Hybrid capture vs. PCR screening of cervical human papilloma virus infections. Cytological and histological associations in 1270 women

<p>Abstract</p> <p>Background</p> <p>We evaluated two molecular methods of HPV detection and their correlation with cytological and histological diagnosis in a large sample of Greek women.</p> <p>Methods</p> <p>All women with liquid-based cytology performed at a University Hospital between 2000 and 2003 were included. The Hybrid Capture 2 (HC2) kit and in house Polymerase Chain Reaction (PCR) were used for HPV DNA detection. Cervical biopsy was performed for women with ASCUS+ cytology, HPV detection, or abnormal colposcopy. Positive (PLR) and negative (NLR) likelihood ratios were calculated for cytology and HPV molecular testing for the prediction of CIN2 and greater histology.</p> <p>Results</p> <p>Of the 1270 women evaluated 241 (18.5%) had abnormal cytology. Cytology diagnosed high-grade squamous intraepithelial lesion (HSIL) or invasive carcinoma in 21(1.7%) cases whereas 26 (2%) women had CIN2+ or greater histology. PCR detected HPV in 397/1270 (31.3%) and HC2 in 260/1270 (20.4%) samples. Both molecular tests exhibited high reproducibility (Cohen's kappa value 0.691, 95% CI: 0.664 - 0.718). Positive likelihood ratios (PLR) of 9.4, 3.8 and 3.4 and negative likelihood ratios of 0.13, 0.21, and 0 were noted for ≥ LSIL, any positive HC2 or any positive PCR-HPV testing, for predicting CIN2+ histology, respectively. All CIN 3+ lesions harbored high risk oncogenic HPV type infections.</p> <p>Conclusions</p> <p>HPV infection was found in a large proportion of this population and was associated with CIN 2/3 lesions and infiltrating carcinomas. Thin prep testing and HPV detection by HC2 or PCR performed very well with regards to identifying high grade lesions in an environment with experienced examiners.</p

DNA flow-cytometric, histological and hormonal analysis of sertoli cell only syndrome (SECOS)

Sertoli cell only syndrome (SECOS) was identified on histology in 21 cases (16,28%) among 129 testicular biopsies performed in our department for azoospermia over the last 5 years. In these patients history, clinical features, hormonal levels, and histological findings were analyzed. In addition DNA flow-cytometric analysis was performed and showed an almost complete absence of haploid cells. All patients presented with elevated serum FSH levels suggesting a Sertoli cell damage or reduced production of inhibin due to the absence of sermatogenic cells. An good correlation was found between histological findings and DNA histograms. In conclusion SECOS is a syndrome of unknown aetiology presenting in men with azoospermia. DNA flow-cytometric analysis is a reliable, rapid and easy method in the diagnosis of SECOS, and can replace histological examination

Deoxyribonucleic acid measurements in transitional cell carcinomas: Comparison of flow and image cytometry techniques

Purpose: We compared the results of deoxyribonucleic acid (DNA) ploidy determinations performed by flow cytometry and image cytometry in transitional cell carcinomas of the bladder. Materials and Methods: In 81 cases of transitional cell carcinoma of the bladder DNA indexes were measured by flow cytometry of bladder washings and tumor tissue samples, and by image cytometry of imprints from tumor tissue samples. Results: There was good correlation between bladder washings and tissue samples analyzed by flow cytometry in 68 cases but aneuploidy was missed with bladder washings in 13 (16%). There was also good correlation between flow and image cytometry in 75 cases (92.59%) regarding the detection of aneuploidy. There was agreement between detection of aneuploidy and DNA index in 45 cases (55.5%), while both methods detected an aneuploid population in 30 (37%) but there was disagreement regarding DNA index. Aneuploid populations were missed by flow cytometry in 6 cases (7.4%). Furthermore, in 10 cases peridiploid peaks were found on the image cytometry histograms, which were not visible on flow cytometry. However, it was not possible to assess accurately if these were true peridiploid populations. Conclusions: There is good overall correlation between DNA content measured by flow and image cytometry but image cytometry has the advantage of visual discrimination, permitting preferential selection and analysis of tumor cells. However, certain problems remain with image cytometry, particularly in the case of peridiploid peaks, which cannot be classified accurately as showing true peridiploid or right shifted diploid populations

Discrimination of benign from malignant breast lesions using statistical classifiers

The objective of this study is to investigate the discrimination of benign from malignant breast lesions using: the linear, the feedforward neural network, the k-nearest neighbor and the boosting classifiers. Nuclear morphometric parameters from cytological smears taken by Fine Needle Aspiration (FNA) of the breast, have been measured from 193 patients. These parameters undergo an appropriate transformation and then, the classifiers are performed on the raw and on the transformed data. The results show that in terms of the raw data set all classifiers exhibit almost the same performance (overall accuracy ≡ 87%), Thus the linear classifier suffices for the discrimination of the present problem, Also, baaed on the previous results, one can conjecture that the use of these classifiers combined with image inorphometry and statistical techniques for feature transformation, may offer useful information towards the improvement of the diagnostic accuracy of breast FNA. © Springer-Verlag Berlin Heidelberg 2006

Deoxyribonucleic acid flow cytometry in the assessment of spermatogenesis

Purpose: We compared deoxyribonucleic acid (DNA) flow cytometric analysis of testicular tissue to quantitative assessment of spermatogenesis. Materials and Methods: We studied 35 infertile men with azoospermia or oligospermia. All patients underwent incisional testicular biopsies. DNA flow cytometric analysis was performed on each specimen to evaluate the ability of the method to quantify alterations in spermatogenesis. The results were compared to quantitative histological examination. At least 100 spermatic tubules were examined on each specimen and the number of spermatids per tubule was counted. All histological specimens were examined by the same pathologist. Results: Of the 35 specimens analyzed with DNA flow cytometry 5 were normal, while the percentage of haploid cells (spermatids and spermatozoa) was decreased (hypospermatogenesis) in 14, complete maturation arrest was noted in 2 and almost complete absence of haploid cells was found in 14. Comparing the findings on histological examination with histograms, excellent correlation was noted in cases of the Sertoli-cell-only syndrome and complete maturation arrest, while 3 of 14 histograms with hypospermatogenesis demonstrated normal spermatogenesis on histological examination. Additionally 1 of 5 histograms with normal spermatogenesis demonstrated hypospermatogenesis on histological examination. Conclusions: DNA flow cytometry of the testicular tissue seems to be an objective and quantified method that can be used to investigate spermatogenesis in infertile men. It is also less time-consuming than any histological examination, permits management decisions within 1.5 hours after biopsy and may replace testicular histopathological study. Flow cytometric diagnoses correlated well with histopathological findings

DNA ploidy as a prognostic factor in muscle invasive transitional cell carcinoma of the bladder

Radical cystectomy represents the treatment of choice for muscle-infiltrative bladder carcinoma; however, about 50% of patients relapse and die from the disease. In the present study, the prognostic significance of the DNA ploidy in transitional cell carcinoma of the urinary bladder (TCCB) is analyzed. The study was carried out on 66 patients with TCCB who underwent radical cystectomy. DNA ploidy was determined by flow cytometry (FCM) on paraffin-embedded specimens, and the results were analyzed and correlated with the tumor malignancy grade and stage and the clinical course. Forty of the 66 tumors studied (63%) were aneuploid. Aneuploid status was correlated with higher tumor T stage (P &lt; 0.001) and grade (P &lt; 0.001). Median follow up was 68 months (range: 12-105). Median survival was significantly longer in patients with diploid tumors (&gt; 60 vs 45 months, P &lt; 0.001). All patients with diploid tumors were alive and free of bladder cancer during follow-up, in contrast to only 30% of patients with aneuploid tumors. DNA ploidy was an independent prognostic factor, as shown by multivariate analysis (P=0.006). All patients with pT &gt;= 3b and diploid tumors were alive at the time of analysis as opposed to none with aneuploid tumors. The results of this study suggest that DNA ploidy can provide prognostic information on patients with muscle invasive carcinoma of the bladder and might represent a means of selection for postoperative management

The role of liquid-based cytology in the investigation of thyroid lesions

Objective: This study investigates the role of liquid-based cytology by ThinPrep® technique in the detection of thyroid lesions. Methods: In all, 252 specimens from 157 patients for pre-operative evaluation of thyroid nodules, prepared by the ThinPrep®, were examined. In all cases thyroidectomy followed the initial cytological evaluation. All cytological diagnoses were correlated to the histological ones. Results: According to our findings, a sensitivity of 87.80%, a specificity of 99.50%, a positive predictive value of 97.30%, a negative predictive value of 97.56% and an overall accuracy of 97.52% were observed in fine needle aspiration cytology in correlation to the histological diagnosis after thyroidectomy. Conclusions: ThinPrep® technique is a valid method for the pre-operative cytological diagnosis of thyroid nodules, offering the possibility of ancillary techniques, such as immunocytochemical and molecular methods and can, therefore, be potentially complementary to histological evaluation for further investigation of follicular lesions. © 2007 The Authors

Report of Sparicotyle (Microcotyle) chrysophrii Van Beneden and Hesse 1863, Atrispinum seminalis Euzet and Maillard 1973 and Polylabris tubicirrus Paperna and Kohn 1964 (Monogenea) on captive sea bream (Sparus aurata) and sharp snout sea bream (Diplodus puntazzo) in coastal Greece and Italy

During the years 2000-2003, caged gilthead sea bream (Sparus aurata L.) and sharp snout sea bream (Diplodus puntazzo C.) from two farms in Greece and two farms in Italy (the Central Thyrrenian Sea) were examined. Sparus aurata from Greece was moderately infected with Furnestinia echeneis and Sparicotyle chrysophrii. Diplodus puntazzo from the Greek farms were infected with three polyopisthocotylean monogeneans: Atrispinum salpae, A. seminalis and S. chrysophrii. In Italy, on the gills of cultivated S. aurata, a polyopisthocotylean parasite was identified as Polylabris tubicirrus which is considered host specific and only affecting breams of the genus Diplodus. Furthermore, D. puntazzo and S. aurata showed the transfer of the monogenean S. chrysophrii from S. aurata (its type host) to D. puntazzo which is never found infected in wild conditions by this parasite. The presence of these parasites, and in particular S. chrysophrii in cultured D. puntazzo, is an important finding for aquaculturists in terms of potential disease problems

Study on the reproducibility of thyroid lesions telecytology diagnoses based upon digitized images

The objective of this study is to evaluate the reproducibility and usefulness of telecytology diagnoses proffered on the basis of digitized images from fine-needle aspiration specimens prepared by means of liquid-based cytology. Representative digital cytological images from a total of 270 thyroid fine-needle aspiration specimens were transferred via file transfer protocol to specific password-protected accounts and were remotely reviewed by five independent board-certified cytopathologists (initial round). Their reports were recorded and classified. After 6 and 12 months, the same representative digital images were transferred in random order to the same cytopathologists and were reviewed again (first and second review rounds). The cytopathologists&apos; 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 initial and review diagnoses. The overall interobserver agreement was almost perfect with K values of 0.869-0.939, whereas intraobserver agreement ranged from almost perfect to perfect with K values of 0.967-1 in all diagnostic rounds. Digitized cytological images transmission and remote evaluation allows reproducible diagnosis of thyroid gland lesions. Diagnoses made by using static telecytology systems can be equally reliable to those made by using conventional microscopy, provided that representative images are utilized and that the standard cytological diagnostic criteria are applied. Diagn. Cytopathol. 2010. © 2010 Wiley-Liss, Inc