Performance Evaluation of Manual and Automated (MagNA Pure) Nucleic Acid Isolation in HPV Detection and Genotyping Using Roche Linear Array HPV Test

Nucleic acids of human papillomavirus (HPV) isolated by manual extraction method (AmpliLute) and automated MagNA pure system were compared and evaluated with cytohistological findings in 253 women. The concordance level between AmpliLute and MagNA was very good 93.3% (κ = 0.864, P < .0001). Overall HPVpositivity detected by AmpliLute was 57.3% (30.4% as single and 27% as multiple infections) in contrast to MagNA 54.5% (32% and 23%, resp.). Discrepant results observed in 25 cases: 11 MagNA(−)/AmpliLute(+), 10 of which had positive histology; 5 MagNA(+)/AmpliLute(−) with negative histology; 8 MagNA(+)/AmpliLute(+): in 7 of which AmpliLute detected extra HPV genotypes and 1 MagNA(invalid)/AmpliLute(+) with positive histology. Both methods performed well when compared against cytological (area under curve (AUC) of AmpliLute 0.712 versus 0.672 of MagNA) and histological diagnoses (AUC of AmpliLute 0.935 versus 0.877 of MagNA), with AmpliLute showing a slightly predominance over MagNA. However, higher sensitivities, specificities, and positive/negative predictive values were obtained by AmpliLute

Radial Basis Function Artificial Neural Network for the Investigation of Thyroid Cytological Lesions

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

The role of biological markers in investigation of ductal carcinomas of the breast processes with liquid based cytology, ThinPrep

Introduction: Several studies suggest that assessment of biological markers ER, PgR, Ki-67, C-ERB2 and p53, as well as ploidy analysis can predict the biologic behavior of breast carcinomas. The aim of this study is the application of these techniques in FNA material, processed with liquid based cytology (ThinPrep®). Material and Methods: FNA was performed on female patients presenting with breast lumps at the outpatient clinic. DNA ploidy and immunocytochemical assessment of ER, PgR, Ki-67, C-ERB2 and p53 was performed, with the technique of image analysis. Results: In the present study a correlation was found between DNA ploidy and histologic grade of the breast carcinomas. A statistical difference was observed among DNA index, ploidy balance, degree of aneuploidy, proliferation index, degree of hyperploidy and histologic grade (p<0,05). Immunocytochemical investigation of breast carcinomas revealed a statistical difference among the parameters calculated with Image analysis, Label index and Quick score, and the histologic grade (p<0,05). Discussion: DNA ploidy and immunocytochemical evaluation of biologic markers with the technique of Image Analysis could be appropriately applied to FNA material, processed with the ThinPrep® method. Investigation of breast carcinomas with these techniques can provide additional information concerning the biologic behavior of the examined carcinomas.Εισαγωγή: Σύμφωνα με τα δεδομένα της βιβλιογραφίας η σύγχρονη εκτίμηση πολλαπλών βιολογικών παραμέτρων, συμπεριλαμβανομένων των ER, PgR, Ki- 67, C-ERB2 και p53, καθώς και η ανάλυση της πλοειδίας, είναι δυνατό να καθορίσουν την βιολογική συμπεριφορά των καρκινωμάτων του μαστού. Σκοπός της μελέτης μας είναι η εφαρμογή των τεχνικών αυτών σε υλικό παρακέντησης δια λεπτής βελόνης (FNA), τα οποία έχουν παρασκευαστεί με την μέθοδο κυτταρολογίας υγρής φάσης ThinPrep®. Υλικό και μέθοδος: Πραγματοποιήθηκε παρακέντηση με λεπτή βελόνη (FNA) σε γυναίκες ασθενείς που προσήλθαν στο νοσοκομείο με ογκίδιο μαστού. Ακολούθησε ανάλυση της πλοειδίας του DNA και ανοσοκυτταροχημική διερεύνηση των νεοπλασμάτων του μαστού ως προς τους υποδοχείς ER, PR και τα αντισώματα έναντι Κi-67, p53 και C-erbB2, με την τεχνική ανοσοκυτταροχημείας με ανάλυση εικόνας. Αποτελέσματα: Στην παρούσα μελέτη παρατηρήθηκε συσχέτιση της πλοειδίας του όγκου με τον ιστολογικό βαθμό διαφοροποίησης. Παρατηρήθηκε στατιστικά σημαντική διαφορά του δείκτη DNA, της διακυμάνσεως της πλοειδίας, του βαθμού ανευπλοειδίας, του δείκτη πολλαπλασιασμού και του βαθμού υπερπλοειδίας μεταξύ καρκινωμάτων βαθμού κακοήθειας I, II και III (p<0,05). Κατά την ανοσοκυτταροχημική διερεύνηση των καρκινωμάτων παρατηρήθηκε στατιστικά σημαντική διαφορά του ποσοστού θετικής ανοσοχρώσης και της παραμέτρου Quick score ως προς στους οιστρογονικούς και προγεστερονικούς υποδοχείς, καθώς και των δεικτών Κi-67 και του p53 μεταξύ των καρκινωμάτων ιστολογικού βαθμού κακοήθειας I, II και ΙΙΙ (p<0,05). Συμπεράσματα: Με την εφαρμογή της κυτταρολογίας υγρής φάσης, ThinPrep®, είναι δυνατή η ανάλυση της πλοειδίας του DNA, καθώς και η ανοσοκυτταροχημική διερεύνηση των νεοπλασμάτων του μαστού με την τεχνική ανοσοκυτταροχημείας με ανάλυση εικόνας. Η διερεύνηση της πλοειδίας, καθώς και η εκτίμηση των παραμέτρων της θετικής ανοσοχρώσης και Quick Score των βιολογικών δεικτών που μελετήθηκαν, με την τεχνική DNA image analysis, μπορούν να προσφέρουν επιπρόσθετες πληροφορίες σχετικά με την βιολογική συμπεριφορά των καρκινωμάτων του μαστού

An Adolescent With a Rare Midline Neck Tumor: Thyroid Carcinoma in a Thyroglossal Duct Cyst

Occurrence of malignancy in a thyroglossal duct cyst (TDC) in children and adolescents is very rare, preoperative diagnosis is a challenge, and appropriate management is still debated. We report a 19-year-old male patient referred for a midline neck mass diagnosed as an atypical TDC after initial and subsequent investigations. Ultrasound-guided fine-needle aspiration (FNA) of the mass was diagnostic for papillary thyroid carcinoma. Sistrunk procedure and total thyroidectomy were performed. Histologic analysis confirmed the presence of papillary thyroid carcinoma within TDC, infiltrating surrounding soft tissues. Postoperatively, radioiodine ablation treatment was administered, followed by TSH suppression therapy

Artificial Neural Networks as Decision Support Tools in Cytopathology: Past, Present, and Future.

OBJECTIVE: This study aims to analyze the role of artificial neural networks (ANNs) in cytopathology. More specifically, it aims to highlight the importance of employing ANNs in existing and future applications and in identifying unexplored or poorly explored research topics. STUDY DESIGN: A systematic search was conducted in scientific databases for articles related to cytopathology and ANNs with respect to anatomical places of the human body where cytopathology is performed. For each anatomic system/organ, the major outcomes described in the scientific literature are presented and the most important aspects are highlighted. RESULTS: The vast majority of ANN applications are related to cervical cytopathology, specifically for the ANN-based, semiautomated commercial diagnostic system PAPNET. For cervical cytopathology, there is a plethora of studies relevant to the diagnostic accuracy; in addition, there are also efforts evaluating cost-effectiveness and applications on primary, secondary, or hybrid screening. For the rest of the anatomical sites, such as the gastrointestinal system, thyroid gland, urinary tract, and breast, there are significantly less efforts relevant to the application of ANNs. Additionally, there are still anatomical systems for which ANNs have never been applied on their cytological material. CONCLUSIONS: Cytopathology is an ideal discipline to apply ANNs. In general, diagnosis is performed by experts via the light microscope. However, this approach introduces subjectivity, because this is not a universal and objective measurement process. This has resulted in the existence of a gray zone between normal and pathological cases. From the analysis of related articles, it is obvious that there is a need to perform more thorough analyses, using extensive number of cases and particularly for the nonexplored organs. Efforts to apply such systems within the laboratory test environment are required for their future uptake

Artificial Neural Networks as Decision Support Tools in Cytopathology: Past, Present, and Future

Objective This study aims to analyze the role of artificial neural networks (ANNs) in cytopathology. More specifically, it aims to highlight the importance of employing ANNs in existing and future applications and in identifying unexplored or poorly explored research topics. Study Design A systematic search was conducted in scientific databases for articles related to cytopathology and ANNs with respect to anatomical places of the human body where cytopathology is performed. For each anatomic system/organ, the major outcomes described in the scientific literature are presented and the most important aspects are highlighted. Results The vast majority of ANN applications are related to cervical cytopathology, specifically for the ANN-based, semiautomated commercial diagnostic system PAPNET. For cervical cytopathology, there is a plethora of studies relevant to the diagnostic accuracy; in addition, there are also efforts evaluating cost-effectiveness and applications on primary, secondary, or hybrid screening. For the rest of the anatomical sites, such as the gastrointestinal system, thyroid gland, urinary tract, and breast, there are significantly less efforts relevant to the application of ANNs. Additionally, there are still anatomical systems for which ANNs have never been applied on their cytological material. Conclusions Cytopathology is an ideal discipline to apply ANNs. In general, diagnosis is performed by experts via the light microscope. However, this approach introduces subjectivity, because this is not a universal and objective measurement process. This has resulted in the existence of a gray zone between normal and pathological cases. From the analysis of related articles, it is obvious that there is a need to perform more thorough analyses, using extensive number of cases and particularly for the nonexplored organs. Efforts to apply such systems within the laboratory test environment are required for their future uptake

Endocervical Carcinogenesis and HPV Vaccination: An Occasional Circumstance or a Gap in the Chain?

As a result of the Human Papillomavirus (HPV) vaccination program, the prevalence of precancerous dysplasia and invasive cervical cancer has substantially decreased. In this brief report, we present a case of a young patient who was diagnosed with in situ adenocarcinoma of the cervix. This 30-year-old female had completed the HPV vaccination after she became sexually active and has been undergoing annual gynecological assessments, including clinical examination and Pap test, all of which had been negative. This year, her Pap test revealed a low grade squamous intraepithelial lesion (LGSIL) and additionally a colposcopy was performed. Given the extent of the lesion and since the colposcopy was inadequate, the patient underwent a type 3 large loop excision of the transformation zone and a curettage of the endocervix under local anesthesia. The pathological diagnosis from cervical biopsy revealed an in situ adenocarcinoma of the endocervix with negative limits. The HPV subtypes 16 and 83 were detected with PCR. After proper consultation she decided to preserve her fertility and to undergo a regular follow-up, postponing hysterectomy after the completion of her family planning. In conclusion, this case report highlights the need for diagnostic surveillance regarding HPV-related cervical cancer even after vaccination

An Intelligent Clinical Decision Support System for Patient-Specific Predictions to Improve Cervical Intraepithelial Neoplasia Detection

Nowadays, there are molecular biology techniques providing information related to cervical cancer and its cause: the human Papillomavirus (HPV), including DNA microarrays identifying HPV subtypes, mRNA techniques such as nucleic acid based amplification or flow cytometry identifying E6/E7 oncogenes, and immunocytochemistry techniques such as overexpression of p16. Each one of these techniques has its own performance, limitations and advantages, thus a combinatorial approach via computational intelligence methods could exploit the benefits of each method and produce more accurate results. In this article we propose a clinical decision support system (CDSS), composed by artificial neural networks, intelligently combining the results of classic and ancillary techniques for diagnostic accuracy improvement. We evaluated this method on 740 cases with complete series of cytological assessment, molecular tests, and colposcopy examination. The CDSS demonstrated high sensitivity (89.4%), high specificity (97.1%), high positive predictive value (89.4%), and high negative predictive value (97.1%), for detecting cervical intraepithelial neoplasia grade 2 or worse (CIN2+). In comparison to the tests involved in this study and their combinations, the CDSS produced the most balanced results in terms of sensitivity, specificity, PPV, and NPV. The proposed system may reduce the referral rate for colposcopy and guide personalised management and therapeutic interventions

A Pyrosequencing Assay for the Quantitative Methylation Analysis of GALR1 in Endometrial Samples: Preliminary Results

Endometrial cancer is the most common malignancy of the female genital tract while aberrant DNA methylation seems to play a critical role in endometrial carcinogenesis. Galanin’s expression has been involved in many cancers. We developed a new pyrosequencing assay that quantifies DNA methylation of galanin’s receptor-1 (GALR1). In this study, the preliminary results indicate that pyrosequencing methylation analysis of GALR1 promoter can be a useful ancillary marker to cytology as the histological status can successfully predict. This marker has the potential to lead towards better management of women with endometrial lesions and eventually reduce unnecessary interventions. In addition it can provide early warning for women with negative cytological result