Is a CIS phenotype apparent in children with Disorders of Sex Development? Milder testicular dysgenesis is associated with a higher risk of malignancy.

All malignant testicular germ cell tumors (TGCT) of adult men are preceded by an in situ stage (CIS) of protracted evolution. The adult CIS is well characterized, but there is debate on the phenotype of infantile CIS, its distinction from delayed maturation of germ cells and prognostic potential. A large series of 43 patients with Disorders of Sex Development (DSD) and dysgenetic testes (90% ranging from neonates to 12 years, mean age 4.7 years), was studied by quantifying dysgenetic features, degree of germ cell abnormalities/atypia (GCA), expression of OCT 3/4 (a pluripotency-undifferentiation marker), germ cell ploidy and evolution to CIS and invasive TGCT. Findings were compared with those of normal testes. The type of gonads present defined three groups of patients: bilateral testes (BT-DSD, n = 21), one testis and one streak gonad (CT-DSD, C for combined, n = 13), and ovarian-testicular combinations (OT-DSD, n = 9). There were 5 boys with infantile CIS, bilateral in 3 (total of 8 infantile CIS) and two patients with adult CIS, bilateral in one (total of 3 adult CIS). Two patients had bilateral seminomas one at 12-17 and the other at 23 years. Histological dysgenesis was significantly higher in CT-DSD (p < 0.05), that had only 1 CIS. The highest frequency of GCA was in BT-DSD (p < 0.05), which coincided with a total of 11CIS + Seminomas. In all patients, aneuploidy was significantly higher (63%) than diploidy (p < 0.02), and GCA were more frequent in aneuploid than in diploid samples (p < 0.02). All CIS and TGCT were OCT 3/4 positive. Finally, there was a significant association between the triad Aneuploidy + GCA + OCT 3/4 positivity and the incidence of CIS (Fisher Exact test p < 0.002, relative risk 7.0). The degree of testicular dysgenesis (derived from abnormal organization of Sertoli cells in fetal testicular cords) is inversely related to the incidence of CIS. Our data demonstrate that the combined use of OCT 3/4 expression, quantification of germ cell abnormalities-atypia and ploidy in dysgenetic testes can satisfactorily identify infantile CIS with high risk of malignant evolution and set it aside from delayed germ cell maturation with lower or nil neoplastic potential.

Usefulness and limitations of E-cadherin and ß-catenin in the classification of breast carcinomas in situ with mixed pattern.

BACKGROUND: The distinction between lobular neoplasia of the breast and ductal carcinoma in situ has important therapeutic implications. In some cases, it is very difficult to determine whether the morphology of the lesion is ductal or lobular. The aim of this study was to evaluate the value of E-cadherin and ß-catenin expression through the immunophenotypical characterization of carcinoma in situ with mixed pattern (CISM). METHODS: A total of 25 cases of CISM were analyzed considering cytology/mixed architecture (ductal and lobular), nuclear pleomorphism, loss of cell cohesion, and presence of comedonecrosis. The immunophenotype pattern was considered E-cadherin positive and ß-catenin positive, or negative. RESULTS: Nineteen (76%) cases presented a mixed cytology and / or architectural pattern, two (8%) presented nuclear pleomorphism, two (8%) presented mixed cytology and nuclear pleomorphism, and two (8%) presented comedonecrosis and nuclear pleomorphism. A complete positivity for E-cadherin and ß-catenin was observed in 11 cases (44%). In one case, the lesion was negative for both markers and showed nuclear pleomorphis. Thirteen lesions showed negative staining in areas of lobular cytology and positive staining in cells presenting the ductal pattern. CONCLUSIONS: The expression of E-cadherin and ß-catenin, combined with cytological and architectural analysis, may highlight different immunophenotypes and improve classification of CISM. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/ 1693384202970681

Rokitansky-Aschoff sinuses mimicking adenocarcinoma of the gallbladder: a study of 8 cases.

We describe 8 cases of cholecystectomy specimens (6 laparoscopic and 2 open cholecystectomies) with Rokitansky-Aschoff (R-A) sinuses that were misinterpreted as adenocarcinomas. They were compared with 8 examples of classical R-A sinuses and 6 cases of R-A sinuses containing foci of adenocarcinoma. Five cases misinterpreted as adenocarcinomas consisted of densely packed, closely opposed R-A sinuses with little intervening stroma or surrounded by a desmoplastic stroma. They were lined by a single layer of cuboidal or columnar cells. There were also pseudostratified columnar cells with mucin-containing cytoplasm and hyperchromatic or vesicular nuclei but without mitotic figures. In 2 cases, the columnar cells had subnuclear vacuoles. Small papillary projections into R-A sinuses were seen in 4 cases, and in 3 others collections of metaplastic pyloric glands, some connected to the epithelium of the sinuses, were recognized. There was focal reactive atypia in both the epithelium of the surface and that of the sinuses. The R-A sinuses resembling gland-like structures had a laminar distribution rather than a disorderly haphazard distribution seen in well-differentiated adenocarcinoma. The remaining 3 cases misinterpreted as adenocarcinomas consisted of numerous deeply penetrating long and short R-A sinuses that branched in different directions and which reach the subserosal or perimuscular connective tissue mimicking invasion. The sinuses were surrounded by hyperplastic smooth muscle bundles and lined by pseudostratified columnar cells mixed with a few goblet cells showing reactive atypia and no mitotic figures. There was focal reactive atypia in both the epithelium of the surface and that of the sinuses. The 2 types of R-A sinuses did not label with carcinoembryonic antigen or p53 and had very low proliferative activity as measured by the MIB1-labeling index. All patients are alive and disease free from 8 months to 17 years (mean follow-up 7 y). In contrast, the foci of invasive adenocarcinoma that arose in R-A sinuses consisted of glands lined by atypical cuboidal or columnar cells with loss of polarity, large hyperchromatic or vesicular nuclei, prominent nucleoli, and mitotic figures, quite different from the cells lining the R-A sinuses. Because of increasing number of laparoscopic cholecystectomies performed annually in the United States, pathologists should become familiar with these gallbladder lesions that are usually incidental findings but can simulate malignant epithelial neoplasms.

Distinctive association of p16INK4a overexpression with penile intraepithelial neoplasia depicting warty and/or basaloid features: a study of 141 cases evaluating a new nomenclature.

From the pathogenic point of view, penile cancers may be grouped in human papillomavirus-related and unrelated tumors, each one of them with distinctive morphologic features. The former are predominantly composed of small, undifferentiated basaloid cells, with more or less prominent koilocytic changes, and the latter of keratinizing differentiated squamous cells. The same cellular types are observed in precancerous lesions. On the basis of these observations, we constructed a novel nomenclature for penile precancerous lesions and classified them as penile intraepithelial neoplasia (PeIN) of differentiated, warty, basaloid, and warty-basaloid types. The aim of this study was to test the usefulness of immunohistochemical p16 overexpression, considered as a surrogate for high-risk human papillomavirus infection, using this classification system. We pathologically evaluated 141 patients with PeIN, associated (123 cases) and unassociated (18 cases) with invasive cancer. Distribution of PeIN types was: differentiated, 72%; basaloid, 9%; warty-basaloid, 7%; warty, 4%; and mixed, 7%. There was a striking similarity in the morphology of in situ and invasive squamous cell carcinomas. Differentiated PeIN was commonly associated with usual, verrucous, papillary, and other low-grade keratinizing variants of squamous cell carcinoma whereas in basaloid and warty carcinomas the presence of in situ lesions with similar morphology was habitual. We evaluated p16 overexpression using a 4-tiered (0, 1, 2, and 3) pattern-based system. To properly distinguish differentiated PeIN from in situ lesions with warty and/or basaloid features only pattern 3, which requires full-thickness staining in all epithelial cells, was considered positive. Using this approach, there was a significant association of the negative patterns and differentiated PeIN and of the positive pattern and warty, basaloid, and warty-basaloid PeIN (P<0.0001). Basaloid variant had the strongest association. The sensitivity rate of p16 positivity for discriminating types of PeIN was of 82%, with a specificity of 100% and an accuracy of 95%. Lichen sclerosus was identified in 42 cases and their epithelial component was p16 negative in all cases. Although more studies are necessary to confirm these observations, p16 overexpression seems to be a useful tool for discriminating differentiated from warty, basaloid, and warty-basaloid PeIN.

Human papillomavirus infection and p53 protein expression in vulvar intraepithelial neoplasia and invasive squamous cell carcinoma.

The etiopathogenesis of vulvar intraepithelial neoplasia (VIN III) and invasive squamous cell carcinoma are largely unknown. Since there are few studies on Brazilian patients, our purpose was to determine the frequency of human papillomavirus (HPV) infection and the expression of p53 in these lesions, and associate them with other factors such as age, morphological subtypes, multicentric and multifocal disease. Thirty-eight cases of VIN III, nine of superficially invasive carcinoma, and 55 of invasive vulvar carcinoma were retrospectively evaluated from 1983 to 1995 for the presence of HPV by immunohistochemistry and in situ hybridization, and for p53 protein expression by immunohistochemistry on paraffin sections. All cases for whom material (slides and paraffin blocks) and clinical data were available were included. HPV and p53 were detected in 57.9 and 21.1% of the VIN III lesions, 33.3 and 66.7% of superficially invasive carcinomas, and 7.3 and 58.2% of invasive squamous cell carcinomas, respectively. HPV infection was associated with younger age in the VIN III and invasive carcinoma groups. In the latter, HPV infection was associated with the basaloid variant. p53 expression rate was higher in superficially invasive and invasive lesions and was not related to HPV infection. Our findings are similar to others and support the hypothesis that there are two separate entities of the disease, one associated with HPV and the other unrelated, with p53 inactivation possibly being implicated in some of the cases.

Changes in retinoblastoma gene expression during cervical cancer progression.

The role of tumour suppressor genes in the development of human cancers has been studied extensively. In viral carcinogenesis, the inactivation of suppressor proteins such as retinoblastoma (pRb) and p53, and cellular oncogenes overexpression, such as c-myc, has been the subject of a number of investigations. In uterine-cervix carcinomas, where high-risk human papillomavirus (HPV) plays an important role, pRb and p53 are inactivated by E7 and E6 viral oncoproteins, respectively. However, little is known about the in situ expression of some of these proteins in pre-malignant and malignant cervical tissues. On the other hand, it has also been demonstrated that c-myc is involved in cervical carcinogenesis, and that pRb participates in the control of c-myc gene expression. By using immunostaining techniques, we investigated pRb immunodetection pattern in normal tissues, squamous intraepithelial lesions (SILs) and invasive carcinomas from the uterine cervix. Our data show low pRb detection in both normal cervical tissue and invasive lesions, but a higher expression in SILs. C-Myc protein was observed in most of the cellular nuclei of the invasive lesions, while in SILs was low. These findings indicate a heterogeneous pRb immunostaining during the different stages of cervical carcinogenesis, and suggest that this staining pattern could be a common feature implicated in the pathogenesis of uterine-cervix carcinoma.

Ki-67 expression in anal intraepithelial neoplasia in AIDS.

CONTEXT: AIDS is one of the most important risk factors for progression and recurrence of anogenital condyloma. In a previous work, we observed that patients with warts and high-grade AIN (HAIN) had recurrences more frequently than did patients with warts without AIN. The mechanisms of this increased incidence of high-grade lesions in AIDS are not known. OBJECTIVE: We studied the expression of the proliferative marker Ki-67 by immunohistochemical methods, in specimens of anal condyloma from HIV+ patients to clarify whether its expression can be associated to the grade of AIN. DESIGN: A retrospective study of histological specimens. SETTING: University referral unit. SAMPLE: 34 patients were divided into two groups: (1) condylomas with low grade AIN (LAIN), with 25 patients; and (2) condylomas with HAIN, with 9 patients. In this latter group we examined two areas: 2A (HAIN area) and 2B (LAIN area). MAIN MEASUREMENTS: The immunohistochemical reaction for Ki-67 was done on histological sections. Slices were lightly stained with hematoxylin, to help us in Ki-67 positive cell counting. The percentage of Ki-67 marked nuclei was calculated. We applied one-way variance analysis for statistics. RESULTS: The mean number of Ki-67 positive cells in group 1 was 19.68 +/- 10.99; in group 2 (area A) it was 46.73 +/- 10.409; and in area B it was 36.43 +/- 14.731. There were statistical differences between groups 1 and 2A and between groups 1 and 2B. Ki-67 positive cells predominated in the lower layer in LAIN. Positive Ki-67 cells were found in all layers in group 2A, and in group 2B they predominated in the two lower or in all layers of the epithelium. CONCLUSIONS: Our results suggest that LAIN areas (using routine staining techniques) in HAIN can have a biological behavior more similar to HAIN.

[Pilot study with the monoclonal antibody IOR-C5 as a potential agent of radioimmunoscintigraphy in colorectal cancer]. / Ensayo piloto con el anticuerpo monoclonal ior-C5 como agente potencial de la inmunocentelleografía del cáncer colorrectal.

UNLABELLED: IOR C-5 is a G1 immunoglobulin type intact murine monoclonal antibody (MAb) that was developed in the Center of Molecular Immunology in Havana City, Cuba. In immunohistochemical studies, this demonstrated a significant affinity for the epithelial tissues so that it was used in a pilot clinical study to perform a radioimmunoscintigraphy of the colorectal primary tumors and their locoregional recurrences. It was labeled with 99mTc using the Schwarz method, with a > 95% performance. Planar images of the chest, abdomen and pelvis were performed at 10 minutes, 4-6 hours and 18-24 hours post-injection in the anterior and posterior projections and the SPECT was performed 4-6 hours and 18-24 hours post-injection of 1.85 GBq 99mTC. This study has aimed to verify in vivo the capacity of ior-C5 MAb to accumulate in the malignant colorectal lesions. ior-C5 accumulated in 5 out of the 7 patients who were studied and who were suffering from colorectal cancer or in whom there was suspicion of recurrence. There was a negative case of primary tumors, which was an adenocarcinoma in situ in a tubular-papillary adenoma. The second case with a negative radioimmunoscintigraphy was a true negative case. CONCLUSIONS: It can be concluded that even though the number of patients is quite low, ior-C5 fulfilled the expectations of recognizing the epitope expressed in colorectal tumors in an in vivo human environment.

[Estradiol receptors in cervical intraepithelial neoplasia and cervico-uterine cancer]. / Receptores a estradiol en neoplasia intraepitelial cervical y cáncer cervicouterino.

The objective of this study was to know if there is a correlation in contents o (R + E) in biopsies from patients pre and post-menopausal, in normal cervix with cervical intraepithelial neoplasia (NIC) and invasive cancer. Thirty four patients with abnormal cytology; colposcopy was carried out, and two biopsies were taken from the suspicious lesion, they were sent for histopathological study, and for estrogenic receptors; both studies were correlated later. It was seen that (R +/- E) concentration is higher in intraepithelial lesions of low and high degree than in control patients; and it was not so in premenopausal with invasive cancer where (R +/- E) are much lower, as to postmenopausal (R + E) concentration is higher specially in invasive cancer. It is concluded that the study on quantification to (R + E) in cervical pathology means a good alternative in hormonal treatment, as it has been seen in mammary pathology, and so, more research has to be done.