High-Content Screening Pipeline for Natural Products Targeting Oncogenic Signaling in Melanoma.

The incidence of melanoma, the most fatal dermatological cancer, has dramatically increased over the last few decades. Modern targeted therapy with kinase inhibitors induces potent clinical responses, but drug resistance quickly develops. Combination therapy improves treatment outcomes. Therefore, novel inhibitors targeting aberrant proliferative signaling in melanoma via the MAPK/ERK and PI3K/AKT pathways are urgently needed. Biosensors were combined that report on ERK/AKT activity with image-based high-content screening and HPLC-based activity profiling. An in-house library of 2576 plant extracts was screened on two melanoma cell lines with different oncogenic mutations leading to pathological ERK/AKT activity. Out of 140 plant extract hits, 44 were selected for HPLC activity profiling. Active thymol derivatives and piperamides from Arnica montana and Piper nigrum were identified that inhibited pathological ERK and/or AKT activity. The pipeline used enabled an efficient identification of natural products targeting oncogenic signaling in melanoma.

Fertility Sparing Treatments in Endometrial Cancer Patients: The Potential Role of the New Molecular Classification.

Endometrial cancer is the most frequent gynecological malignancy, and, although epidemiologically it mainly affects advanced age women, it can also affect young patients who want children and who have not yet completed their procreative project. Fertility sparing treatments are the subject of many studies and research in continuous evolution, and represent a light of hope for young cancer patients who find themselves having to face an oncological path before fulfilling their desire for motherhood. The advances in molecular biology and the more precise clinical and prognostic classification of endometrial cancer based on the 2013 The Cancer Genome Atlas classification allow for the selection of patients who can be submitted to fertility sparing treatments with increasing oncological safety. It would also be possible to predict the response to hormonal treatment by investigating the state of the genes of the mismatch repair.

Towards Personalized Medicine: Non-Coding RNAs and Endometrial Cancer.

Endometrial cancer (EC) is the most frequent female cancer associated with excellent prognosis if diagnosed at an early stage. The risk factors on which clinical staging is based are constantly updated and genetic and epigenetic characteristics have recently been emerging as prognostic markers. The evidence shows that non-coding RNAs (ncRNAs) play a fundamental role in various biological processes associated with the pathogenesis of EC and many of them also have a prognosis prediction function, of remarkable importance in defining the therapeutic and surveillance path of EC patients. Personalized medicine focuses on the continuous updating of risk factors that are identifiable early during the EC staging to tailor treatments to patients. This review aims to show a summary of the current classification systems and to encourage the integration of various risk factors, introducing the prognostic role of non-coding RNAs, to avoid aggressive therapies where not necessary and to treat and strictly monitor subjects at greater risk of relapse.

Non-Coding RNAs as Prognostic Markers for Endometrial Cancer.

Endometrial cancer (EC) has been classified over the years, for prognostic and therapeutic purposes. In recent years, classification systems have been emerging not only based on EC clinical and pathological characteristics but also on its genetic and epigenetic features. Noncoding RNAs (ncRNAs) are emerging as promising markers in several cancer types, including EC, for which their prognostic value is currently under investigation and will likely integrate the present prognostic tools based on protein coding genes. This review aims to underline the importance of the genetic and epigenetic events in the EC tumorigenesis, by expounding upon the prognostic role of ncRNAs.

Recurrent Bilateral Salpingitis in a Sexually Inactive Adolescent: Don't Forget about the Appendix.

BACKGROUND: Salpingitis is an acute inflammation of the fallopian tubes and is extremely uncommon in patients who are sexually inactive or premenarchal. CASE: We describe a 15-year-old sexually inactive patient with recurrent bilateral salpingitis. After the second episode, she underwent an exploratory laparoscopy and was diagnosed with chronic appendicitis. SUMMARY AND CONCLUSION: Appendicular involvement should be kept in mind in recurrent salpingitis episodes, especially if previous imaging studies do not show signs of appendicitis. Imaging studies should be repeated in experienced centers in case of recurrent episodes. Laparoscopy is the gold standard for the diagnosis of salpingitis and can confirm appendicular or other abdominal involvement. Early diagnosis can help to prevent recurrent episodes of salpingitis and thus reduce the risk of sequelae.

Deep Infiltrating Endometriosis and Endometrial Adenocarcinoma Express High Levels of Myostatin and Its Receptors Messenger RNAs.

Myostatin is a growth factor member of the transforming growth factor ß superfamily, which is known to play major roles in cell proliferation and differentiation. The present study investigated the messenger RNA (mRNA) expression of myostatin and myostatin receptors (activin receptor-like kinase 4 [ALK4], transforming growth factor (TGF)-ß type I receptor kinase [ALK5] and activin receptor type IIB [ActRIIB]) in endometrium of healthy women during menstrual cycle as well as in benign (endometriosis, polyps) and malignant (endometrial adenocarcinoma) conditions. Endometrial specimens were collected by hysteroscopy, whereas endometriotic lesions were collected by laparoscopy, and adenocarcinomas were sampled after hysterectomy. Total RNA was extracted from tissue homogenates, and gene expression was assessed by quantitative real-time polymerase chain reaction. Myostatin and myostatin receptors mRNAs were expressed by healthy endometrium throughout the menstrual cycle, with no differences between the proliferative and secretory phase. The highest myostatin mRNA expression was found in patients with deep infiltrating endometriosis (DIE) and in endometrial carcinoma; expression was also found in ovarian endometrioma (OMA ) and endometrial polyps. Myostatin receptors mRNA expression was higher in DIE and adenocarcinomas compared to control endometrium. The expression of ALK5 and ActRIIB in OMA was higher than in controls, whereas polyps had an increased expression of ALK5 mRNA. In conclusion, the present data showed for the first time the expression of myostatin in healthy endometrium and a higher expression in endometriosis and endometrial cancer, suggesting myostatin involvement in human endometrial physiology and related pathologies.

Adenomyosis: What the Patient Needs.

A panel of experts in the field of endometriosis expressed their opinions on management options in a 28-year-old patient, attempting pregnancy for 1 year, with severe cyclic pelvic pain and with clinical examination and imaging techniques suggestive of adenomyosis. Many questions this paradigmatic patient may pose to the clinician are addressed, and all clinical scenarios are discussed. A decision algorithm derived from this discussion is also proposed.

Abdominal Wall Endometriosis on the Right Port Site After Laparoscopy: Case Report and Literature Review.

BACKGROUND: Endometriosis can be intrapelvic or, rarely, extrapelvic. Endometriosis involving the rectus abdominis muscle on the trocar port site is a rare event; until now, only 16 cases have been reported in the literature. The majority of cases were associated with previous abdominal surgery such as diagnostic laparoscopy, cyst excision, appendectomy, myomectomy, or cholecystectomy. We review all the reported cases of this unusual form of extrapelvic endometriosis. CASE REPORT: We report a new case of abdominal wall endometriosis at the trocar port site in the rectus abdominis muscle in a woman who had undergone 2 laparoscopies for endometriosis in the 3 years before coming to our attention. The diagnosis was made by sonography. We performed a surgical resection of the lesion with a free macroscopic margin of 5-10 mm. CONCLUSION: Endometriosis should be considered in the differential diagnosis of any abdominal swelling. In our experience, surgery is the treatment of choice.

How to Manage Bowel Endometriosis: The ETIC Approach.

A panel of experts in the field of endometriosis expressed their opinions on management options in a 35-year-old patient desiring pregnancy with a history of previous surgery for endometrioma and bowel obstruction symptoms. Many questions that this paradigmatic patient may pose to the clinician are addressed, and various clinical scenarios are discussed. A decision algorithm derived from this discussion is proposed as well.

The role of progesterone in maternal and fetal medicine.

Progesterone is an essential hormone in the process of reproduction. It has been extensively studied in the treatment of different gynecological pathologies, as a contraceptive and in assisted reproductive technologies. However, the use of progesterone in the pathophysiology of pregnancy remains controversial. Progesterone, and its synthetic form 17 α-hydroxyprogesterone caproate (17 OHP-C), offer an effective intervention when the continuation of pregnancy is at risk from immunological factors, luteinic and neuroendocrine deficiencies, and myometrial hypercontractility. Progesterone has been successfully used as prophylaxis in the prevention of spontaneous miscarriage, with treatment beginning from the first trimester of pregnancy. There is substantial evidence, too, to indicate that women with idiopathic recurrent miscarriage may benefit from the immunomodulatory properties of progesterone in early pregnancy. The use of progesterone and 17 OHP-C has been extensively studied in the prevention of preterm birth in a variety of settings. Transvaginal ultrasound measurement of cervical length in singleton pregnancies between 19 and 24 weeks' gestation has been deemed the best way to identify women (approximately 2% of the pregnant population) who would benefit from prophylactic progesterone treatment for the prevention of spontaneous preterm birth. This paper reviews the evidence for the safety and efficacy of the use of progesterone in each of these indications.

Techniques of laparoscopic myomectomy.

Myomectomy is one of the commonest gynaecological operations. Laparoscopic myomectomy has emerged over the last two decades as a possible alternative to the traditional laparotomy. Most studies have revealed that the laparoscopic procedure is at least as safe as the open procedure as to the rate of complications and may retain relevant advantages in terms of shorter admission and recovery times. Currently laparoscopic myomectomy is still a challenging operation that requires a well-trained surgical team, adequate instrumentation and accurate patient selection; the increasing slant of gynaecologists towards laparoscopic techniques, along with the advances in surgical instrumentation and suturing materials, will hopefully contribute to keep laparoscopic myomectomy no longer confined to tertiary care centres.

Use of a No. 11 blade scalpel and reusable blunt trocar to establish pneumoperitoneum: description of a safe and inexpensive technique.

First access is crucial in laparoscopic surgery because of its potentially life-threatening complications. A number of procedures using a variety of instruments have been previously described; however, the safest approach remains uncertain. Herein, we describe a simple and inexpensive method for direct trocar insertion using reusable instruments that was developed over 10 years in a series of 4721 consecutive gynecologic laparoscopic procedures. Observed data revealed that the technique is feasible, rapidly performed, and safe, with a likely cost savings, using a small set of reusable instruments. This procedure should be compared with other access methods in randomized studies to confirm the observed advantages.

The presence of trophoblastic cells in intrauterine lavage samples: lack of correlation with maternal and obstetric characteristics.

OBJECTIVES: To investigate the correlation between maternal, obstetric and sample characteristics and the quality (i.e. yield of trophoblastic cells) of intrauterine lavage (IUL) samples. METHODS: We collected 202 IUL samples from women scheduled for first trimester termination of pregnancy (TOP). Trophoblastic cells were isolated from IUL samples and used for DNA analysis by a multiplex quantitative fluorescent polymerase chain reaction (QF-PCR) assay. A multivariate logistic regression analysis was performed, and a p<0.05 was considered statistically significant. RESULTS: The presence of trophoblastic cells in IUL samples was documented in 151/202 cases (74.7%). Blood contamination of IULs was the only characteristic to positively correlate with the presence of trophoblasts (p=0.039; OR: 1.99; 95% CI: 1.03-3.82). CONCLUSIONS: The correlation between the presence of contaminating blood and trophoblastic cells would indirectly confirm the hypothesis that IUL might act as a mini-CVS. The high yield rate of trophoblasts irrespective of maternal characteristics and past obstetric history would support the clinical use of this sampling technique, provided that its safety is clearly defined.

A multicenter randomized, controlled study comparing laparoscopic versus minilaparotomic myomectomy: reproductive outcomes.

OBJECTIVE: To assess the reproductive outcomes after minilaparotomic and laparoscopic myomectomy in patients wishing to conceive. DESIGN: Randomized controlled trial. SETTING: Departments of obstetrics and gynecology of the universities of Catanzaro, Rome, and Florence, Italy. PATIENT(S): One hundred thirty-six women with symptomatic uterine leiomyomas or unexplained infertility. INTERVENTION(S): Laparoscopic and minilaparotomic myomectomy. MAIN OUTCOME MEASURE(S): Pregnancy, abortion, and live-birth rates. RESULT(S): Between the laparoscopic and minilaparotomic groups no difference was observed in cumulative pregnancy, live-birth, and abortion rates, whereas pregnancy and live-birth rates per cycle, and time to first pregnancy and live-birth were significantly higher in the laparoscopic than in the minilaparotomic group. Categorizing the patients according to surgical indication for myomectomy, cumulative pregnancy rate, pregnancy, and live-birth rates per cycle, and time to first pregnancy and live-birth were significantly better after laparoscopic myomectomy in symptomatic patients, whereas all reproductive outcomes were similar between the two groups in patients with unexplained infertility. CONCLUSION(S): Minilaparotomic and laparoscopic myomectomy improves in a similar manner the reproductive outcomes in patients with unexplained infertility, whereas the laparoscopic approach provides the best benefits in fertile patients with symptomatic leiomyomas.

A multicenter randomized, controlled study comparing laparoscopic versus minilaparotomic myomectomy: short-term outcomes.

OBJECTIVE: To compare the laparoscopic and minilaparotomic approaches for symptomatic uterine leiomyomas treatment in terms of safety and feasibility. DESIGN: Randomized controlled trial. SETTING: Three university departments of obstetrics and gynecology of Catanzaro, Rome, and Florence, Italy. PATIENT(S): One hundred thirty-six women wishing to conceive and candidate for myomectomy due to symptomatic uterine leiomyomas or unexplained infertility. INTERVENTION(S): Myomectomy through laparoscopic or minilaparotomic access. MAIN OUTCOME MEASURE(S): Surgical outcomes. RESULT(S): Leiomyoma enucleation and hysterotomy suturing times were significantly shorter after minilaparotomic myomectomy, whereas the degree of surgical difficulty was significantly higher for the laparoscopic myomectomy. Intraoperative blood loss, variation in hemoglobin levels, quantity of pain control drugs used postoperatively, and hospitalization were significantly lower in the laparoscopic group than in the minilaparotomic one. Our surgical outcomes were significantly influenced by specific investigational centers involved, and by leiomyoma dimensions and localizations. This last variable is the strongest predictor of surgical outcome. CONCLUSION(S): Laparoscopic and minilaparotomic approaches to myomectomy are two safe and minimally invasive surgical procedures. A careful evaluation of the dimensions and localizations of fibroids are needed to address to the right choice to the best approach.

Presurgical assessment of intraabdominal visceral fat in obese patients with early-stage endometrial cancer treated with laparoscopic approach: relationships with early laparotomic conversions.

STUDY OBJECTIVE: To evaluate the relationship between intraabdominal visceral fat (IVF) and early conversion from laparoscopy to laparotomy in obese patients. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: Department of obstetrics and gynecology, University "Magna Graecia" of Catanzaro. PATIENTS: One hundred fifty-one obese patients with endometrial cancer and laparoscopic surgery. INTERVENTIONS: Laparoscopic treatment for early-stage endometrial cancer. MEASUREMENT AND MAIN RESULTS: In all patients, demographic, anthropometric (including body mass index, waist-to-hip ratio, and waist circumference), and IVF assessments (using ultrasonography [IVF-USG, cm] and computed tomography [IVF-CT, cm(2)]), were evaluated. In 122 patients (122/151, 80.8%), the laparoscopies were successfully completed (group A), whereas 29 patients (29/151, 19.2%) had a laparotomic conversion. Specifically, in 19 (19/29, 65.5%) and in 10 patients (10/29, 34.5%) the conversion was early (group B1) and late (group B2), respectively. A significant (p <.05) difference in surgical stage IC and in postoperative complications rate was detected between group A and groups B1 and B2, without difference between these 2 last groups. IVF-USG and IVF-CT were significantly (p <.01) higher in group B1 in comparison with groups A and B2, without difference between these 2 last groups. A strong and significant (p <.001) association between IVF-USG and IVF-CT was detected in all groups by Spearman bivariate correlation (r = 0.78, 0.918, and 0.978 for group A, B1, and B2, respectively). Multivariate analysis showed the IVF-CT as the main predictor for early laparotomic conversion (OR 2.184; 95% CI 1.227-3.886). CONCLUSIONS: Intraabdominal visceral fat is a predictor for early laparotomic conversion in obese patients with endometrial cancer. Further studies are necessary to assess the efficacy of the IVF evaluation for reducing the laparotomic conversion risk in obese women undergoing laparoscopic surgery for gynecologic malignancy.

Laparoscopic uterine nerve ablation versus vaginal uterosacral ligament resection in postmenopausal women with intractable midline chronic pelvic pain: a randomized study.

OBJECTIVE: To assess and compare the laparoscopic uterine nerve ablation (LUNA) and the vaginal uterosacral ligament resection (VUSR) in postmenopausal women with chronic pelvic pain (CPP). STUDY DESIGN: Eighty postmenopausal women with intractable and severe midline CPP were randomized to undergo LUNA or VUSR. Costs of two surgical procedures were assesses. Cure rate, severity of CPP, and deep dyspareunia were also evaluated after 6 and 12 months from surgery. RESULTS: The mean cost of LUNA resulted significantly higher in comparison with VURS (2078+/-637 versus 1497+/-297, P<0.001). The cure rate was not significantly different between the two groups at 6 (33/40, 82.5% versus 35/40, 87.5% for groups A and B, respectively; P=0.530; RR 0.94, 95% CI 0.78-1.13), and 12 months (27/36, 75.0% versus 28/38, 73.7%, for groups A and B, respectively; P=0.901; RR 0.90, 95% CI 0.78-1.33) of follow-up. At same times, a significant (P<0.01) decrease in severity of CPP and deep dyspareunia was observed in both groups with no difference between them. CONCLUSIONS: Both LUNA and VUSR are equally effective surgical treatments in postmenopausal women with central CPP but VUNR is significantly cheaper than LUNA.

Progesterone and pregnancy.

PURPOSE OF REVIEW: Progesterone is an essential hormone in the process of reproduction. Although the pharmacokinetics and pharmacodynamics of progesterone have been well studied, its use in the pathophysiology of pregnancy remains controversial. One of these concerns is the way in which the hormone is administered. RECENT FINDINGS: In obstetrics the most frequent uses of progesterone are in the treatment of threatened abortion, prevention of recurrent miscarriage, or in the support of the luteal phase in assisted reproduction programmes, and in threatened preterm labour. Randomized, controlled trials showed that women who received progesterone were statistically significantly less likely to have recurrent miscarriages before 34 weeks, to have an infant with birth weight of 2.5 kg or lower, or to have an infant diagnosed with intraventricular haemorrhage. There is currently, however, insufficient information to allow recommendations regarding the optimal dose, route and timing of administration of progesterone supplementation. SUMMARY: Progesterone has shown to be efficacious when continuation of pregnancy is hampered by immunological factors, luteinic and neuroendocrine deficiencies and myometrial hypercontractility. This may explain the reduction in the incidence of preterm birth in high-risk pregnant women using high-dosage prophylactic progesterone.