two modes (pulse and surge) of GnRH/gonadotrophin secretion to regulate puberty onset and normal reproductive performance. In addition, the present study suggests that kisspeptin neurones play a critical role as a hub integrating major stimulatory neural inputs to GnRH neurones, using newly established Kiss1 KO rats, which serve as a useful model for detailed analysis of hormonal profiles.

Surgical outcomes of robotic radical hysterectomy after neoadjuvant chemotherapy for locally advanced cervical cancer: comparison with early stage disease.

AIMS: To evaluate the feasibility and safety of robotic radical hysterectomy (RRH) with pelvic lymphadenectomy for locally advanced cervical cancer (LACC) after neoadjuvant chemotherapy (NACT). METHODS: Starting from 04/2009, consecutive patients with LACC were submitted to robotic surgical staging after NACT. Surgical outcomes were compared to those achieved by women undergoing robotic surgery for an early stage disease during the same temporal interval. RESULTS: Overall 25 (Group 1) and 21 (Group 2) patients had an early stage and a LACC, respectively. Among women with LACC, 18 achieved best tumor responses to NACT and therefore they were addressed to RRH. Outcomes resulted comparable between Groups in terms of operative time, blood loss, hospitalization and complications. No differences were found in terms of nodal yield, parametrial and vaginal cuff length. CONCLUSIONS: RRH is feasible and safe also in patients previously submitted to NACT for LACC. Larger series with longer follow-up are mandatory to establish survival outcomes.

Robotic radical parametrectomy with pelvic lymphadenectomy: our experience and review of the literature.

AIMS: To evaluate the feasibility and safety of robotic radical parametrectomy (RRP) and pelvic lymphadenectomy for the management of occult invasive cervical cancer or local recurrence of endometrial cancer and to compare our outcomes with the evidence available in the literature. METHODS: Starting from 07/2008 consecutive patients submitted to RRP have been included in this study. A comprehensive literature review of published papers about this subject was carried out. RESULTS: During the study period 11 patients were managed; 7 and 4 patients had an occult cervical cancer and a vaginal recurrence of endometrial cancer, respectively. One intra-operative and one post-operative complications were recorded. Neither conversion to laparotomy, nor blood transfusions occurred. Three women required further adjuvant therapies. After a median follow-up of 19 months (range 8-36) one recurrence has been detected. The outcomes of other 200 women from 15 different papers have been collected and compared to our findings. CONCLUSIONS: Robotic surgery represents an effective alternative to accomplish radical parametrectomy with comparable results of those reported in the literature in terms of feasibility and safety. RRP is certainly a demanding procedure which however avoids radiotherapy in more than 80% of cases.

A phase II randomised clinical trial comparing cisplatin, paclitaxel and ifosfamide with cisplatin, paclitaxel and epirubicin in newly diagnosed advanced epithelial ovarian cancer: long-term survival analysis.

To test the feasibility and efficacy of epirubicin and ifosfamide added to first-line chemotherapy with cisplatin and paclitaxel in a phase II randomised clinical trial. Patients with histologically proven epithelial ovarian cancer were randomly assigned to receive first-line polychemotherapy with cisplatin/paclitaxel/epirubicin (CEP) or cisplatin/paclitaxel/ifosfamide (CIP) for six cycles every 21 days. Two hundred and eight patients were randomised between the two treatment arms and the median number of cycles per patient was six. Toxicity was predominantly haematological with both regimens; however, anaemia, leucopaenia, neutropaenic fever and use of granulocyte colony-stimulating factors and transfusion were significantly more frequent in the CIP treatment arm. Response rates were 85% (95% confidence interval (CI) 77-93%) in the CIP arm and 90% (95% CI 84-96%) in the CEP arm; complete response rates were 48 and 52%. After a median follow-up of 82 months, median overall survival (OS) was 51 and 65 months; 5-year survival rates were respectively 43 and 50%. In this clinical trial, both regimens showed good efficacy, but toxicity was heavier with the CIP regimen. Considering that more than 50% of patients were suboptimally debulked after the first surgery, OS seems to be longer than is commonly reported. This unexpected finding might be a consequence of the close surgical surveillance and aggressive chemotherapeutic approach.

[Fertility after surgery for rAFS stage III and IV endometriosis]. / Fertilità dopo chirurgia per endometriosi stadio III e IV rAFS.

AIM: The aim of this paper was to assess the impact on fertility of surgery to eradicate endometriosis. METHODS: One-hundred and twenty-six patients aged between 20 and 40 were observed. All wished to have offspring after the operation and were subjected to conservative surgery for stage III and IV endometriosis (rAFS score > 16) in the period 1992-2002. The type of surgical approach was chosen in consideration of the patient's clinical condition and on the basis of the experience of the surgeon, with the aim of radically removing the disease and, where necessary, restoring fertility. RESULTS: Fifty-six of 126 patients (44.4%) conceived after the operation; 55 spontaneously and 1 with assisted fecundation. Of the patients observed who became pregnant, about 1/3 (32%) conceived within 6 months of the operation and 1/3 (31%) after 12 months. Forty-four (78.5%) pregnancies reached term with a positive outcome, 7 (12.5%) were in progress at the moment of follow-up, 4 (7.1%) suffered a miscarriage and 1 (1.7%) was extrauterine; 48.2% (27/56 patients) of the pregnancies were stage III, 40% (28/70 patients) were stage IV. CONCLUSIONS: It emerges clearly from the data extrapolated from our series that surgery to eradicate endometriosis increases the fertility of the patients affected, without being resolutive: more than half the patients in fact remained sterile in spite of the operation.

Clinical relevance of retroperitoneal involvement from epithelial ovarian tumors of borderline malignancy.

Rota SM, Zanetta G, Ieda N, Rossi R, Chiari S, Perego P, Mangioni C. Clinical relevance of retroperitoneal involvement from epithelial ovarian tumors of borderline malignancy. Ovarian tumors of borderline malignancy have an outstanding prognosis. The need for aggressive surgical staging is questionable and the need for retroperitoneal node sampling is debated. From 1982 to 1996, 81 women underwent surgical staging including retroperitoneal sampling. Three patients (3.7%) with serous tumor had microscopic nodal involvement. Retroperitoneal metastases were found in two intraperitoneal stage I tumors and in one stage IIIA tumor. Positive nodes were found in 1/31 (3.2%) women undergoing sampling of para-aortic nodes and in 2/69 (2.8%) women undergoing sampling of pelvic nodes. With a median follow-up of 79 months we observed five recurrences, but none involved the retroperitoneum. The three patients with positive nodes remain alive without disease. Among 236 patients with diagnosis of borderline tumor but without sampling of the nodes, we observed one retroperitoneal recurrence (0.4%) in a serous tumor. There are no indications for retroperitoneal sampling of mucinous borderline tumors. For serous tumors this procedure should only be performed as a part of prospective trials. The clinical relevance of retroperitoneal involvement in borderline tumors appears minimal and does not justify routine aggressive surgery.

Beneficial effects of iloprost on acute myocardial ischemia in dogs.

The effects of intravenous administration of iloprost, a prostacyclin analogue, on myocardial energy metabolism and myocardial blood flow (MBF) were examined in anesthetized open-chest dogs subjected to 60 min of myocardial ischemia by coronary ligation. Iloprost administration at levels of 0.1 or 0.2 micrograms/kg/min was started 30 min before the commencement of ischemia and continued throughout the 90 min observation period. Since systolic aortic pressure in the iloprost 0.2 micrograms/kg/min group showed significantly lower values than that in the control group, whereas no clear effect was observed with the lower concentration (0.1 micrograms/kg/min), this latter group was further investigated. This 0.1 microgram/kg/min dose of iloprost lacked influence on MBF in both ischemic and nonischemic areas but did result in a significantly higher value for high energy phosphate contents in the ischemic myocardium. Moreover, myocardial mitochondrial respiratory function in the ischemic area was significantly improved. These results indicate that iloprost brought about preservation of myocardial energy metabolism without alteration of coronary perfusion, suggesting that it may exert a direct cardioprotective effect.

[Tricuspid regurgitation evaluated by intraoperative epicardial pulsed Doppler echocardiography: investigation of patients with combined valvular diseases].

To evaluate the grade of tricuspid regurgitation (TR) associated with mitral valve disease and to ascertain the operative procedure for the involved tricuspid valve, epicardial pulsed Doppler echocardiography (PDE) was performed during cardiac surgery. Thirty-two patients with mitral valve disease were studied, 17 of whom had only mitral valve lesion; the remaining 15 had combined mitral and aortic valve disease. The patients' ages ranged from 24 to 63 years and averaged 48.3 years. There were nine men and 23 women. Echocardiographic examinations were performed using a Toshiba SSH-60A for parasternal study and a SSH-11A combined with a SDS-10A with a specially-devised flat transducer for intraoperative use. Intraoperatively, the PDE performed was from the right side of the right atrium (RA), referenced by a four-chamber view and a long-axis view of the right ventricular inflow. The sampling volumes were positioned in the inflow of the right ventricle, immediately above the tricuspid valve, the middle and upper areas of the RA, and adjacent to the interatrial septum. PDE was performed before and immediately after the operative procedure and before chest closure. By severity, TR was classified as non -, mild +/-, moderate +, and severe ++, according to the distances attained by the TR signals from the tricuspid valve orifice, and the velocities and durations of the TR signals during systole. The TR signal was recorded in 23 of 32 patients before surgery, whereas it was determined more adequately in 28 patients by intraoperative epicardial PDE. The gradings of TR via the parasternal approach before surgery were as follows: no TR, in nine cases; mild TR, in three; moderate, in 13; and severe, in seven. Intraoperatively, four patients had none; eight had mild TR; 14, moderate TR; six, severe TR before surgical intervention, respectively. In cases with mild or no TR before surgery, TR was rarely detected by contrast echocardiography using saline solution injected into the right ventricle during surgery. The moderate or severe cases before surgery had moderate or severe TR according to the contrast method during surgery, except for one case not operated on for tricuspid valve disease. Tricuspid valve replacement was performed for two patients, and tricuspid annuloplasty or valvuloplasty for eight.(ABSTRACT TRUNCATED AT 400 WORDS)