Leonardo Micheletti

Increased activity of 6-phosphogluconate dehydrogenase and glucose-6-phosphate dehydrogenase in purified cell suspensions and single cells from the uterine cervix in cervical intraepithelial neoplasia

The activities of 6-phosphogluconate dehydrogenase and glucose-6-phosphate dehydrogenase have been measured in squamous epithelial cells of the uterine cervix from normal patients and cases of cervical intraepithelial neoplasia (CIN). A biochemical cycling method, which uses only simple equipment and is suited to routine use and to automation, was applied to cells separated by gradient centrifugation. In addition, cells were examined cytochemically, and the intensity of staining in the cytoplasm of single whole cells was measured using computerised microcytospectrophotometry. Twenty per cent of cells in samples from normal patients (n=61) showed staining intensities above an extinction of 0.15 at 540 nm, compared to 71% of cases of CIN 1 (n=14), 91% of cases of CIN 2 (n=11) and 67% of cases of CIN 3 (n=15). The cytochemical data do not allow definitive distinctions to be made between different grades of CIN whereas the biochemical assay applied to cell lysates shows convincing differences between normal samples and cases of CIN. There are no false negatives for CIN 3 (n=14) and CIN 2 (n=10) and 11% false negatives for CIN 1 (n=9) and 14% of false positives for normal cases (n=21). The results of this preliminary study with reference to automation are discussed [corrected].

Vulvar intraepithelial neoplasia

Vulvar intraepithelial neoplasia (VIN) is a high-grade intraepithelial squamous lesion and precursor of invasive squamous cell carcinoma (SCC). The 2004 International Society for the Study of Vulvovaginal Disease (ISSVD) classification distinguished two types of VIN: usual type (human papillomavirus (HPV)-related) and differentiated type (not HPV-related). The incidence of usual-type VIN is higher in younger women, while differentiated-type VIN is more common in older patients with chronic dermatologic conditions. Differentiated-type VIN has a greater invasive potential and shorter time between diagnosis and SCC than usual-type VIN. The diagnosis of VIN is carried out by identifying a lesion by visual inspection and confirming by performing a biopsy. Screening tests are not available. Patients with usual-type VIN are at a higher risk of developing another HPV-related malignancy of the anogenital tract; therefore, examination from the cervix to the perianal area is mandatory. The therapeutic approach to VIN balances the invasive potential with the need to be as conservative as possible. Current prophylactic HPV vaccines offer protection against usual-type VIN and related invasive carcinoma.

Groin lymphadenectomy with Preservation of Femoral Fascia: Total Inguinofemoral Node Dissection for Treatment of Vulvar Carcinoma

This article describes a new technique for groin lymphadenectomy with preservation of the femoral fascia based on correct embryologic and anatomic knowledge of inguinofemoral lymph node disposition and their exact relation with the fascial structures of Scarpa’s triangle. Scarpa’s triangle dissection follows a three-step procedure: development of the side starting from the inguinal ligament; development of the angles proceeding from the apex where the saphenous vein is resected; and dissection of the fossa ovalis by grasping and elevating the entire block of adipose tissue containing the superficial inguinofemoral nodes and the stump of the great saphenous vein. This surgical step allows us to expose and remove en bloc the deep femoral nodes lying medial to the portion of the femoral vein located within the fossa ovalis. The total number of inguinofemoral nodes removed from a series of 156 patients operated on during 1981–2002 ranged from 8 to 35 (mean 20) bilaterally and from 4 to 18 (mean 10) unilaterally. The 5-year survivals by stage were, respectively, 86.2% for stage Ib, 69.2% for stage II, 49.3% for stage III, and 13.3% for stage IVa; these figures are comparable to the survival rates reported by those performing the classic groin lymphadenectomy. Groin lymphadenectomy with preservation of the femoral fascia is thus an oncologically sound conservative procedure that can replace the classic Way’s technique, which involves femoral vessel skeletonization, and can be useful for treating malignant diseases requiring groin dissection.

Development and validation of a nomogram for predicting outcome of patients with vulvar cancer.

OBJECTIVE: To construct and validate a nomogram to predict relapse-free survival of patients treated for vulvar cancer. METHODS: Data from 244 patients treated for vulvar cancer at a single institution (Creteil, France) were used as a training set to develop and calibrate a nomogram for predicting relapse-free survival and local relapse-free survival. We used bootstrap resampling for the internal validation and we tested the nomogram on an independent validation set of patients (Torino, Italy) for the external validation. RESULTS: The nomograms were based on a Cox proportional hazards regression model. Covariates for the relapse-free survival model included age, T stage, number of metastatic nodes, bilateral lymph node involvement, omission of the lymphadenectomy, margin status, lymphovascular space invasion, and depth of invasion. The concordance indices were 0.85 and 0.83 in the training set before and after bootstrapping, respectively, and 0.83 in the validation set. The predictions of our nomogram discriminated better than did the International Federation of Gynecology and Obstetrics stage (0.83 compared with 0.78, P = .01). The calibration of our nomogram was good. In the validation set, 2-year and 5-year relapse-free survival were well predicted with less than 5% difference between the predicted and observed survivals for each quartile. A nomogram for predicting local relapse was also developed. CONCLUSION: We have developed nomograms for predicting distant and local relapse of vulvar cancer at 2 and 5 years and validated them both internally and externally. These nomograms will be freely available on the International Society for the Study of Vulvovaginal Disease Web site. LEVEL OF EVIDENCE: III

Vulvar paget disease: one century after first reported.

Objectives. To provide a critical assessment of the published literature on vulvar Paget disease and to allow individualized approaches to affected patients. Materials and Methods. A computerized search for studies published in the literature up to June 2002 was carried out using Ovid© and Medline databases. We excluded single case reports, letters to editors, and abstracts. Results. Historical and epidemiological aspects of vulvar Paget disease are summarized. Clinical and histopathological data support a recent proposal to classify vulvar Paget disease into two categories, primary and secondary, with significant clinical and prognostic implications. The treatment for primary vulvar Paget disease is wide and deep surgical excision. Inguinofemoral lymphadenectomy is added in the management of invasive neoplasms. In the presence of secondary Paget disease, therapy must be directed toward treatment of associated carcinoma. Conclusions. The subclassification of vulvar Paget disease is essential for correct clinical management and treatment. Immunohistochemistry may assist in this important distinction.

VIN usual type—from the past to the future

Usual vulvar intraepithelial neoplasia (uVIN) is the most common VIN type, generally related to a human papillomavirus (HPV) infection, predominantly type 16. The incidence of uVIN has been increasing over the last decades, and a bimodal peak is observed at the age of 40–44 and over 55 years. Almost 40% of patients with uVIN have a past, concomitant or future HPV-associated lesion of the lower genital tract. HPV-related malignancies are associated with a persistent HPV infection. The host immune response is of crucial importance in determining clearance or persistence of both HPV infections and HPV-related VIN. About 60% of the patients present with symptoms. Clinical features of uVIN vary in site, number, size, shape, colour, and thickness of lesions. Multicentric disease is often present. Most uVIN lesions are positive at immunohistochemistry to p16ink4a and p14arf, but negative to p53. Irrespective of surgical treatment used, uVIN recurrence rates are high. Positive margins do not predict the development of invasive disease and the need to re-excide the tissue around the scare remains to be demonstrated. Therefore, considering the low progression rate of uVIN and psycosexual sequelae, treatments should be as conservative as possible. Medical treatments available are mainly based on immunotherapy to induce normalisation of immune cell count in uVIN. None are approved by the food and drug administration (FDA) for the treatment of uVIN. If medical treatment is performed, adequate biopsies are required to reduce the risk of unrecognised invasive disease. Some studies suggest that failure to respond to immunotherapy might be related to a local immunosuppressive microenvironment, but knowledge of the uVIN microenvironment is limited. Moreover, our knowledge of the potential mechanisms involved in the escape of HPV-induced lesions from the immune system has many gaps. HPV vaccines have been demonstrated to be effective in preventing uVIN, with 94.9% efficacy in the HPV-naive population, while studies on therapeutic vaccines are limited. The low incidence of VIN requires large multicentre studies to determine the best way to manage affected patients and to investigate the immunological characteristics of the ‘vulvar microenviroment’ which leads to the persistence of HPV.

Surgery of the vulva in vulvar cancer

The standard radical mutilating surgery for the treatment of invasive vulval carcinoma is, today, being replaced by a conservative and individualised approach. Surgical conservative modifications that are currently considered safe, regarding vulval lesion, are separate skin vulval-groin incisions, drawn according to the lesion diameter, and wide local radical excision or partial radical vulvectomy with 1-2 cm of clinically clear surgical margins. Regarding inguinofemoral lymph nodes management, surgical conservative modifications not compromising patient survival are omission of groin lymphadenectomy only when tumour stromal invasion is ≤ 1 mm, unilateral groin lymphadenectomy only in well-lateralised early lesions and total or radical inguinofemoral lymphadenectomy with preservation of femoral fascia when full groin resection is needed. Sentinel lymph node dissection is a promising technique but it should not be routinely employed outside referral centres. Pelvic nodes are better managed by radiation. Locally advanced vulval carcinoma can be managed by ultraradical surgery, exclusive radiotherapy or chemoradiation.

Provoked vestibulodynia: Inflammatory, neuropathic or dysfunctional pain? A neurobiological perspective

Abstract This paper aims to clarify the nature of the pain in provoked vestibulodynia (PV). It reviews published data about the nature of the pain in PV, employing a recent pain classification, which divides pain from a neurobiological perspective, into nociceptive, inflammatory and pathological pain, with the latter subdivided into neuropathic and dysfunctional pain. Nociceptive pain is high-threshold pain provoked by noxious stimuli; inflammatory pain is adaptive, low-threshold pain associated with peripheral tissue inflammation; pathological pain is maladaptive, low-threshold pain caused by structural damage to the nervous system (neuropathic) or by its abnormal function (dysfunctional). Most of the published data show that in PV, there is no active peripheral tissue inflammation. Similarly, no neural damage has been demonstrated. It is reasonable to consider PV as dysfunctional pain induced by exposure to acute physical or psychological precipitating events in the presence of an individual predisposition to produce or maintain abnormal central sensitisation.

Vulvar Lichen Sclerosus and Neoplastic Transformation: A Retrospective Study of 976 Cases.

ObjectiveThe aim of the study was to estimate the neoplastic potential of vulvar lichen sclerosus (VLS). Materials and MethodsThis was a retrospective study of 976 women with VLS. We recorded age at diagnosis of VLS, length of follow-up, and type of neoplasia, categorized as the following: (1) vulvar intraepithelial neoplasia (VIN), further subdivided in differentiated VIN (dVIN) and high-grade squamous intraepithelial lesion; (2) superficially invasive squamous cell carcinoma; and (3) frankly invasive squamous cell carcinoma. Neoplasia incidence risk, neoplasia incidence rate, and cumulative probability of progression to neoplasia according to the Kaplan-Meier method were estimated. Log-rank test was used to compare the progression-free survival curves by age at diagnosis of VLS. ResultsThe mean age at diagnosis of VLS was 60 (median = 60; range = 8–91) years. The mean length of follow-up was 52 (median = 21; range = 1–331) months. The following 34 patients developed a neoplasia: 8 VIN (4 dVIN, 4 high-grade squamous intraepithelial lesions), 6 keratinizing superficially invasive squamous cell carcinoma (5 with adjacent dVIN), and 20 keratinizing invasive squamous cell carcinoma (1 with adjacent dVIN). The neoplasia incidence risk was 3.5%. The neoplasia incidence rate was 8.1 per 1,000 person-years. The cumulative probability of progression to neoplasia increased from 1.2% at 24 months to 36.8% at 300 months. The median progression-free survival was significantly shorter in older women (≥70 years) when compared with that in younger women (p = .003). ConclusionsVulvar lichen sclerosus has a nonnegligible risk of neoplastic transformation and requires a careful and lifelong follow-up in all patients, particularly in elderly women. Early clinical and histological detection of preinvasive lesions is essential to reduce the risk of vulvar cancer.

Is the 2003 ISSVD terminology and classification of vulvodynia up-to-date? A neurobiological perspective.

This paper aims to determine if the 2003 International Society for the Study of Vulvovaginal Disease (ISSVD) terminology and classification of vulval pain is up-to-date, according to a current and widely accepted neurobiological pain classification, which divides pain into nociceptive, inflammatory and pathological pain with the latter subdivided into neuropathic and dysfunctional pain. Nociceptive pain is protective, adaptive, high-threshold pain provoked by noxious stimuli. Inflammatory pain is protective, adaptive, low-threshold pain associated with peripheral tissue damage and inflammation. Pathological pain is non-protective, maladaptive, low-threshold pain caused by structural damage to the nervous system (neuropathic pain) or by its abnormal function (dysfunctional pain). The 2003 ISSVD vulval pain classification should be revised in terms of current neurobiological pain information. Inflammatory vulval pain occurs as a result of specific infectious, inflammatory and neoplastic disorders. Neuropathic vulval pain arises following a specific neurological disorder, responsible for structural damage to the nervous system. Vulvodynia is dysfunctional vulval pain, caused by abnormal function of the nervous system itself.