Marker molecolari relativi all'infezione da Papillomavirus umani: significato diagnostico e prognostico in lesioni virus-correlate

Introduzione: Il cervico-carcinoma è la seconda neoplasia maligna per incidenza e mortalità nelle donne in tutto il mondo dopo il cancro al seno. L’infezione persistente da Papillomavirus Umani (HPV) è causa necessaria dell’insorgenza del cervico-carcinoma e delle sue lesioni pre-cancerose. L’infezione da HPV si associa anche ad altri carcinomi del distretto ano-genitale (a livello anale, vulvare, vaginale e del pene) e a circa il 25% dei carcinomi squamosi dell’orofaringe. I circa 40 genotipi di HPV che infettano la mucosa genitale vengono suddivisi in alto rischio (HR-HPV) e basso rischio (LR-HPV) oncogeno a seconda della alta e bassa associazione con la neoplasia cervicale. I 13 genotipi a più alto rischio oncogeno sono 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 e 68. Di questi, otto (16, 18, 31, 33, 35, 45, 52 e 58) sono associati alla maggior parte dei carcinomi cervicali (circa 89%) e i genotipi 16 e 18 da soli sono riscontrati nel 70% circa delle neoplasie. L’insorgenza e progressione delle lesioni preneoplastiche cervicali è, però, associata non solo alla presenza di HPV ad alto rischio, ma, soprattutto, alla persistenza virale (> 18 mesi), alla capacità di integrazione degli HPV ad alto rischio e alla conseguente sovraespressione delle oncoproteine E6/E7. Inoltre, l’integrazione è spesso favorita da un’alta carica virale soprattutto per quanto riguarda alcuni genotipi (HPV16 e 18). L’infezione da HPV non interessa solo la cervice uterina ma tutto il distretto ano-genitale e quello testa-collo. L’HPV, in particolare il genotipo 16, è implicato, infatti, nell’insorgenza delle lesioni preneoplastiche della vulva (VIN) classificate come VIN classiche e nei carcinomi ad esse associati. L’incidenza del carcinoma vulvare in Europa è di 1.5/100.000 di cui circa il 45% è dovuto a HR-HPV (80% ad HPV16). Nonostante l’associazione tra HPV16 e carcinoma vulvare sia alta, ancora poco si conosce sul ruolo della carica virale e dell’integrazione in tali lesioni. Le lesioni che possono presentarsi nella regione testa-collo possono essere sia di natura benigna che maligna. I genotipi più frequentemente riscontrati in associazione a lesioni benigne (papillomi) sono HPV 6 e 11, quelli associati a forme tumorali (HNSCC) sono il genotipo 18 ma soprattutto il 16. Molti aspetti del coinvolgimento di HPV in queste patologie non sono ancora perfettamente conosciuti e spesso studi su tale argomento hanno mostrato risultati contraddittori, soprattutto perché vengono utilizzate metodiche con gradi diversi di sensibilità e specificità. Recenti dati di letteratura hanno tuttavia messo in evidenza che i pazienti affetti da HNSCC positivi ad HPV hanno una elevata risposta al trattamento chemioradioterapico rispetto ai pazienti HPV-negativi con un notevole impatto sul controllo locale e sulla sopravvivenza ma soprattutto sulla qualità di vita di tali pazienti, evitando di sottoporli a chirurgia sicuramente demolitiva. Scopo del lavoro: Sulla base di queste premesse, scopo di questo lavoro è stato quello di valutare l’importanza di marker quali la presenza/persistenza di HPV, la carica virale, la valutazione dello stato fisico del genoma virale e l’espressione degli mRNA oncogeni nella gestione di pazienti con lesioni preneoplastiche e neoplastiche di diverso grado, associate a papillomavirus mucosi. Per la valutazione dei markers virologici di progressione neoplastica abbiamo sviluppato dei saggi di real time PCR qualitativi e quantitativi studiati in modo da poter fornire, contemporaneamente e a seconda delle esigenze, risposte specifiche non solo sulla presenza e persistenza dei diversi genotipi di HPV, ma anche sul rischio di insorgenza, progressione e recidiva delle lesioni mediante lo studio di markers virologici quali carica virale, integrazione ed espressione degli mRNA. Abbiamo pertanto indirizzato la nostra attenzione verso tre popolazioni specifiche di pazienti: - donne con lesioni vulvari preneoplastiche (VIN) e neoplastiche, allo scopo di comprendere i complessi meccanismi patogenetici di tali patologie non sempre associate ad infezione da HPV; - pazienti con lesioni maligne a livello della regione testa-collo allo scopo di fornire informazioni utili all’elaborazione di un percorso terapeutico mirato (radiochemioterapico o chirurgico) a seconda o meno della presenza di infezione virale; - donne con lesioni cervicali di alto grado, trattate chirurgicamente per la rimozione delle lesioni e seguite nel follow-up, per stabilire l’importanza di tali marker nella valutazione della persistenza virale al fine di prevenire recidive di malattia.

The unspoken disease: symptom experience in women with vulval neoplasia and surgical treatment: a qualitative study

Women with vulval neoplasia often experience severe post-surgical complications. This study focuses on symptom experience of women during the first 6 months following surgical treatment for vulval neoplasia considering their socio-cultural context. In this qualitative study using a critical hermeneutic approach, narrative interviews were conducted. A purposeful sample of 20 patients was recruited from one Swiss and two German university hospitals. Content analysis was employed to analyse the transcribed interviews considering women's experiences and social perceptions. Narratives showed eight interrelated themes: delayed diagnosis, disclosed disease, disturbed self-image, changed vulva care, experienced wound-related symptoms, evoked emotions, affected interpersonal interactions and feared illness progression. The women experienced a general lack of information pertaining to above themes and all described strategies used to handle their situation, which affected their distress. The communication, assessment and treatment of symptoms were hampered by the society's and the health system's tendency to overlook these symptoms and leave them in the realm of the unspeakable. Health professionals need new strategies to support these women to recognise, assess and evaluate the seriousness of symptoms, and to communicate their symptom experience so that timely medical treatment is sought. This support may minimise potentially preventable complications and symptom-related distress.

Prevalence, motivations, and adverse effects of vaginal practices in Africa and Asia: findings from a multicountry household survey

Background: Women worldwide use various vaginal practices to clean or modify their vulva and vagina. Additional population-level information is needed on prevalence and motivations for these practices, characteristics of users, and their adverse effects. Methods: This was a household survey using multistage cluster sampling in Tete, Mozambique; KwaZulu-Natal, South Africa; Yogyakarta, Indonesia; and Chonburi, Thailand. In 2006–2007, vaginal practices and their motivations were examined using structured interviews with women 18–60 years of age (n=3610). Results: Prevalence, frequency, and motivations varied markedly. Two thirds of women in Yogyakarta and Chonburi reported one or more practices. In Yogyakarta, nearly half ingest substances with vaginal effects, and in Chonburi, external washing and application predominate. In Tete, half reported three or four current practices, and a quarter reported five or more practices. Labial elongation was near universal, and 92% of those surveyed cleanse internally. Two third's in KwaZulu-Natal practiced internal cleansing. Insertion of traditional solid products was rare in Chonburi and Yogyakarta, but one tenth of women in KwaZulu-Natal and nearly two thirds of women in Tete do so. Multivariate analysis of the most common practice in each site showed these were more common among less educated women in Africa and young urban women in Asia. Explicit sexual motivations were frequent in KwaZulu-Natal and Tete, intended for pleasure and maintaining partner commitment. Practices in Chonburi and Yogyakarta were largely motivated by femininity and health. Genital irritation was common at African sites. Conclusions: Vaginal practices are not as rare, exotic, or benign as sometimes assumed. Limited evidence of their biomedical consequences remains a concern; further investigation of their safety and sexual health implications is warranted.

[The mandatory diaper peek in newborn girls]

We present a 3 day old girl born at term presenting with an interlabial, cystic mass. Pregnancy, delivery and routine antenatal screening were unremarkable. The smooth lesion was located in the anterior half of the vulva, covered with thin vessels. The medially displaced urethra and the medio-posteriorly displaced hymen were identified. Voiding was not impaired. We discuss the differential diagnosis of vulvar masses in the newborn girl. A thorough clinical examination must be the standard of care of pediatric examinations in infants.

Comparison of outcome and recurrence-free survival after sentinel lymph node biopsy and lymphadenectomy in vulvar cancer

OBJECTIVES: Lymph node status is an important prognostic factor in patients with squamous cell carcinoma (SCC) of the vulva. Complete inguinofemoral lymph node dissection (ILND) is accompanied by a high morbidity. Sentinel lymph node biopsy (SLNB) was established for less invasive lymph node (LN) staging. The aim of this study was to evaluate safety of SLNB in terms of accuracy and outcome in a clinical routine setting. METHODS: We retrospectively reviewed the data of patients who underwent SLNB and/or ILND for vulvar SCC in the years 1990-2007. Clinical follow-up was evaluated for histological nodal-negative patients with tumor stage T1 or T2. The false negative rate of SLNB was determined in patients who underwent both SLNB and ILND. RESULTS: Preoperative sentinel lymph node (SLN) visualization by scintigraphy was successful in 95% of all patients. SLNB was false negative in 1/45 inguinae (2.2%). All SLN were detected intraoperatively. During the follow-up period (median 24 months for SLNB and 111 months for ILND), no groin recurrences in initially nodal negative patients occurred (n=34, 59 inguinae). Transient lymph edema occurred in 7/18 patients after ILND (39%) and 2/16 patients (13%) after SLNB. No persistent edemas were found after SLNB and ILND. CONCLUSION: According to our experience SLNB is feasible and accurately predicts LN status of vulvar SCC under clinical routine conditions. SLNB in vulvar cancer seems to be a safe alternative to ILND in order to reduce morbidity of surgical treatment.

Chronic bovine besnoitiosis: intra-organ parasite distribution, parasite loads and parasite-associated lesions in subclinical cases.

Bovine besnoitiosis caused by Besnoitia besnoiti is a chronic and debilitating disease. The most characteristic clinical signs of chronic besnoitiosis are visible tissue cysts in the scleral conjunctiva and the vagina, thickened skin and a generally poor body condition. However, many seropositive animals remain subclinically infected, and the role that these animals may play in spreading the disease is not known. The aim of the present study was to assess the intra-organ parasite distribution, the parasite load and the parasite-associated lesions in seropositive but subclinically infected animals. These animals were seropositive at the time of several consecutive samplings, had visible tissue cysts in the past and, at time of slaughter, had detectable specific anti-Besnoitia spp. antibody levels, but they did not show evident clinical signs at culling. Thus, histopathological, immunohistochemical and molecular analyses of several samples from the respiratory tract, reproductive tract, other internal organs and skin from six cows were performed. The tissue cysts were located primarily in the upper respiratory tract, i.e., in the rhinarium and larynx/pharynx (four cows), followed by the distal genital tract (vulva/vagina) and the skin of the neck (three and two cows, respectively, out of the four cows with cysts in the respiratory tract). We were unable to detect any parasites in the two remaining cows. Cysts were associated with a significant non-purulent inflammatory infiltrate consisting predominantly of T lymphocytes and activated monocytes/macrophages in two cows. The parasite burden, estimated by quantitative real-time PCR, was very low. It is noteworthy that the only animal that showed a recent increase in the antibody titre had the highest parasite burden and the most conspicuous inflammatory reaction against the cysts. In conclusion, although these cows no longer displayed any visible signs of besnoitiosis, they remained infected. Therefore, cows without visible signs of disease may still be able to transmit the parasite.

Between anxiety and hope: the experiences of women with vulval intraepithelial neoplasia during their illness trajectory - a qualitative approach

The vulvar intraepithelial neoplasia (VIN) is a rare chronic skin condition that may progress to an invasive carcinoma of the vulva. Major issues affecting women's health were occurring symptoms, negative influences on sexuality, uncertainty concerning the illness progression and changes in the body image. Despite this, there is little known about the lived experiences of the illness trajectory. Therefore, the aim of this study was to describe the experiences of women with VIN during the illness trajectory. In a secondary data analysis of the foregoing qualitative study we analysed eight narrative interviews with women with VIN by using thematic analysis in combination with critical hermeneutics. Central for these women during their course of illness was a sense of "Hope and Fear". This constitutive pattern reflects the fear of recurrence but also the trust in healing. The eight narratives showed women's experiences during their course of illness occurred in five phases: "there is something unknown"; "one knows, what IT is"; "IT is treated and should heal"; "IT has effects on daily life"; "meanwhile it works". Women's experiences were particularly influenced by the feeling of "embarrassment" and by "dealing with professionals". Current care seems to lack adequate support for women with VIN to manage these phases. We suggest, based on our study and the international literature, that new models of counselling and providing information need to be developed and evaluated.

Disease progression and recurrence in women treated for vulvovaginal intraepithelial neoplasia

OBJECTIVE The malignant potential of intraepithelial neoplasia of the vulva and vagina after treatment is not well defined. Our objective was to examine risk factors for recurrence and invasive disease. METHODS Four hundred sixty-four women with biopsy proven high-grade intraepithelial neoplasia of the vulva and vagina were identified in the electronic databases of four colposcopy clinics. Inclusion criteria were a follow-up of more than one year, no history of invasive cancer and no invasive cancer within the first year after initial treatment. We investigated the potential factors associated with recurrence and progression using a logistic regression analysis to estimate odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS Of the 411 eligible patients, 123 patients (29.9%) recurred later than one year after initial treatment and 24 patients (5.8%) progressed to invasive disease. According to multivariate analyses, the risk factors associated with recurrence were multifocality (OR, 3.33; 95% CI, 2.02 to 5.51), immunosuppression (OR, 2.51; 95% CI, 1.09 to 5.81), excision as initial treatment (vs. laser evaporation; OR, 1.79; 95% CI, 1.11 to 2.91) and smoking (OR, 1.61; 95% CI, 1.02 to 2.55). Risk factors for progression to invasive disease were immunosuppression (OR, 4.00; 95% CI, 1.30 to 12.25), multifocality (OR, 3.05; 95% CI, 1.25 to 7.43) and smoking (OR, 2.97; 95% CI, 1.16 to 7.60), but not treatment modality. CONCLUSION Laser evaporation combined with extensive biopsy is at least as efficacious as initial treatment of intraepithelial neoplasia with excision. Smoking is a risk factor for both recurrence and progression to invasive disease. Hence, smoking cessation should be advised and maintaining a long follow-up period due to late relapses is necessary.

AMERICAN TRYPANOSOMIASIS (CHAGAS') DISEASE - A STUDY OF FETO-MATERNAL MORBIDITY AND MORTALITY

The possibility of a relationship between American Trypanosomiasis (Chagas') disease and pregnancy outcome was analyzed measuring feto-maternal morbidity and mortality in a sample of 604 pregnant women and their offspring seen at the Hospital Universitario de Maternidad y Neonatologia in Cordoba, Argentina during 1979.^ A cross-sectional, "case-comparison" investigation was employed to determine the degree of risk between having a reactive chagasic serologic test and a negative pregnancy outcome as determined by abortion, stillbirth, and infant death prior to one week of age. Patients were selected using a dichotomous, 0-1 scale with either the presence or the absence of a reactive Machado-Guerreiro complement fixation serologic blood test result.^ The data obtained were analyzed using appropriate statistical techniques for measuring the comparisons between the case and control groups under various demographic and socioeconomic variables such as, age, marital status, educational attainment, and residence. Similarly, additional biological variables of birth order, maternal and fetal complications, and prematurity were examined.^ From the analysis of the data obtained in this investigation, no definite conclusions can be reached regarding the risk of having an unsuccessful pregnancy outcome in the presence of a reactive serologic finding because the study design was a cross-sectional one and the number of events were too few for an adequate analysis. Notwithstanding these limitations, the results obtained, after statistical adjustments were employed, demonstrated that women with a reactive test result were older, were of a higher parity, and were less educated. Marital status and residence were not significant variables. The risk of pregnancy wastage, however, was almost twice as frequent in the reactive group as in the non-reactive group of women. Statistically significant differences in maternal morbidity involved two complications, polyhydramnios and varicosities of the lower extremities and vulva; while in the newborn, infection was higher in infants whose mothers exhibited a reactive serologic test result.^ In summary, what this research study has shown is the need for engaging in a larger, longitudinal study for an in-depth exploration of feto-maternal morbidity and mortality--an investigation that would corraborate or refute the findings of this study.^

Carcinoma escamoso vulvar : revisión de la literatura

Fil: Parra, Viviana Gladys. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas. Cátedra de Dermatología

Morphological descriptions from the recent bivalve of the genus Calyptogena (Vesicomyidae)

The genus Calyptogena (Bivalvia: Vesicomyidae) comprises highly specialized bivalves living in symbiosis with sulphur-oxidizing bacteria in reducing habitats. In this study, the genus is revised using shell and anatomical features. The work is based on type material, as well as on the extensive collection of vesicomyids obtained during twelve expeditions to the Pacific and Indian Oceans. Nine Recent species are ascribed to the genus Calyptogena, four of which are new: C. pacifica Dall, 1891, C. fausta Okutani, Fujikura & Hashimoto, 1993, C. rectimargo Scarlato, 1981, C. valdiviae (Thiele & Jaeckel, 1931), C. gallardoi Sellanes & Krylova, 2005, C. goffrediae n. sp., C. starobogatovi n. sp., C. makranensis n. sp. and C. costaricana n. sp. The characteristic features of Calyptogena are: shell up to 90 mm in length, elongate-elliptical or elongate; presence of escutcheon; presence of broad posterior ramus (3b) of right subumbonal cardinal tooth as well as right posterior nymphal ridge; absence of pallial sinus as a result of attachment of intersiphonal septal retractor immediately adjacent to ventral surface of posterior adductor; absence of processes on inner vulva of inhalant siphon; presence of inner demibranch only, with descending and ascending lamellae with interlamellar septa not divided into separate tubes. The most closely related taxa to Calyptogena are probably the genus Isorropodon Sturany, 1896, and the group of species represented by 'Calyptogena' phaseoliformis Métivier, Okutani & Ohta, 1986. These groups have several characters in common, namely absence of pallial sinus, presence of single inner pair of demibranchs and absence of processes on inner vulva of inhalant siphon. The worldwide distribution of the genus Calyptogena suggests that methane seeps at continental margins are the major dispersal routes and that speciation was promoted by geographical isolation. Recent species diversity and fossil records indicate that the genus originated in the Pacific Ocean. Sufficient data to discuss the distribution at species level exist only for C. pacifica, which has a remarkably narrow bathymetric range. Published studies on the physiology of C. pacifica suggest that adaptation to a specific geochemical environment has led to coexisting vesicomyid genera. The bacteria-containing gill of C. pacifica and other Calyptogena species is one of the most specialized in the family Vesicomyidae and may reflect these ecological adaptations.

sqv-3, -7, and -8, a set of genes affecting morphogenesis in Caenorhabditis elegans, encode enzymes required for glycosaminoglycan biosynthesis

sqv (squashed vulva) genes comprise a set of eight independent loci in Caenorhabditis elegans required zygotically for the invagination of vulval epithelial cells and maternally for normal oocyte formation and embryogenesis. Sequencing of sqv-3, sqv-7, and sqv-8 suggested a role for the encoded proteins in glycolipid or glycoprotein biosynthesis. Using a combination of in vitro analysis of SQV enzymatic activities, sqv+-mediated rescue of vertebrate cell lines, and biochemical characterization of sqv mutants, we show that sqv-3, -7, and -8 all affect the biosynthesis of glycosaminoglycans and therefore compromise the function of one specific class of glycoconjugates, proteoglycans. These findings establish the importance of proteoglycans and their associated glycosaminoglycans in epithelial morphogenesis and patterning during C. elegans development.

Coordinated morphogenesis of epithelia during development of the Caenorhabditis elegans uterine-vulval connection.

Development of the nematode egg-laying system requires the formation of a connection between the uterine lumen and the developing vulval lumen, thus allowing a passage for eggs and sperm. This relatively simple process serves as a model for certain aspects of organogenesis. Such a connection demands that cells in both tissues become specialized to participate in the connection, and that the specialized cells are brought in register. A single cell, the anchor cell, acts to induce and to organize specialization of the epidermal and uterine epithelia, and registrates these tissues. The inductions act via evolutionarily conserved intercellular signaling pathways. The anchor cell induces the vulva from ventral epithelial cells via the LIN-3 growth factor and LET-23 transmembrane tyrosine kinase. It then induces surrounding uterine intermediate precursors via the receptor LIN-12, a founding member of the Notch family of receptors. Both signaling pathways are used multiple times during development of Caenorhabditis elegans. The outcome of the signaling is context-dependent. Both inductions are reciprocated. After the anchor cell has induced the vulva, it stretches toward the induced vulval cells. After the anchor cell has induced specialized uterine intermediate precursor cells, it fuses with a subset of their progeny.

Caso Dermatológico

Menina de cinco anos com antecedentes de obstipação e enurese noturna primária, medicada com desmopressina. Referenciada pelo médico assistente à consulta de Pediatria por despigmentação e erosões vulvares associada a prurido intenso, com dois meses de evolução