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INTRODUCTION: Pleural mesothelioma (PM) is an aggressive malignancy with no identified predictive biomarkers. We assessed whether tumor BAP1 status is a predictive biomarker for survival in patients receiving first-line combination platinum and pemetrexed therapy.METHODS: PM cases (n = 114) from Aalborg, Denmark, were stained for BAP1 on tissue microarrays. Demographic, clinical, and survival data were extracted from registries and medical records. Surgical cases were excluded. BAP1 status was associated with overall survival (OS) by Cox regression and Kaplan-Meier methods. Results were validated in an independent cohort from Perth, Australia (n = 234).RESULTS: BAP1 loss was found in 62% and 60.3% of all Danish and Australian samples, respectively. BAP1 loss was an independent predictor of OS in multivariate analyses corrected for histological subtype, performance status, age, sex, and treatment (hazard ratio = 2.49, p < 0.001, and 1.48, p = 0.01, respectively). First-line platinum and pemetrexed-treated patients with BAP1 loss had significantly longer median survival than those with retained BAP1 in both the Danish (20.1 versus 7.3 mo, p < 0.001) and Australian cohorts (19.6 versus 11.1 mo, p < 0.01). Survival in patients with BAP1 retained and treated with platinum and pemetrexed was similar as in those with best supportive care. There was a higher OS in patients with best supportive care with BAP1 loss, but it was significant only in the Australian cohort (16.8 versus 8.3 mo, p < 0.01).CONCLUSIONS: BAP1 is a predictive biomarker for survival after first-line combination platinum and pemetrexed chemotherapy and a potential prognostic marker in PM. BAP1 in tumor is a promising clinical tool for treatment stratification.

INTRODUCTION: Serous effusion is often the first sign of mesothelioma. Diagnosis based on cytologic material from the effusions remains controversial and complementary biopsy is usually required. However, obtaining representative tissue sample may be challenging, while obtaining cytologic material is a minimally invasive procedure, providing potential for an earlier diagnosis. Loss of BRCA1-associated protein (BAP1), combined with loss of methylthionadenosine phosphorylase (MTAP) detected by immunohistochemistry, have shown to be reliable markers in the diagnosis of mesothelioma on histologic sections. Here we evaluate the value of these biomarkers in cytologic specimens.MATERIALS AND METHODS: The BAP1 and MTAP expression in specimens of 162 mesothelioma patients (156 pleural, 6 peritoneal)-71 cytologic, 91 histologic (44 epithelioid, 31 biphasic, 16 sarcomatoid)-and 20 patients with reactive mesothelial proliferations were investigated.RESULTS: The loss of BAP1 and/or MTAP was highly sensitive and specific in differentiating mesothelioma from reactive mesothelial proliferations, with no significant difference between pleural effusions and biopsies, specificity of 100% in both and a sensitivity of 78.9% and 80.2%, respectively (P = 0.3). There was a 100% concordance of the expression of BAP1 and MTAP in cytologic and corresponding histopathologic samples. Loss of BAP1 and/or MTAP in histologic sections discriminated sarcomatoid, biphasic, and epithelioid mesothelioma from reactive mesothelial proliferations with a sensitivity of 81.2%, 83.9%, and 77.3% respectively.CONCLUSION: Loss of expression of BAP1 and/or MTAP differentiated mesothelioma from reactive mesothelial proliferations with excellent specificity and high sensitivity in cytologic samples, comparable to histopathologic sections.

BACKGROUND: The study investigated the association of the relative dose-intensity (RDI) of cisplatin and timing of adjuvant platinum-based chemotherapy (APC) with survival for stage I-III non-small cell lung cancer (NSCLC) patients.MATERIAL AND METHODS: Real-life data of patients treated with APC (four cycles of cisplatin and vinorelbine) between 2007 and 2014 was included to analyse the association between disease-free survival (DFS) and overall survival (OS) with RDI (ratio of received to planned dose-intensity). High RDI was defined as cisplatin RDI of > 75% and low RDI ≤ 75%.RESULTS: Out of 198 patients, 166 were eligible. Low RDI was administered to 72 (43%) patients. In multivariate analysis, those patients had a significantly higher risk of recurrence (HR: 1.87, 95%CI 1.13-3.09, p = 0.01) and death (HR: 1.91, 95%CI 1.32-3.23, p = 0.01) versus patients in the high RDI group. The risk of death was significantly higher in patients with PS 1 treated with low versus high RDI (HR: 2.72, 95%CI: 1.22-6.09, p = 0.014). The risk of recurrence was higher for patients with squamous cell carcinoma of low versus high RDI (HR: 3.82, 95%CI: 1.01-14.4, p = 0.048). No impact of delayed APC beyond six weeks from surgery on neither DFS (HR: 0.78, 95%CI: 0.46-1.33, p = 0.36) nor OS (HR 0.67, 95%CI: 0.40-1.15, p = 0.15) was observed.CONCLUSION: Low cisplatin RDI ≤ 75% of APC, but not extended time from surgery to APC onset > six weeks, was associated with significantly shorter survival in NSCLC patients.

Objective: This study investigates the potential of an artificial intelligence (AI) methodology, the radial basis function (RBF) artificial neural network (ANN), in the evaluation of thyroid lesions. Study Design. The study was performed on 447 patients who had both cytological and histological evaluation in agreement. Cytological specimens were prepared using liquid-based cytology, and the histological result was based on subsequent surgical samples. Each specimen was digitized; on these images, nuclear morphology features were measured by the use of an image analysis system. The extracted measurements (41,324 nuclei) were separated into two sets: the training set that was used to create the RBF ANN and the test set that was used to evaluate the RBF performance. The system aimed to predict the histological status as benign or malignant.Results: The RBF ANN obtained in the training set has sensitivity 82.5%, specificity 94.6%, and overall accuracy 90.3%, while in the test set, these indices were 81.4%, 90.0%, and 86.9%, respectively. Algorithm was used to classify patients on the basis of the RBF ANN, the overall sensitivity was 95.0%, the specificity was 95.5%, and no statistically significant difference was observed.Conclusion: AI techniques and especially ANNs, only in the recent years, have been studied extensively. The proposed approach is promising to avoid misdiagnoses and assists the everyday practice of the cytopathology. The major drawback in this approach is the automation of a procedure to accurately detect and measure cell nuclei from the digitized images.

BRCA1-associated protein 1 (BAP1) immunohistochemistry (IHC) was assessed as a predictive biomarker for pemetrexed-based treatment in non-surgically treated malignant pleural mesothelioma (MM) patients.IHC was performed on 168 MM patient slides to assess BAP1 loss (BAP1-) or retain (BAP1+). BAP1 mouse monoclonal antibody (C-4) was used. Gender, age, performance status (PS), subtype, types of treatment and overall survival (OS) was retrieved from medical records and registries. Clinical endpoint was OS. Data were analyzed by Fisher´s exact test, Kaplan-Meier estimate, log rank test and cox regression.In total, 91 MM patients that received best supportive care (BSC) and 77 treated with pemetrexed were included in the study (Table 1). Each group was analyzed according to BAP1- or BAP1+. A significantly higher OS was observed in the pemetrexed group for BAP1- versus BAP1+ (OS=20 vs 10 months, p

Background To evaluate reproducibility of a reporting system for endometrial cytology. Methods Cytologic slides from 49 patients, prepared via liquid based cytology, were blindly examined by five cytopathologists of various experience levels, applying a recently introduced reporting system as previously reported. The agreement among cytopathologists was evaluated via Kappa (κ) statistics and the Kendall's Coefficient of Variation (W); cytologic results were compared with the relevant histologic report. Results Substantial agreement among all five raters was found in the benign, ACE-L and malignant categories, fair agreement in inadequate and ACE-H categories, whereas only slight agreement in ACE-U. For the three more experienced cytopathologists, an almost perfect agreement was found in inadequate, benign, and ACE-L categories, substantial agreement in ACE-H and malignant categories and fair agreement in ACE-U category. Overall agreement for all five cytopathologists and for all categories was moderate, whereas it was very high for the three senior raters. Using the Kendall's test, both five cytopathologists (W = 0.81) and the three senior ones (W = 0.93) had very high agreement. Sensitivity: 83.33–92.59%, specificity: 83.33–94.74%, ROC area: 71.72–90.3%. Conclusion Application of appropriate statistical tests shows that integration of a new reporting cytologic system is effective with an overall accuracy around 90%. Both statistical tests applied disclosed lower agreement rates among both all five raters and the three most experienced ones in the intermediate categories constituting the gray zone, thus delineating the need for better training of cytopathologists to correctly identify diagnostic criteria for classification of a given case into these categories.

Objectives Diffuse malignant mesothelioma (MM) is mainly caused by asbestos inhalation. The malignancy is rare among women and studies of the prevalence and causative role of non-occupational asbestos exposure among women with MM are scarce. This observational study aimed to elucidate the asbestos exposure patterns among women with MM. Methods All histological and cytological specimens from women diagnosed with MM between 1974–2015 at the Institute of Pathology, Aalborg University Hospital in Denmark, were re-evaluated. Occupational and habitation information were obtained from Danish registries and medical journals based on record linkage via the unique person ID. The number of MM cases in each parish in the region of North Jutland was determined and the incidence density in parishes was used to calculate the spatial relative risk (RR) of MM among women. Results Diagnosis of MM was confirmed in 91 women. Exposure types were classified as occupational (9%), domestic (10%), environmental (22%), combination of domestic and environmental (34%) and unknown (25%). Twenty continuous parishes formed a MM “hotspot” around the asbestos-consuming industries in the city of Aalborg. Of these, the maximum RR was found in a parish housing an asbestos factory [RR 10.5, 95% confidence interval (CI) 5.5–19.4, environmental exposure in particular RR 2.9, 95% CI 0.7–9.1]. Conclusion Non-occupational asbestos exposure is the main cause of MM and may account for up to 66% of MM cases among women in North Jutland, Denmark.

Familial hypocalciuric hypercalcemia (FHH) is an autosomal dominant disorder, associated with inactivating mutations of the calcium-sensing receptor (CaSR). To evaluate the functional significance of a CaSR mutation, identified in a young infant who presented with hypercalcemia and hypocalciuria. The CaSR gene coding sequences were analyzed by polymerase chain reaction amplification and direct sequencing analysis. The mutation identified was introduced by site-directed mutagenesis into a wild-type (WT) CaSR plasmid, and human embryonic kidney 293 T cells were transfected with either the WT or mutant CaSR. The function of the mutated CaSR protein was analyzed by evaluating the free intracellular calcium [(Ca2+)i] response after challenge with extracellular calcium (Ca2+). We identified a heterozygous mutation c.772_773delGTinsA in exon 4 resulting in the substitution of amino acid valine (Val) with amino acid arginine (Arg) and the premature pause of the translation 46 amino acids later (Val258ArgfsTer47). Functional assay showed that cells transfected with the mutant CaSR had a significantly poorer response to extracellular Ca2+ stimulation compared with the WT. We have shown that the c.772_773delGTinsA mutation causes a significant alteration of CaSR function leading to features of FHH in an affected young infant since the first months of life.

Temephos is a major organophosphate (OP) larvicide that has been used extensively for the control of Aedes albopictus and Aedes aegypti, the major vectors for viral diseases, such as dengue fever, zika and chikungunya. Resistance to temephos has been recently detected and associated with the upregulation of carboxylesterases (CCEs) through gene amplification, in both species. Here, we expressed the CCEae3a genes which showed the most striking up-regulation in resistant Aedes strains, using the baculovirus system. All CCEae3a variants encoded functional enzymes, with high activity and preference for p-nitrophenyl butyrate, a substrate that was shown capable to differentiate temephos resistant from susceptible Aedes larvae. Enzyme kinetic studies showed that CCEae3as from both Ae. aegypti and Ae. albopictus (CCEae3a_aeg and CCEae3a_alb, respectively) strongly interact with temephos oxon and slowly released the OP molecule, indicating a sequestration resistance mechanism. No difference was detected between resistant and susceptible CCEae3a_aeg variants (CCEae3a_aegR and CCEae3a_aegS, respectively), indicating that previously reported polymorphism is unlikely to play a role in temephos resistance. HPLC/MS showed that CCEae3as were able to metabolize temephos oxon to the temephos monoester [(4-hydroxyphenyl) sulfanyl] phenyl O,O-dimethylphosphorothioate. Western blot and immunolocalization studies, based on a specific antibody raised against the CCEae3a_alb showed that the enzyme is expressed at higher levels in resistant insects, primarily in malpighian tubules (MT) and nerve tissues.

Fine tuning of c-MYC expression is critical for its action and is achieved by several regulatory mechanisms. The contribution of c-mycmRNA regulatory sequences on its translational control has been investigated individually. However, putative interactions have not been addressed so far. The effect of these interactions upon the translatability of monocistronic and bicistronic chimaeric mRNAs, carrying combinations of the c-mycmRNA 5′-untranlated region (UTR), 3′-UTR, and coding region instability element (CRD) was investigated on this study. The presence of the 5′-UTR induced an increase in translatability of 50%. The presence of the CRD element, when in frame, reduced translatability by approximately 50%, regardless of the expression levels of the wild type CRD- binding protein (CRD-BP/IMP1). Conversely, overexpression of a mutated CRD-BP/IMP1 (Y396F) further impeded translation of the chimaeric mRNAs carrying its cognate sequences. The presence of the c-myc3′-UTR increased translatability by approximately 300% affecting both cap and c-mycinternal ribosome entry site (IRES) mediated translation. In addition, 3′-UTR rescued the cap mediated translation in the presence of the polyadenylation inhibitor cordycepin. Furthermore, the 3′-UTR rescued cap mediated translation under metabolic stress conditions and this was enhanced in the absence of a long poly (A) tail.

'The objective of this study is to investigate the potential of classification and regression trees (CARTs) in discriminating benign from malignant endometrial nuclei and lesions. The study was performed on 222 histologically confirmed liquid based cytological smears, specifically: 117 benign cases, 62 malignant cases and 43 hyperplasias with or without atypia. About 100 nuclei were measured from each case using an image analysis system; in total, we collected 22783 nuclei. The nuclei from 50% of the cases (the training set) were used to construct a CART model that was used for knowledge extraction. The nuclei from the remaining 50% of cases (test set) were used to evaluate the stability and performance of the CART on unknown data. Based on the results of the CART for nuclei classification, we propose two classification methods to discriminate benign from malignant cases. The CART model had an overall accuracy for the classification of endometrial nuclei equal to 85%, specificity 90.68%, and sensitivity 72.05%. Both methods for case classification had similar performance: overall accuracy in the range 94-95%, specificity 95%, and sensitivity 91-94%. The results of the proposed system outperform the standard cytological diagnosis of endometrial lesions. This study highlights interesting diagnostic features of endometrial nuclear morphology and provides a new classification approach for endometrial nuclei and cases. The proposed method can be a useful tool for the everyday practice of the cytological laboratory.

Objective: Thyroid fine needle aspiration (FNA) contributes to the appropriate management of nodular thyroid lesions. The introduced categories in the Bethesda system for reporting thyroid cytopathology (TBSRTC) are associated with an implied cancer risk, providing a clinical management guideline. This study aims to evaluate the reproducibility of this implied risk and to compare the results from two different cytopathology departments. Methods: Five hundred histologically confirmed FNAs, studied since the introduction of TBSRTC, were obtained from 4208 and 3587 FNAs performed in a large regional hospital in Herakleion, Crete (group A) and a university hospital in Athens (group B), respectively. Reports were issued according to TBSRTC. Aspirates were prepared with ThinPrep and evaluated by two experienced cytopathologists. The reproducibility and accuracy were evaluated. Results: The proportion test for suspicious for malignancy (SFM) and malignant (M) cytology reports (P < 0.0001), and the number of malignancies on histology (P < 0.0001), were significantly higher in group A than in group B, consistent with a higher incidence of thyroid carcinomas in southern Greece. Although the malignancy rates were higher in group A than in group B for all categories, except M (A, 99.3%; B, 100%), the difference was only significant for benign aspirates (P = 0.0303). Malignancy rates for all categories in group A were above the TBSRTC recommended range, but were consistent with an increased prevalence of malignancy in that centre, differences in reporting practice and the variable ranges reported in the literature. There was lower sensitivity (P = 0.019) and overall accuracy (P = 0.003) in group A relative to group B, but no difference in specificity. Conclusions: TBSRTC provides valuable information for the appropriate management of nodular thyroid lesions, both in a university and a large regional hospital.

Poly(A)polymerase-alpha (PAPOLA) has been the most extensively investigated mammalian polyadenylating enzyme, mainly in regard to its multifaceted post-translational regulation. The possibility of translational regulation of this enzyme was addressed. The transcription start site was mapped and two uORFs, highly conserved among several species, were identified in the 211-bp long, GC-rich, 5' UTR of the PAPOLA mRNA. Mutation of the 5' proximal AUG resulted in increased translational efficiency of the adjacent coding sequence, whereas no significant effect was observed after mutation of the second AUG. These observations imply that translational regulation is among the conserved mechanisms regulating PAPOLA expression.

Malignant pleural mesothelioma (MPM) is an asbestos-related cancer with a median survival of 12months. The MPM incidence is 1-6/100,000 and is increasing as a result of historic asbestos exposure in industrialized countries and continued use of asbestos in developing countries. Lack of accurate biomarkers makes diagnosis, prognostication and treatment prediction of MPM challenging. The aim of this review is to identify the front line of MPM biomarkers with current or potential clinical impact. Literature search using the PubMed and PLoS One databases, the related-articles function of PubMed and the reference lists of associated publications until April 26th 2015 revealed a plethora of candidate biomarkers. The current gold standard of MPM diagnosis is a combination of two positive and two negative immunohistochemical markers in the epithelioid and biphasic type, but sarcomatous type do not have specific markers, making diagnosis more difficult. Mesothelin in serum and pleural fluid may serve as adjuvant diagnostic with high specificity but low sensitivity. Circulating proteomic and microRNA signatures, fibulin-3, tumor cell gene-ratio test, transcriptomic, lncRNA, glycopeptides, pleural fluid FISH assay, hyaluronate/N-ERC mesothelin and deformability cytometry may be important future markers. Putative predictive markers for pemetrexed-platinum are tumor TS and TYMS, for vinorelbine the ERCC1, beta-tubuline class III and BRCA1. Mutations of the BAP1 gene are potential markers of MPM susceptibility. In conclusion, the current status of MPM biomarkers is not satisfactory but encouraging as more sensitive and specific non-invasive markers are emerging. However, prospective validation is needed before clinical application.

8560 Background: Malignant Mesothelioma (MM) is an asbestos-related malignancy that presents mainly in the pleura (MPM) and peritoneum (MAM). In a densely populated area of Aalborg city in North Jutland, Denmark, a large Eternit asbestos factory that was active for 60 years until 1986 and two shipyards were situated. The Region of North Jutland, Denmark has a high mm incidence in women of 1.0/100,000. Methods: From 1974-2015, 101 histological and cytological samples of women diagnosed with mm in Aalborg University Hospital were identified re-evaluated by modern immunohistochemistry. Patient information regarding asbestos exposure was retrieved from medical records and selected Danish registries. Asbestos exposure was classified as primary for asbestos workers; domestic, for women living with an asbestos worker; environmental, when living or working within 10.000 meters from an asbestos emitting location; unknown, where no source of asbestos exposure could be identified. Results: Clinical and histopathological mm diagnosis was certain for 91 women. Potential asbestos exposure is summarized in Table 1. The employment of the women and their relatives include work at the asbestos cement factory, shipyard, construction, laboratory, pipe factory, electrician and insulator. The women with domestic and combined domestic and environmental (secondary) exposure to asbestos were most prone to develop MPM rather than MAM while women primary exposed to asbestos developed MAM rather than MPM (p=0.016). Conclusions: This study showed that the vast majority (64%) of the women diagnosed with mm had documented non-occupational asbestos exposure and almost 1/5 had a sole environmental exposure, by living in proximity to asbestos industry. Environmental asbestos exposure is a serious risk factor for mm in women. Primary asbestos exposure, inferring more intense exposure through occupation, may predispose to peritoneal mesothelioma in women. [Table: see text]

The present review aims at providing an assessment of the clinical significance of Biphosphonates (BPs) in the treatment of patients with cancer.A systematic literature review was performed based on database search in PubMed/Medline and included articles up to August 2013.BPs can reduce, delay, and prevent complications related to bone metastases. They improve mobility, functionality, pain, and quality of life. They limit survival of any inactive cancer cells in the microenvironment of the bone marrow, contributing to their death from anti-neoplastic treatments. Moreover, they limit and delay bone morbidity due to osteoporosis related to hormonotherapy in breast and prostate cancer. Finally, benefits can be derived from the combination of BPs with radiotherapy in bone density, recalcification, opioid use, and patient's quality of life and performance status.The contribution of BPs in the course of certain neoplasms is preventive and synergistic to other treatments.

To compare the clinical efficiency of two commercially available HPV DNA detection and typing tests. The CLART HPV2 test, a novel HPV test based on DNA microarrays that can identify 35 HPV genotypes, was compared to the Linear Array HPV Genotyping (LA) test, a more widely used test able to identify 37 HPV genotypes.The CLART test was evaluated by comparing the genotyping results of 538 ThinPrep Pap tests with the LA test as well as with the cytological and histological findings.The exact same types and results were identified in 86.1% of the samples (kappa = 0.74). The tests showed excellent agreement in HPV positivity and identification of single and multiple infections (concordance rate 88.7%, kappa = 0.827).The CLART test demonstrated results comparable to those of the LA test in clinical sensitivity as measured by the positive predictive value of CIN2+ in ASCUS (67.3% vs. 57.1%), while overall it exhibited higher sensitivity, specificity, positive and negative predictive values and area under curves than the LA test in all cytological and histological subgroups analyzed.

'The objective of this study is to investigate the potential of classification and regression trees (CARTs) in discriminating benign from malignant endometrial nuclei and lesions. The study was performed on 222 histologically confirmed liquid based cytological smears, specifically: 117 benign cases, 62 malignant cases and 43 hyperplasias with or without atypia. About 100 nuclei were measured from each case using an image analysis system; in total, we collected 22783 nuclei. The nuclei from 50% of the cases (the training set) were used to construct a CART model that was used for knowledge extraction. The nuclei from the remaining 50% of cases (test set) were used to evaluate the stability and performance of the CART on unknown data. Based on the results of the CART for nuclei classification, we propose two classification methods to discriminate benign from malignant cases. The CART model had an overall accuracy for the classification of endometrial nuclei equal to 85%, specificity 90.68%, and sensitivity 72.05%. Both methods for case classification had similar performance: overall accuracy in the range 94-95%, specificity 95%, and sensitivity 91-94%. The results of the proposed system outperform the standard cytological diagnosis of endometrial lesions. This study highlights interesting diagnostic features of endometrial nuclear morphology and provides a new classification approach for endometrial nuclei and cases. The proposed method can be a useful tool for the everyday practice of the cytological laboratory.

Fine-needle aspiration (FNA) cytology is an important diagnostic tool in patients with thyroid lesions. Several systems have been proposed for the cyropathologic diagnosis of the thyroid nodules. However cases with indeterminate cytological findings still remain a matter of debate. In this review we analyze all literature regarding Thyroid Cytopathology Reporting systems trying to identify the most suitable methodology to use in clinical practice for the preoperative diagnosis of thyroid nodules. A review of the English literature was conducted, and data were analyzed and summarized and integrated from the authors' perspective. The main purpose of thyroid FNA is to identify patients with higher risk for malignancy, and to prevent unnecessary surgeries for benign conditions. The Bethesda System for Reporting Thyroid Cytopathology is the most widely used system for the diagnosis of thyroid FNA specimens. This system also contains guidelines for the diagnosis and treatment of indeterminate or suspicious for malignancy cases. In conclusion, patients who require repeated FNAs for indeterminate diagnoses will be resolved by repeat FNA in a percentage of 72%-80%.

To investigate capability of combination of learning vector quantizer (LVQ) neural networks (NNs) in discrimination of benign from malignant thyroid lesions.The study included 335 liquid-based cytology, fine needle aspiration (FNA), Papanicolaou-stained specimens. All cases were compared to the histologic diagnosis. Features describing size, shape, and texture of -100 nuclei per case were extracted from cytologic images using a custom image analysis system. These features were used to classify each nucleus by LVQ type NNs. The nucleus classification results were used to classify individual lesions with a second LVQ NN. Cases were distributed by histologic diagnosis. Data from -50% from each category were used for training LVQ classifiers. Remaining data were used to test classifier performance. The system was used to discriminate to individual cellular level and individual patient level between benign and malignant nuclei.Application of the proposed algorithm combining two LVQ NNs allows discrimination between benign and malignant cell nuclei and lesions.Results indicate that use of NNs, combined with image morphometry, can provide information on thyroid lesion malignancy potential. The system could improve FNA diagnostic accuracy of the thyroid gland, especially in follicular neoplasms suspicious for malignancy and in Hürthle cell tumors.

Objective of this study is to investigate the potential of the learning vector quantizer neural network (LVQ-NN) classifier on various diagnostic variables used in the modern cytopathology laboratory and to build an algorithm that may facilitate the classification of individual cases. From all women included in the study, a liquid-based cytology sample was obtained; this was tested via HPV DNA test, E6/E7 HPV mRNA test, and p16 immunostaining. The data were classified by the LVQ-NN into two groups: CIN-2 or worse and CIN-1 or less. Half of the cases were used to train the LVQ-NN; the remaining cases (test set) were used for validation. Out of the 1258 cases, cytology identified correctly 72.90% of the CIN-2 or worst cases and 97.37% of the CIN-1 or less cases, with overall accuracy 94.36%. The application of the LVQ-NN on the test set allowed correct classification for 84.62% of the cases with CIN-2 or worse and 97.64% of the cases with CIN-1 or less, with overall accuracy of 96.03%. The use of the LVQ-NN with cytology and the proposed biomarkers improves significantly the correct classification of cervical precancerous lesions and/or cancer and may facilitate diagnosis and patient management.

Introduction: B lymphocyte, macrophage and mast cell densities in the stroma underlying cervical low and high grade squamous intraepithelial lesions (LSIL and HSIL) and squamous cell carcinoma (SCC) in 200 tissue samples, with concomitant HPV typing, was assessed, in order to examine their relationship to disease status and progression. Methods: Sections from 215 cervical specimens (149 LSIL, 38 HSIL, 13 SCC and 15 normal cervical mucosae) were immunostained for B lymphocytes (CD20), macrophages (CD68) and mast cells (CD117). The number of cells per high power field (henceforth called density) in the stroma underlying epithelial lesions was assessed. Statistical analysis was performed using four ordinal scale groups of increasing severity (normal, LSIL, HSIL and SCC). Results: Densities of all three cell types had a statistically significant, proportional correlation to disease severity, more so for B lymphocytes. Increased density of any cell type is linked to an increase in the densities of the remaining two. A statistically significant difference in B lymphocyte and mast cell density was found between LSIL and HSIL. No cell type density was found to be predictive of the outcome of LSIL. No relationship with HPV type was found. Discussion: Increased B lymphocyte and mast cell density in the stroma underlying cervical HSIL and SCC compared to LSIL suggests a possible relationship of both cell types to progression of cervical SIL. No predictive value of the density of any cell type was found concerning the outcome of LSIL. Immunohistochemistry may contribute to elucidate the relationship of local immunity effector cells to cervical epithelial lesions.

To investigate the potential value of tree classifiers for the triage of high-grade squamous intraepithelial lesions.The dataset comprised 808 histologically confirmed cases having a complete range of the cytologic sample assessments--liquid-based cytology, reflex human papillomavirus (HPV) DNA test, E6/E7 HPV mRNA test, and p16 immunocytochemical examinations. Data include 488 histologically negative (cervical intraepithelial neoplasia [CIN] 1 and below) or clinically negative cases and 320 with histologic diagnosis of CIN 2 or worse. Cytologic diagnosis was made according to the criteria of the Bethesda System. Cases were classified in two groups according to histology: those with CIN 2 or worse and those with CIN 1 and below. Fifty percent were randomly selected as a training set and the remaining were as a test set.Application of tree classifier on the test set gave correct classification of 66.9% for CIN 2 and above cases and 97.3% for CIN 1 and below, producing overall accuracy of 91.5%, outperforming cytologic diagnosis alone.Application of tree classifiers, based on standard cytologic diagnosis and expression of studied biomarkers, produces improved classification results for cervical precancerous lesions and cancer diagnosis and

Cervical cancer is a common cancer inflicting women worldwide. Even though, persistent infection with oncogenic Human Papillomavirus (HPV) types is considered the most important risk factor for cervical cancer development, less than 5% of women with HPV will eventually develop cervical cancer supporting that other molecular events, like methylation-dependent inactivation of tumor suppressor genes, may cocontribute in cervical carcinogenesis. We analyzed promoter methylation of three candidate genes (p16, MGMT, and hMLH1) in 403 liquid-based cytology samples. Methylation was commonly identified in both benign and pathologic samples and correlated with higher lesion grade determined by cytological, colposcopical, or histological findings, with HPV DNA and mRNA positivity of specific HPV types and p16INK4Aprotein expression. Overall accuracy of methylation is much lower than traditional diagnostic tests ranking it as an ancillary technique with more data needed to identify the exact value of methylation status in cervical carcinogenesis.

Simultaneous occurrence of papillary carcinoma and lymphoma in the thyroid is extremely rare, only three cases having so far been reported. In a 51-year-old woman with thyroid nodules, papillary carcinoma was diagnosed in an ultrasound-guided fine needle aspiration of a right lobe nodule. Subsequent thyroidectomy revealed a 1 cm large, Warthin-like papillary carcinoma (figure 1), whereas a 0.7 cm focus of classical type papillary carcinoma was found in the lower third of the right lobe. Hashimoto thyroiditis was also seen (figure 2). Moreover, a 1.2 cm area with scant thyroid follicles and large lymphoid follicles surrounded by small lymphocytes was found in the left lobe. The lymphoid infiltrate showed immunopositivity for both CD20 and CD43; …

ObjectiveHPV infection is a common finding, especially in young women while the majority of infections are cleared within a short time interval. The aim of this study was to examine the efficacy of HPV DNA and mRNA testing in a population attending colposcopy units of two University hospitals.Methods1173 liquid based cervical samples from two colposcopy clinics were tested for HPV DNA positivity using a commercial typing kit and HPV E6/E7 mRNA positivity with a flow cytometry based commercial kit. Statistic measures were calculated for both molecular tests and morphological cytology and colposcopy diagnosis according to histology results.ResultsHPV DNA, high-risk HPV DNA, HPV16 or 18 DNA and HPV mRNA was detected in 55.5%, 50.6%, 20.1% and 29.7% of the cervical smears respectively. Concordance between the DNA and the mRNA test was 71.6% with their differences being statistically significant. Both tests' positivity increased significantly as lesion grade progressed and both displayed higher positivity rates in samples from women under 30 years old. mRNA testing displayed similar NPV, slightly lower sensitivity but significantly higher specificity and PPV than DNA testing, except only when DNA positivity for either HPV16 or 18 was used.ConclusionsOverall mRNA testing displayed higher clinical efficacy than DNA testing, either when used as a reflex test or as an ancillary test combined with morphology. Due to enhanced specificity of mRNA testing and its comparable sensitivity in ages under 25 or 30 years old, induction of mRNA testing in young women could be feasible if a randomized trial verifies these results.

The molecular epidemiology of human papillomavirus (HPV) infection in a sample of Greek women (n = 2952, mean age 42.2 +/- 13.3 years) was examined. HPV prevalence was 50.7% (95% confidence interval, 48.8-52.6). The most frequent HPV types were HPV 53, 51 and 66 (10.2%, 9.4% and 9.3%, respectively). HPV positivity was associated with age, age of sexual debut, number of sexual partners and duration of sexual relationship, while marriage or multiparity protected against infection (all p

This study evaluated the accuracy and reproducibility of telecytological diagnoses proffered on the basis of digitized images from cervical smears prepared by means of liquid-based cytology.Representative digital cytological images from a total of 404 cervical smears (benign, 135; atypical squamous cells of undetermined significance, 92; low-grade squamous intraepithelial lesion, 62; high-grade squamous intraepithelial lesion, 87; squamous cell carcinoma, 26; and adenocarcinoma, 2) were uploaded to the CytoTrainer e-learning telecytology platform (developed in the Department of Cytopathology, "ATTIKON" University General Hospital, Athens, Greece) with specific password-protected accounts and were reviewed remotely by four independent board-certified cytopathologists (checking round). Their reports were recorded and classified. After 12 and 24 months, the same representative digital images were transferred in random order to the same cytopathologists and were reviewed again (first and second review rounds, respectively). The cytopathologists' first and second round diagnoses were recorded and compared with their initial ones.Statistical evaluation of cytological diagnoses detected no significant difference in diagnostic accuracy among checking and review diagnoses. The overall interobserver agreement was almost perfect with κ values of 0.79-0.97, whereas intraobserver agreement ranged from almost perfect to perfect with κ values of 0.76-1 in all diagnostic rounds.Digital images of cervical smears can be used for rapid and accurate diagnosis, by diminishing turnaround times and improving small cytology departments' quality indices. Diagnoses made by using static telecytological systems can be as reliable as those made by using conventional microscopy, under the conditions that representative images are taken and that standard cytological diagnostic criteria are applied. Telecytology can be used as an alternative method for the cytologic diagnosis of cervical smears, particularly in quality assurance programs.