Atypical Squamous Metaplasia Research Articles

Interobserver reproducibility of cervical histology interpretation with and without p16 immunohistochemistry.

Histopathological diagnosis of colposcopically identified cervical lesions is a critical step for the recognition of cervical cancer precursors requiring treatment. Although there have been efforts to standardize the histologic diagnosis of cervical biopsy specimens, in terms of terminology and use of biomarkers, there is no uniform approach in the pathology community. Adjunctive p16 immunohistochemistry (IHC) can highlight precancer diagnoses, with use recommendations outlined by the Lower Anogenital Squamous Terminology project. We assessed the diagnostic reproducibility of cervical histopathological biopsy specimens with and without p16 staining among 2 expert pathologists. Interpretation of p16 IHC as positive vs negative was highly reproducible (92.5% agreement, κ = 0.85); greater variation was seen in the choice of which biopsy specimens required adjunctive p16 staining (78.0% agreement, κ = 0.43). Adjunctive p16 IHC did not significantly increase diagnostic agreement under multitiered grading systems (benign vs cervical intraepithelial neoplasia [CIN] 1/low-grade squamous intraepithelial lesion vs atypical squamous metaplasia vs CIN2/high-grade squamous intraepithelial lesion [HSIL] vs CIN3/HSIL-CIN3 vs cancer) (65.5% agreement, κ = 0.56 without p16; 70.0% agreement, κ = 0.58 with p16). However, when dichotomizing diagnoses based on clinical management (less than HSIL vs HSIL+), diagnostic agreement increased with p16 IHC (90.5% agreement, κ = 0.79 without p16; 92.0% agreement, κ = 0.84 with p16). For biopsy specimens taken from women positive for human papillomavirus (HPV) type 16, agreement was similar with or without adjunctive p16 (κ = 0.80 without p16; κ = 0.78-0.80 with p16). In contrast, p16 IHC substantially improved diagnostic agreement for cervical biopsy specimens taken from women positive for other high-risk HPV strains, producing improvements in κ from 0.03 to 0.24. Adjunctive p16 immunostaining provides useful information in the evaluation of cervical biopsies for precancer. In our study, we have demonstrated that it is highly reproducible between 2 pathologists, although the decision of which biopsies warrant its use is less so. Furthermore, although p16 IHC showed a limited increase in diagnostic reproducibility for all biopsies included in our study, it did demonstrate a more sizable gain in biopsies negative for HPV 16 but positive for other high-risk genotypes. Further studies are needed to clarify the role of p16 IHC and how it can be optimized for the detection of cervical precancer, particularly in HPV-vaccinated populations where types other than HPV 16 are relatively more important.

Performance of specific morphologic features in distinguishing low-grade squamous intraepithelial lesions from high-grade squamous intraepithelial lesions in borderline cases: a College of American Pathologists Cytopathology Committee multiobserver study.

Distinguishing between low-grade squamous intraepithelial lesions (LSIL) and high-grade squamous intraepithelial lesions (HSIL) can be difficult on certain Papanicolaou (Pap) tests, hindering interobserver concordance. We investigated the variables influencing the interpretation of LSIL versus HSIL in Pap test slides rejected from the College of American Pathologists PAP education program. Eleven cytologists, who were unaware of the reference interpretation, examined 21 Pap slides (11 submitted as LSIL and 10 as HSIL) rejected from the PAP education program and recorded the number of LSIL cells, HSIL cells, keratinized dysplastic cells, LSIL clusters with mixed HSIL cells, atypical squamous metaplasia, atypical glandular cells, the presence of inflammation or infectious organisms, and the overall interpretation (LSIL or HSIL). We evaluated the significance of these 11 variables using a nonlinear mixed model analysis. LSIL had greater concordance (92 of 121 responses; 76.0% concordance) than HSIL (68 of 110 responses; 61.8% concordance; P < 0.001). The only predictors of misclassified cases were the number of atypical squamous metaplastic cells and the number of HSIL cells (P < 0.001). The more of these cells identified, the more likely the reviewers were to classify the slide as HSIL. The reproducibility of the diagnosis was fair (Gwet's agreement coefficient, 0.33). Interobserver reproducibility is a challenge for a subset of cases with features intermediate between LSIL and HSIL. Atypical squamous metaplasia and dysplastic nuclei with a nuclear/cytoplasmic ratio greater than one half of the cell volume (HSIL) present on a Pap test influenced the likelihood that a reviewer would interpret the case as HSIL rather than LSIL.

Expression of P63, P16 and CK17 in Atypical Squamous Metaplasia and Cervical Intraepithelial Neoplasia.

Background & Objective:Cervical intraepithelial neoplasia (CIN) is a dysmaturation process in squamous cells in epithelial layer, which highly increases the risk of developing cervical cancer. The aim of this study was to compare the expression of three biomarkers, p16, p63, and CK17 in patients with CIN and in those with atypical squamous metaplasia (ASM).Methods:In this study, 100 patients underwent a colposcopy-guided cervix biopsy. Immunostaining for the biomarkers was undertaken on tissue samples presented with ASM (n=50) and CIN (n=50).Results:A significant increase in immunostaining for CK7, P63, and P16 in patients with CIN was found compared to ASM subjects.Conclusion:Expression of CK17, P63, and P16 in CIN varied from those in ASM. Those biomarkers could be reliable factors to distinguish ASM from CIN; however, all the biomarkers could differentiate CIN from its mimics due to their high degree of sensitivity and specificity.

Stratified Mucin Producing Intraepithelial Lesion (SMILE): Report of Two Cases

Abstract Introduction/Objective Stratified mucin producing intraepithelial lesion (SMILE) is an uncommon variant of adenocarcinoma in-situ (AIS), often confused with immature squamous metaplasia or high-grade intraepithelial lesion (HG-SIL). This premalignant lesion is often found in proximity to HG-SIL, AIS and invasive carcinomas. SMILE is also a precursor for Invasive Stratified Mucinous Carcinoma. As was first described in the original article, SMILE lesions are considered derived from the reserve cells at the transformation zone where HPV infections lead to varied phenotypes. Methods Two cases of SMILE were collected from our files. A 31-year-old female with a previous abnormal Pap smear result. She underwent cervical biopsy and subsequent LEEP.The second patient, a 26-year-old female, underwent a cervical biopsy after an abnormal Pap smear. Results Histologically, stratified cells with presence of conspicuous mucin production with abundant cytoplasm separating cells in mid and lower layer of epithelium accompanied with conspicuous spacing of nuclei in the lower to middle epithelial layer are seen. Additionally, there is nuclear pleomorphism with absence of gland formation. Mucicarmine stain was performed and showed positive staining in mucin producing dysplastic cells throughout the epithelium. Immunoperoxidase stains demonstrated positivity for p16 (nuclear and cytoplasmic) and p63 in the basal cells. The Ki67 proliferation index was high (approximately 90%). These results supported the diagnostic interpretation of SMILE. Conclusion SMILE is a rare variant of glandular precursor lesion in the cervix. It is frequently associated with HG-SIL and/or AIS, but may rarely appear as an isolated finding. The differential diagnosis include adenocarcinoma in-situ, atypical immature squamous metaplasia and squamous intraepithelial lesion. Since these lesions are cervical glandular intraepithelial neoplasms, the morphologic recognition of a SMILE in a biopsy is important for patient follow up, particularly in cases were only squamous intraepithelial lesion is present in the sample.

P16 Positive Histologically Bland Squamous Metaplasia of the Cervix

With increasing use of p16 immunohistochemistry (IHC) in diagnosis of premalignant lesions of cervix, we occasionally encounter p16 positivity in squamous metaplasia that lacks morphologic characteristics of "atypical squamous metaplasia" or of squamous intraepithelial lesion (SIL). Our study aims to investigate if transcriptionally active human papilloma virus (HPV) can be identified in such foci and if they have any relationship with squamo-columnar junction (SCJ) cells. Twenty-two cases of cervical specimens with at least a focus of p16 positive bland squamous metaplasia, were selected. HPV E6/E7 mRNA in situ hybridization followed by IHC for CK7 (SCJ biomarker), Ki67, and HPV16 E2, were performed. Follow-up information was obtained. Four cases were excluded due to insufficient tissue. Of the final 18 cases, HPV E6/E7 mRNA in situ hybridization was positive in all. Nine cases showed positivity in >50% cells and the epithelial thickness involved was ≥lower two-thirds in 13 cases. Of the further evaluable 15 cases, CK7 was positive in 14, Ki67 was positive in 10, and HPV16 E2 was negative in all. Concomitant high-grade squamous intraepithelial lesion was identified in 10 cases. On follow-up (duration: 1 to 19 mo), 6 patients showed histologic high-grade squamous intraepithelial lesion. Our study demonstrates that p16 positivity in squamous metaplasia of cervix is associated with the presence of transcriptionally active high-risk HPV even when there are no clear morphologic features of dysplasia. Our results suggest that these lesions are early SILs or SILs that are not yet morphologically evident, most of which arise from SCJ and should be closely followed.

Demographic and Clinical Study on Patients Referred to the National Cancer Research Center for Pap Smears

Background: Cervical cancer ranks the fourth most common cancer among women worldwide and the eighth cancer overall. It is considered as one of the most preventable malignancies, and can be controlled through screening and Pap smears.&#x0D; Objectives: To explore the demographic and clinical characteristics of a sample of Iraqi patients who were referred to the National Cancer Research Center of the University of Baghdad for Pap smear test. &#x0D; Patients and methods: The study enrolled 175 female participants who were subjected to clinical examination and Pap smear tests. A questionnaire was filled for each patient on which demographic and clinical variables were recorded. Cellular specimens were collected, fixed, stained by Papanicolaou stain and examined cytologically. The corresponding cytological findings were correlated with the studied variables and tabulated accordingly.Results: 60% of the 175 women were housewives, 40% got married before the age of 20 years, 35.4% had used contraceptive pills and 10.3% were nulliparous. The mean ages at marriage, pregnancy and first child birth were 20.2, 22.0 and 23.0 years respectively. On clinical examination, cervical erosions were obviously apparent in 41.7% of the cases and genital warts were noted in only one case (0.6%), while koilocytotic cellular atypia was observed in 4.0%. Pap smears revealed infections with Moniliasis and Trichomonas Vaginalis in 16.5% and 1.1% respectively. Atypical metaplastic changes, AGUS (atypical glandular cells of uncertain significance) and ASCUS (atypical squamous cells of undetermined significance) were displayed in 21.1%, 5.7% and 22.3% respectively. Diagnoses of CIN I (LSIL) (cervical squamous intraepithelial neoplasia 1 - low-grade squamous intraepithelial lesion), CIN II and CIN III (HSIL) (high-grade squamous intraepithelial lesion) were reported in 26.8% and 2.3% respectively. Among patients with cervical erosions, the Pap smear findings illustrated more pronounced cellular changes consistent with atypical squamous metaplasia, AGUS and ASCUS (43.8%, 19.2% and 45.2% respectively). On the other hand, LSIL, kiolocytotic atypia and HSIL in these cases were registered in 15.1%, 6.8% and 1.3% respectively.&#x0D; Conclusions: The demonstrated young ages at marriage, pregnancy and childbirth of the examined patients, reflecting the early onset of sexual reproductive activities, and the associated cellular changes urge the necessity for promoting public awareness on the significance of Pap smear screening among Iraqi females. The Bethesda system facilitates easier interpretation of the results; thus fostering the communications between the examining pathologists and the gynecologists.

دراسة ديموغرافية وسريرية للمريضات المحالات الى المركز الوطني لبحوث السرطان في بغداد لاجراء مسحات عنق الرحم

Background: Cervical cancer ranks the fourth most common cancer among women worldwide and the eighth cancer overall. It is considered as one of the most preventable malignancies, and can be controlled through screening and Pap smears. Objectives: To explore the demographic and clinical characteristics of a sample of Iraqi patients who were referred to the National Cancer Research Center of the University of Baghdad for Pap smear test. Patients and methods: The study enrolled 175 female participants who were subjected to clinical examination and Pap smear tests. A questionnaire was filled for each patient on which demographic and clinical variables were recorded. Cellular specimens were collected, fixed, stained by Papanicolaou stain and examined cytologically. The corresponding cytological findings were correlated with the studied variables and tabulated accordingly.Results: 60% of the 175 women were housewives, 40% got married before the age of 20 years, 35.4% had used contraceptive pills and 10.3% were nulliparous. The mean ages at marriage, pregnancy and first child birth were 20.2, 22.0 and 23.0 years respectively. On clinical examination, cervical erosions were obviously apparent in 41.7% of the cases and genital warts were noted in only one case (0.6%), while koilocytotic cellular atypia was observed in 4.0%. Pap smears revealed infections with Moniliasis and Trichomonas Vaginalis in 16.5% and 1.1% respectively. Atypical metaplastic changes, AGUS (atypical glandular cells of uncertain significance) and ASCUS (atypical squamous cells of undetermined significance) were displayed in 21.1%, 5.7% and 22.3% respectively. Diagnoses of CIN I (LSIL) (cervical squamous intraepithelial neoplasia 1 - low-grade squamous intraepithelial lesion), CIN II and CIN III (HSIL) (high-grade squamous intraepithelial lesion) were reported in 26.8% and 2.3% respectively. Among patients with cervical erosions, the Pap smear findings illustrated more pronounced cellular changes consistent with atypical squamous metaplasia, AGUS and ASCUS (43.8%, 19.2% and 45.2% respectively). On the other hand, LSIL, kiolocytotic atypia and HSIL in these cases were registered in 15.1%, 6.8% and 1.3% respectively. Conclusions: The demonstrated young ages at marriage, pregnancy and childbirth of the examined patients, reflecting the early onset of sexual reproductive activities, and the associated cellular changes urge the necessity for promoting public awareness on the significance of Pap smear screening among Iraqi females. The Bethesda system facilitates easier interpretation of the results; thus fostering the communications between the examining pathologists and the gynecologists.

Utility of p16 Immunohistochemistry in Evaluating Negative Cervical Biopsies Following High-risk Pap Test Results.

The Lower Anogenital Squamous Terminology (LAST) Standardization Project for human papilloma virus (HPV)-associated lesions specifically recommends the use of p16 immunohistochemistry (IHC) as an adjunct to morphologic assessment of cervical biopsies interpreted as negative or low-grade squamous intraepithelial lesion (LSIL) from patients with prior high-risk Pap test results (high-grade squamous intraepithelial lesion [HSIL], atypical squamous cells cannot exclude HSIL, atypical glandular cells [AGC], or HPV16 atypical squamous cells of undetermined significance [ASC-US]). The impetus for this recommendation is to increase detection of missed high-grade disease. However, the quality of evidence supporting this recommendation was lower than that for the other LAST recommendations addressing improved consistency in the diagnosis of HSIL with the use of p16. A database search spanning 10 years identified 341 cases (encompassing 736 discrete biopsy specimens) interpreted as negative for dysplasia from 330 patients with a prior high-risk Pap result (atypical squamous cells cannot exclude HSIL, HSIL, atypical glandular cells, not otherwise specified [AGC-NOS], atypical endocervical cells--NOS [AEC-NOS], and AEC-favor neoplastic). p16 IHC was performed and detected missed abnormalities in 11/341 (3.2%) cases. The abnormalities corresponded to missed foci of HSIL (cervical intraepithelial neoplasia [CIN] 2) (n=1), SIL-indeterminate grade (n=7), atypical squamous metaplasia (n=2), and LSIL [CIN1]) (n=1). Subsequent histologic follow-up identified HSIL or greater in 6/8 (75%) p16 cases versus 20/79 (25.3%) p16 cases (P=0.0079). p16 IHC performed on biopsies interpreted as negative from patients with prior high-risk Pap test results increased the detection rate of missed SIL. A p16 result also significantly increased the likelihood of HSIL on subsequent biopsy. Although further studies are required to determine what percentage of missed HSIL justifies the additional cost, improved detection of HSIL in high-risk patients may lead to fewer diagnostic procedures and fewer patients lost to follow-up.

Role of p16, Ki67 and CK17 in differentiating benign lesions, cervical intraepithelial neoplasia(CIN) and atypical immature squamous metaplasia(AIM) of uterine cervix

Cervical intraepithelial neoplasia (CIN) is a premalignant lesion capable of progressing to cervical cancer. Despite the existing well-defined criteria, the histopathological diagnosis is subject to high rates of discordance among pathologists. Aim: To study the role of p16, Ki67 and CK17 in differentiating benign lesions, cervical intraepithelial lesions(CIN) and atypical immature squamous metaplasia (AIM)and to improve intra and interobserver reproducibility of diagnosis of cervical neoplasia. Material and Methods: In a cross sectional study, a total of 75 cervical biopsies including benign lesions (n=24), AIM (n=28), CIN (n=23) were studied and analyzed immunohistochemically using p16, Ki67 and CK17 immunomarkers. Data was evaluated using chi-square test. Results: p16 and Ki 67positivity were observed in 91.3% and 78.26% of CIN and 28.57% of AIM respectively. None of the benign lesions expressed p16 and Ki67while CK17 positivity was observed in 46.42% of CIN and 100% of AIM with 12.5% of benign lesions. Conclusion: The three biomarkers (p16, CK17 and Ki67) had a high degree of sensitivity and specificity and appear to be a useful and reliable diagnostic adjunct to improve the routine diagnosis and reduce interobserver variability in cervical biopsy specimens. Immunohistochemical markers such as p16 alone or with Ki67 represents important tool for the pathologists in distinguishing high grade cervical dysplasia from its benign mimics such as AIM and reactive inflammatory lesion thus avoiding overtreatment. Cervical intraepithelial neoplasia (CIN) is a premalignant lesion capable of progressing to cervical cancer. Despite the existing well-defined criteria, the histopathological diagnosis is subject to high rates of discordance among pathologists. Aim: To study the role of p16, Ki67 and CK17 in differentiating benign lesions, cervical intraepithelial lesions(CIN) and atypical immature squamous metaplasia (AIM)and to improve intra and interobserver reproducibility of diagnosis of cervical neoplasia. Material and Methods: In a cross sectional study, a total of 75 cervical biopsies including benign lesions (n=24), AIM (n=28), CIN (n=23) were studied and analyzed immunohistochemically using p16, Ki67 and CK17 immunomarkers. Data was evaluated using chi-square test. Results: p16 and Ki 67positivity were observed in 91.3% and 78.26% of CIN and 28.57% of AIM respectively. None of the benign lesions expressed p16 and Ki67while CK17 positivity was observed in 46.42% of CIN and 100% of AIM with 12.5% of benign lesions. Conclusion: The three biomarkers (p16, CK17 and Ki67) had a high degree of sensitivity and specificity and appear to be a useful and reliable diagnostic adjunct to improve the routine diagnosis and reduce interobserver variability in cervical biopsy specimens. Immunohistochemical markers such as p16 alone or with Ki67 represents important tool for the pathologists in distinguishing high grade cervical dysplasia from its benign mimics such as AIM and reactive inflammatory lesion thus avoiding overtreatment. Normal 0 false false false EN-IN X-NONE X-NONE /* Style Definitions */ table.MsoNormalTable {mso-style-name:Table Normal; mso-tstyle-rowband-size:0; mso-tstyle-colband-size:0; mso-style-noshow:yes; mso-style-priority:99; mso-style-qformat:yes; mso-style-parent:; mso-padding-alt:0cm 5.4pt 0cm 5.4pt; mso-para-margin-top:0cm; mso-para-margin-right:0cm; mso-para-margin-bottom:10.0pt; mso-para-margin-left:0cm; line-height:115%; mso-pagination:widow-orphan; font-size:11.0pt; font-family:Calibri,sans-serif; mso-ascii-font-family:Calibri; mso-ascii-theme-font:minor-latin; mso-fareast-font-family:Times New Roman; mso-fareast-theme-font:minor-fareast; mso-hansi-font-family:Calibri; mso-hansi-theme-font:minor-latin;}

Functional association of oestrogen receptors with HPV infection in cervical carcinogenesis.

Repeated parity and usage of oral contraceptives have demonstrated an increased risk of cervical cancer (CC) in HPV-infected women. These lifestyle observations raise the likelihood that oestrogens and HPV infection might act synergistically to affect cancers of the cervix. In vivo studies have indicated the requirement of oestrogens and ERα in the development of atypical squamous metaplasia followed by cervical intraepithelial neoplasia (CIN) I, II and III. CIN II and III are precancerous cervical lesions that can progress over time to CC as an invasive carcinoma. Recently, there has been evidence suggesting that ERα signalling in the tumour epithelium is a preliminary requisite during cancer initiation that is subsequently lost during tumorigenic progression. Conversely, continued expression of stromal ERα gains control over tumour maintenance. This review summarises the current information on the association between oestrogens and HPV infection in contributing to CC and the possibility of SERMs as a therapeutic option.

Human Papillomavirus (HPV) Associated Atypical Squamous Metaplasia in Diffuse Alveolar Damage (DAD) Mimicking Squamous Cell Carcinoma (SCC)

Human Papillomavirus (HPV) Associated Atypical Squamous Metaplasia in Diffuse Alveolar Damage (DAD) Mimicking Squamous Cell Carcinoma (SCC)

Role of deltaNp63(pos)CD44v(pos) cells in the development of N-nitroso-tris-chloroethylurea-induced peripheral-type mouse lung squamous cell carcinomas.

The role of cells expressing stem cell markers deltaNp63 and CD44v has not yet been elucidated in peripheral‐type lung squamous cell carcinoma (pLSCC) carcinogenesis. Female A/J mice were painted topically with N‐nitroso‐tris‐chloroethylurea (NTCU) for induction of pLSCC, and the histopathological and molecular characteristics of NTCU‐induced lung lesions were examined. Histopathologically, we found atypical bronchiolar hyperplasia, squamous metaplasia, squamous dysplasia, and pLSCCs in the treated mice. Furthermore, we identified deltaNp63pos CD44vpos CK5/6pos CC10pos clara cells as key constituents of early precancerous atypical bronchiolar hyperplasia. In addition, deltaNp63pos CD44vpos cells existed throughout the atypical bronchiolar hyperplasias, squamous metaplasias, squamous dysplasias, and pLSCCs. Overall, our findings suggest that NTCU induces pLSCC through an atypical bronchiolar hyperplasia–metaplasia–dysplasia–SCC sequence in mouse lung bronchioles. Notably, Ki67‐positive deltaNp63pos CD44vpos cancer cells, cancer cells overexpressing phosphorylated epidermal growth factor receptor and signal transducer and activator of transcription 3, and tumor‐associated macrophages were all present in far greater numbers in the peripheral area of the pLSCCs compared with the central area. These findings suggest that deltaNp63pos CD44vpos clara cells in mouse lung bronchioles might be the origin of the NTCU‐induced pLSCCs. Our findings also suggest that tumor‐associated macrophages may contribute to creating a tumor microenvironment in the peripheral area of pLSCCs that allows deltaNp63pos CD44vpos cancer cell expansion through activation of epidermal growth factor receptor signaling, and that exerts an immunosuppressive effect through activation of signal transducer and activator of transcription 3 signaling.

Efficacy of p16 and Ki-67 immunostaining in the detection of squamous intraepithelial lesions in a high-risk HPV group.

p16 and Ki-67 immunohistochemistry can be used as an ancillary method for the diagnosis of squamous intraepithelial lesion (SIL) versus atrophic change and atypical squamous metaplasia. The aim of the present study was to evaluate the efficacy of these two immunohistochemical markers in the accurate interpretation of cervical biopsies and correlate this data with human papilloma virus (HPV) infection status. The study included 103 formalin-fixed cervical punch and cone biopsy samples, with corresponding HPV DNA test data. Histopathological staining with hematoxylin and eosin, and immunohistochemical staining for p16 and Ki-67 were reviewed by two pathologists. The positivity of p16 and Ki-67 increased significantly with the severity of the cervical lesion in patients with a high-risk-HPV (HR-HPV) infection status (P<0.001). However, there was discordance in the HPV-negative group. Furthermore, concomitant diffuse, strong and block positive staining of p16, and a high Ki-67 index were implicated in high-grade SIL in the HR-HPV group. Thus, the two markers were efficient in advancing the diagnostic accuracy of cervical biopsies in the HR-HPV group; however, application of immunohistochemical results should be carefully considered in the HPV-negative group.

Histomorphologic Features of Biopsy Sites Following Excisional and Core Needle Biopsies of the Breast.

Mammographic studies have documented a number of architectural changes occurring around breast biopsy sites. These changes are well described in the radiological literature, but similar studies do not appear to be present in the pathology literature. We reviewed 100 consecutive mastectomy specimens from women who had undergone prior core needle or excisional biopsies. Multiple sections of the needle tract or excisional biopsy site were reviewed and morphologic findings reported. Hemorrhage, fat necrosis, granulation tissue, necrosis of fibrous tissue, and epithelium along with fibrosis and foreign body type giant cells were common features. Less frequent were areas of synovial metaplasia, atypical spindle cells, atypical duct-like structures, single atypical cells, squamous metaplasia, proliferations of abnormal blood vessels, and hemosiderin deposition. The misinterpretation of atypical spindle cells, single atypical cells, atypical duct-like structures and squamous metaplasia could result in the false-positive diagnosis of residual malignancy. Careful attention to the reactive nature of these changes aids in their distinction from carcinoma.

Immunohistochemistry and in situ hybridization for the diagnosis and classification of squamous lesions of the anogenital region.

Distinguishing anogenital squamous intraepithelial lesions from benign conditions and mimics may be problematic. Immunohistochemistry for surrogate markers of HPV infection, such as Ki-67, p16, and ProEx™ C, may aid the diagnosis in equivocal cases. The main diagnostic pitfall in the diagnosis of LSIL is the occurrence of "pseudokoilocytes" in benign squamous mucosa, which may lead to overdiagnosis. When interpreted correctly, Ki-67 is a sensitive and specific marker for dysplasia in mature squamous epithelium and is therefore useful for confirmation of LSIL and condyloma. A Ki-67 positive result is defined as the presence of a cluster of at least two strongly stained epithelial nuclei in the upper two-thirds of the epithelial thickness. With such a definition, there is almost complete concordance between consensus diagnosis of LSIL/condyloma confirmed by detection of HPV DNA and positive Ki-67. A related proliferation marker, ProEx™ C, has similar staining patterns and utility for the diagnosis of low grade dysplasia. The differential diagnosis of HSIL includes atypical immature squamous metaplasia and atrophy. A marker with high sensitivity and specificity for the detection of HSIL in cervical, vulvar, and anal mucosa is p16. A 2-tier scoring system is used to evaluate p16 staining. No staining or a discontinuous, patchy nuclear and cytoplasmic staining pattern is considered as a negative result. A positive result is defined as diffuse and strong staining of cells of the basal and parabasal layers of the squamous epithelium, with or without staining of superficial cell layers. New markers that are undergoing evaluation for their clinical utility include stathmin-1, phosphorylated S6, and SOX2. Confirmation of the diagnosis of dysplasia by HPV detection in tissue sections using HPV capsid protein immunohistochemistry, HPV DNA or HPV RNA in situ hybridization offers lower sensitivity as compared to immunohistochemistry for surrogate markers and therefore has more limited utility in this context.

Superovulation does not affect the endocrine activity nor increase susceptibility to carcinogenesis of uterine and mammary glands of female offspring in mice.

To evaluate the dual effects of superovulation on the endocrine activity and susceptibility to carcinogenesis of uterine and mammary glands of female offspring in mice The mice were superovaluted. The relative uterine weight, ERα protein expression, and endocrine activity of female offspring (F1 generation and F2 generation) were measured. Furthermore, proliferative lesion of uterine and mammary glands of female offspring (F1 generation and F2 generation) was assessed by histopathologic examinations. There were no significant differences in relative uterine weight, ERα protein expression, incidence of proliferative lesion in mammary glands, and incidence of atypical hyperplasia, adenocarcinoma, and squamous metaplasia in uterine among the offspring (F1 generation and F2 generation) in each group. Likewise, there were no significant intergroup differences in the serum levels of sex related hormones. No significant alterations were found in the endocrine activity and susceptibility to carcinogenesis of uterine and mammary glands of female offspring in mice produced by superovaluted oocytes compared with those of naturally conceived offspring.

P16INK4a Immunoprofiles of Squamous Lesions of the Uterine Cervix–Implications for the Reclassification of Atypical Immature Squamous Metaplasia

p16(INK4a) immunoprofiles of non-precancerous and dysplastic squamous cervical lesions were defined and applied to the reclassification of atypical immature squamous metaplasia (AIM). The immunoexpression of cytokeratin 17 (CK 17) in AIM was also evaluated. Totally, 295 cervical cone biopsies representing squamous metaplasia, reactive changes, koilocytosis, flat condyloma, CIN I, CIN II, CIN III and AIM were subjected to p16(INK4a) immunohistochemistry. AIM cases were analyzed using CK 17 antibody. Typical p16(INK4a) immunoprofiles for the metaplastic, LSIL/HPV and HSIL phenotypes were recorded and used for the categorization of AIM into particular phenotype groups. Results were correlated with CK 17 immunoexpression. All CIN II and CIN III lesions, all but one case of CIN I and all flat condylomas overexpressed p16(INK4a). Other non-precancerous lesions, including koilocytosis, were predominantly negative. Contrary to the sporadic and focal immunostaining, diffuse positivity was associated with the dysplastic features of the lesion. CIN II and CIN III were characterized by a diffuse, strong/weak, full-thickness staining, whereas CIN I showed a heterogeneous diffuse/focal, weak/strong, lower half positivity. One third of AIM lesions may be reclassified as HSIL, one third as LSIL/HPV and one third shows metaplastic phenotype. All AIM cases with metaplastic and LSIL/HPV phenotypes expressed CK 17 diffusely, whereas focal positivity slightly prevailed in AIM with HSIL phenotype. We conclude that p16(INK4a) immunohistochemistry is a supporting method for the differential diagnosis of cervical lesions, which may be especially useful for the reclassification of AIM. The efficacy of CK 17 immunohistochemistry seems to be controversial for these purposes.

Chlamydial infection in female lower genital tract and its correlation with cervical smear abnormalities

It is widely known that infection by Chlamydia trachomatis (CT) is the most common curable sexually transmitted infection (STI) among the young population.Chlamydial cervicitis has been suspected in the past to be a risk factor for squamous intraepithelial neoplasia (SIL) and cervical cancer on the basis of case-control comparisons of serological tests. A total of 110 women with symptomatic cervicitis were examined and samples for microbiologic detection of Chlamydia trachomatis antigen were obtained. Colposcopic, cytologic and microbiologic analyses were performed. All results were statistically analysed and correlated. 41 patients (37.2 %) were found positive for Chlamydia trachomatis antigen (Group I) and 69 patients (62.8 %) were negative for Chlamydia trachomatis antigen (Group II). When compared to group II, the chlamydia-infected group I of women presented a larger percentage of dyspareunia (60 %), and the dysuric complaints were more common. The chlamydia-positive group revealed a higher incidence of atypical squamous metaplasia (52.5 % vs 27.9 %). Among the women with positive ELISA for chlamydia, a higher percentage of low-grade SIL was observed (6.3 % vs 3.6 %). Chlamydial infection, especially chlamydial cervicitis occurs often among young women of reproductive age. In these women, a good clinical assessment is necessitated including previous clinical history record, as well as detailed microbiologic, cytologic and colposcopic evaluations (Tab. 5, Ref. 19).

Atypical squamous metaplasia in a benign cystic thyroid nodule mimicking high‐grade carcinoma

Squamous metaplasia (SM) occurs in a variety of thyroid conditions, both neoplastic and non-neoplastic. In a small subset of benign thyroid lesions, SM can create a diagnostic pitfall by mimicking a malignant process. Here, we describe the case of a 52-year-old male with a 1.5 cm right thyroid nodule. Ultrasound-guided fine-needle aspiration biopsy showed markedly atypical, mitotically active epithelial cells which were suspicious for high-grade carcinoma. Resection of the nodule demonstrated a follicular adenoma with cystic changes and atypical SM of the cyst-lining component. As illustrated here, SM of the thyroid represents an important diagnostic pitfall for the cytopathologist, and should be included in the differential diagnosis of thyroid nodules, particularly those with potentially benign cystic changes.

Is the Expression Pattern of BD ProExC the Same as Ki-67? A Comparative Analysis in Cervical Biopsies

BD ProExC (ProExC) and Ki-67, both used as biomarkers for high-grade cervical intraepithelial neoplasia (HG CIN), yield discordant staining in some cervical biopsies. This study compared ProExC and Ki-67 expression in 197 cervical biopsies with consensus diagnoses (55 negative, 21 atypical squamous metaplasia, 61 low-grade CIN, and 60 HG CIN). Percentages of immunostained nuclei were evaluated by 2 pathologists yielding 68 (35%) cases with discordant ProExC/Ki-67 immunostaining for analysis. In 78% of cases the difference in staining involved <25% of lesional cells. This was noted across all morphologic diagnoses, being most frequent in HG CIN. Discordant staining involving >50% of lesional cells occurred in 22% of discrepant cases, being most frequent in atypical squamous metaplasia. Using staining in >50% of lesional nuclei as a positive result, positive/negative discordance occurred in 25 cases (13% of all cases) including 18% of HG CIN cases. Fourteen cases were ProExC+ (7 of which were strongly p16+) and 11 were Ki-67+ (6 of which were strongly p16+).