Immunohistochemical Staining For Cytokeratin Research Articles

Clinical values of nuclear morphometric analysis in fibroepithelial lesions

BackgroundFibroepithelial lesions (FELs) of the breast encompass a broad spectrum of lesions, ranging from commonly encountered fibroadenomas (FAs) to rare phyllodes tumors (PTs). Accurately diagnosing and grading these lesions is crucial for making management decisions, but it can be challenging due to their overlapping features and the subjective nature of histological assessment. Here, we evaluated the role of digital nuclear morphometric analysis in FEL diagnosis and prognosis.MethodsA digital nuclear morphometric analysis was conducted on 241 PTs and 59 FAs. Immunohistochemical staining for cytokeratin and Leukocyte common antigen (LCA) was used to exclude non-stromal components, and nuclear area, perimeters, calipers, circularity, and eccentricity in the stromal cells were quantified with QuPath software. The correlations of these features with FEL diagnosis and prognosis was assessed.ResultsAll nuclear features, including area, perimeter, circularity, maximum caliper, minimum caliper and eccentricity, showed significant differences between FAs and benign PTs (p ≤ 0.002). Only nuclear area, perimeter, minimum caliper and eccentricity correlated significantly with PT grading (p ≤ 0.022). For differentiation of FAs from benign PTs, the model integrating all differential nuclear features demonstrated a specificity of 90% and sensitivity of 70%. For PT grading, the nuclear morphometric score showed a specificity of 78% and sensitivity of 96% for distinguishing benign/borderline from malignant PTs. In addition, a relationship of nuclear circularity was found with PT recurrence. The Kaplan-meier analysis, using the best cutoff determined by ROC curve, showed shorter event free survival in benign PTs with high circularity (chi-square = 4.650, p = 0.031).ConclusionsOur data suggested the digital nuclear morphometric analysis could have potentials to objectively differentiate different FELs and predict PT outcome. These findings could provide the evidence-based data to support the development of deep-learning based algorithm on nuclear morphometrics in FEL diagnosis.

The potential role of SNHG16/ miRNA-146a/ TRAF6 signaling pathway in the protective effect of zoledronate against colorectal cancer and associated osteoporosis in mouse model

Bone fracture as a consequence of colorectal cancer (CRC) and associated osteoporosis (OP) is considered a risk factor for increasing the mortality rate among CRC patients. SNHG16/ miRNA-146a/ TRAF6 signaling pathway is a substantial contributor to neoplastic evolution, progression, and metastasis. Here, we investigated the effect of zoledronate (ZOL) on the growth of CRC and associated OP in a mouse model. Thirty Balb/c mice were divided into Naïve, azoxymethane (AOM)/dextran sodium sulfate (DSS), and ZOL groups. Body weight and small nucleolar RNA host gene 16 (SNHG16) expression, microRNA-146a, and TRAF6 in bone, colon, and stool were investigated. Samples of colon and bone were collected and processed for light microscopic, immunohistochemical staining for cytokeratin 20 (CK20), nuclear protein Ki67 (pKi-67), and caudal type homeobox transcription factor 2 (CDx2) in colon and receptor activator of nuclear factor kB (RANK) and osteoprotegerin (OPG) in bone. A computerized tomography (CT) scan of the femur and tibia was studied. ZOL produced a significant decrease in the expression of SNHG16 and TRAF6 and an increase in miRNA-146a in the colon and bone. ZOL administration improved the histopathological changes in the colon, produced a significant decrease in CK20 and Ki-67, and increased CDx2 expressions. In bone, ZOL prevented osteoporotic changes and tumour cell invasion produced a significant decrease in RANK and an increase in OPG expressions, alongside improved bone mineral density in CT scans. ZOL could be a promising preventive therapy against colitis-induced cancer and associated OP via modulation expression of SNHG16, miRNA-146a, and TRAF6.

Multimodal Prediction of Cervical Lymph Node Metastasis and Recurrence in Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is the most common malignancy in the head/neck region, and cervical lymph node (CLN) metastasis is a strong poor-prognosis factor. In addition, many patients with OSCC experience recurrence despite multidisciplinary treatment. We sought to identify factors associated with CLN metastasis and recurrence in patients with OSCC. We evaluated a total of 45 patients and 233 target CLNs. The longest diameter of the target CLN, the shortest diameter of the target CLN (LS), the area of the target CLN, and the relative computed tomography (CT) values of the target CLNs calculated based on the CT values of the internal jugular vein (LCT) were obtained from preoperative CT images, and the maximum standardized uptake values of the primary tumor (pSUV) and target CLN (nSUV) were obtained from preoperative 18F-fluorodeoxyglucose-positron emission tomography/CT images. We performed immunohistochemical staining for cytokeratin 13 (CK13) and 17 (CK17) on neck dissection tissues. A discrimination equation was used that can predict CLN metastasis with a 92.2% discrimination rate using LS, LCT, pSUV, and nSUV. The CLNs were divided into discrimination and non-discrimination groups based on discriminant equations and CK13 and CK17 were used as the objective variables. A significantly higher recurrence rate was observed in the non-discrimination group (CK13: 5-year recurrence rate 28.6% vs. 64.3%, p<0.01; CK17: 5-year recurrence rate 28.0% vs. 76.0%, p<0.01). CLN metastases in OSCC can be assessed by combining preoperative imaging. The combined use of CK13 and CK17 expression with imaging findings offers an integrated approach to predict OSCC recurrence.

Ultrastructural Profile Combined with Immunohistochemistry of a Hepatic Progenitor Cell Line in Pediatric Autoimmune Hepatitis: New Insights into the Morphological Pattern of the Disease.

Considering that the heterogenic population of a hepatic progenitor cell line (HPCL) can play a vital role in autoimmune hepatitis (AIH), we decided to conduct pioneering retrospective evaluation of these cells in pediatric AIH by means of transmission electron microscopy (TEM). The aim of the study was to assess the ultrastructure of the HPCL in children with untreated AIH. Ultrastructural analysis of the HPCL population, preceded by immunohistochemical staining for cytokeratin 7 (CK7), was performed using pretreatment liver biopsies from 23 children with clinicopathologically diagnosed AIH. Immunohistochemical assessment for CK7 allowed detection of proliferating immature epithelial cells differentiating towards periportal and intralobular intermediate hepatocytes without marked formation of ductular reactions in AIH children. Using TEM, we distinguished three morphological types of HPCs: I—the most undifferentiated progenitor cells; III—intermediate hepatocyte-like cells; II—intermediate bile duct cells. Most frequent were the cells differentiating towards hepatocytes, most rare—those differentiating towards cholangiocytes. The results indicate that an HPCL may be an important source of hepatocyte regeneration. Ultrastructural analyses of the HPCL population, combined with immunohistochemistry for CK7, might be a useful tool to evaluate liver cell regeneration, including fibrogenesis, and may help better understand the morphological pattern of the disease, in pediatric AIH. Frequent appearance of an HPCL in the vicinity of fibrotic foci, often accompanied by hyperactive Kupffer cells and transitional hepatic stellate cells, may indicate their significant involvement in liver fibrogenesis.

Evaluating Three Elastic-Fiber Staining Methods for Detecting Visceral Pleural Invasion in Lung Adenocarcinoma Patients.

The aim of this study was to evaluate the accuracy of three combination staining approaches, each composed of pancytokeratin immunostaining with an elastic-fiber staining method of choice, in diagnosing visceral pleural invasion (VPI) in lung adenocarcinoma patients. Tissue samples were resected from 23 lung adenocarcinoma patients, for whom neither hematoxylin and eosin nor Gomori's aldehyde-fuchsin (GAF) staining accurately detected the presence of VPI. Three slices were prepared from each sample and examined by immunohistochemical staining of cytokeratin AE1/AE3 and one of the following methods for elastic-fiber staining, including Victoria blue (VB), GAF, or Weigert's resorcin-fuchsin (WRF). The staining scores and slider view time were compared among the three staining protocols with tumor extension beyond the elastic layer of the visceral pleura as the diagnostic criterion for VPI. Conclusive diagnoses were achieved for all 23 tissue samples stained with VB, 18 with GAF, and 17 with WRF. VB staining was the fastest of the three methods, and produced significantly brighter, clearer color, and better contrast between the elastic fibers and tumor cells, thus facilitating slide review. The combination of VB elastic fiber and cytokeratin AE1/AE3 staining is a user-friendly, fast, and highly effective method that produces bright stains, favorable color contrast against the background, and high tumor localization accuracy. Hence, this method can improve the accuracy of VPI diagnosis and the corresponding staging of lung adenocarcinoma. We recommend the combined use of VB and pancytokeratin immunostaining when VPI is suspected.

Diagnostic utility of Cytokeratin 13 and Cytokeratin 17 in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma

Objective: To determine the immunohistochemical expression of CK 13 and CK 17 in Oral Epithelial Dysplasia and Oral Squamous Cell Carcinoma. Methods: A total of 170 cases were retrieved from record files of Histopathology Department, to conduct a cross sectional study at Armed Forces Institute of Pathology, Rawalpindi, over a period of one year from June 2018 to June 2019 along with their formalin-fixed, paraffin embedded blocks comprising 85 cases of each oral squamous cell carcinoma and oral epithelial dysplasia. Blocks were trimmed and cut into very thin sections of 5 microns using microtome and mounted on slides. Tissue mounted slides were stained with routine haematoxylin and eosin followed by immunohistochemical staining of cytokeratin 13 and cytokeratin 17. New histological diagnosis of each case was made. Mean and standard deviation were calculated for quantitative variables. Frequency and percentages were calculated for qualitative variables. Chi-square test was employed to assess the significance of difference. The P-value &lt;0.05 was considered significant. Results: In this study, 101 (59.4%) male and 69 (40.6%) female patients with the mean age of 60.63 ± 13.814 (mean ± SD) were collected and buccal mucosa was the most common site of presentation. In a total of 170 cases, 34 (20%) cases showed positive expression of cytokeratin 13 whereas 136 (80%) cases showed negative expression of cytokeratin 13. In comparison, out of 170 cases, 133 (78.2%) cases showed positive expression of cytokeratin 17 whereas 37 (21.8%) cases showed negative expression of cytokeratin 17. The P-value was found to be &lt; 0.001 and 0.001 for expression of cytokeratin 13 and cytokeratin 17 respectively. Conclusion: Opposite expression of cytokeratin 13 and cytokeratin 17 was seen in this study, in the form of loss of cytokeratin 13 and over expression of cytokeratin 17 with increase in the degree of dysplasia and invasive carcinoma. Correct assessment, diagnosis and management with the help of these markers can lead to early diagnosis and favorable treatment outcome.

Oncocytic variant, a novel subtype of chromophobe renal cell carcinoma: a report of two cases and a literature review

A novel variant of chromophobe renal cell carcinoma showing an oncocytic phenotype is proposed. Two new cases of this rare entity are presented and discussed along with six previous cases from our colleagues. A 76-year-old man and a 78-year-old man had a 3.4-cm and a 3.2-cm-diameter renal mass, respectively. On histopathological examination of surgical specimens, uniform eosinophilic cuboidal cells without a perinuclear halo growing in a tubular pattern were seen, and differential diagnosis from oncocytoma was necessary. Immunohistochemical staining for cytokeratin 7 and E-cadherin showed diffusely positive patterns in both, as in the previous reports. Although monosomy of chromosomes 7, 10, 13, and 17 was commonly observed in the previous reports, gains of chromosome 19 were observed in the two present cases. Immunohistochemical and cytogenetic approaches lead to exclusion of oncocytoma and the diagnosis of an oncocytic variant of chromophobe renal cell carcinoma.

Interobserver Variability in the Assessment of Tumor Budding in pT 3/4 Colon Cancer: Improvement by Supporting Immunohistochemistry?

The prognostic significance of tumor budding in colon cancer is unequivocally documented, and the recommendations of the International Tumor Budding Consensus Conference (ITBCC) are currently the accepted basis for its assessment. Up to now, it is unknown whether the general use of a supporting cytokeratin immunohistochemistry can improve the interobserver variability and prognostic significance. Six investigators with different levels of experience reassessed 229 cases of colon carcinoma (pT3/4, N+/−, M0) with a supporting cytokeratin immunohistochemistry. The results were compared to previous assessments, which have been performed only on H & E. Bd3 was significantly associated with the occurrence of distant metastases according to the assessments of three out of six investigators (p < 0.05). Only one single investigator reached significant results concerning the cancer specific survival (p = 0.01). The pairwise kappa values range between a poor and moderate level of agreement (range 0.17–0.45; median 0.21). In conclusion, the results show no superiority of the use of an additional cytokeratin immunohistochemistry compared to the conventional analysis on sole H & E slides. Therefore, the general supporting use of a cytokeratin immunohistochemical staining seems to be inadvisable in colon cancer in consideration of necessary resources and costs.

Low recurrence rates for challenging squamous cell carcinomas using Mohs micrographic surgery with AE1/AE3 cytokeratin immunostaining

To the Editor: Local recurrence rates of high-risk cutaneous squamous cell carcinoma (SCC) are lower after Mohs micrographic surgery compared with conventional excision.1 Although local recurrence rates of primary cutaneous SCC after Mohs micrographic surgery are low,1,2 local recurrence can reach 15% or higher for tumors with high-risk factors.3 Adjuvant AE1/AE3 cytokeratin immunohistochemical stains ease visualization of SCC in challenging cases by helping to identify single-cell spread, neoplastic cells masked by inflammation, and perineural invasion.

Eccrine and Apocrine Carcinoma in Dogs

Background: Sweat gland carcinomas divide into eccrine, apocrine, mixed origin (eccrine and apocrine). Eccrine carcinoma (EC) is a rare malignant neoplasm of the sweat glands that can affect dogs, cats, and humans. EC can present itself as a solitary swelling in the pads, digits, or distal limbs. EC is more common in elder animals, and exhibits no predisposition according to breed. In humans, EC is more frequent in the head and neck, and is more likely to occur in mid-aged people; metastases can develop in any site of the body. A diagnosis of this type of tumor can be determined by histopathological examination. Apocrine carcinomas occur most frequently in the axillary areas even though they may occur in other regions of the body, and affect mainly elder animals.Case: A 13-year-old male mongrel dog with a history of presence of smooth reddish infiltrative nodule in the skin of the thorax, but with no history of progression, was examined. After running complementary tests, a fragment of the skin on the thorax was taken for biopsy. The second animal was an 8-year-old female Golden Retriever, which was presented with a history of presence of a nodule on the right digital pad. After running complementary tests, a fragment of this nodule was collected. In both cases, the excised fragments were placed in 10% buffered formalin and routinely processed for the preparation of histological slides, which were stained with hematoxylin and eosin, and subjected to histopathological evaluation. Light microscopy analysis revealed, in both cases, the presence of a poorly delimited, non-encapsulated mass infiltrating the muscle and adipose tissue. Necrotic areas, and presence of eosinophilic material in the nucleus were observed. Additionally, the samples from both cases were subjected to immunohistochemical staining for cytokeratin (CK Pan).Discussion: A definitive diagnosis of sweat gland carcinoma was achieved by means of the histopathological analysis. Sweat gland neoplasms are uncommon and difficult to diagnose; distinguishing eccrine carcinoma from apocrine carcinoma requires knowledge on the site of origin of the tumor. Some authors assert that, in spite the growing number of reports on tumors of cutaneous glands in dogs, there are only few immunohistochemical studies on cutaneous gland neoplasms in these animals. There is no specific immunohistochemical marker to distinguish eccrine carcinoma from apocrine carcinoma. Consequently, investigation on the anatomical origin of the tumor is important. In the cases describe herein, involvement of the thorax (apocrine) and digital pad (eccrine) are reported. However, there are accounts of the occurrence of sweat gland tumors in axillary and inguinal regions, as well as in the limbs. Both biopsy samples tested positive for cytokeratin (CK) after immunohistochemical examination. This marker is specific for simple epithelium and, according to some authors, can distinguish an eccrine carcinoma from basal cell carcinoma. The antibody used did not stain the innermost are of the tumor, where cells may be negative for CK. Some tumors may not have specific markers, which makes a combination of clinical data and complementary histopathological and immunohistochemical exams necessary for the determination of a diagnosis, as is the case for eccrine and apocrine carcinomas. In view of the scarcity of reports on eccrine carcinoma in the literature, and considering the small number of immunohistochemical studies on cutaneous gland tumors in dogs and cats, this work can help the determination of the diagnosis of cutaneous neoplasms, which are frequently observed in the clinical routine.

Open-Top Light-Sheet Microscopy Image Atlas of Prostate Core Needle Biopsies

Ex vivo microscopy encompasses a range of techniques to examine fresh or fixed tissue with microscopic resolution, eliminating the need to embed the tissue in paraffin or produce a glass slide. One such technique is light-sheet microscopy, which enables rapid 3D imaging. Our pathology-engineering collaboration has resulted in an open-top light-sheet (OTLS) microscope that is specifically tailored to the needs of pathology practice. To present an image atlas of OTLS images of prostate core needle biopsies. Core needle biopsies (N = 9) were obtained from fresh radical prostatectomy specimens. Each biopsy was fixed in formalin, dehydrated in ethanol, stained with TO-PRO3 and eosin, optically cleared, and imaged using OTLS microscopy. The biopsies were then processed, paraffin embedded, and sectioned. Hematoxylin-eosin and immunohistochemical staining for cytokeratin 5 and cytokeratin 8 was performed. Benign and neoplastic histologic structures showed high fidelity between OTLS and traditional light microscopy. OTLS microscopy had no discernible effect on hematoxylin-eosin or immunohistochemical staining in this pilot study. The 3D histology information obtained using OTLS microscopy enabled new structural insights, including the observation of cribriform and well-formed gland morphologies within the same contiguous glandular structures, as well as the continuity of poorly formed glands with well-formed glands. Three-dimensional OTLS microscopy images have a similar appearance to traditional hematoxylin-eosin histology images, with the added benefit of useful 3D structural information. Further studies are needed to continue to document the OTLS appearance of a wide range of tissues and to better understand 3D histologic structures.

Pyogenic granuloma associated with conjunctival epithelial neoplasia: report of nine cases

AimsTo systematically describe the clinical and histopathological features of a case series of conjunctival carcinomatous lesions underlying as—and also masquerading—pyogenic granuloma.MethodsNine cases of conjunctival carcinomatous lesions underlying a pyogenic granuloma...

A Report of Bone Marrow Metastasis of Colon Cancer as a Primary Diagnosis, Supported by Cytokeratin Immunohistochemical Staining

A Report of Bone Marrow Metastasis of Colon Cancer as a Primary Diagnosis, Supported by Cytokeratin Immunohistochemical Staining

Abstract 475: Characterization of miR-21 expression in budding colon cancer cells using digital images of multiplex fluorescence stained slides obtained by confocal scanning microscopy

Abstract This study was undertaken to analyze the presence and prevalence of microRNA-21 (miR-21) positive budding cancer cells in colon cancer using confocal scanning microscopy. Budding cancer cells are locally invasive cancer cells with increased metastatic properties and characteristics of epithelial to mesenchymal transition (EMT). Expression of miR-21 in stromal fibroblastic cells in colorectal cancers is well documented (1,2), whereas miR-21 expression in budding cells is poorly described. In order to characterize miR-21 positive budding cells, we first developed a multiplex fluorescence staining method by combining miR-21 in situ hybridization with immunohistochemical staining for cytokeratin and budding cell marker laminin-γ2, and stained 16 colon cancer cases (stage II, n=5, stage III, n=11). We then employed a confocal scanning microscope to obtain 15-55 GB digital images covering areas of 10-40 mm2 of the invasive front. The high resolution of the confocal digital images allowed detailed examination of the 4-fluorophore-stained slides using extended focus and z-stack images, e.g. in the discrimination of epithelial cells from adjacent stromal cells. All cytokeratin-positive budding cells were evaluated for the presence of miR-21 and laminin-γ2. Five out of the 16 cases had more than 10% miR-21 positive budding cells, a few of which were also laminin-γ2 positive. All five cases were stage III cancers. The presence of miR-21 in the budding cancer cells was not associated with the level of tumor budding. These observations suggest that the miR-21 expression in tumor budding cells increases with the progression of the cancer and is independent of laminin-γ2.

Postoperative radiotherapy is dispensable for OSCC patients with micrometastases in lymph nodes.

Lymph node metastasis is a decisive factor for performing postoperative radiotherapy for oral squamous cell carcinoma (OSCC). However, whether OSCC patients with only micrometastasis need postoperative radiotherapy is unclear. In this study, OSCC patients (n = 311) with negative (n = 247), only micrometastasis (n = 44) and macrometastasis (n = 20) were detected and selected by HE staining. Micrometastasis was re-assessed using immunohistochemical staining of cytokeratin (CK) in HE-negative patients to find out the false negative cases. The results indicated that, among the negative lymph node cases (n = 247), the positive rate of CK was 4.94% (n = 12). Besides, the clinical features of the primary tumor in relation to the only micrometastatic status and the value of the postoperative radiotherapy on the only micrometastasis patients were evaluated. Patients with only micrometastasis had higher T stage and inferior worst pattern of invasion (WPOI) than patients without micrometastasis, but they had longer overall survival (OS), metastasis-free survival (MFS), and disease-free survival (DFS) than macrometastasis patients. However, the survival time of only micrometastasis patients with or without postoperative radiotherapy was comparable, even in patients with inferior WPOI. Radiotherapy, however, may only benefit patients with IV/V levels of micrometastasis. These data indicated that postoperative radiotherapy is dispensable for only micrometastasis OSCC patients.

Small cell carcinoma of the gastric remnant: a case report

Abstract Objective Small cell carcinoma (SCC) is mostly found in the lungs. It is extremely rare in the gastric remnant. Here, we report a case and review the literature in order to improve the diagnosis and treatment of SCC of the gastric remnant. Methods We report a case of SCC of the gastric remnant in a 71-year-old male Chinese patient who presented with epigastric pain, acid regurgitation, and belching and who underwent Billroth II gastrectomy more than 38 years ago. Results Physical examination showed no obvious abnormalities. Laboratory data were within normal limits, except for anemia. Pathology of the mass showed a protruded tumor measuring 5.0 × 5.0 × 2.5 cm at the anastomotic edge of the gastric remnant that infiltrated through the full wall of the stomach; this was confirmed by immunohistochemical staining for cytokeratin [CK (-)], leukocyte common antigen (LCA) (+), synaptophysin (+), CD56 (+), and Ki-67 (+ &gt; 50%). Conclusion SCC of the gastric remnant is extremely rare, although the pathology, symptoms, diagnosis, treatment, and prognosis of SCC are similar to those of gastric SCC. Although the standard treatment of SCC of the gastric remnant remains unclear, effective surgical resection and subsequent multiagent chemotherapy should be performed for long-term survival. Our case shows the efficacy of tegafurgimeracil-oteracil-potassium capsule chemotherapy. Examination of a large series is required to determine the optimal treatment strategy for SCC of the gastric remnant.

Tumor budding as a novel predictor of occult metastasis in cT2N0 tongue squamous cell carcinoma

Occult neck metastasis is an important prognostic factor in patients with tongue squamous cell carcinoma (TSCC) who are deemed clinically negative for neck metastasis. The purpose of this study was to identify predictive factors for occult neck metastasis arising from TSCC and to determine patient prognosis. Ninety-seven patients with cT2N0 TSCC who underwent surgical resection of their primary lesion as initial therapy were enrolled in this retrospective study. Cutoff values for depth of invasion (≥3.3 mm) and the tumor budding score (≥4) were determined using receiver operator characteristic analyses. Univariate and multivariate analyses revealed that a tumor budding score ≥4 is a significant independent predictive factor for the occurrence of occult neck metastasis, which in turn is a significant independent prognostic factor. When evaluating tumor budding, we demonstrated greater interobserver and intraobserver agreement when using immunohistochemical staining for cytokeratin AE1/AE3 than with hematoxylin and eosin staining (HE). We conclude that the evaluation of tumor budding is effective for identifying populations at high risk of occult neck metastasis, which will enable the planning of appropriate therapeutic strategies for patients with cT2N0 TSCC. Furthermore, cytokeratin staining is recommended over HE staining for simpler and more accurate evaluation of tumor budding.

Immunohistochemical staining of cytokeratin 20 and cytokeratin 7 in colorectal carcinomas: Four different immunostaining profiles.

Background/Aim:Aberrant expression of CK20/CK7 is reported in a percentage of colorectal carcinomas (CRC); however, its relation to clinicopathological variables and survival data is still unclear. The objective of this study is to explore patterns of CK20/CK7 immunostaining in CRC and to analyse the diagnostic, prognostic, and predictive role of patterns of CK20/CK7 immunostaining.Materials and Methods:A total of 144 CRC cases were retrieved from the archives at the Department of Pathology, King Abdulaziz University, Jeddah, Saudi Arabia. Immunohistochemistry was performed using antibody to CK7 and CK20. Immunostaining was defined as low and high by using the extent of staining. The association of CK7 and CK20 with clinicopathological characteristics and survival.Results:CK20 was expressed in a higher percentage of CRC and nodal metastasis than CK7. No difference in CK7 and CK20 immunostaining in primary and metastasis carcinomas was found. Four patterns of CK20/CK7 were identified; CK20+/CK7− (60.4%), CK20+/CK7+ (2.1%), CK20−/CK7− (35.4%), and CK20−/CK7+ (2.1%). There was no statistically significant correlation between CK20/CK7 immunohistochemical profile and clinicopathological characteristics, prognosis, and survival was determined.Conclusions:Our results may support the heterogeneity of CRC. CRC showed four different subclasses following patterns of relative CK20/CK7 immunostaining. A considerable number of CRC expressed aberrant immune profile of CK20/CK7, which should be considered during diagnosing CRC in metastatic regions. Further studies on larger cohorts correlating different immunohistochemical cytokeratin profiles to molecular subtypes of CRC are recommended for better understanding of pathogenesis and behaviour of CRC.

Determination of Proliferative Activity of Neoplastic Cells by Ki67 and AgNOR Staining in Ovine Pulmonary Adenocarcinoma Affected Lungs

Ovine Pulmonary Adenocarcinoma (OPA) is a natural lung cancer of the sheep caused by Jaagsiekte Sheep Retrovirus (JSRV). In the present study, 17 OPA affected lung samples were subjected to histopathological, histochemical and immunohistochemical studies. Grossly, lungs revealed diffuse areas of consolidation or tumor nodules and histologically lung sections showed proliferation of alveolar epithelial cells that were arranged in papillary or acinar pattern. The lung sections were further subjected to immunohistochemical staining for cytokeratin and Ki-67. The proliferative activity of tumor cells was determined by Ki67 and AgNor staining and the mean value of Ki-67 index was 9.53 + 1.88. The mean number of AgNORs for all the cases ranged from 6.5 to 11.9 with a mean + standard deviation values of 9.2 + 1.2. The mean values of Ki-67 index and AgNOR staining were increased in the sections of lung tumor than the normal lung sections indicating the increased proliferative activity of lung tumor epithelial cells.

Relationship of Lymph Node Micrometastasis and Micropapillary Component and Their Joint Influence on Prognosis of Patients With Stage I Lung Adenocarcinoma.

This study aimed to investigate the relationship between lymph node micrometastasis and histologic patterns of adenocarcinoma, with a particular focus on their joint effect on prognosis. We retrospectively reviewed 235 patients with stage I adenocarcinoma from January 2009 to December 2009. Lymph node micrometastasis was evaluated by immunohistochemical staining for cytokeratin (AE1/AE3) and thyroid transcription factor-1. A logistic regression model was applied to confirm the predictive factors of micrometastasis. Survival analysis was performed to evaluate the effect of micrometastasis on prognosis. Lymph node micrometastasis was observed in 35 patients (15%). Patients with micrometastasis had significantly worse recurrence-free survival (P<0.001) and overall survival (P<0.001) compared with those without micrometastasis. Micropapillary component was confirmed as an independent predictor of increased frequency of micrometastasis (P<0.001). Among 62 patients with adenocarcinoma with a micropapillary component, 23 (37%) had lymph node micrometastasis. Micropapillary-positive/micrometastasis-positive patients had significantly worse survival compared with micropapillary-positive/micrometastasis-negative patients (RFS, P=0.039; OS, P=0.002) and micropapillary-negative patients (recurrence-free survival, P<0.001; overall survival, P<0.001). Moreover, the presence of micrometastasis correlated with a higher risk of locoregional recurrence (P=0.031) rather than distant recurrence (P=0.456) in micropapillary-positive patients. In summary, lymph node micrometastasis was more frequently observed in adenocarcinoma with a micropapillary component. Moreover, lymph node micrometastasis could provide helpful prognostic information in patients with resected stage I lung adenocarcinoma with a micropapillary component; thus, immunohistochemical detection of micrometastatic tumor cells in lymph nodes should be recommended.