Investigating The Frequency of Serrated Polyps/Adenomas and Their Subtypes in Colonic Polyp Samples.

BACKGROUND: The purpose of this study was to determine the frequency of Serrated polyps of colonic polyps samples in Hazrat_e Rasoul_e Akram Hospital over ten years. MATERIALS: The target group in this study was patients with colonic polyps in Hazrat_e Rasoul_e Akram Hospital. Pathologic evaluation of these patients was done. Serrated polyps, by location, gender, age and type of polyps were divided and frequency of them were determined separately. RESULTS: Of 381 patients studied, 224 (58.79%) and 157(41.20%) were males and females, respectively. Mean age of patients was 59.25 years. In initial diagnosis, frequency of Adenomatous polyp, Hyperplastic polyp and Mixed polyp were 92.44% and 5.33%, and 2.22%, respectively. In final diagnosis (Second evaluation), frequency of Adenomatous polyp, Hyperplastic polyp, Mixed polyp, Sessile Serrated Adenoma/ Polyp, Traditional Serrated Adenoma and SPU (Serrated Polyp Unclassifiable) were 90.44%, 4.88%, 2.44%, 1.11%, 0.66% and 0.44%, respectively. 72.13% and 27.86% of polyps were low grade dysplasia and high grade dysplasia, respectively. According to the results of this study, the incidence of all types of polyps detected was more in men than women. Rectum and sigmoid were most abundant in the area polyp in both initial and final diagnosis. CONCLUSION: Despite the low prevalence of Serrated polyps in patients, early diagnosis is the best action to reduce morbidity and mortality. Probability of the risk of progression from low grade to high grade dysplasia and transforming into Adenocarcinoma is high in Serrated polyps.

Electronic expert consultation using digital still images for evaluation of atypical small acinar proliferations of the prostate: a comparison with immunohistochemistry.

This study was performed on a series of prostate needle biopsies with diagnosis of atypical small acinar proliferation (ASAP) to verify to what extent the application of immunohistochemistry (IHC) for p504s and p63 markers as well as expert consultation by still images could affect the diagnosis. The results of these 2 methods were compared. Immunohistochemistry staining for p504s and p63 was performed on sections from 42 patients with a primary diagnosis of ASAP. Meanwhile, digital still images were taken from hematoxylin and eosin-stained slides of cases and were sent to an expert uropathologist, blind to IHC staining interpretations. The results of IHC staining were compared with diagnostic interpretations of the consultant pathologist. In 13 cases, the focus of concern was not detectable on IHC slides. In the remaining 29 cases, IHC showed a benign and malignant expression pattern in 17 and 9 patients, respectively. In 3 cases, IHC findings were inconclusive and retained the diagnosis of ASAP. The consultant pathologist diagnosed 11 cases of benign and 7 cases of malignant processes. He retained the diagnosis of ASAP in 11 cases. There was high concordance between the results of IHC and electronic consultation in the group of benign cases. All 11 cases with the diagnosis of benignancy by electronic consultation showed a benign IHC pattern. Among 7 cases with the diagnosis of malignancy by the consultant pathologist, 5 were classified as malignant, 1 as benign, and 1 as inconclusive IHC groups. Considering problems with IHC staining of prostate needle biopsy, including loss of focus of interest, expert consultation using still images can provide very useful diagnostic information. This approach can be used as an adjunct to other diagnostic activities like IHC or even as an independent source of information to reach more accurate diagnoses in ASAP cases, particularly in institutions with limited resources.

Breast pathology after cryotherapy. Histological regression of breast cancer after cryotherapy.

A breast saving treatment is contemporary the preferred method of treatment with comparable results in comparing with mastectomy. In this study were evaluated the effects of cryotherapy by histological verification of changes in post treatment resection specimens. Fifty-three patients in age of 38-81 year with histologically confirmed breast cancer in needle biopsies were managed by cryotherapy between 1999 and 2007. The patients were operated between day 1 and 35 after cryotherapy. The histologic examination of operation materials showed in all cases at least partial tumor destruction. In general in 54.7% of all handled cases (29 patient) there was no residual tumor. In 6 cases (22.2%) from group 1 and in 23 cases (88.5%) of group 2 no tumor rest was found. Cryotherapy can lead to complete destruction of tumoral tissue. In our study all 29 (54.7%) of tumor-free cases after cryotherapy were those with cT1 stage. The experience of operator and the correct selection of appropriate patients (primarily taking the tumor size into account) play the most important role for achieving the best results.

The diagnostic value of cytokeratin 5/6, 14, 17, and 18 expression in human non-small cell lung cancer.

OBJECTIVES: The expression patterns of cytokeratin (CK) filaments in human epithelial neoplasms are complex and distinctive. The aims of this study were to analyze CK expression and to evaluate the diagnostic application of CKs in human non-small cell lung cancer (NSCLC). METHODS: mRNA expression of CK5, CK6, CK14, CK15, CK17, and CK19 was analyzed by Northern blotting. Protein expression of CK5/6, CK7, CK14, CK17, and CK18 was evaluated by immunohistochemistry on tissue microarrays. RESULTS: Northern blotting showed that CKs were highly expressed in human bronchial epithelial cells and/or small airway epithelial cells. In NSCLC cell lines, the expression pattern of CKs was heterogeneous. Regarding protein expression of CKs in 95 primary lung tumors, expression of CK5/6, CK14, and CK17 proteins was increased in squamous cell carcinomas compared to adenocarcinomas (ADC; p = 0.001, p = 0.030, and p = 0.001, respectively), and higher expression was significantly associated with lower grading (p = 0.006, p = 0.002, and p = 0.001, respectively), while increased expression of CK7 and CK18 was observed in ADC (p = 0.001, respectively). CONCLUSIONS: Our data suggest that CK5/6, CK7, CK14, CK17, and CK18 have diagnostic value in the subclassification of NSCLC.

Morphologically 'invisible' proinsulin - secreting adenoma detected by Ga-68 Exendin-4 (GLP-1 Receptor) positron emission tomography/CT.

This case report of a young man suffering from recurring hypoglycaemia illustrates a rare condition of a neuroendocrine tumour, predominantly secreting proinsulin and invisible to conventional imaging approaches. Only a GLP-1 receptor PET/CT using Exendin-4 visualized the pancreatic lesion and enabled curative therapy, confirming the diagnostic value of this tracer for detection of neuroendocrine tumours. As only few publications on this topic are available, an overview of the available data is also given. The known cut-off value of 60% for proinsulin level indicating malignancy is critically discussed.

EGFR immunohistochemistry as biomarker for antibody-based therapy of squamous NSCLC - Experience from the first ring trial of the German Quality Assurance Initiative for Pathology (QuIP®).

BACKGROUND: EGFR and its downstream signaling pathway are important targets for cancer therapy. Recently, the monoclonal anti-EGFR antibody Necitumumab in combination with gemcitabine and cisplatin was approved (EMA/14106/2016) for first-line treatment of squamous non-small cell carcinoma (SqNSCLC). Eligibility was restricted to cases with positive EGFR expression. In this context, a ring trial of the Quality Assurance Initiative for Pathology (QuIP®) was launched to prepare the German pathology community for a reliable and reproducible, immunohistochemically based biomarker test. MATERIALS AND METHODS: The trial was set up by a three-step approach. Two lead institutes were nominated to organize the trial process and to select appropriate cancer samples. These were first tested by the H-score (range 0-300) to identify positive and negative cases. Seven additional pathology institutes with experience in EGFR immunohistochemistry each tested the selected panel of identical cases (internal ring trial) to confirm the suitability of samples and scoring criteria. Then the open ring trial for all institutes of pathology in German speaking countries was announced. RESULTS: For the internal trial 8 EGFR-positive and 2 negative lung sqNSCLC samples were selected. A cut-off value of cell membranous staining in≥1% of tumor cells was introduced to define a case as EGFR negative or positive. Two points were attainable per correctly assessed sample leading to a maximum of 20 points,≥18 points were required for a successful participation. All 7 panel institute passed this barrier, 5 with the maximum of 20 points and two with one error (18 points) being related to one case with incorrect interpretation of cytoplasmic versus membranous staining and one case with an H-score of 2 as being considered EGFR positive. A second cut-off value (H-score≥3) was therefore introduced. In the open ring trial, 34 institutions participated of which 28 were successful according to the above criteria. The trial revealed a high reproducibility despite the use of different EGFR antibodies and detection systems. There was no association between technical parameters and trial failure. Again, one participant misinterpreted the subcellular EGFR localization. CONCLUSIONS: The first nationwide ring test for determination of EGFR IHC expression in sqNSCLC could be successfully performed in a very tight time frame. By this, the national pathology community was prepared to incorporate this marker in the panel of predictive cancer tests in a quality assessed manner and to initiate and accompany future studies on EGFR pathway pathology.

Systematic evaluation of the biological variance within the Raman based colorectal tissue diagnostics.

Being among the most common cancers worldwide screening and early diagnosis of colorectal cancer is of high interest for the health system, the patients and for research. Raman microspectroscopy as a label-free, non-invasive and non-destructive technique is a promising tool for an early diagnosis. However, to ensure a reliable diagnosis specially designed statistical analysis workflows are required. Several statistical approaches have been introduced leading to varying results in the overall accuracy, sensitivity and specificity. In this study a systematic evaluation of different statistical analysis approaches has been performed using a colon cancer mouse model with genotypic identical individuals. Based on the inter-individual Raman spectral variances a measure for the biological variance can be estimated. By applying a leave-one-individual-out cross-validation a clinically relevant discrimination of healthy tissue versus adenoma and carcinoma with an accuracy of 95% is shown. Furthermore, the transfer of a model from tissue to biopsy specimen is demonstrated.

Expression of desmogleins 1-3 and their clinical impacts on human lung cancer.

AIMS: Desmogleins (DSGs) are components of the cell-cell connecting desmosomes. Desmosomal proteins have been found dysregulated in various cancers. Here we studied the role of DSGs in human lung cancer. METHODS: Expression of DSG1-3 mRNA in lung cancer cell lines and human bronchial epithelial cells (HBEC) was examined by real time RT-PCR. Methylation status of DSG1-2 was evaluated by demethylation test and bisulfite sequencing (BS). Moreover, DSG1-3 protein expression was analysed in 112 primary lung tumor samples by immunohistochemistry (IHC) on tissue microarrays. RESULTS: It turned out that DSG1-3 was downregulated in most of the lung cancer cell lines. Reexpression of DSG2 and DSG3 was found in several cancer cell lines after demethylation treatment with 5-aza-2'-deoxycytidine (DAC), a DNA methyltransferase inhibitor. Complete or partial methylation of DSG2 promoter region was detected in 5 out of 6 cancer cell lines by BS. In primary lung tumors, higher protein expression of DSG2 and DSG3 correlated to the diagnosis of squamous cell lung carcinoma (SCC) (P=0.009 and P<0.001, respectively), additionally, a lower expression of DSG3 was significantly linked to higher tumor grade (P=0.012). CONCLUSIONS: Our data suggest that downregulation of DSG2 and DSG3 could be partially explained by DNA methylation. DSG2 and DSG3 might be potential diagnostic markers for SCC, and DSG3 could be a potential differentiation marker for lung cancer.

Teleconsultation in diagnostic pathology: experience from Iran and Germany with the use of two European telepathology servers.

Eighty pathology cases were sent independently to each of two telepathology servers. Cases were submitted from the Department of Pathology at the University of Kerman in Iran (40 cases) and from the Institute of Pathology in Berlin, Germany (40 cases). The telepathology servers were located in Berlin (the UICC server) and Basel in Switzerland (the iPATH server). A scoring system was developed to quantify the differences between the diagnoses of the referring pathologist and the remote expert. Preparation of the cases, as well as the submission of images, took considerably longer from Kerman than from Berlin; this was independent of the server system. The Kerman delay was mainly associated with a slower transmission rate and longer image preparation. The diagnostic gap between referrers' and experts' diagnoses was greater with the iPATH system, but not significantly so. The experts' response time was considerably shorter for the iPATH system. The results showed that telepathology is feasible for requesting pathologists working in a developing country or in an industrialized country. The key factor in the quality of the service is the work of the experts: they should be selected according to their diagnostic expertise, and their commitment to the provision of telepathology services is critical.

Association of small foci of diffuse idiopathic pulmonary neuroendocrine cell hyperplasia (DIPNECH) with adenocarcinoma of the lung.

DIPNECH is regarded as a precursor lesion of neuroendocrine lung tumors, specifically carcinoids. A relationship with lung adenocarcinomas has not been clearly established so far. We present a series of four cases with a concomitant presence of adenocarcinoma and DIPNECH in the lung. The cases were retrieved from the archives of the Institutes of Pathology of the Jena University Hospital and the Charité, Berlin. The clinical data were collected from the hospital information system. The microscopic findings of adenocarcinoma and DIPNECH were reviewed. A panel of neuroendocrine and epithelial markers was analyzed immunohistochemically. In addition, the H&E slides of a series of 82 lung carcinomas were reevaluated for the presence of DIPNECH foci and the parameters of the IASLC/ATS/ERS classification for lung adenocarcinoma. DIPNECH foci were composed of small intramucosal nests of proliferating pulmonary neuroendocrine cells alongside or at the periphery of terminal airways. All detected foci measured less than 5mm in maximal diameter and showed a consistent reactivity against Synaptophysin. They did not express epithelial markers of squamous cell carcinoma and adenocarcinoma. In three cases, the DIPNECH foci were clearly associated with the adenocarcinoma, while in one case, they were observed in the non-neoplastic lung tissue. The adenocarcinoma with DIPNECH inside mainly showed low grade histology, while the fourth case was intermediate to high grade. The histologic evaluation of the HE slides of the other 82 lung cancer cases showed no suspected or definite DIPNECH foci. Within this series, we could confirm the prognostic significance of the IASLC/ATS/ERS classification of lung adenocarcinoma. Our series suggest that a subset of lung adenocarcinoma is characterized by the concomitant presence of DIPNECH within the tumor, suggesting a causal relationship. These adenocarcinomas seem to be low grade ones, and may have a particular tumorigenesis and clinical behavior. These observations need to be confirmed in larger tumor collectives. We could confirm the prognostic relevance of the new adenocarcinoma classification.

FGFR1 expression and gene copy numbers in human lung cancer.

FGFR1 is a receptor tyrosine kinase of which the ligands belong to the fibroblast growth factor family. To evaluate the significance of FGFR1 in lung cancer, we analysed tumours by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Tissue microarrays were constructed containing 380 lung cancer samples including squamous cell carcinomas (SCC), adenocarcinomas (ADC), non-small cell lung cancer not otherwise specified, metastases, neuroendocrine tumours, large cell lung cancer and small cell lung cancer. FGFR1 expression was analysed by IHC and scored semi-quantitatively by a four-tier approach (0, 1, 2, 3). Using dual-colour interphase FISH with probes specific for the locus on 8p12 and the centromere of chromosome 8 (CEN8), copy numbers of FGFR1 were determined. High expression of FGFR1 was associated with increased FGFR1 gene copy numbers in squamous cell carcinoma (p < 0.001). The FGFR1 locus was equally affected by copy number losses and gains. The higher FGFR1 gene copy numbers in SCC compared to ADC did not reach statistical significance. High copy number amplification of FGFR1 was a very rare event, the FGFR1/CEN8 signal ratio reaching a maximum value of 2.75. There were no significant associations between FGFR1 and clinicopathological parameters. Fibroblast growth factor signalling represents an interesting therapeutic target in lung cancer. However, the pathways are complex with potential oncogenic and anti-oncogenic activities. Our data may help to define criteria for selecting patients that may benefit from these new therapeutic options.

Diagnostic and prognostic impact of desmocollins in human lung cancer.

OBJECTIVE: Desmosomes are intercellular junctions that confer strong cell-cell adhesion. Two main members of desmosomal cadherins, desmogleins (DSGs) and desmocollins (DSCs), are involved in carcinogenesis. However, their role in human lung cancer remained elusive. The aims of this study were to analyse the expression of DSCs and to evaluate their clinical application in lung cancer. METHODS: The expression of DSC1-3 mRNAs was analysed by RT-PCR. The methylation status of DSCs was analysed by demethylation tests and bisulphite sequencing. Protein expression of DSCs in primary lung cancer was evaluated by immunohistochemistry on tissue microarrays. RESULTS: DSC1-3 mRNAs were downregulated in lung cancer cells, and the expression was restored in four out of seven cell lines, respectively, after 5-aza-2'-deoxycytidine treatment. A heterogeneous methylation pattern was detected by bisulphite sequencing in exon 1 of DSC2 and DSC3. In 199 patients with primary lung cancer, we found that lower protein expression of DSC1 was significantly linked to worse tumour differentiation (p=0.017), DSC3 proteins were more expressed in squamous cell carcinoma (SCC) compared with adenocarcinoma (ADC) (p<0.001), and reduced expression of DSC1 and DSC3 was significantly correlated with poor clinical outcome (p=0.045 and p=0.007, respectively). CONCLUSIONS: Our data suggest that downregulation of DSC1-3 may be explained by DNA methylation, DSC1 may be a marker for tumour differentiation, DSC3 has a potential diagnostic value in subclassification of non-small cell lung carcinoma into SCC and ADC, and furthermore, DSC1 and DSC3 may be prognostic markers for lung cancer.

Measurement of serum levels of mannose-binding lectin in hemodialysis patients: a comparison with healthy individuals.

INTRODUCTION: Mannose-binding lectin (MBL) is a part of the innate immune system. Many studies showed an association of low serum MBL levels with decreased host defense against various infectious agents. Considering paradoxical reports about the serum level of MBL in hemodialysis patients, this study aimed to measure and compare serum MBL levels in hemodialysis patients and healthy individuals. MATERIALS AND METHODS: In a cross-sectional study, 70 hemodialysis patients and 70 volunteers with normal routine laboratory tests and physical examination were assessed for serum MBL level (measured by an enzyme-linked immunosorbent assay). In addition, serum C-reactive protein levels in hemodialysis patients were measured to rule out correlation of increased serum MBL level with inflammation. RESULTS: In hemodialysis patients, 32 (45.7%) were men and 38 (54.3%) were women. In the control group, 34 (48.6%) were men and 36 (51.4%) were women (P = .87). The mean age showed no significant difference in hemodialysis (44.5 ± 13.5 year) and control (46.4 ± 12.4 years) groups. Serum level of MBL was significantly higher in hemodialysis patients (2.12 ± 1.49 microg/mL) than that in the controls (1.49 ± 2.12 microg/mL; P < .001). No significant correlation was found between serum MBL and C-reactive protein levels (r = 0.002, P = .98) among the hemodialysis patients. CONCLUSIONS: Serum MBL level in hemodialysis patients was significantly higher than that in the control group of healthy individuals. This may have some implications in management of patients and prediction of kidney allograft survival.

How do surgical pathologists evaluate critical diagnoses (critical values)?

BACKGROUND: After introduction of the concept of critical value (CV) in laboratory medicine, some efforts were performed to define possible critical values in surgical pathology. Critical diagnosis (critical value) is a concept recently established in surgical pathology and is a challenging issue among pathologists and clinical specialists. The concept may be the subject of variation according to the geographical or work setting differences. The current study was performed to bring the contribution of the Iranian pathologists to the evolving concept of critical diagnoses (critical values) in surgical pathology. MATERIALS AND METHODS: During annual meeting of Iranian Pathologist Society, November 2006, Tehran, Iran, anonymous questionnaires were distributed among participants. They were requested to openly name conditions in which a pathologist should communicate the results immediately with clinicians. RESULTS: 147 pathologists completed the questionnaire. They were varied in their level of experience and setting of workplace. Each participant referred to 1-7 (mean 3) conditions as CV. About 90 different conditions which were considered as CV by participants were extracted from the questionnaires. DISCUSSION: The list of conditions obtained through this survey as CVs in surgical pathology covered most items previously described in literature. Major differences are low number (or lack) of refers to some relatively routine and potentially important conditions and considering many unimportant conditions as CV by participants of present survey. Almost all conducted surveys have been performed on this issue so far (including the present survey) suffer from lack of supportive scientific evidences and based mainly on experience and common sense of participants in survey. Potential problems with application of CV concept in daily routine work flow of pathology, particularly in developing countries like Iran, were discussed.

How to measure image quality in tissue-based diagnosis (diagnostic surgical pathology).

BACKGROUND: Automated image analysis, measurements of virtual slides, and open access electronic measurement user systems require standardized image quality assessment in tissue-based diagnosis. AIMS: To describe the theoretical background and the practical experiences in automated image quality estimation of colour images acquired from histological slides. THEORY, MATERIAL AND MEASUREMENTS: Digital images acquired from histological slides should present with textures and objects that permit automated image information analysis. The quality of digitized images can be estimated by spatial independent and local filter operations that investigate in homogenous brightness, low peak to noise ratio (full range of available grey values), maximum gradients, equalized grey value distribution, and existence of grey value thresholds. Transformation of the red-green-blue (RGB) space into the hue-saturation-intensity (HSI) space permits the detection of colour and intensity maxima/minima. The feature distance of the original image to its standardized counterpart is an appropriate measure to quantify the actual image quality. These measures have been applied to a series of H&E stained, fluorescent (DAPI, Texas Red, FITC), and immunohistochemically stained (PAP, DAB) slides. More than 5,000 slides have been measured and partly analyzed in a time series. RESULTS: Analysis of H&E stained slides revealed low shading corrections (10%) and moderate grey value standardization (10 - 20%) in the majority of cases. Immunohistochemically stained slides displayed greater shading and grey value correction. Fluorescent stained slides are often revealed to high brightness. Images requiring only low standardization corrections possess at least 5 different statistically significant thresholds, which are useful for object segmentation. Fluorescent images of good quality only possess one singular intensity maximum in contrast to good images obtained from H&E stained slides that present with 2 - 3 intensity maxima. CONCLUSION: Evaluation of image quality and creation of formally standardized images should be performed prior to automatic analysis of digital images acquired from histological slides. Spatial dependent and local filter operations as well as analysis of the RGB and HSI spaces are appropriate methods to reproduce evaluated formal image quality.

Texture- and object-related automated information analysis in histological still images of various organs.

OBJECTIVE: To create algorithms and application tools that can support routine diagnoses of various organs. MATERIALS: A generalized algorithm was developed that permits the evaluation of diagnosis-associated image features obtained from hematoxylin-eosin-stained histopathologic slides. The procedure was tested for screening of tumor tissue vs. tumor-free tissue in 1,442 cases of various organs. Tissue samples studied include colon, lung, breast, pleura, stomach and thyroid. The algorithm distinguishes between texture- and object-related parameters. Texture-based information-defined as gray value per pixel measure--is independent from any segmentation procedure. It results in recursive vectors derived from time series analysis and image features obtained by spatial dependent and independent transformations. Object-based features are defined as gray value per biologic object measured. RESULTS: The accuracy of automated crude classification was between 95% and 100% based upon a learning set of 10 cases per diagnosis class. Results were independent from the analyzed organ. The algorithm can also distinguish between benign and malignant tumors of colon, between epithelial mesothelioma and pleural carcinomatosis or between different common pulmonary carcinomas. CONCLUSION: Our algorithm distinguishes accurately among crude histologic diagnoses of various organs. It is a promising technique that can assist tissue-based diagnosis and be expanded to virtual slide evaluation.

Lack of CD117 and rare bcl-2 expression in stomach cancer by immunohistochemistry. An immunohistochemical study with review of the literature.

BACKGROUND: Gastric adenocarcinoma is one of the most frequent malignancies worldwide including Iran. This study was designed to immunohistochemically evaluate the CD117 and bcl-2 expression in gastric carcinomas and their potential use as therapeutic targets in the treatment of patients with advanced stage gastric cancer. MATERIALS AND METHODS: Representative paraffin blocks obtained from 38 operated gastric adenocarcinoma patients were retrieved from Afzalipour Hospital pathology department archive, Kerman, Iran. Immunohistochemical analysis (IHC) for CD117 was carried out in all cases including negative (normal gastric epithelium) and positive (Gastrointestinal Stromal Tumor) controls. In addition, the cases were evaluated immunohistochemically for apoptosis-related protein (bcl-2), to evaluating a potential association of CD117 expression with the cell proliferation regulatory pathways. RESULTS: No positive reaction for CD117 was seen in gastric carcinoma tumor cells irrespective to the cell type, grade, and stage, proliferation and apoptosis rate. Expression of bcl-2 was observed in only one case. CONCLUSION: We conclude that CD117 overexpression detectable by immunohistochemistry does not play a significant role in gastric carcinoma pathways and development, although overexpression at the gene level and/or mutated CD117 expression might exist. Thus, it is unlikely that the CD117 pathway is of clinical significance in gastric carcinoma patients.