Role of endoscopic ultrasound-guided fine needle aspiration biopsies in diagnosing pancreatic neoplasms in the paediatric population: experience from a tertiary center and review of the literature.

BACKGROUND: Endoscopic ultrasound-guided fine needle aspiration biopsy (EUS FNAB) is a well established diagnostic method in adult patients, but is rarely used in the paediatric population. The Clinical Department of Gastroenterology at the University Clinical Centre Ljubljana and the Department of Cytopathology at the Institute of Pathology, Faculty of Medicine, University of Ljubljana, Slovenia, have been closely collaborating on EUS FNAB since the introduction in 2010. The aim of the study was to review the cases of EUS FNAB of pancreatic neoplasms in children. PATIENTS AND METHODS: In the digital archive of the Institute of Pathology (IP), Faculty of Medicine (FM), University of Ljubljana (UL), we found 6 cases of EUS FNAB in children, 3 had EUS FNAB of the pancreas, 2 of whom had a cytopathologic diagnosis of a tumour. In the first case, the lesion was ultrasonographically solid, and the cell sample contained branching papillary structures surrounded by aggregates of small cells with nuclear grooves. In the second case, the lesion was ultrasonographically cystic, and predominantly necrosis was seen, with only single preserved cells. Positive nuclear reaction for ß-catenin was found in both cases by immunohistochemical staining. RESULTS: In both cases, the cytopathological diagnosis of solid pseudopapillary neoplasm of the pancreas was made, the cases represent the totality of paediatric cases of pancreatic neoplasms from the Children's Hospital Ljubljana since 2010. There were no adverse events during and after EUS FNAB. A histopathological examination of the tumour resection specimens confirmed the cytopathological diagnosis. CONCLUSIONS: Our experience indicates that EUS FNAB is a safe and effective method for diagnosing pancreatic neoplasms in the pediatric population, as supported by the findings in the literature.

Dose-Modifying Factor of Radiation Therapy with Concurrent Cisplatin Treatment in HPV-Positive Squamous Cell Carcinoma: A Preclinical Study.

Human papillomavirus (HPV) is an important etiological factor in oropharyngeal squamous cell carcinoma (SCC). Compared to HPV-negative tumors, HPV-positive oropharyngeal SCC has shown a better response to nonsurgical treatments. In this study, we determined the dose-modifying factors for HPV-positive tumors with single-dose irradiation, with or without low radiosensitizing doses of cisplatin. In vitro, we determined an increased radiosensitivity of HPV-positive SCC, which might be a consequence of HPV-induced changes in the cell cycle regulation and DNA damage response, leading to increased cell death. Additionally, compared to HPV-negative tumors, 30% higher radiosensitivity of HPV-positive tumors was determined by tumor growth delay monitoring in immunodeficient mice in vivo. Concurrent cisplatin treatment had an additive effect in both HPV-negative and HPV-positive tumors, resulting in 20% better response in HPV-positive tumors than in HPV-negative tumors.

Microinvasive cervical squamous cell carcinoma in Slovenia during the period 2001-2007.

BACKGROUND: Microinvasive squamous cell carcinoma (MISCC) comprises a significant portion of all cervical cancers in Slovenia. Criteria of carcinomatous invasion are well described in the literature, however histopathological assessment of MISCC is difficult, because morphological characteristics can overlap with cervical intraepithelial neoplasia grade 3 (CIN 3) and other pathological changes. The aim of our study was to evaluate the reliability of the histopathological diagnosis of MISCC in Slovenia during the period from 2001 to 2007. MATERIALS AND METHODS: Data on patients with a histopathological diagnosis of cervical MISCC (FIGO stage IA) in the period of 2001 to 2007 were obtained from the Cancer Registry of Slovenia. Histological slides were obtained from the majority of pathology laboratories in Slovenia. We received 250 cases (69% of all MISCC) for the review; 30 control cases with CIN 3 and invasive squamous cell carcinoma FIGO stage IB were intermixed. The slides were coded and reviewed. RESULTS: Among 250 cases originally diagnosed as MISCC, there was an agreement with MISCC diagnosis in 184 (73.6%) cases (of these 179/184 (97.3%) cases were FIGO stage IA1 and 5/184 (2.7%) cases were FIGO stage IA2). Among 179 FIGO stage IA1 cases 117 (65.4%) showed only early stromal invasion. CONCLUSIONS: The retrospective review of cases diagnosed as MISCC during the period 2001-2007 in Slovenia showed a considerable number of overdiagnosed cases. Amongst cases with MISCC confirmed on review, there was a significant proportion with early stromal invasion (depth of invasion less than 1 mm).

Vegetable cells in urinary samples of patients with bricker ileal conduit.

During routine cytopathological evaluation of urines for malignant cells we have occasionally noticed vegetable cells that were only present in patients with Bricker ileal conduit. We wanted to identify the means and sources of contamination of urinary samples from these patients. During the period between May and November 2010, 637 urinary samples were routinely evaluated for malignant cells. Among them were 13 urinary samples from Bricker ileal conduit which we rescreened. We prepared all urinary samples by membrane filtration and stained them according to Papanicolaou. Subsequently, we prepared samples from ostomy adhesives made by Coloplast and by ConvaTec which are used to secure the ostomy bag onto urostomy. We also took samples from different constituents (hydrocolloids) of ostomy adhesives. On the cytopathological review, we found vegetable cells along with intestinal mucosa cells in urinary samples of seven patients with Bricker ileal conduit. With the light microscopic examination of the samples prepared from different ostomy adhesives, we found vegetable cells only in Coloplast adhesives. In preparations of hydrocolloids, we found vegetable cells only in guar gum. They were morphologically identical to those found in urine samples of patients with Bricker ileal conduit and in Sensura and Sensura Xpro (Coloplast) ostomy adhesives. We determined that the origin of vegetable cells in urines from Bricker ileal conduit is the ostomy adhesive. The vegetable cells differ from human intestinal epithelial cells regarding size, shape, and color so it is difficult to misinterpret them as dysplastic cells.

Impact of different sectioning of excision biopsies of the cervical transformation zone for detection of invasive cervical cancer in 2 European centers related to national cervical screening programs.

BACKGROUND: The impact of the number of sections used in cervical excision biopsies of transformation zone on sensitivity of histological detection of cervical cancer is poorly documented. AIM: To assess whether different techniques of sectioning cervical excision biopsies in 2 European laboratories influence the finding of cervical cancer by histopathological examination. MATERIALS AND METHODS: The routine assessment at the Department of Histopathology and Cytology, Sheffield Teaching Hospitals NHS Foundation Trust, Sheffield, UK (Sheffield), encompassed sectioning a cone in 3-mm tissue blocks and cutting 3 levels per block. At the Institute of Pathology, Faculty of Medicine, University of Ljubljana, Slovenia (Ljubljana), cones were cut in a 3- to 4-mm-thick tissue block, and 10 levels were cut per block, spaced every 100 microm. The number of blocks and levels per block were assessed for each cone. Histopathological detection of cervical carcinoma between the laboratories was compared. RESULTS: Among 820 cones in the Sheffield laboratory, we detected 35 invasive carcinomas, whereas 6 invasive carcinomas were detected among 94 cones in the Ljubljana laboratory. Although the Slovenian laboratory examined a significantly larger number of levels per cone biopsy (64 vs 24 in Sheffield), this was not associated with a larger proportion of invasive cervical carcinoma, especially foci of stage IA1 or high-grade cervical intraepithelial neoplasia diagnosed on excision biopsies: 6.4% of invasive cervical carcinomas (5 [83.3%] of stage IA1) in Ljubljana and 4.3% (30 [85.7%] of stage IA1) in Sheffield laboratory (P > 0.05); 71.2% of high-grade cervical intraepithelial neoplasia in Ljubljana and 75.5% in Sheffield (P > 0.05). CONCLUSIONS: Our assessment showed that a method with a large number of levels per cervical cone or large loop excision of the transformation zone biopsy did not increase sensitivity for the detection of cervical cancer. However, extensive sectioning substantially affects the pathologist's workload, and this may need to be reconsidered.

Urine and bladder washing cytology for detection of urothelial carcinoma: standard test with new possibilities.

BACKGROUND: Light microscopic evaluation of cell morphology in preparations from urine or bladder washing containing exfoliated cells is a standard and primary method for the detection of bladder cancer and also malignancy from other parts of the urinary tract. The cytopathologic examination is a valuable method to detect an early recurrence of malignancy or new primary carcinoma during the follow-up of patients after the treatment of bladder cancer. CONCLUSIONS: Characteristic cellular and nuclear signs of malignancy indicate invasive or in situ urothelial carcinoma or high-grade papillary urothelial carcinoma. However, low sensitivity of the method reflects the unreliable cytopathologic diagnosis of low-grade urothelial neoplasms as cellular and nuclear signs of malignancy in these neoplasms are poorly manifested. Many different markers were developed to improve the diagnosis of bladder carcinoma on urinary samples. UroVysion™ test is among the newest and most promising tests. By the method of in situ hybridization one can detect specific cytogenetic changes of urothelial carcinoma.

Radiosensitivity of squamous cell carcinoma metastases to the neck assessed by immunocytochemical profiling of fine-needle aspiration biopsy cell specimens: a pilot study.

PURPOSE: To assess radiosensitivity of neck metastases of squamous cell carcinoma of the head and neck (SCCHN) by immunocytochemical profiling of fine-needle aspiration biopsy (FNAB) cell specimens. PATIENTS AND METHODS: Immunocytochemical reactions to p53, cyclin D1, stefin A and Ki-67 were determined in FNAB cell samples of neck metastases from 21 patients treated with concomitant chemoradiotherapy and correlated to clinical characteristics and response to therapy. RESULTS: Six (28.6%), eight (38.1%), 15 (71.4%) and nine (42.9%) FNAB cell samples were classified as p53, cyclin D1, stefin A and Ki-67 positive, respectively. Statistically significant predictors of favorable nodal response to chemoradiation were p53 (P=0.025) and cyclin D1 (cytoplasmic fraction, P=0.048) negativity and Ki-67 positivity (P=0.045). Regional recurrence correlated with low Ki-67 immunoreactivity. A favorable profile of cyclin D1 and Ki-67 (one or both of the two) further improved the predictive strength of these markers. CONCLUSIONS: FNAB is a non-invasive, simple and cheap procedure, which could serve simultaneously for diagnostic purposes and for radiosensitivity testing. Immunocytochemical determination of cyclin D1 and Ki-67 in FNAB cell samples from neck metastases of SCCHN seems to be a valuable marker for predicting regional response to radiotherapy and might assist when deciding on appropriate primary therapy.

Image cytometric evaluation of nuclear texture features and DNA content of the reticular form of oral lichen planus.

OBJECTIVE: To analyze image cytometric chromatin changes reflected in nuclear texture features and DNA ploidy of oral lichen planus in relation to the normal buccal mucosa and buccal mucosa expressing malignancy-associated changes in cancer patients. STUDY DESIGN: Twenty-eight patients with the reticular form of oral lichen planus, with a follow-up period of 25 years, 50 healthy controls and 50 lung cancer patients were included in the study. Scrapings of buccal mucosa were suspended in transport medium. Monolayer filter preparations were Feulgen-thionin stained. Image cytometric analysis was performed by Cyto-Savant. RESULTS: All oral lichen planus specimens in our study were diploid. In univariate analysis, differences between the normal buccal mucosa and oral lichen planus were found in several nuclear texture features, which gave an 80% correct classification rate in multivariate analysis. In the second part of the study, the classifier that recognizes malignancy-associated changes on the buccal mucosa of patients with lung cancer correctly recognized > 80% of oral lichen planus samples as normal buccal mucosa. CONCLUSION: Our results indicate that chromatin changes in oral lichen planus exist compared to normal cells; however, the chromatin structure of the reticular form of oral lichen planus does not express malignancy-associated changes and is more similar to normal squamous cells.

Flow and image cytometric DNA ploidy, including 5c exceeding cells, of serous borderline malignant ovarian tumors. Correlation with clinicopathologic characteristics.

OBJECTIVE: To analyze DNA ploidy of serous borderline ovarian tumors by flow cytometry (FCM) and image cytometry (ICM), with 5c exceeding cells also analyzed, and to evaluate their correlation with clinicopathologic characteristics of patients and tumors. STUDY DESIGN: Cell suspensions were prepared according to a modified Hedley method from formalin-fixed, paraffin-embedded tissue blocks of 43 tumors. One part of the suspension was used for flow cytometric measurement; from the other part, filter slides were prepared for ICM. RESULTS: FCM and ICM found 2 aneuploid (peridiploid) serous borderline ovarian tumors, and FCM found 1. ICM found 3 tumors with 5c exceeding cells and 2 tumors with octaploid cells. There was no correlation between DNA aneuploidy and presence of 5c exceeding cells with tumor size, International Federation of Gynecology and Obstetrics stage or survival. CONCLUSION: The results confirm a good correlation between FCM and ICM DNA ploidy and the ability of ICM to detect 5c exceeding cells. The prognostic value of DNA ploidy and 5c exceeding cells in serous borderline malignant ovarian tumors warrant further evaluation.