Journal: Frontiers in Oncology

Article Title: Whole-exome sequencing of calcitonin-producing pancreatic neuroendocrine neoplasms indicates a unique molecular signature

doi: 10.3389/fonc.2023.1160921

Figure Lengend Snippet: Representative images of immunohistochemistry stainings (IHC) of the calcitonin- producing pancreatic neuroendocrine neoplasms tissue (CT-pNENs), isolated from 5 patients (P1–P5), for Mucin 4 ( MUC4 ), Mucin 16 ( MUC16 ), human Insulinoma associated 1 ( INSM1 ), Calcitonin, Chromogranin A ( Chrgr A ), Synaptophysin and proliferation Marker Protein ( Ki-67 ). The staining was performed in primary resected tumor tissue. Magnification 100x (except MUC4 and MUC16 : magnification 40x). Patient P1 shows the double positive liver metastasis for staining MUC4 and MUC16 . Patient P2 shows MUC4 cytoplasmic and nuclear staining in the same case. Patient P4 is positive for MUC16 and Patient P5 for MUC4 . Patient P3 and P4 are negative for MUC4 and patient P2, P3 and P5 are negative for MUC16 . Patient P2 and P3 show a negative staining for INSM1 .

Article Snippet: In this study, 2-μm FFPE tissue slices were stained with primary antibodies against human calcitonin 1:1,000 (A0576, Dako Agilent, Santa Clara, CA, USA), human synaptophysin 1:600 (M7315, Dako Agilent), human chromogranin A 1:2,000 (503-1524 ZYTOMED SYSTEMS, Berlin, Germany), human Ki-67 1:200 (M7240, Dako Agilent), human insulin 1:200 (Mob234 Diagnostic BioSystems, Pleasanton, CA, USA), human adrenocorticotropin (ACTH) 1:1,000 (M3501, Dako Agilent), and human insulinoma-associated 1 ( INSM1 ) 1:100 (BSB-123 Bio SB, Santa Barbara, CA, USA).

Techniques: Immunohistochemistry, Isolation, Marker, Staining, Negative Staining

Journal: Frontiers in Oncology

Article Title: Whole-exome sequencing of calcitonin-producing pancreatic neuroendocrine neoplasms indicates a unique molecular signature

doi: 10.3389/fonc.2023.1160921

Figure Lengend Snippet: Germline and missense single nucleotide variations (SNV) in genes mutated in calcitonin- producing pancreatic neuroendocrine neoplasms tissue (CT-pNEN) tumors previously described in other pancreatic neuroendocrine neoplasms tissue (pNEN) ( 28 - 33 ).

Article Snippet: In this study, 2-μm FFPE tissue slices were stained with primary antibodies against human calcitonin 1:1,000 (A0576, Dako Agilent, Santa Clara, CA, USA), human synaptophysin 1:600 (M7315, Dako Agilent), human chromogranin A 1:2,000 (503-1524 ZYTOMED SYSTEMS, Berlin, Germany), human Ki-67 1:200 (M7240, Dako Agilent), human insulin 1:200 (Mob234 Diagnostic BioSystems, Pleasanton, CA, USA), human adrenocorticotropin (ACTH) 1:1,000 (M3501, Dako Agilent), and human insulinoma-associated 1 ( INSM1 ) 1:100 (BSB-123 Bio SB, Santa Barbara, CA, USA).

Techniques: Mutagenesis

Journal: Frontiers in Oncology

Article Title: Whole-exome sequencing of calcitonin-producing pancreatic neuroendocrine neoplasms indicates a unique molecular signature

doi: 10.3389/fonc.2023.1160921

Figure Lengend Snippet: Pathogenic, germline and missense single nucleotide variations (SNV) in genes mutated in calcitonin- producing pancreatic neuroendocrine neoplasms tissue (CT-pNEN) tumors not previously described in other pancreatic neuroendocrine neoplasms tissue (pNEN) ( 28 - 33 ).

Article Snippet: In this study, 2-μm FFPE tissue slices were stained with primary antibodies against human calcitonin 1:1,000 (A0576, Dako Agilent, Santa Clara, CA, USA), human synaptophysin 1:600 (M7315, Dako Agilent), human chromogranin A 1:2,000 (503-1524 ZYTOMED SYSTEMS, Berlin, Germany), human Ki-67 1:200 (M7240, Dako Agilent), human insulin 1:200 (Mob234 Diagnostic BioSystems, Pleasanton, CA, USA), human adrenocorticotropin (ACTH) 1:1,000 (M3501, Dako Agilent), and human insulinoma-associated 1 ( INSM1 ) 1:100 (BSB-123 Bio SB, Santa Barbara, CA, USA).

Techniques: Mutagenesis

Journal: Frontiers in Oncology

Article Title: Whole-exome sequencing of calcitonin-producing pancreatic neuroendocrine neoplasms indicates a unique molecular signature

doi: 10.3389/fonc.2023.1160921

Figure Lengend Snippet: Potentially deleterious, somatic and missense single nucleotide variations (SNV) of calcitonin- producing pancreatic neuroendocrine neoplasms tissue (CT-pNEN) tumors.

Article Snippet: In this study, 2-μm FFPE tissue slices were stained with primary antibodies against human calcitonin 1:1,000 (A0576, Dako Agilent, Santa Clara, CA, USA), human synaptophysin 1:600 (M7315, Dako Agilent), human chromogranin A 1:2,000 (503-1524 ZYTOMED SYSTEMS, Berlin, Germany), human Ki-67 1:200 (M7240, Dako Agilent), human insulin 1:200 (Mob234 Diagnostic BioSystems, Pleasanton, CA, USA), human adrenocorticotropin (ACTH) 1:1,000 (M3501, Dako Agilent), and human insulinoma-associated 1 ( INSM1 ) 1:100 (BSB-123 Bio SB, Santa Barbara, CA, USA).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Role of Epstein-Barr Virus and Human Papillomavirus Coinfection in Cervical Intraepithelial Neoplasia in Chinese Women Living With HIV

doi: 10.3389/fcimb.2021.703259

Figure Lengend Snippet: Sequences of RT-PCR primers used for validation.

Article Snippet: Briefly, sections were blocked for 1 h, incubated overnight at 4°C with a rabbit primary antibody against human CACNG4 (Immunoway, YM3404), and subsequently incubated with a secondary Alexa Fluor 568-conjugated goat anti-rabbit IgG antibody (Abcam, ab175696) for 1 h. Slides were mounted with mounting medium containing DAPI and imaged by a Leica SP8 laser scanning confocal microscope system.

Techniques: Biomarker Discovery

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Role of Epstein-Barr Virus and Human Papillomavirus Coinfection in Cervical Intraepithelial Neoplasia in Chinese Women Living With HIV

doi: 10.3389/fcimb.2021.703259

Figure Lengend Snippet: Selected mRNAs with significant changes and overexpression of CACNG4 in EBV-HPV-associated CIN lesions. Expression levels of the selected, significantly altered mRNAs of the mRNA-seq analysis (A) and RT-PCR validation (B) . Each value was normalized to the GAPDH value and is presented as the mean ± SD, * p <0.05, compared with the HPV group. (C) A red fluorescence signal indicating the presence of CACNG4 was detected in cervical tissues. (D) Protein expression of CACNG4 relative to β-actin expression in cervical tissues. (E) Densitometric analysis of the immunoblots. The intensity of each band was normalized to the β-actin intensity. Due to the limited amount of clinical material, the EBV-HPV sample consisted of 4 pooled biological replicates and the HPV sample consisted of 3 pooled biological replicates in (D, E) . (F) The expression of CACNG4 between cervical cancer samples and normal tissue samples taken from TCGA and the GTEx databases. TPM, transcripts per million.

Article Snippet: Briefly, sections were blocked for 1 h, incubated overnight at 4°C with a rabbit primary antibody against human CACNG4 (Immunoway, YM3404), and subsequently incubated with a secondary Alexa Fluor 568-conjugated goat anti-rabbit IgG antibody (Abcam, ab175696) for 1 h. Slides were mounted with mounting medium containing DAPI and imaged by a Leica SP8 laser scanning confocal microscope system.

Techniques: Over Expression, Expressing, Reverse Transcription Polymerase Chain Reaction, Biomarker Discovery, Fluorescence, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Regional Differences in the Permeability Barrier of the Skin—Implications in Acantholytic Skin Diseases

doi: 10.3390/ijms221910428

Figure Lengend Snippet: Different barrier functions and gene expression profiles of permeability barrier components characterize healthy GP, AGR, and SGR skin regions. ( a ) Transepidermal water loss levels in healthy GP, AGR, and SGR skin areas. Measurements were carried out with Tewameter TM300 (Courage and Khazaka, Cologne, Germany) on the flexural forearm, axilla, and forehead of healthy individuals ( n = 30). Gene expression of the components of ( b ) cornified envelope formation, ( c ) corneocyte desquamation, ( d ) intercellular lipid lamellae, ( e ) desmosome organization, and ( f ) tight junction formation in healthy GP, AGR, and SGR skin regions were examined by qRT-PCR. The graphs show the mean ± 95% confidence interval of measured mRNA transcript levels (* p < 0.05, ** p < 0.01, *** p < 0.001, determined by one-way ANOVA followed by Sidac post hoc test). Abbreviations: ABCA12, ABC transporter 12B; AGR, apocrine gland-rich; CDH, cadherin; CDSN, corneodesmosin; CLDN, claudin; DSC, desmocollin; DSG, desmoglein; GP, gland-poor; KLK, kallikrein-related peptidase; KRT, keratin; LCE, late cornified envelope; OCLN, occludin; PKP, plakophilin; SGR, sebaceous gland-rich; SPRR, small proline-rich protein; TEWL, transepidermal water loss; TGM, transglutaminase.

Article Snippet: Sections were stained with primary antibodies against human ABCA12 (rabbit polyclonal IgG, GTX51202, GeneTex), human CDSN (rabbit polyclonal IgG, HPA054184, Sigma-Aldrich, St. Louis, MO, USA), human CLDN1 (rabbit polyclonal IgG, ab15098, Abcam, Cambridge, UK), human DSG1 (rabbit polyclonal IgG, NBP1-84567, Novus Biologicals, Centennial, CO, USA), human KLK5 (rabbit polyclonal IgG, ab7283, Abcam), human KRT1 (rabbit monoclonal IgG, ab185628, Abcam), human OCLN (rabbit monoclonal IgG, ab216327, Abcam), and human TGM5 (rabbit polyclonal IgG, ab133786, Abcam).

Techniques: Gene Expression, Permeability, Quantitative RT-PCR

Journal: International Journal of Molecular Sciences

Article Title: Regional Differences in the Permeability Barrier of the Skin—Implications in Acantholytic Skin Diseases

doi: 10.3390/ijms221910428

Figure Lengend Snippet: Prominently weakened cell junction components characterize SGR and AGR skin regions compared with GP areas under steady-state. Representative images for immunostaining and quantification of epidermal levels of KRT1, TGM5, CDSN, DSG1, CLDN1, OCLN, ABCA12, KLK5, and KLK7 in healthy GP, AGR, and SGR skin sections. Bar = 100 μm. Means ± 95% confidence intervals for protein levels are shown. (* p < 0.05, ** p < 0.01, *** p < 0.001, determined by one-way ANOVA followed by Sidac post hoc test) Abbreviations: ABCA12, ABC transporter 12B; AGR, apocrine gland-rich; CDSN, corneodesmosin; CLDN, claudin; DSG1, desmoglein 1; GP, gland-poor; KLK, kallikrein-related peptidase; KRT, keratin; OCLN, occludin; SGR, sebaceous gland-rich; TGM, transglutaminase.

Article Snippet: Sections were stained with primary antibodies against human ABCA12 (rabbit polyclonal IgG, GTX51202, GeneTex), human CDSN (rabbit polyclonal IgG, HPA054184, Sigma-Aldrich, St. Louis, MO, USA), human CLDN1 (rabbit polyclonal IgG, ab15098, Abcam, Cambridge, UK), human DSG1 (rabbit polyclonal IgG, NBP1-84567, Novus Biologicals, Centennial, CO, USA), human KLK5 (rabbit polyclonal IgG, ab7283, Abcam), human KRT1 (rabbit monoclonal IgG, ab185628, Abcam), human OCLN (rabbit monoclonal IgG, ab216327, Abcam), and human TGM5 (rabbit polyclonal IgG, ab133786, Abcam).

Techniques: Immunostaining