scholarly journals Expression of “Connexin 43” in Colorectal Carcinomas: Histopathological and Immunohistochemical Study

2020 ◽  
Vol 8 (A) ◽  
pp. 354-359
Author(s):  
Essam Ayad ◽  
Ahmed Naeem Eesa ◽  
Rabab Radi ◽  
Lubna Omer El Farouk Abdel-Salam
Keyword(s):  
Connexin 43 ◽  
Cancer Chemoprevention ◽  
Colorectal Carcinogenesis ◽  
Formalin Fixed Paraffin ◽  
Gap Junction Intercellular Communication ◽  
Formalin Fixed Paraffin Embedded ◽  
Histologic Evaluation ◽  
Cell Growth And Differentiation ◽  
Hematoxylin And Eosin ◽  
Cx 43

BACKGROUND: Colorectal cancer is one of the most common cancers worldwide and leading cause of cancer related deaths. Connexins are integral membrane proteins that form channels between adjacent cells. Gap junction intercellular communication plays essential roles in tissue homoeostasis and regulation of cell growth and differentiation. Connexins can act as either tumor suppressors or tumor promoters. The human connexin protein family contains 21 members, of which the most widely studied is connexin 43 (Cx 43). OBJECTIVES: Investigation of immunohistochemical expression of Cx 43 in cases of colorectal adenoma and carcinoma and correlation of this expression with the clinico-pathological aspects of the tumors. MATERIALS AND METHODS: Seventy formalin fixed paraffin embedded BC tissue sections were randomly collected. All the available data were collected from the patients’ reports. The paraffin blocks were sectioned and stained with hematoxylin and eosin stains for histologic evaluation. Additional sections were immunostained with Cx 43. RESULTS: Cx 43 expression was negative in all studied cases. CONCLUSION: Cx 43 is a tumor suppressor that is lost early in colorectal carcinogenesis and can be considered as potential target for cancer chemoprevention and chemotherapy aiming at restoration of normal connexin expression and functional gap junctions.

Canine Hepatitis Associated with Intrahepatic Bacteria in Three Dogs

2018 ◽  
Vol 54 (1) ◽  
pp. 65-70 ◽  
Author(s):  
Joon Im ◽  
Derek P. Burney ◽  
Sean P. McDonough ◽  
Brigid Nicholson ◽  
Adam Eatroff ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Case Report ◽  
Fluorescence In Situ Hybridization ◽  
Antimicrobial Therapy ◽  
Granulomatous Inflammation ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hematoxylin And Eosin ◽  
Formalin Fixed

ABSTRACT This case report describes the detection of intrahepatic bacteria in formalin-fixed paraffin-embedded histopathological sections from three dogs with neutrophilic, pyogranulomatous, or lymphoplasmacytic hepatitis and cholangiohepatitis. In each of these cases, eubacterial fluorescence in situ hybridization enabled colocalization of intrahepatic bacteria with neutrophilic and granulomatous inflammation in samples that were negative for bacteria when evaluated by routine hematoxylin and eosin histopathology augmented with histochemical stains. Positive responses to antimicrobial therapy were observed in of 2 out of 2 patients that were treated with antimicrobials. These findings suggest that eubacterial fluorescence in situ hybridization analysis of formalin-fixed paraffin-embedded histopathological sections is more sensitive than conventional histochemical stains for the diagnosis of bacteria-associated canine hepatitis.

Glioblastoma Contains Topologically Distinct Proliferative and Metabolically Defined Subpopulations of Nestin- and Glut1-Expressing Cells

2021 ◽  
Author(s):  
Michael Prosniak ◽  
Lawrence C Kenyon ◽  
D Craig Hooper
Keyword(s):  
Glucose Transporter ◽  
Intermediate Filament Protein ◽  
Protein Markers ◽  
Ki 67 ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hematoxylin And Eosin ◽  
Glial Lineage ◽  
Filament Protein ◽  
Von Willebrand

Abstract The difficulty in treatment of glioblastoma is a consequence of its natural infiltrative growth and the existence of a population of therapy-resistant glioma cells that contribute to growth and recurrence. To identify cells more likely to have these properties, we examined the expression in tumor specimens of several protein markers important for glioma progression including the intermediate filament protein, Nestin (NES), a glucose transporter (Glut1/SLC2A1), the glial lineage marker, glial fibrillary acidic protein, and the proliferative indicator, Ki-67. We also examined the expression of von Willebrand factor, a marker for endothelial cells as well as the macrophage/myeloid markers CD163 and CD15. Using a multicolor immunofluorescence and hematoxylin and eosin staining approach with archival formalin-fixed, paraffin embedded tissue from primary, recurrent, and autopsy IDH1 wildtype specimens combined with high-resolution tissue image analysis, we have identified highly proliferative NES(+)/Glut1(–) cells that are preferentially perivascular. In contrast, Glut1(+)/NES(–) cells are distant from blood vessels, show low proliferation, and are preferentially located at the borders of pseudopalisading necrosis. We hypothesize that Glut1(+)/NES(–) cells would be naturally resistant to conventional chemotherapy and radiation due to their low proliferative capacity and may act as a reservoir for tumor recurrence.

scholarly journals Parvovirus-induced encephalitis in a juvenile raccoon

2020 ◽  
Vol 33 (1) ◽  
pp. 140-143
Author(s):  
Arno Wünschmann ◽  
Robert Lopez-Astacio ◽  
Aníbal G. Armién ◽  
Leslie Reed ◽  
Colin R. Parrish
Keyword(s):  
Cerebral Cortex ◽  
Cell Loss ◽  
Canine Parvovirus ◽  
Vp2 Gene ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Histologic Evaluation ◽  
Ns1 Gene ◽  
Gross Lesions ◽  
Pcr Targeting

A juvenile raccoon was euthanized because of severe neurologic signs. At postmortem examination, no significant gross lesions were present. Histologic evaluation demonstrated nonsuppurative encephalitis in thalamus, brainstem, and hippocampus, cerebellar Purkinje cell loss, as well as poliomyelitis and demyelination of the spinal cord. Parvovirus antigen–specific immunohistochemistry revealed immunopositive neurons in the brainstem, cerebral cortex, and hippocampus. A few Purkinje cells were also immunopositive. DNA extracted from formalin-fixed, paraffin-embedded brain tissue (thalamus, hippocampus, cerebral cortex) yielded a positive signal using PCR targeting both feline and canine parvovirus. Sequencing analyses from a fragment of the NS1 gene and a portion of the VP2 gene confirmed the presence of DNA of a recent canine parvovirus variant (CPV-2a–like virus) in the cerebellum. Our case provides evidence that a recent canine parvovirus (CPV) strain ( Carnivore protoparvovirus 1) can infect cerebral and diencephalic neurons and cause encephalitis in an otherwise healthy raccoon. Parvovirus-induced encephalitis is a differential diagnosis of rabies and canine distemper in raccoons with neurologic signs.

scholarly journals Clinicopathologic Significance of Histologic Grade, Pgp, and P53 Expression in Canine Lymphoma

2013 ◽  
Vol 49 (3) ◽  
pp. 175-184 ◽  
Author(s):  
Ravinder S. Dhaliwal ◽  
Barbara E. Kitchell ◽  
EJ Ehrhart ◽  
Victor E. Valli ◽  
Nikolaos G. Dervisis
Keyword(s):  
Histologic Grade ◽  
Canine Lymphoma ◽  
P53 Expression ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Histologic Evaluation ◽  
Chemotherapy Protocol ◽  
First Remission ◽  
Resistance Markers ◽  
Significant Expression

To characterize the expression of P-glycoprotein (Pgp) and p53 in different histologic grades of canine multicentric lymphosarcoma (LSA), 31 cases of LSA without prior treatment were studied. The expression levels of the Pgp and p53 proteins were evaluated for their clinicopathologic significance among standard histologic evaluation. Immunohistochemistry (IHC) was performed on formalin-fixed, paraffin-embedded archival samples of 31 previously untreated LSA cases to detect the expression of Pgp and p53. All dogs were subsequently treated with a combination chemotherapy protocol. Remission and survival durations were evaluated for correlation with histologic grade and presence of drug resistance markers. Of the 31 cases, 24 (80%) and 7 (22%) were positive for Pgp and p53, respectively. Overall, the median survival and duration of remission in the study was 246 days and 137 days, respectively. The National Cancer Institute working formulation histologic grade was not associated with either survival or duration of first remission (DOR). The Pgp protein expression and DOR and survival was not statistically significant. Expression of p53 was statistically correlated with survival.

Phase III Randomized Trial Assessing Rofecoxib in the Adjuvant Setting of Colorectal Cancer: Final Results of the VICTOR Trial

2010 ◽  
Vol 28 (30) ◽  
pp. 4575-4580 ◽  
Author(s):  
Rachel S. Midgley ◽  
Christopher C. McConkey ◽  
Elaine C. Johnstone ◽  
Janet A. Dunn ◽  
Justine L. Smith ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Colorectal Carcinogenesis ◽  
Phase Iii ◽  
Adjuvant Setting ◽  
Case Control Studies ◽  
Curative Surgery ◽  
Tissue Samples ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Cox 2

Purpose Laboratory and case-control studies suggest a pivotal role for the cyclooxygenase-2 (COX-2) pathway in colorectal carcinogenesis. The purpose of this study was to test whether the COX-2 inhibitor rofecoxib could reduce recurrence and improve survival when administered in the adjuvant setting of colorectal cancer (CRC). Patients and Methods Patients who had undergone potentially curative surgery and completion of adjuvant therapy for stage II and III CRC were randomly assigned to receive rofecoxib (20 mg daily) or placebo. The primary end point was overall survival (OS). Where formalin-fixed paraffin-embedded tumor tissue samples were available, COX-2 expression was evaluated by immunohistochemistry and correlated with clinical outcome. Results Two thousand four hundred thirty-four patients were entered onto the study. The trial was terminated early because of the worldwide withdrawal of rofecoxib. At this point, 1,167 patients had received rofecoxib and 1,160 patients had received placebo for median treatment durations of 7.4 and 8.2 months, respectively. For the rofecoxib and placebo arms, median follow-up times were 4.84 and 4.85 years, with 241 and 246 deaths and 297 and 329 recurrences, respectively. No difference was demonstrated in OS (hazard ratio [HR] = 0.97; 95% CI, 0.81 to 1.16; P = .75) or recurrence (HR = 0.89; 95% CI, 0.76 to 1.04; P = .15) comparing the two groups. Tumor COX-2 expression by immunohistochemistry was assessed for 871 patients, but neither prognostic nor predictive effects were observed. Conclusion In this study of abbreviated therapy in the adjuvant setting of CRC, rofecoxib did not improve OS or protect from recurrence in unselected patients. In addition, COX-2 expression did not correlate with prognosis overall or predict effectiveness of COX-2 inhibitors.

scholarly journals Anatomopathological Assessment of the Diaphragm in Formalin-Fixed, Paraffin-Embedded Sections

2018 ◽  
Vol 35 (03) ◽  
pp. 173-176 ◽  
Author(s):  
Ricardo Nucci ◽  
Wilson Jacob-Filho ◽  
Alexandre Busse ◽  
Laura Maifrino ◽  
Romeu de Souza
Keyword(s):  
Diaphragm Muscle ◽  
Tissue Samples ◽  
Muscle Tissues ◽  
Formalin Fixed Paraffin ◽  
Paraffin Block ◽  
Formalin Fixed Paraffin Embedded ◽  
Embedded Blocks ◽  
Health And Disease ◽  
Hematoxylin And Eosin ◽  
Formalin Fixed

Introduction The analysis of frozen muscle biopsies has become a routine method in the evaluation of muscle structure in health and disease. However, the technique for frozen muscle specimens is not widely available in countries with limited medical facilities. The present study aimed to elucidate a reproducible formalin-fixed and paraffin-embedded (FFPE) method for this type of analysis in postmortem muscles. Methods Diaphragm muscle was obtained within 1 hour of sudden death. Diaphragm strips were washed in saline solution, fixed in 10% formalin, frozen at 4°C in a refrigerator, and stored for 24 hours. Then, the tissue samples were processed into paraffin-embedded blocks. Transversal sections were cut from each paraffin block and stained with hematoxylin and eosin, Picrosirius red, Verhoeff-Van Gieson, and Congo red for the qualitative analysis. Results Our analysis indicated a well-preserved muscle. Conclusion In summary, we demonstrate a simple technique for a reproducible FFPE method in postmortem muscle tissues.

scholarly journals p16 Expression Correlates with Invasive Ocular Surface Squamous Neoplasms in HIV-Infected Mozambicans

2019 ◽  
Vol 6 (2) ◽  
pp. 123-128 ◽  
Author(s):  
Carla Carrilho ◽  
Chau Miu ◽  
Yeji Kim ◽  
Susan Karki ◽  
Alexandra Balmaceda ◽  
...  
Keyword(s):  
Ocular Surface ◽  
P Value ◽  
P16 Immunohistochemistry ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Intraepithelial Neoplasms ◽  
Hematoxylin And Eosin ◽  
Tissue Specimens ◽  
Formalin Fixed ◽  
P16 Expression

Background: p16 immunohistochemistry is widely used to diagnose human papillomavirus (HPV)-related squamous neoplasms of cervix, anogenital, head, and neck tissues. The incidence of these HPV-related squamous neoplasms is markedly increased in the HIV-infected population. Ocular surface squamous neoplasia (OSSN) is also more common in HIV-infected patients. However, the expression pattern of p16 in OSSN among HIV-infected patients is unclear. Here, we examined the expression of p16 in OSSN surgical excisions collected from a large HIV-infected cohort from ­Mozambique. Methods: OSSN surgical tissue specimens were collected from 75 Mozambican patients. Formalin-fixed, paraffin-embedded tissue blocks from these OSSNs were sectioned, stained with hematoxylin and eosin (H&E), and p16 expression by immunohistochemistry. H&E slides were reviewed to determine if OSSNs were noninvasive conjunctival intraepithelial neoplasms or invasive squamous cell carcinomas (SCC). Cases were classified as p16 positive or negative based on diffuse nuclear and cytoplasmic expression of p16 in neoplastic cells. Results: p16 positivity was found in a minority of OSSN cases (14/75). p16 positivity was significantly associated with the invasive SCC type of OSSN in HIV-infected patients (p value of 0.026). Conclusions: The majority of OSSNs in our HIV-infected cohort do not express p16. However, those cases that are p16-positive are significantly more likely to be the invasive SCC form of OSSN. We propose that p16 expression may identify more aggressive OSSNs in HIV-infected populations.

scholarly journals Alternative to xylene as a clearing agent in histopathology

2018 ◽  
Vol 10 (02) ◽  
pp. 189-193
Author(s):  
Nasar Alwahaibi ◽  
Shaima Aljaradi ◽  
Horiyah Alazri
Keyword(s):  
Cell Morphology ◽  
Nuclear Staining ◽  
Cytoplasmic Staining ◽  
Essential Step ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hematoxylin And Eosin ◽  
Formalin Fixed ◽  
Different Tissues

ABSTRACT INTRODUCTION: Clearing is an essential step in processing tissue for light microscopy. Xylene is the clearing agent used most commonly worldwide. Xylene is toxic and therefore a threat to personnel working in histopathology laboratories. We evaluated a safer alternative clearing agent for use in the histopathology laboratory. MATERIALS AND METHODS: We used 230 formalin-fixed, paraffin-embedded tissue blocks from 19 different tissues. Half of the specimens were processed using xylene and half were processed using UltraClear™. Tissues were evaluated for eight parameters: sectioning, nuclear staining, cytoplasmic staining, overall cell morphology, clarity of staining, uniformity of staining, quality of immunohistochemistry (IHC), and cost. RESULTS: Both UltraClear™ and xylene processed sections scored 100% for IHC. Sections processed using UltraClear™ were easy to cut (81.7%) as were xylene processed sections (96.5%). UltraClear™ processed sections showed 67%, 60.9%, 52.2%, 63.5%, and 67% for nuclear staining, cytoplasmic staining, cell morphology, clarity of staining, and uniformity of staining, respectively. UltraClear™ is twice as expensive as xylene. We found that tissues processed using UltraClear™ were easy to cut and worked well for both hematoxylin and eosin and IHC staining. CONCLUSION: UltraClear™ is less toxic, less flammable, friendlier to the environment, and easy to handle, but it is two times expensive than xylene. The findings of this study recommend the use of UltraClear™ solution as a routine clearing agent in histopathology laboratories. However, further studies are required.

scholarly journals A Method to Reuse Archived H&E Stained Histology Slides for a Multiplex Protein Biomarker Analysis

2019 ◽  
Vol 2 (4) ◽  
pp. 86
Author(s):  
James P. Hinton ◽  
Katerina Dvorak ◽  
Esteban Roberts ◽  
Wendy J. French ◽  
Jon C. Grubbs ◽  
...  
Keyword(s):  
Human Tumor ◽  
Clinical Value ◽  
Tissue Samples ◽  
Analysis Techniques ◽  
Formalin Fixed Paraffin ◽  
Biomarker Analysis ◽  
Formalin Fixed Paraffin Embedded ◽  
Hematoxylin And Eosin ◽  
Image Analysis Techniques ◽  
Formalin Fixed

Archived Hematoxylin and Eosin (H&E) stained pathology slides are routinely stored to index formalin-fixed paraffin-embedded (FFPE) sample tissue blocks. FFPE blocks are clinically annotated human tumor specimens that can be valuable in studies decades after the tissue is collected. If stored properly, they have the potential to yield a valuable number of serial sectioned slides for diagnostic or research purposes. However, some retrospective studies are limited in scope because the tissue samples have been depleted or not enough material is available in stored blocks for serial sections. The goal of these studies was to determine if archived H&E-stained slides can be directly reutilized by optimizing methods to de-stain and then re-stain the H&E stained slides to allow the detection of several biomarkers of interest using a conjugated antibody with chromogen multiplex immunohistochemistry procedure. This simple but innovative procedure, combined with image analysis techniques, demonstrates the ability to perform precise detection of relevant markers correlated to disease progression in initially identified tumor regions in tissue. This may add clinical value in retaining H&E slides for further use.

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