PD-L1 Immunohistochemistry on Cell Block Preparations From Different Fixatives: A Pilot Study

2019 ◽  
Vol 152 (Supplement_1) ◽  
pp. S95-S95
Author(s):  
Victoria Costa ◽  
David Kim ◽  
Abha Goyal ◽  
Bing He ◽  
Yifang Liu ◽  
...  
Keyword(s):  
Pilot Study ◽  
Personalized Medicine ◽  
Cell Block ◽  
Fixation Methods ◽  
Tonsillar Tissue ◽  
Fixed Cell ◽  
Specific Fixation ◽  
Formalin Fixed

Abstract Objectives Immunohistochemistry especially for biomarkers such as PD-L1 in personalized medicine is increasingly being performed on cell blocks from cytopathology samples. We evaluated the effects of staining cell blocks (CBs) from different cytologic fixatives using four different commercially available PD-L1 antibody clones. Methods PD-L1 immunohistochemistry (IHC) using four different commercially available clones was performed on eight cell blocks processed from two different fixatives and also on eight formalin-fixed tissues. The clones used were 22C3 at 1:50 dilution and 1:100 dilution, SP263, SP142, and E1L3N. The cell block (CB) samples contained cells that strongly express PD-L1 and either fixed in CytoLyt (methanol-based fixative, n = 4) or CytoRich (ethanol-formalin-based fixative, n = 4). Formalin-fixed control tissues (tonsillar tissue, n = 4; stomach, n = 4) were also included. Results Expression for PD-L1 was noted on all the CytoRich fixed cell blocks (n = 4) for all four antibody clones, with the intensity and distribution of expression comparable to the formalin fixed tissues (n = 8). However, consistent absence of expression for PD-L1 was noted on all the CytoLyt fixed cell blocks (n = 4) for all four antibody clones. Conclusion The results of this pilot study demonstrate that PD-L1 IHC on cell blocks is feasible. However, there is a need to validate IHC protocols according to specific fixation methods.

Validation of PD-L1 Immunohistochemical Testing on Formalin Fixed Paraffin Embedded (FFPE) Cell Block (CB) Preparations

2020 ◽  
Vol 9 (6) ◽  
pp. S53-S54
Author(s):  
Priyanka Karam ◽  
Yonah Ziemba ◽  
Sean Hacking ◽  
Karen Chau ◽  
Suganthi Soundararajan ◽  
...  
Keyword(s):  
Cell Block ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed ◽  
Immunohistochemical Testing

scholarly journals Statin dose reduction with complementary diet therapy: A pilot study of personalized medicine

2018 ◽  
Vol 11 ◽  
pp. 137-144 ◽  
Author(s):  
Bianca Scolaro ◽  
Marina S. Nogueira ◽  
Aline Paiva ◽  
Adriana Bertolami ◽  
Lucia P. Barroso ◽  
...  
Keyword(s):  
Pilot Study ◽  
Personalized Medicine ◽  
Dose Reduction ◽  
Diet Therapy ◽  
Statin Dose ◽  
Complementary Diet

scholarly journals CellBlockistry: Chemistry and art of cell-block making – A detailed review of various historical options with recent advances

CytoJournal ◽  
2019 ◽  
Vol 16 ◽  
pp. 12 ◽  
Author(s):  
Vinod B Shidham
Keyword(s):  
Ffpe Tissue ◽  
Cell Block ◽  
Molecular Tests ◽  
Recent Advances ◽  
Pattern Approach ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Technical Advances ◽  
Ancillary Studies ◽  
Formalin Fixed

Cell-blocks are paraffin-embedded versions of cytology specimens comparable to the formalin-fixed paraffin-embedded (FFPE) tissue from surgical pathology specimens. They allow various elective ancillary studies on a variety of specimens with enhanced cytopathologic interpretation, including opportunity to perform molecular tests. However, different dictionaries and internet search engines primarily project “cellblock” and “cell block” definition in relation to prisons. Most of the top searches lead to information related to “prison cells” followed by a few cytopathology-related searches. Due to this in the current review, it is recommended that the word for cytopathology purposes should be hyphenated and spelled as “cell-block.” Cell-blocks have been increasingly indicated on most cytology specimens. Its role is growing further with the ongoing addition of new immunohistochemistry (IHC) markers with technical advances including multicolor IHC and the SCIP (subtractive coordinate immunoreactivity pattern) approach. In addition, it is an important source of tissue for many ancillary studies even as archived material retrospectively at later stage of management if the cell-blocks are improved qualitatively and quantitatively. Because of this, the significance of cell-block is critical with the increasing number of molecular markers standardized predominantly on FFPE tissue. As compared to core biopsies, high-quality cell-blocks prepared with enhanced methodologies predominantly contain concentrated diagnostic tumor cells required for the molecular tests without significant stromal contamination. This review introduces the terminology of CellBlockistry as the science of studying chemistry and the art of achieving quantitatively and qualitatively improved cell-blocks from different types of specimens. The review addresses the cell-block making process as “cell-blocking” and discusses different historical limitations with emphasis on recent advances.

scholarly journals Mycobiome analysis in fungal infected formalin-fixed and paraffin-embedded tissues for identification of pathogenic fungi: a pilot study

F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 758
Author(s):  
Taebum Lee ◽  
Hee Young Na ◽  
Sun-ju Byeon ◽  
Kyoung-Mee Kim ◽  
Hey Seung Lee ◽  
...  
Keyword(s):  
Pilot Study ◽  
Dna Sequences ◽  
Pathogenic Fungi ◽  
Median Number ◽  
Fungal Organisms ◽  
Ffpe Tissues ◽  
Ncbi Refseq ◽  
Fungal Dna ◽  
Multiplex Sequencing ◽  
Formalin Fixed

Background: Fungal organisms are frequently observed in surgical pathological diagnosis. In order to more accurately identify fungi in formalin-fixed and paraffin-embedded (FFPE) tissues, it is necessary to use genomic information. The purpose of our pilot study is to identify the factors to be considered for the identification of pathogenic fungi using mycobiome analysis in FFPE tissues. Methods: We selected 49 cases in five hospitals. In each case, FFPE tissue was cut into 50 µm and DNA was extracted. Multiplex PCR with four primers (ITS1, ITS2, ITS3 and ITS4) was performed. Multiplex sequencing was performed using a MinION device according to the manufacturer’s protocol. Sequences of each case were searched using BLASTN with an ITS database from NCBI RefSeq Targeted Loci Project with default parameters. Results: A total of 2,526 DNA sequences were sequenced. We were able to identify 342 fungal sequences in 24 (49.0%, 24/49) cases. The median number of detected fungal sequences per case was 3 (1Q: 1 and 3Q: 14.25). Of the fungal DNA sequences, 215 (62.87%) contained the entire region of ITS1 or ITS2. The remaining 127 fungal DNA sequences were identified as fungi using a partial sequence of ITS1, ITS2, 5.8S, LSU or SSU. Conclusion: In conclusion, we have identified the possibility of finding pathogenic fungi through mycobiome analysis in fungal infected FFPE tissues using nanopore sequencing. However, we have also found several limitations to be solved for further studies. If we develop a method to characterize pathogenic fungi in FFPE tissues in a follow-up study, we think it will help patients to use appropriate antifungal agents.

scholarly journals Differential expression of FGFRs signaling pathway components in bladder cancer: A step toward personalized medicine

2017 ◽  
Vol 20 (2) ◽  
pp. 75-81 ◽  
Author(s):  
Z Ousati Ashtiani ◽  
J Tavakkoly-Bazzaz ◽  
SA Salami ◽  
MR Pourmand ◽  
F Mansouri ◽  
...  
Keyword(s):  
Bladder Cancer ◽  
Pilot Study ◽  
Growth Factor ◽  
Personalized Medicine ◽  
Fibroblast Growth Factor ◽  
Real Time ◽  
Expression Patterns ◽  
Fibroblast Growth ◽  
Tissue Samples ◽  
Over Expression

AbstractVariations Improper activation and inappropriate expression of fibroblast growth factor receptors (FGFRs) in cancer suggests that they can act as therapeutic targets. Fibroblast growth factor receptor inhibitors are currently employed in clinical trials of different cancers. Regarding the essence and the importance of the personalized medicine, mainly mirrored by remarkable inter-individual variations in different populations, we aimed to perform a pilot study to addressFGFR1andFGFR3expression levels and their correlation with the clinicopathological features in Iranian patients with bladder cancer (BC). Paired tumor and adjacent non tumor tissue samples along with their clinico-pathological parameters were obtained from 50 cases diagnosed with BC in different stages and grades. The mRNA expressions ofFGFR1andFGFR3in tissue samples were determined by real-time polymerase chain reaction (real-time PCR). The expression levels ofFGFR3were significantly higher in tumor tissues when compared to adjacent normal tissues (p= 0.007), regardless of the stages and grades of the tumor. Over expression was associated with cigarette smoking (p= 0.037) and family history for cancer (p= 0.004). Decreased expression ofFGFR1was observed, remarkably evident in high-grade tumors (p= 0.047), while over expression was detected in low-grade samples. This pilot study clearly suggests that in Iranian BC patientsFGFR1andFGFR3expression patterns are different, and also highly distinctive with regard to the tumor’s stage and grade. Such particular expression patterns may indicate their special values to be employed for interventional studies aiming targeted therapy. Further studies with a larger sample size are needed to validate our results.

scholarly journals Mycobiome analysis in fungal Infected formalin-fixed and paraffin-embedded tissues for identification of pathogenic fungi: A pilot study

2020 ◽  
Author(s):  
Taebum Lee ◽  
Hee Young Na ◽  
Kyoung-Mee Kim ◽  
Hey Seung Lee ◽  
Sung-Hye Park ◽  
...  
Keyword(s):  
Pilot Study ◽  
Pathogenic Fungi ◽  
Sequencing Method ◽  
Fungal Organisms ◽  
Ffpe Tissues ◽  
Ncbi Refseq ◽  
Fungal Dna ◽  
Multiplex Sequencing ◽  
Formalin Fixed

ABSTRACTBackgroundFungal organisms are frequently observed in surgical pathological diagnosis. In order to more accurately identify fungi in formalin-fixed and paraffin-embedded (FFPE) tissues, it is necessary to use genomic information. The purpose of our pilot study is to identify the factors to be considered for the identification of pathogenic fungi using mycobiome analysis in FFPE tissues.MethodsWe selected 49 cases in five hospitals. In each case, FFPE tissue was cut into 50 µm and DNA was extracted. Multiplex PCR with four primers (ITS1, ITS2, ITS3 and ITS4) was performed. Multiplex sequencing was performed using MinION device according to the manufacturer’s protocol. Sequences of each case were searched using BLASTN with an ITS database from NCBI RefSeq Targeted Loci Project with default parameter.ResultsA total of 2,526 DNA nucleotides were sequenced. We were able to identify 342 fungal nucleotides in 24 (49.0%, 24/49) cases. The median value of the detected fungal DNA per case was 3 (1Q: 1 and 3Q: 14.25). The 215 (62.87%) fungal DNA contained the entire region of ITS1 or ITS2. The remaining 127 fungal DNAs were identified as fungi using partial sequence of ITS1, ITS2, 5.8S, LSU or SSU.ConclusionIn conclusion, we have identified the possibility of finding pathogenic fungi through mycobiome analysis in fungal infected FFPE tissues using nanopore sequencing method. However, we have also found several limitations to be solved for further studies. If we develop a method to characterize pathogenic fungi in FFPE tissues in a follow-up study, we think it will help patients to use appropriate antifungal agents.

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