scholarly journals Postmortem Cardiopulmonary Pathology in Patients with COVID-19 Infection: Single-Center Report of 12 Autopsies from Lausanne, Switzerland

Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1357
Author(s):  
Sabina Berezowska ◽  
Karine Lefort ◽  
Kalliopi Ioannidou ◽  
Daba-Rokhya Ndiaye ◽  
Damien Maison ◽  
...  
Keyword(s):  
Diffuse Alveolar Damage ◽  
Virus Detection ◽  
Symptom Duration ◽  
Center Report ◽  
Formalin Fixed Paraffin ◽  
High Prevalence ◽  
Formalin Fixed Paraffin Embedded ◽  
Viral Copy ◽  
High Virus ◽  
High Level

We report postmortem cardio-pulmonary findings including detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in formalin-fixed paraffin embedded tissue in 12 patients with COVID-19. The 5 women and 7 men (median age: 73 years; range 35–96) died 6–38 days after onset of symptoms (median: 14.5 days). Eight patients received mechanical ventilation. Ten patients showed diffuse alveolar damage (DAD), 7 as exudative and 3 as proliferative/organizing DAD. One case presented as acute fibrinous and organizing pneumonia. Seven patients (58%) had acute bronchopneumonia, 1/7 without associated DAD and 1/7 with aspergillosis and necrotic bronchitis. Microthrombi were present in 5 patients, only in exudative DAD. Reverse transcriptase quantitative PCR detected high virus amounts in 6 patients (50%) with exudative DAD and symptom-duration ≤14 days, supported by immunohistochemistry and in-situ RNA hybridization (RNAscope). The 6 patients with low viral copy levels were symptomatic for ≥15 days, comprising all cases with organizing DAD, the patient without DAD and one exudative DAD. We show the high prevalence of DAD as a reaction pattern in COVID-19, the high number of overlying acute bronchopneumonia, and high-level pulmonary virus detection limited to patients who died ≤2 weeks after onset of symptoms, correlating with exudative phase of DAD.

PD-L1 expression, tumor mutational burden, and microsatellite instability status in 746 pancreas ductal adenocarcinomas.

2020 ◽  
Vol 38 (4_suppl) ◽  
pp. 757-757
Author(s):  
Amanda Hemmerich ◽  
Claire I. Edgerly ◽  
Daniel Duncan ◽  
Richard Huang ◽  
Natalie Danziger ◽  
...  
Keyword(s):  
Microsatellite Instability ◽  
Mutational Burden ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Comprehensive Genomic Profiling ◽  
Tumor Mutational Burden ◽  
Positive Score ◽  
Combined Positive Score ◽  
High Level ◽  
Positive Groups

757 Background: Pancreas ductal adenocarcinomas (PDA) has a 5-year survival rate of 6% with a need for new therapeutic options. The approval of pembrolizumab for some gastrointestinal cancers shows the potential of immunotherapy (IMT) in PDA. We evaluated the IMT-associated biomarkers of PD-L1 expression, tumor mutational burden (TMB), microsatellite instability (MSI) and PD-L1 amplification in PDAs. Methods: 746 formalin-fixed paraffin embedded samples were evaluated for PD-L1 IHC using the Dako 22C3 pharmDx assay and scored using tumor proportion score (TPS). The cases had comprehensive genomic profiling (CGP) via DNA sequencing, using a hybrid-capture next-generation sequencing assay (FoundationOne and FoundationOneCDx) for genomic alterations (GAs), TMB, and MSI. Results: PD-L1 was positive (TPS ≥ 1%) in 29% (214/746) and negative in 71% (532/746). 43/214 (20%) of positive cases were high positive (TPS ≥ 50%). TMB (590 cases) had a mean of 3.20, 3.46, and 3.61 mutations/Mb for PD-L1 negative, positive, and high positive groups. 3 hypermutated (TMB ≥ 20) were negative for PD-L1 expression. 3/581 cases were MSI-high with a high TMB score (average 23.53 mutations/Mb). 2 MSI-high cases were negative for PD-L1 and 1 was high positive. PD-L1 amplification was not detected (0/746). Only BCOR was significantly different between PD-L1 high positive and PD-L1 negative tumors (Table). Conclusions: Of 729 PDA cases, 29% were positive (TPS ≥ 1%) for PD-L1 expression while only 6% of all cases showed a high level of PD-L1 expression on tumor cells. TMB high (3/729) and MSI-High (3/729) cases were rare. Only 2 of the TMB high cases were also MSI-high. PD-L1 amplification was not detected. Comparing GAs in PD-L1 high positive vs negative cases was only significantly different for BCOR. Further investigation is needed to see if a combined positive score of PD-L1 expression may identify a subset of patients with PDA who are more likely to respond to IMT. [Table: see text]

scholarly journals High Prevalence of Human Polyomavirus 7 in Cholangiocarcinomas and Adjacent Peritumoral Hepatocytes: Preliminary Findings

2020 ◽  
Vol 8 (8) ◽  
pp. 1125 ◽  
Author(s):  
Faisal Klufah ◽  
Ghalib Mobaraki ◽  
Emil Chteinberg ◽  
Raed A. Alharbi ◽  
Véronique Winnepenninckx ◽  
...  
Keyword(s):  
Molecular Techniques ◽  
Human Polyomavirus ◽  
Biliary Duct ◽  
Formalin Fixed Paraffin ◽  
High Prevalence ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues ◽  
Rna In Situ Hybridization ◽  
Hepatobiliary Diseases

Cholangiocarcinoma (CCA) is a rare biliary-duct malignancy with poor prognosis. Recently, the presence of the human polyomavirus 6 (HPyV6) has been reported in the bile of diverse hepatobiliary diseases, particularly in the bile of CCA patients. Here, we investigated the presence of novel HPyVs in CCA tissues using diverse molecular techniques to assess a possible role of HPyVs in CCA. Formalin-Fixed Paraffin-Embedded (FFPE) tissues of 42 CCA patients were included in this study. PCR-based screening for HPyVs was conducted using degenerated and HPyV-specific primers. Following that, we performed FISH, RNA in situ hybridization (RNA-ISH), and immunohistochemistry (IHC) to assess the presence of HPyVs in selected tissues. Of all 42 CCAs, 25 (59%) were positive for one HPyV, while 10 (24%) CCAs were positive for 2 HPyVs simultaneously, and 7 (17%) were negative for HPyVs. Of the total 35 positive CCAs, 19 (45%) were positive for HPyV7, 4 (9%) for HPyV6, 2 (5%) for Merkel cell polyomavirus (MCPyV), 8 (19%) for both HPyV7/MCPyV, and 2 (5%) for both HPyV6/HPyV7 as confirmed by sequencing. The presence of viral nucleic acids was confirmed by specific FISH, while the RNA-ISH confirmed the presence of HPyV6 on the single-cell level. In addition, expression of HPyV7, HPyV6, and MCPyV proteins were confirmed by IHC. Our results strongly indicate that HPyV7, HPyV6, and MCPyV infect bile duct epithelium, hepatocytes, and CCA cells, which possibly suggest an indirect role of these viruses in the etiopathogenesis of CCA. Furthermore, the observed hepatotropism of these novel HPyV, in particular HPyV7, might implicate a role of these viruses in other hepatobiliary diseases.

scholarly journals Molecular Detection of SARS-CoV-2 Infection in FFPE Samples and Histopathologic Findings in Fatal SARS-CoV-2 Cases

2020 ◽  
Vol 154 (2) ◽  
pp. 190-200 ◽  
Author(s):  
Miroslav Sekulic ◽  
Holly Harper ◽  
Behtash G Nezami ◽  
Daniel L Shen ◽  
Simona Pichler Sekulic ◽  
...  
Keyword(s):  
Diffuse Alveolar Damage ◽  
Ffpe Tissue ◽  
Qrt Pcr ◽  
Morphological Findings ◽  
Formalin Fixed Paraffin ◽  
Ffpe Samples ◽  
Formalin Fixed Paraffin Embedded ◽  
Control And Prevention ◽  
Next Generation Sequencing Ngs ◽  
Viral Sequencing

Abstract Objectives To report methods and findings of 2 autopsies with molecular evaluation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positive individuals. Methods Postmortem examination was completed following Centers for Disease Control and Prevention public guidelines. Numerous formalin-fixed paraffin-embedded (FFPE) tissue types from each case were surveyed for SARS-CoV-2 RNA by quantitative reverse transcription polymerase chain reaction (qRT-PCR). SARS-CoV-2 viral genome was sequenced by next-generation sequencing (NGS) from FFPE lung tissue blocks. Results Postmortem examinations revealed diffuse alveolar damage, while no viral-associated hepatic, cardiac, or renal damage was observed. Viral RNA was detected in lungs, bronchi, lymph nodes, and spleen in both cases using qRT-PCR method. RNA sequencing using NGS in case 1 revealed mutations most consistent with Western European Clade A2a with ORF1a L3606F mutation. Conclusions SARS-CoV-2 testing and viral sequencing can be performed from FFPE tissue. Detection and sequencing of SARS-CoV-2 in combination with morphological findings from postmortem tissue examination can aid in gaining a better understanding of the virus’s pathophysiologic effects on human health.

scholarly journals High prevalence of Epstein-Barr virus in the Reed-Sternberg cells of Hodgkin's disease occurring in Peru

Blood ◽  
1993 ◽  
Vol 81 (2) ◽  
pp. 496-501 ◽  
Author(s):  
KL Chang ◽  
PF Albujar ◽  
YY Chen ◽  
RM Johnson ◽  
LM Weiss
Keyword(s):  
In Situ Hybridization ◽  
Epstein Barr Virus ◽  
Lower Percentage ◽  
Double Labeling ◽  
Barr Virus ◽  
Formalin Fixed Paraffin ◽  
High Prevalence ◽  
Formalin Fixed Paraffin Embedded ◽  
Epstein Barr

Abstract The Epstein-Barr virus (EBV) has been implicated in the pathogenesis of Hodgkin‧s disease (HD). This study was undertaken to determine whether the association of EBV with HD showed geographical variation, as in Burkitt‧s lymphoma. We studied 32 formalin-fixed, paraffin-embedded cases of HD occurring in Peru. EBV DNA-RNA in situ hybridization was performed using a 30-base biotinylated antisense oligonucleotide complementary to the EBER1 gene of EBV. EBV immunohistochemistry was also performed, using a monoclonal antibody (MoAb) to the latent membrane protein (LMP1) of EBV. Identification of the precise cellular subset staining with EBV was accomplished via double-labeling with MoAbs directed against Reed-Sternberg cells (LeuM1/CD15) and B cells (L26/CD20). EBV RNA was identified in all or virtually all of the Reed- Sternberg cells and variants in 30 of the 32 (94%) cases of HD by in situ hybridization. LMP1 expression was identified in 83% of the EBER1- positive cases. Double-labeling studies confirmed the localization of EBV RNA to CD15-expressing Hodgkin‧s cells. This study found an extremely high prevalence of EBV in Peruvian HD, in contrast to the much lower percentage of EBV-associated cases of HD occurring in “Western” patients.

scholarly journals Prevalence of Somatic Mutations in Aldosterone-Producing Adenomas in Japanese Patients

2020 ◽  
Vol 105 (11) ◽  
Author(s):  
Kazutaka Nanba ◽  
Yuto Yamazaki ◽  
Nolan Bick ◽  
Kei Onodera ◽  
Yuta Tezuka ◽  
...  
Keyword(s):  
Racial Differences ◽  
Somatic Mutations ◽  
Japanese Patients ◽  
University Hospital ◽  
Female Patients ◽  
Targeted Next Generation Sequencing ◽  
Formalin Fixed Paraffin ◽  
High Prevalence ◽  
Formalin Fixed Paraffin Embedded ◽  
Male Patients

Abstract Context Results of previous studies demonstrated clear racial differences in the prevalence of somatic mutations among patients with aldosterone-producing adenoma (APA). For instance, those in East Asian countries have a high prevalence of somatic mutations in KCNJ5, whereas somatic mutations in other aldosterone-driving genes are rare. Objectives To determine somatic mutation prevalence in Japanese APA patients using an aldosterone synthase (CYP11B2) immunohistochemistry (IHC)-guided sequencing approach. Method Patients with a unilateral form of primary aldosteronism who underwent adrenalectomy at the Tohoku University Hospital were studied. Based on CYP11B2 immunolocalization of resected adrenals, genomic DNA was isolated from the relevant positive area of 10% formalin-fixed, paraffin-embedded tissue of the APAs. Somatic mutations in aldosterone-driving genes were studied in APAs by direct Sanger sequencing and targeted next-generation sequencing. Results CYP11B2 IHC-guided sequencing determined APA-related somatic mutations in 102 out of 106 APAs (96%). Somatic KCNJ5 mutation was the most frequent genetic alteration (73%) in this cohort of Japanese patients. Somatic mutations in other aldosterone-driving genes were also identified: CACNA1D (14%), ATP1A1 (5%), ATP2B3 (4%), and CACNA1H (1%), including 2 previously unreported mutations. KCNJ5 mutations were more often detected in APAs from female patients compared with those from male patients [95% (36/38) vs 60% (41/68); P < 0.0001]. Conclusion IHC-guided sequencing defined somatic mutations in over 95% of Japanese APAs. While the dominance of KCNJ5 mutations in this particular cohort was confirmed, a significantly higher KCNJ5 prevalence was detected in female patients. This study provides a better understanding of genetic spectrum of Japanese APA patients.

scholarly journals Chromogenic In Situ Hybridization and p16/Ki67 Dual Staining on Formalin-Fixed Paraffin-Embedded Cervical Specimens: Correlation with HPV-DNA Test, E6/E7 mRNA Test, and Potential Clinical Applications

2013 ◽  
Vol 2013 ◽  
pp. 1-11 ◽  
Author(s):  
Roberta Zappacosta ◽  
Antonella Colasante ◽  
Patrizia Viola ◽  
Tommaso D’Antuono ◽  
Giuseppe Lattanzio ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Dna Test ◽  
Hpv Dna ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Chromogenic In Situ Hybridization ◽  
High Level ◽  
Formalin Fixed ◽  
Hpv Dna Test

Although HPV-DNA test and E6/E7 mRNA analyses remain the current standard for the confirmation of human papillomavirus (HPV) infections in cytological specimens, no universally adopted techniques exist for the detection of HPV in formalin-fixed paraffin-embedded samples. Particularly, in routine laboratories, molecular assays are still time-consuming and would require a high level of expertise. In this study, we investigated the possible use of a novel HPV tyramide-based chromogenic in situ hybridization (CISH) technology to locate HPV on tissue specimens. Then, we evaluate the potential usefulness ofp16INK4a/Ki-67 double stain on histological samples, to identify cervical cells expressing HPV E6/E7 oncogenes. In our series, CISH showed a clear signal in 95.2% of the specimens and reached a sensitivity of 86.5%. CISH positivity always matched with HPV-DNA positivity, while 100% of cases with punctated signal joined with cervical intraepithelial neoplasia grade 2 or worse (CIN2+). p16/Ki67 immunohistochemistry gave an interpretable result in 100% of the cases. The use of dual stain significantly increased the agreement between pathologists, which reached 100%. Concordance between dual stain and E6/E7 mRNA test was 89%. In our series, both CISH andp16INK4a/Ki67 dual stain demonstrated high grade of performances. In particular, CISH would help to distinguish episomal from integrated HPV, in order to allow conclusions regarding the prognosis of the lesion, whilep16INK4a/Ki67 dual stain approach would confer a high level of standardization to the diagnostic procedure.

scholarly journals Tissue distribution and biochemical properties of an ocular melanoma-associated antigen.

1985 ◽  
Vol 33 (12) ◽  
pp. 1190-1196 ◽  
Author(s):  
L A Donoso ◽  
R Folberg ◽  
K Edelberg ◽  
V Arbizo ◽  
B Atkinson ◽  
...  
Keyword(s):  
Screening Method ◽  
Biochemical Properties ◽  
Ocular Melanoma ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
High Prevalence ◽  
Formalin Fixed Paraffin Embedded ◽  
Protein Rich ◽  
Formalin Fixed ◽  
Uveal Melanomas

A screening method is described to select monoclonal antibodies (Mabs) that bind to ocular melanoma-associated antigens (MAAs) retained in formalin-fixed, paraffin-embedded tissue sections. Small sections of epithelioid or spindle-cell-type uveal melanomas were cut into 2 mm cubes and reembedded in one block. Microslides were cut from this block and used to screen hybridoma supernatant fluid. Using this screening method, three MAbs were selected from two separate fusions of mouse myeloma cells with spleen cells of mice immunized previously with either ocular melanoma cells obtained fresh at enucleation or cells of a cutaneous melanoma cell line. Although all three MAbs showed similar specificities, MAb8-1H showed the strongest immunohistochemical reaction and was studied further in detail. MAb8-1H bound to 91% (71/79) of the choroidal or ciliochoroidal melanomas tested, indicating a high prevalence of this antigen in uveal melanomas. The antigen defined by MAb8-1H was isolated, purified, and partially characterized as a 40,000-50,000 dalton, highly glycosylated protein rich in glycine, serine, and glutamic acid, as is typical of a mucin-type glycoprotein.

scholarly journals Clinicopathological and Immunohistochemical Analysis of 21 cases of Traumatic Ulcerative Granuloma with Stromal Eosinophilia Using CD30, CD68 and TGF-β1

2018 ◽  
Vol 30 (4) ◽  
pp. 45-53
Author(s):  
Mustafa B Al-Talqani ◽  
Bashar H Abdullah ◽  
Ameer D Hameedi
Keyword(s):  
Lymphoproliferative Disorder ◽  
Mononuclear Cells ◽  
Immunohistochemical Analysis ◽  
Excisional Biopsy ◽  
Clinicopathological Parameters ◽  
Ulcerative Lesion ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
High Level ◽  
Tgf Β1

Background: Traumatic ulcerative granuloma with stromal eosinophilia is an impressive benign chronic ulcerative lesion of the oral mucosa with vague etiopathogenesis. It was supposed to represent an oral counterpart of primary cutaneous CD30+ lymphoproliferative disorder. Histopathologically, it is characterized by mixed inflammatory infiltrate predominated by histiocytes, lymphocytes and eosinophils along with presence of scattered large atypical mononuclear cells. It has worrisome clinical presentation. It may heal spontaneously, but in most occasions it persists and never heal unless removed surgically (incisional or excisional biopsy). A rare subset may show worrisome immunohistochemical features. Follow up is highly recommended. Materials and methods: Formalin fixed - paraffin embedded tissue blocks of twenty-one cases were cut and mounted on positively charged slides and stained by primary antibodies (CD30, CD68 and TGF-β1). A statistical analysis was performed between the immunohistochemical scores for markers with each other and with clinicopathological parameters (age, sex, size of ulcer, number of eosinophils and mitoses). Results: The age of the patients ranged from 20 to 72 years, with a higher female propensity. Immunohistochemical positive expression for CD30 (16 case) mainly involved round small lymphocytes, while all cases were positive for CD68 and TGF-β1. Statistically, there was no significant relation between the scores of CD30, CD68 and TGF-β1 with each other and with the aforementioned parameters, (P<0.05). The eosinophils count showed a significant positive correlation with age (P=0.008), size of ulcer (P=0.007) and mitoses (P=0.004). Conclusion: Traumatic ulcerative granuloma with stromal eosinophilia is a benign and reactive chronic oral ulcerative lesion rather than being CD30+ lymphoproliferative disorder; this conclusion is supported by heterogeneous, focal and nonspecific staining for CD30 and being typically infiltrated by CD68+ macrophages. Whereas, a high level of expression for TGF-β1 indicated that the aforementioned factor was not associated with the delayed healing of this lesion

scholarly journals Multisystemic cellular tropism of SARS-CoV-2 in autopsies of COVID-19 patients

2021 ◽  
Author(s):  
Dickson W.L. Wong ◽  
Barbara M. Klinkhammer ◽  
Sonja Djudjaj ◽  
Sophia Villwock ◽  
M. Cherelle Timm ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
In Situ Hybridization ◽  
Virus Detection ◽  
Viral Rna ◽  
Transmission Electron ◽  
Formalin Fixed Paraffin ◽  
Organ Systems ◽  
Formalin Fixed Paraffin Embedded ◽  
Ffpe Tissues

Background: Multiorgan tropism of SARS-CoV-2 has previously been shown for several major organs. Methods: We have comprehensively analyzed 25 different formalin-fixed paraffin-embedded (FFPE) tissues/organs from autopsies of fatal COVID-19 cases (n=8), using detailed histopathological assessment, detection of SARS-CoV-2 RNA using polymerase chain reaction and RNA in situ hybridization, viral protein using immunohistochemistry, and virus particles using transmission electron microscopy. Finally, we confirmed these findings in an independent external autopsy cohort (n=9). Findings: SARS-CoV-2 RNA was mainly localized in epithelial cells, endothelial and mesenchymal cells across all organs. Next to lung, trachea, kidney, heart, or liver, viral RNA was also found in tonsils, salivary glands, oropharynx, thyroid, adrenal gland, testicles, prostate, ovaries, small bowel, lymph nodes, skin and skeletal muscle. Viral RNA was predominantly found in cells expressing ACE2, TMPRSS2, or both. The SARS-CoV-2 replicating RNA was also detected in these organs. Immunohistochemistry and electron microscopy were not suitable for reliable and specific SARS-CoV-2 detection in autopsies. The findings were validated using in situ hybridization on external COVID-19 autopsy samples. Finally, apart from the lung, correlation of virus detection and histopathological assessment did not reveal any specific alterations that could be attributed to SARS-CoV-2. Conclusion: SARS-CoV-2 could be observed in virtually all organs, colocalizing with ACE2 and TMPRSS2 mainly in epithelial but also in mesenchymal and endothelial cells, and viral replication was found across all organ systems. Apart from the respiratory tract, no specific (histo-)morphologic alterations could be assigned to the SARS-CoV-2 infection.

scholarly journals High prevalence of Epstein-Barr virus in the Reed-Sternberg cells of Hodgkin's disease occurring in Peru

Blood ◽  
1993 ◽  
Vol 81 (2) ◽  
pp. 496-501 ◽  
Author(s):  
KL Chang ◽  
PF Albujar ◽  
YY Chen ◽  
RM Johnson ◽  
LM Weiss
Keyword(s):  
In Situ Hybridization ◽  
Epstein Barr Virus ◽  
Lower Percentage ◽  
Double Labeling ◽  
Barr Virus ◽  
Formalin Fixed Paraffin ◽  
High Prevalence ◽  
Formalin Fixed Paraffin Embedded ◽  
Epstein Barr

The Epstein-Barr virus (EBV) has been implicated in the pathogenesis of Hodgkin‧s disease (HD). This study was undertaken to determine whether the association of EBV with HD showed geographical variation, as in Burkitt‧s lymphoma. We studied 32 formalin-fixed, paraffin-embedded cases of HD occurring in Peru. EBV DNA-RNA in situ hybridization was performed using a 30-base biotinylated antisense oligonucleotide complementary to the EBER1 gene of EBV. EBV immunohistochemistry was also performed, using a monoclonal antibody (MoAb) to the latent membrane protein (LMP1) of EBV. Identification of the precise cellular subset staining with EBV was accomplished via double-labeling with MoAbs directed against Reed-Sternberg cells (LeuM1/CD15) and B cells (L26/CD20). EBV RNA was identified in all or virtually all of the Reed- Sternberg cells and variants in 30 of the 32 (94%) cases of HD by in situ hybridization. LMP1 expression was identified in 83% of the EBER1- positive cases. Double-labeling studies confirmed the localization of EBV RNA to CD15-expressing Hodgkin‧s cells. This study found an extremely high prevalence of EBV in Peruvian HD, in contrast to the much lower percentage of EBV-associated cases of HD occurring in “Western” patients.

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