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The clinical utility of the proliferation marker Ki67 in breast cancer treatment and prognosis is an active area of research. Studies have suggested that differences in pre-analytic and analytic factors contribute to low analytical validity of the assay, with scoring methods accounting for a large proportion of this variability. Use of standard scoring methods is limited, in part due to the time intensive nature of such reporting protocols. Therefore, use of digital image analysis tools may help to both standardize reporting and improve workflow. In this study, digital image analysis was utilized to quantify Ki67 indices in 280 breast biopsy and resection specimens during routine clinical practice. The supervised Ki67 indices were then assessed for agreement with a manual count of 500 tumor cells. Agreement was excellent, with an intraclass correlation coefficient of 0.96 for the pathologist-supervised analysis. This study illustrates an example of a rapid, accurate workflow for implementation of digital image analysis in Ki67 scoring in breast cancer.

Brenner tumors is a very uncommon ovarian epithelial neoplasm, morphologically resembling the transitional cell neoplasm of urinary tract. It is further classified into three subtypes as a disease spectrum based on histological examination and tumorigenesis: benign Brenner tumors, borderline Brenner tumors (BBTs), and malignant Brenner tumors (MBTs). The etiology of these tumors is not well understood, and literature is limited due to the rarity of this entity, but recent advances, particularly in molecular alterations, have emerged. The scope of this review is to provide an update on the clinical, histopathological, and most recently, molecular characterizations of ovarian Brenner tumors.

Differentiated vulvar intraepithelial neoplasia (dVIN) is a non-human papilloma virus (HPV)-related high-grade precursor lesion to vulvar squamous cell carcinoma (vSCCa). Although TP53gene mutations have been identified in 80% of dVIN, its role in dVIN pathogenesis as well as malignant transformation is still being poorly understood. Poor reproducible diagnostic criteria and ambiguous p53 immunostaining patterns, along with morphologic discordance still pose a diagnostic challenge.

We investigated the prevalence and carcinogenic risks of individual high-risk human papillomavirus (HR-HPV) in all types of cervical cytology specimens in the Shanghai population.

Intraoperative consultation includes gross evaluation and frozen section, which is an integral component of surgical oncology for many reasons. First, it allows appropriate surgical procedures to be undertaken. Second, it can potentially protect the patient from a second procedure. Third, it helps towards counselling of patients and their family members and lastly, it facilitates early management planning. Utilization of frozen section is justified by the reported sensitivity, specificity, positive and negative predictive values all of which are as high as 90-100% in some studies. It is very important for gynecologists to have good communication with pathologists, especially for cases with an indeterminate, frozen result. Discussion with the pathologist can help the gynecologist understand the diagnostic dilemma and the differential diagnosis, which can help them make best surgical decisions.

•Discuss ASCCP guideline.•CIN3 reliable surrogates for cervical cancer?•The Pittsburgh Cervical Cancer Screening Model.

High risk (hr) human papillomavirus (HPV) testing has been proposed as a possible replacement for Papanicolaou (Pap) cytology for cervical screening. The aim of the present study was to assess the hrHPV detection rates using 3 available Food and Drug Administration–approved HPV assays in patients with high-grade squamous intraepithelial lesion (HSIL) cytology results and to correlate the cervical screening test results with the immediate histopathologic findings.

The risk of developing cervical squamous lesions in women with multiple high-risk human papillomavirus (hrHPV) infections is uncertain. The aim of this retrospective study was to investigate the type-specific attribution and phylogenetic effects of single and multiple hrHPV subtypes in cervical squamous lesions. All cases with cervical histopathologic diagnosis and human papillomavirus (HPV) genotyping results in the 6 months preceding biopsy from October 2018 to December 2022 were studied and analyzed. Over the study period, 70,361 cases with histopathologic follow-up and prior HPV genotyping were identified. The hrHPV-positive rate was 55.6% (39,104/70,361), including single hrHPV detected in 27,182 (38.6%), 2 types of hrHPV detected in 8158 (11.6%), and 3 types of hrHPV detected in 2486 (3.5%). Among 16,457 cases with a histologically diagnosed squamous lesion (cervical intraepithelial neoplasia 1: 11411; cervical intraepithelial neoplasia 2/3: 4192; squamous cell carcinoma: 854 cases), the prevalence of single hrHPV infection increased, but the rate of multiple concomitant hrHPV infections showed negative association as the degree of squamous lesions increased. Among women with a single HPV16 infection, cervical intraepithelial neoplasia 2/3 and squamous cell carcinoma (CIN2+) diagnostic rate was 30.6%, and it increased to 47.6% when coinfected with HPV33 (P< .001) but significantly decreased when coinfected with all other hrHPV types (P< .05). By comparing CIN2+ diagnostic rates in 40 most common 2 types of hrHPV infections with related single hrHPV infection, CIN2+ rates were decreased in 12 combinations (30.0%), equivalent in 26 combinations (65.0%), and increased in 2 combinations (5.0%). The cases with 3 types of HPV infections reduced the risk for CIN2+ compared with related single HPV infections. HPV16+52+53, HPV16+52+68, HPV16+52+51, HPV16+39+52, and HPV16+58+53 significantly decreased the risk of CIN2+ compared with HPV16 single infection (P< .05). This study demonstrates that multiple hrHPV infections are not associated with cumulatively higher risk for CIN2+ development, suggesting that oncogenic progression of multiple hrHPV-associated cervical squamous lesions is neither synergistic nor a cumulative effect at the phylogenetic level, possibly a way of competitive interference.

Cervical cancer rates in China remain high, with only limited opportunistic screening in urban centers and large mostly unscreened rural areas. Cervical cytology practices in China have been changing over the last decade with introduction of The Bethesda System reporting terminology, liquid-based cytology (LBC), and programs for cervical cytology screening of underserved rural populations. An effort was undertaken for the first time to collect nationwide data on cervical cytology laboratory practices in China, a possible first step toward increased standardization and potential development of nationwide cytology quality benchmarks.

Cervical cancer remains an important public health problem in Chinese women owing to the lack of a national screening program. The aim of the present study was to evaluate human papillomavirus (HPV) and Papanicolaou (Pap) test results preceding the histologic diagnosis of cervical intraepithelial neoplasia 2/3 (CIN2/3) in China’s largest College of American Pathologists-certified clinical laboratory.

The role of Artificial intelligence (AI) in pathology is one that offers many exciting new possibilities for improving patient care. This study contributes to this development by identifying the viability of AICyte Assistive System for cervical screening, and to investigate the utility of the system in assisting with workflow and diagnostic capability. In this study, a novel scanner was developed using a Ruiqian WSI-2400, trademarked AICyte Assistive system, to create AI-generated gallery of the most diagnostically relevant images, objects of interest (OOI), and provide categorical assessment, according to Bethesda category, for cervical ThinPrep Pap slides. For validation purposes, two pathologists reviewed OOIs from 32,451 cases of ThinPrep Paps independently, and their interpretations were correlated with the original ThinPrep interpretations (OTPI). The analysis was focused on the comparison of reporting rates, correlation between cytological results and histological follow-up findings, and the assessment of independent AICyte screening utility. Pathologists using the AICyte system had a mean reading time of 55.14 seconds for the first 3,000 cases trending down to 12.90 seconds in the last 6,000 cases. Overall average reading time was 22.23 seconds per case as compared to a manual reading time approximation of 180 seconds. Usage of AICyte compared to OTPI had similar sensitivity (97.89% vs 97.89%) and a statistically significant increase in specificity (16.19% vs 6.77%). When AICyte was run alone at a 50% negative cut-off value, it was able to read slides with a sensitivity of 99.30% and specificity of 9.87%. When AICyte was run independently at this cut-off value, no sole case of HSIL/SCC squamous lesion was missed. AICyte can provide a potential tool to help pathologists in both diagnostic capability and efficiency, which remained reliable as compared to baseline standard. Also unique for AICyte is the development of a negative cutoff value for which AICyte can categorize cases as “not needed for review” to triage cases and lower pathologist workload. This is the largest case number study that pathologists reviewed OOI with AI assistive system. The study demonstrates that AI assistive system can be broadly applied for cervical cancer screening.

There is no national cervical screening program or national standards for cervical cytology quality control in China. Since 2013, systematic training and quality control programs were implemented in the Papanicolaou testing process at Jinan KingMed Diagnostics. Pathologists were required to complete 1 year of cytology study in the KingMed Diagnostics Cytology School, including 6 months of a diagnostic course and 6 months of practical training in the clinical laboratory. In this study, we compared the Papanicolaou abnormal reporting rates before and after the implementation systematic training and quality control programs.

Atypical glandular cells are gynecologic cytology abnormalities representing glandular cells with morphologic changes that exceed obvious reactive/reparative changes, but fall short of an interpretation of adenocarcinoma either quantitatively or qualitatively. This cytological diagnosis occurs in less than 1% of the results of gynecologic cytology tests (0.2%–0.8%) but potentially reflects a wide range of conditions, from benign changes and cervical squamous/glandular precursor lesions, to invasive cervical or other gynecologic cancers. In this article, we aim at reviewing different subcategories, potential ancillary studies, histological follow-up results, and clinical management of atypical glandular cells.

Coinfection with multiple high-risk human papillomavirus (hrHPV) is frequently observed in cervical specimens; however, the clinical significance of concomitant multiple hrHPV infections is poorly understood, and the published results remain inconsistent. A retrospective study at a tertiary care institution was performed, evaluating Tellgenplex human papillomavirus (HPV) 27 genotyping or YanengBio HPV 23 genotyping results and immediate cervical histologic diagnosis (within 6 months after HPV genotyping), between November 2015 and October 2022. Among 49,299 cases with hrHPV genotyping and histologic diagnosis, 24,361 cases were diagnosed as cervical intraepithelial neoplasia (CIN) and squamous cell carcinoma. Among women with cervical squamous lesions, 86.5% (21,070/24,361) had hrHPV infections, and concomitant multiple hrHPV infections accounted for 24.7% of hrHPV-positive cases (5210/21,070). The hrHPV-positive rates in these cervical squamous lesions increased progressively with disease severity; however, the percentages of concomitant multiple hrHPV infection rates among hrHPV-positive cases decreased significantly with increasing degree of squamous abnormalities. There was no increased detection rate of CIN3+ (CIN3 and squamous cell carcinoma) in cases with concomitant 2 or 3 hrHPV genotype infections when compared with those with corresponding single hrHPV infections. Conversely, some combinations of multiple hrHPV infections demonstrated a decrease in the detection rates of CIN3+ lesions. In this large cohort, our results demonstrated that multiple hrHPV infections do not carry an increased risk for developing CIN3+ lesions when compared to the corresponding single-genotype infection. The reduced risk of CIN3+ in women infected with some combinations of hrHPV genotypes compared to those with single-genotype infections supports the concept of intergenotypic competition of hrHPV genotypes in cervical squamous lesions.

Recently the Chinese government has introduced support for cervical screening in rural areas. The College of American Pathologists (CAP)-certified Guangzhou Kingmed Diagnostics laboratory supported rural screening program in Hainan Providence utilizing low-cost conventional Papanicolaou smears (CPS).

Dedifferentiated endometrioid adenocarcinoma (DEAC) of the uterus or ovary is characterized by the coexistence of low-grade endometrioid adenocarcinoma and an undifferentiated carcinoma (UC) with solid sheets of medium-sized monotonous epithelial cells. This admixed carcinoma has not been widely recognized, because the solid areas of UC have usually been misdiagnosed as a solid form of FIGO grade 3 endometrioid adenocarcinoma. These tumors have been shown to be clinically aggressive; therefore, accurate diagnosis is necessary for proper patient management. We reviewed our experience with DEACs and compared them with grade 3 endometrioid carcinomas regarding their clinicopathologic, morphologic, and immunohistochemical features. Our results indicate that DEACs are clinically aggressive tumors presented at advanced stages with vascular invasions in 73% and lymph node metastases in 46%. Thirty-eight percent of cases also showed distal metastases. Clinical follow-up data revealed that all patients had either recurrent or metastatic diseases within 3 years of diagnosis, except 1 patient who remained disease free for 3 years after diagnosis. Morphologically, UC components of DEACs were composed of diffuse sheets/solid nests of medium-sized epithelial cells with scant to moderate cytoplasm, uniform vesicular nuclei, and inconspicuous nucleoli. Although UC components of DEACs are variably positive for cytokeratin, EMA, and ER, they are mostly negative for PAX8, except 1 case. Instead, well-differentiated components of DEACs and solid grade 3 endometrioid carcinoma retained all these markers. Our results indicate that DEACs exhibit significantly different clinicopathologic features from grade 3 endometrioid adenocarcinoma, and a combination of immunohistochemical stains can be helpful to differentiate them from each other.

Prior screening results in women diagnosed with cervical carcinoma are only reported in the literature in small numbers. We wish to examine a larger number of prior testing results in women with cervical carcinoma from China to better understand the strengths and weaknesses of Pap (Papanicolaou) cytology and human papillomavirus (HPV) testing.

In the past 2 decades, cervical cancer screening guidelines in the United States have undergone numerous revisions with recent greater emphasis on primary high-risk human papillomavirus (hrHPV) testing.

Management of human papillomavirus (HPV)-positive/cytology-negative patients has represented a special challenge ever since the introduction in 2003 of routine cytology and HPV cotesting of women 30 years and older. Since the U.S. Food and Drug Administration’s approval in 2009 of an HPV 16/18 genotyping test, guidelines have included an HPV 16/18 genotyping triage option for identifying high-risk HPV-positive/cytology-negative patients who would benefit from immediate colposcopic referral.

ABSTRACTScreening for cervical cancer with cytology testing has been very effective in reducing cervical cancer in the United States. For decades, the approach was an annual Pap test. In 2000, the Hybrid Capture 2 human papillomavirus (HPV) test was approved by the U.S. Food and Drug Administration (FDA) for screening women who have atypical squamous cells of underdetermined significance (ASCUS) detected by Pap test to determine the need for colposcopy. In 2003, the FDA approved expanding the use of the test to include screening performed in conjunction with a Pap test for women over the age of 30 years, referred to as “cotesting.” Cotesting allows women to extend the testing interval to 3 years if both tests have negative results. In April of 2014, the FDA approved the use of an HPV test (the cobas HPV test) for primary cervical cancer screening for women over the age of 25 years, without the need for a concomitant Pap test. The approval recommended either colposcopy or a Pap test for patients with specific high-risk HPV types detected by the HPV test. This was based on the results of the ATHENA trial, which included more than 40,000 women. Reaction to this decision has been mixed. Supporters point to the fact that the primary-screening algorithm found more disease (cervical intraepithelial neoplasia 3 or worse [CIN3+]) and also found it earlier than did cytology or cotesting. Moreover, the positive predictive value and positive-likelihood ratio of the primary-screening algorithm were higher than those of cytology. Opponents of the decision prefer cotesting, as this approach detects more disease than the HPV test alone. In addition, the performance of this new algorithm has not been assessed in routine clinical use. Professional organizations will need to develop guidelines that incorporate this testing algorithm. In this Point-Counterpoint, Dr. Stoler explains why he favors the primary-screening algorithm, while Drs. Austin and Zhao explain why they prefer the cotesting approach to screening for cervical cancer.

Cervical cancer remains a major health problem in China, where no uniform national standards for cervical cytology quality control exist. This study extends previously reported findings on cervical cytology performance in China's largest independent laboratory to have achieved accreditation under the international Laboratory Accreditation Program of the College of American Pathologists.

Napsin A and α-methylacyl-coenzyme A racemase (AMACR, P504S) have recently been described as being frequently expressed in clear cell carcinomas (CCC) of the gynecological tract. The present study was conducted to assess the test performance of these newer markers relative to the more traditional marker, hepatocyte nuclear factor 1β (HNF1β), in a large and histotypically diverse dataset. A total of 279 ovarian tumours in tissue microarrays were immunohisto-chemically assessed for the expression of Napsin A, AMACR and HNF1β. HNF1β, Napsin A and AMACR were expressed in 92%, 82% and 63% of 65 CCC, 7%, 1% and 1% of 101 serous carcinomas, 37%, 5.3% and 0% of 19 endometrioid carcinomas, 60%, 0% and 0% of 45 mucinous tumours, 100%, 0% and 0% of seven yolk sac tumours, and 0%, 16.7% and 16.7% of six steroid cell tumours NOS, respectively. All other tumours, including 18 adult-type granulosa cell tumours, eight dysgerminomas and nine other miscellaneous tumour types were negative for all three markers. Using a benchmark of ≥1% of tumour cells for positivity and CCC as the diagnostic end-point, the sensitivity, specificity, negative predictive value and positive predictive value of Napsin A expression were 0.82, 0.99, 0.94, and 0.98, respectively (odds ratio 439, p< 0.0001). Respective parameters were 0.92, 0.79, 0.97, and 0.58 (odds ratio 44, p< 0.0001) for HNF1β and 0.63, 0.99, 0.89, and 0.5 (odds ratio 112, p< 0.0001) for AMACR. The combination of any two positive markers, irrespective of the staining pattern of the third, significantly predicted the CCC histotype in every analytic scenario. In summary, HNF1 β is highly sensitive but is suboptimally specific in isolation, whereas AMACR is highly specific but is suboptimally sensitive. Napsin A is specific but of intermediate sensitivity. Napsin A, AMACR and HNF1β are all viable markers of CCC that can be deployed as components of larger panels when CCC is a diagnostic consideration.

Uterine and ovarian undifferentiated carcinomas (UCs) are often associated with low-grade endometrioid carcinomas (EMCs) and are characterized by a solid growth pattern and a lack of appreciable features of differentiation. As compared with pure EMC, UC is highly malignant, and the molecular pathogenesis that leads to disease aggressiveness remains largely unknown. This study interrogates the molecular pathogenesis of UCs by comparing the molecular alterations between the UC and the EMC components. A total of 20 UCs were studied, 12 of which contained both UC and EMC components. Mutation analysis was performed for the genes commonly mutated in EMC, and immunohistochemistry was used to determine the expression pattern of -catenin and PTEN. Sequencing analysis revealed that UCs harbored somatic mutations in PIK3CA(50), CTNNB1(30), TP53(30), FBXW7(20), and PPP2R1A(20). All somatic mutations detected in EMCs were also present in concurrent UCs. Moreover, additional somatic mutations were detected in the UC component in 5 (42) cases with concurrent EMC and UC. Concordance of immunostaining pattern for -catenin and PTEN was recorded in all 12 matched EMCs and UCs, except 4 cases in which nuclear accumulation of -catenin staining was detected in one of the components but not in the other. Our findings support a clonal relationship between EMCs and their associated UCs. Additional molecular genetics alteration, including mutations of CTNNB1, PPP2R1A, and TP53, may contribute to tumor progression from EMC to UC.

Recently published guidelines now specifically recommend cytology and HPV cotesting as follow-up after high-grade cervical intraepithelial neoplasia (CIN 2/3) excision.

Papanicolaou test litigation in the United States reflects widespread societal expectations that even occasional cervical screening should prevent nearly all cervical cancers in screened women. Scientific evidence clearly shows that these expectations are unfounded, especially for increasingly prevalent glandular cervical cancers and for more rapidly progressive cervical cancers in younger women. Reviews of Papanicolaou test slides involved in litigation repeatedly show 1 or more of 3 cytologic patterns difficult-to-interpret abnormal glandular cell groups, difficult-to-interpret hyperchromatic crowded cell groups, and/or difficult-to-locate or -interpret abnormal immature metaplastic squamous cells. Multiple-slide blinded reviews, supported by professional organization guidelines, should be used to identify and differentiate culpable medical errors from factors primarily associated with the inherent limitations of human screening and interpretation. In the authors' opinion, remaining US cases of cervical cancer are most likely to be prevented or minimized by more widespread utilization of cotesting with US Food and Drug Administration-approved methods of liquid-based cytology and high-risk human papillomavirus DNA testing.

BACKGROUND:The fineneedle aspiration FNA diagnosis of proliferative breast lesion is an indeterminate category. The aim of this correlative study was to determine whether a subcategory of “proliferative breast lesion with atypia” was achievable and whether this subcategory has management utility.METHODS:Breast FNA cases from 2000 through 2005 diagnosed as proliferative breast lesion and proliferative breast lesion with atypia were retrieved. Both cytologic and surgical slides of these cases were reviewed blindly. A cytologic diagnosis of proliferative breast lesion without atypia or proliferative breast lesion with atypia was used if the findings of the proliferative breast lesion did not fit a more specific category.RESULTS:Of the 3934 breast FNAs performed on palpable breast masses from January 2000 to December 2005 at the LAC USC Medical Center, 317 8.1 were diagnosed cytologically as proliferative breast lesion with atypia, without atypia or without mention of atypia. There was subsequent histopathology on 201 of these cases. After the cytologic smears were reviewed, 29 cases were excluded from this study. Of the 172 remaining cases, 21 12.2 were found to be malignant and the remaining 151 87.8 were found to be benign on histology. Of the malignant cases, 90 had an FNA diagnosis of proliferative breast lesion with atypia; of the benign cases, 78 were interpreted as proliferative breast lesion without atypia.CONCLUSIONS:Proliferative breast lesion with atypia was clinically significant because it was associated with a significantly increased likelihood of malignancy compared with proliferative breast lesion without atypia. Most of the malignancies had hypocellularity or low nuclear grade on the FNA smears. Fibroadenoma accounted for most of the benign lesions in both proliferative breast lesion and proliferative breast lesion with atypia. Cancer Cancer Cytopathol 2009. © 2009 American Cancer Society.

Immunohistochemistry can be an important part of the diagnosis of Sertoli cell tumor of the ovary, including distinction from non-sex cord-stromal tumors such as the sertoliform variant of endometrioid carcinoma and carcinoid. Several good markers for this differential diagnosis have been identified, particularly inhibin, Wilms tumor 1 gene product (WT1), epithelial membrane antigen, and chromogranin; however, many available markers have limitations to some degree. Steroidogenic factor 1 (SF-1; adrenal 4-binding protein; Ad4BP) is a nuclear transcription factor involved in gonadal and adrenal development. In the testes, SF-1 is expressed in Sertoli cells. Immunohistochemical expression of this marker in ovarian sex cord-stromal tumors, including utility for differential diagnosis, has not been rigorously evaluated. As an extension of our previous immunohistochemical studies of ovarian Sertoli cell tumor, expression of SF-1 and comparison with WT1 and inhibin were assessed in 111 primary ovarian tumors 27 Sertoli cell tumors, 60 endometrioid tumors (including borderline tumors, conventional well-differentiated carcinomas, and sertoliform variants of carcinoma), and 24 carcinoids. SF-1 was expressed in 100 of Sertoli cell tumors but not in endometrioid tumors or carcinoid. WT1 was expressed in 100 of Sertoli cell tumors and 17 of endometrioid tumors; all carcinoids were negative. Inhibin was expressed in 96 of Sertoli cell tumors and 2 of endometrioid tumors (4 of conventional well-differentiated carcinomas); all carcinoids were negative. The extent of expression of all 3 markers was similar in Sertoli cell tumor but greatest for WT1 63, 96, and 78 of cases showed expression of SF-1, WT1, and inhibin, respectively, in more than 50 of tumor cells. Immunohistochemical composite scores combining both extent and intensity of staining in positive cases were calculated for Sertoli cell tumor (possible range 1–12). Combined extentintensity of immunostaining was similar for all 3 markers, but WT1 showed the most robust immunoreactivity in positive cases (mean immunohistochemical composite scores for SF-1, WT1, and inhibin 6.1, 10.8, and 7.8, respectively). We conclude that for the differential diagnosis with endometrioid tumors and carcinoid of the ovary, SF-1 is a sensitive and specific immunohistochemical marker for Sertoli cell tumor and that SF-1 is diagnostically comparable with other good sex cord-stromal markers.

BACKGROUND.Current guidelines recommend that women with negative Papanicolaou Pap test results and no endocervicaltransformation zone ECTZ sample return for screening within 12 months. For some women, this represents earlier followup than advocated in several routine screening guidelines. Controversy remains with regard to the correlation between sampling of the ECTZ, Pap test quality, and disease risk assessment.METHODS.A retrospective study was conducted reviewing the results from 143,438 liquidbased cervical Pap tests performed at a large academic womens hospital between July 2005 and December 2006. Vaginal Pap tests were excluded from the study. Women with any Pap result, women with low–grade squamous intraepithelial lesions LSILs, and patients with high–grade squamous intraepithelial lesion HSIL Pap test results were stratified by 10year age groups and according to the presence or absence of an ECTZ sample ECTZS. Women with LSIL and HSIL Pap test results with and without an ECTZS were also compared for rates of high–risk human papillomavirus hrHPV DNA detection.RESULTS.Of the total of 143,438 cervical Pap tests performed, 27,359 19.1 were reported to be lacking an ECTZS. The absence of an ECTZS was found to be highest in adolescents and in mature women aged ≥50 years. The overall detection rate of LSIL was 4.29 and that of HSIL was 0.64. Both the LSIL and HSIL rates were found to be significantly higher in Pap tests with an ECTZS compared with Pap tests without an ECTZS LSIL: 4.51 vs 3.37 and HSIL: 0.72 vs 0.29. However, when women with LSILs and HSILs were divided into a group in which ECTZS was present and a group in which ECTZS was absent, no significant differences were found to be present with regard to hrHPV DNA rates between the 2 groups.CONCLUSIONS.Adjunctive hrHPV DNA testing is effective in stratifying risk for the presence of SIL in women with and without an ECTZS. This finding is consistent with recently reported data from >9000 patients with negative Pap results, which found that hrHPV DNA–positive test rates are independent of the presence or absence of an ECTZS. hrHPV DNA results provide a useful new optional adjunctive tool for the objective stratification of disease risk in women with negative Pap tests and no ECTZS. Cancer Cancer Cytopathol 2008. © 2008 American Cancer Society.

The prevalence of high‐risk Human Papillomavirus DNA (hrHPV DNA) in women with negative Papanicolaou (Pap) test results provides a measure of residual risk for cervical neoplasia after cytology screening. The purpose of this study was to document the prevalence of hrHPV DNA in several thousand women ages ≥30 years with negative ThinPrep Imaging System (TIS)‐imaged Pap test results in a large academic hospital cytology laboratory.All cytology‐negative TIS‐imaged ThinPrep Pap tests (TPPT) with hrHPV DNA tests that were performed by the United States Food and Drug Administration (FDA)‐approved Hybrid Capture 2 (HC2) method from May 1, 2005 to November 20, 2006 were identified and reviewed. Imaged‐negative Pap test slides associated with a positive hrHPV DNA test result were rescreened manually. Variation in hrHPV DNA prevalence was assessed for different age and ethnic groups.Of 8070 imaged cytology‐negative TPPT from women ages 11 to 90 years, hrHPV DNA test results were also available. Among 7426 women ages ≥30 years with a cytology‐negative, TIS‐imaged, Pap test, a significant age‐associated decline in hrHPV DNA prevalence was noted, 3.4% in 3050 women ages 30–45 years, 2.4% in 7426 women ages 30–90 years, and 1.8% in 5491 women ages 40–90 years. The hrHPV DNA‐positive rate was 2.3% in 6012 imaged cytology‐negative white women and 4.1% in 739 imaged cytology‐negative black women.Very low HC2 hrHPV DNA rates in 7426 women ages ≥30 years with cytology‐negative, TIS‐imaged, ThinPrep, Pap tests were similar to recently published data from 1 other academic center and lower than rates reported in previous studies on cytology‐negative North American or European women screened manually with conventional or liquid‐based Pap tests. These data may impact assessments of how best to combine cytology and HPV testing. Cancer (Cancer Cytopathol) 2007. © 2007 American Cancer Society.

The distinction of ovarian Sertoli cell tumor from other tumors in the histological differential diagnosis, particularly endometrioid carcinoma and carcinoid tumor, may be difficult. Many immunohistochemical markers have been studied for this differential diagnosis, but currently available markers are neither 100% sensitive nor specific. Sox9is a transcription factor involved in Sertoli cell differentiation in the testis. The role that this molecule plays in the pathogenesis of ovarian Sertoli cell tumors and the potential use as an immunohistochemical marker for differential diagnosis have not been investigated. Immunohistochemical staining for Sox9was performed in 152 ovarian tumors pure Sertoli cell tumor (n = 36), endometrioid borderline tumor (n = 38), well-differentiated endometrioid carcinoma (n = 26), sertoliform endometrioid carcinoma (n = 13), and carcinoid tumor (n = 39). Nuclear expression was considered positive. Extent and intensity of staining were semiquantitatively scored. In addition, immunohistochemical composite scores in positive cases (ranging from 1 to 12) were calculated based on the extent score multiplied by the intensity score. Sox9was expressed in 44% of Sertoli cell tumors, 55% of endometrioid borderline tumors, 65% of well-differentiated endometrioid carcinomas, 39% of sertoliform endometrioid carcinomas, and 10% of carcinoid tumors. The mean Sox9immunohistochemical composite scores in positive cases were 6.3 for Sertoli cell tumor, 5.3 for endometrioid borderline tumor, 8.0 for well-differentiated endometrioid carcinoma, 2.8 for sertoliform endometrioid carcinoma, and 6.8 for carcinoid tumor. The differences in the mean Sox9composite scores between Sertoli cell tumor and the other tumor categories were not statistically significant (p values ranged from 0.092 to 0.523). We conclude that Sox9is variably expressed in ovarian Sertoli cell tumor and other tumors that are in the differential diagnosis and, thus, is not helpful for immunohistochemical distinction. Understanding the role of Sox9in the pathogenesis of ovarian Sertoli cell tumor requires further study.

ABSTRACTRhesus monkey bone marrow expresses a cathelicidin whose C-terminal domain comprises a 37-residue alpha-helical peptide (RL-37) that resembles human LL-37. Like its human counterpart, RL-37 rapidly permeabilized the membranes of Escherichia coliML-35p and lysed liposomes that simulated bacterial membranes. When tested in media whose NaCl concentrations approximated those of extracellular fluids, RL-37 was considerably more active than LL-37 against staphylococci. Whereas human LL-37 contains five acidic residues and has a net charge of +6, rhesus RL-37 has only two acidic residues and a net charge of +8. Speculating that the multiple acidic residues of human LL-37 reduced its efficacy against staphylococci, we made a peptide (LL-37 pentamide) in which each aspartic acid of LL-37 was replaced by an asparagine and each glutamic acid was replaced by a glutamine. LL-37 pentamide's antistaphylococcal activity was substantially greater than that of LL-37. Thus, although the precursor of LL-37 is induced in human skin keratinocytes by injury or inflammation, its insufficiently cationic antimicrobial domain may contribute to the success of staphylococci in colonizing and infecting human skin.

We purified two new minidefensins (RTD‐2 and RTD‐3) from the bone marrow of rhesus monkeys. Both were circular octadecapeptides that contained three intramolecular disulfide bonds and were homologous to RTD‐1, a circular (θ) defensin previously described by Tang et al. (Science, 286, 498–502, 1999). However, whereas the 18 residues of RTD‐1 represent spliced nonapeptide fragments derived from two different demidefensin precursors, RTD‐2 and ‐3 comprise tandem nonapeptide repeats derived from only one of the RTD‐1 precursors. Thus, circular minidefensins are products of a novel posttranslational system that generates effector molecule diversity without commensurate genome expansion. A system wherein two demidefensin genes can produce three circular minidefensins might allow nsuch genes to produce (n/2)(n+1) peptides.

ABSTRACTGallinacin-3 and gallopavin-1 (GPV-1) are newly characterized, epithelial β-defensins of the chicken (Gallus gallus) and turkey (Meleagris gallopavo), respectively. In normal chickens, the expression of gallinacin-3 was especially prominent in the tongue, bursa of Fabricius, and trachea. It also occurred in other organs, including the skin, esophagus, air sacs, large intestine, and kidney. Tracheal expression of gallinacin-3 increased significantly after experimental infection of chickens with Haemophilus paragallinarum, whereas its expression in the tongue, esophagus, and bursa of Fabricius was unaffected. The precursor of gallinacin-3 contained a long C-terminal extension not present in the prepropeptide. By comparing the cDNA sequences of gallinacin-3 and GPV-1, we concluded that a 2-nucleotide insertion into the gallinacin-3 gene had induced a frameshift that read through the original stop codon and allowed the chicken propeptide to lengthen. The striking structural resemblance of the precursors of β-defensins to those of crotamines (highly toxic peptides found in rattlesnake venom) supports their homology, even though defensins are specialized to kill microorganisms and crotamines are specialized to kill much larger prey.

Secretory lipophilins are "lipid-loving" proteins that are major constituents of several mammalian secretions, including the prostatic fluid of rats and the tears of humans and rabbits. These proteins form covalent heterodimers that are stabilized by three intramolecular cystine disulfide bonds. The heterodimers, some of which are glycosylated, may undergo additional noncovalent assembly to form tetramers. The peptide components found in secretory lipophilins are from two subfamilies: lipophilins AB and lipophilin C. The C subfamily members described in this report are three rabbit and one human lipophilin, plus human mammaglobin and the C3 subunit of rat prostatein. Human AB and C lipophilins are expressed by many tissues and are especially prominent in endocrine-responsive organs. The gene for human lipophilin B resides at chromosome 10q22-23. This region harbors the PTENMMAC1 gene and is believed to contain additional tumor suppressor genes. Although the functions of secretory lipophilins are imperfectly understood, their abundance in glandular secretions and in hormone-responsive tissues suggests that they deserve considerably more attention than they have received to date.

Lipophilin components A, B and C are human homologues of prostatein, the major secreted protein of rat prostate. This report describes their cDNA sequences, tissue expression and chromosomal localization. Lipophilin gene products were widely expressed in normal tissues, especially in endocrine-responsive organs. The gene for lipophilin C (also called mammaglobin b) is located on chromosome 11q12-q13.1, near the mammaglobin gene, a homologue overexpressed in many breast cancers. The lipophilin B gene resides on chromosome 10q23, a region deleted in many tumors, and the lipophilin A gene is on chromosome 15q12-q13.

ABSTRACTWe identified two novel β-defensin precursors, preproGBD-1 and preproGBD-2, in the tissues of a goat. Although the precursors were identical in 96.8% of their bases and 88.2% (60 of 68) of their amino acids, preproGBD-1 was expressed principally in the tongue and respiratory tract, whereas preproGBD-2 expression predominated throughout the intestine. These findings exemplify the phenomenon of tissue-specific expression in a family of host defense peptides that arose before the avian and mammalian lineages diverged.

ABSTRACTWe purified three proline-rich antimicrobial peptides from elastase-treated extracts of sheep and goat leukocytes and subjected two of them, OaBac5α and ChBac5, to detailed analysis. OaBac5α and ChBac5 were homologous to each other and to bovine Bac5. Both exhibited potent, broad-spectrum antimicrobial activity under low-concentration salt conditions. While the peptides remained active againstEscherichia coli, Pseudomonas aeruginosa,Bacillus subtilis, and Listeria monocytogenesin 100 mM NaCl, they lost activity against Staphylococcus aureusand Candida albicansunder these conditions. ChBac5 was shown to bind lipopolysaccharide, a property that could enhance its ability to kill gram-negative bacteria. Proline-rich Bac5 peptides are highly conserved in ruminants and may contribute significantly to their innate host defense mechanisms.

Hemocytes from the invertebrate Styela clava, a solitary tunicate, contained a family of four α‐helical antimicrobial peptides that were purified, sequenced and named clavanins A, B, C and D. Each clavanin contained 23 amino acid residues and was C‐terminally amidated. The tunicate peptides resembled magainins in size, primary sequence and antibacterial activity. Synthetic clavanin A was prepared and displayed comparable antimicrobial activity to magainins and cecropins. The presence of α‐helical antimicrobial peptides in the hemocytes of a urochordate suggests that such peptides are primeval effectors of innate immunity in the vertebrate lineage.

We cloned precursors of three new antimicrobial peptides, Styelins C, D and E, from a pharyngeal cDNA library of a tunicate, Styela clava. Preprostyelins resembled dipteran preprocecropins, while the mature domain of Styelin C resembled Cecropin P1, an antimicrobial peptide purified from the porcine intestine. Beginning with the last 6 residues of their signal sequences, Styelin C and Cecropin 1 from Drosophila virilishad 8/11 identical amino acids (72.7%). Moreover, 4 of the last 6 residues of their mature peptide domains were also identical. Styelins were shorter, by 8 residues, than dipteran cecropins and preprostyelins contained a conserved, polyanionic C‐terminal extension that was absent in preprocecropins. Delineation of cecropin‐like antimicrobial peptides in a protochordate supports the antiquity of this family as effectors of innate immunity in animals and it increases the likelihood that additional cecropin‐like peptides will be found among other evolutionary descendants of protochordates — vertebrates.

We purified and characterized an unusual antimicrobial peptide, prophenin‐1 (PF‐1), from porcine leukocytes. The peptide had a mass of 8,683 and contained 79 residues, including 42 (53.2%) prolines and 15 (19.0%) phenylalanines. Its N‐terminal 60 residues consisted of three perfect and three nearly perfect repeats of a decamer, FPPPNFPGPR. Prophenin‐1 was encoded on a cathelin‐containing precursor and showed substantially more activity against E. coli, a Gram‐negative bacterium, than against Listeria monocytogenes, a Gram‐positive organism, in vitro.

We characterized genes for prophenin (PF)-2 and PR-39, two cathelin-associated antimicrobial peptides found in porcine leukocytes. Both contained 4 exons and 3 introns and were compact, contiguous and highly homologous. Exons I–III encoded most of their cathelin domains. Exon IV specified the final few cathelin residues, including its conserved C-terminal valine, followed by the mature PR-39 peptide or a PF-2 precursor. The highly conserved 5′ flanking sequences of this gene family contained NF-κB, IL-6, GM-CSF and NF-1 binding motifs and the introns were unusually conserved. These data suggest that the panoply of porcine cathelin-associated antimicrobial peptides arose relatively recently via gene reduplications and exon shuffling, and that in vivo expression of cathelin-associated antimicrobial peptides may respond to mediators generated early during infection.

The porcine leukocyte protegrins are a family of cysteine-rich antimicrobial peptides the primary structures of which combine features of defensins and tachyplesins. We cloned three protegrins from porcine bone marrow mRNA by PCR, including one (PG-4) that was previously unknown. The 691 bp protegrin cDNAs were > 98.8% identical, and each was surrounded by highly conserved 5′ and (in some instances) 3′ sequences present in structurally dissimilar antimicrobial and LPS-binding peptides of animal leukocytes.

We compared the expression of human α‐ and β‐defensins by various human tissues. mRNA for α‐defensins HNP1‐3, abundant in bone marrow, was detected in peripheral blood leukocytes, spleen and thymus by RT‐PCR, which revealed α‐defensins HD5 and HD6 only in the small intestine. In contrast, the pancreas and kidney expressed high levels of hBD‐1 and lower levels of this β‐defensin were found in many organs by RT‐PCR (salivary gland > trachea > prostate and placenta > thymus, testis, small intestine). hBD‐1 mRNA was produced constitutively by cultured normal human epithelial cells derived from the trachea, bronchi, small airways and the mammary gland. These largely non‐overlapping tissue distributions of human α‐ and β‐defensins suggest that hBD‐1 may be positioned to defend epithelial cells and mucosae from infection, whereas expression of HNP1‐3 in neutrophils and HD5 and HD6 in Paneth cells allows these α‐defensins to participate in systemic and small intestinal host defenses, respectively.

We cloned a novel human β-defensin gene and determined its full-length cDNA sequence. The entire gene spanned more than 7 kb and included a large 6962-bp intron. The 362-bp cDNA encoded a prepropeptide that corresponded precisely to the recently identified human β-defensin HBD-1, an antimicrobial peptide implicated in the resistance of epithelial surfaces to microbial colonization. By two-color fluorescencein situhybridization on both metaphase chromosome and released chromatin fiber, HBD-1 gene (DEFB1 in HUGO/GDB nomenclature) mapped to chromosomal region 8p23.1–p23.2 in close proximity (within 100–150 kb) to the gene for the human neutrophil α-defensin HNP-1 (DEFA1). Thus, despite a complete lack of DNA sequence similarity and despite differences in their disulfide-pairing pattern, the α- and β-families appear to have evolved from a common premammalian defensin gene.