Journal: Frontiers in Endocrinology

Article Title: Differential Morphological Diagnosis of Various Forms of Congenital Hyperinsulinism in Children

doi: 10.3389/fendo.2021.710947

Figure Lengend Snippet: Histological and immunohistochemical examination of pancreatic tissue in patients with congenital hyperinsulinism and in controls. Hematoxylin and eosin staining, insulin, NeuroD1 and Nkx2.2, Isl1, chromogranin A, somatostatin and DR1, DR2, DR5, SSTR2 and SSTR5; x50. Note: HEO – hematoxylin and eosin, Somat – somatostatin, ChrA – chromogranin A.

Article Snippet: Immunohistochemical analysis of pancreatic samples from 35 patients with CHI and 10 patients from the control group was carried out according to a standard method ( ) using antibodies to: chromogranin A (rabbit polyclonal antibody, dilution 1:400, Diagnostic BioSystems, The Netherlands); insulin (mouse monoclonal antibody, clone K36aC10, 1:50 dilution, Diagnostic BioSystems, The Netherlands); Isl1 (rabbit polyclonal antibody, 1:1000 dilution, ThermoFisher, USA); Nkx 2.2 (rabbit monoclonal antibody, clone EPR14638, dilution 1:20, Abcam, UK); somatostatin (SST, rabbit polyclonal antibody, dilution 1:1000, Dako, Denmark); NeuroD1 (mouse monoclonal antibody, clone 3H8, dilution 1:1000, Novus Biologicals, USA); somatostatin receptor type 2 (SSTR2, rabbit monoclonal antibody, clone EP149, dilution 1:200, Abcam, UK); somatostatin receptor type 5 (SSTR5, rabbit monoclonal antibody, clone UMB4, dilution 1:100, Abcam, UK); type 1 dopamine receptor (DR1, rabbit polyclonal antibody, dilution 1:50, Novus Biologicals, USA); type 2 dopamine receptor (DR2, mouse monoclonal antibody, clone B10, dilution 1:100, Santa Cruz Biotechnology, USA); and type 5 dopamine receptor (DR5, rabbit polyclonal antibody, 1:100 dilution, ThermoFisher, USA).

Techniques: Immunohistochemical staining, Staining

Journal: Frontiers in Endocrinology

Article Title: Differential Morphological Diagnosis of Various Forms of Congenital Hyperinsulinism in Children

doi: 10.3389/fendo.2021.710947

Figure Lengend Snippet: Confocal laser scanning microscopy of the pancreas in congenital hyperinsulinism. (A–D) focal form, (E–H) diffuse form. (A, E) : blue fluorescence of cell nuclei (DAPI). (B, F) : green fluorescence of insulin in cytoplasm. (C, G) : red fluorescence of SSTR2. (D, H) : coexpression of insulin and SSTR2 observed as orange fluorescence; x400.

Article Snippet: Immunohistochemical analysis of pancreatic samples from 35 patients with CHI and 10 patients from the control group was carried out according to a standard method ( ) using antibodies to: chromogranin A (rabbit polyclonal antibody, dilution 1:400, Diagnostic BioSystems, The Netherlands); insulin (mouse monoclonal antibody, clone K36aC10, 1:50 dilution, Diagnostic BioSystems, The Netherlands); Isl1 (rabbit polyclonal antibody, 1:1000 dilution, ThermoFisher, USA); Nkx 2.2 (rabbit monoclonal antibody, clone EPR14638, dilution 1:20, Abcam, UK); somatostatin (SST, rabbit polyclonal antibody, dilution 1:1000, Dako, Denmark); NeuroD1 (mouse monoclonal antibody, clone 3H8, dilution 1:1000, Novus Biologicals, USA); somatostatin receptor type 2 (SSTR2, rabbit monoclonal antibody, clone EP149, dilution 1:200, Abcam, UK); somatostatin receptor type 5 (SSTR5, rabbit monoclonal antibody, clone UMB4, dilution 1:100, Abcam, UK); type 1 dopamine receptor (DR1, rabbit polyclonal antibody, dilution 1:50, Novus Biologicals, USA); type 2 dopamine receptor (DR2, mouse monoclonal antibody, clone B10, dilution 1:100, Santa Cruz Biotechnology, USA); and type 5 dopamine receptor (DR5, rabbit polyclonal antibody, 1:100 dilution, ThermoFisher, USA).

Techniques: Confocal Laser Scanning Microscopy, Fluorescence

Journal: Clinical Cancer Research

Article Title: ERG Status Is Unrelated to PSA Recurrence in Radically Operated Prostate Cancer in the Absence of Antihormonal Therapy

doi: 10.1158/1078-0432.ccr-11-1251

Figure Lengend Snippet: Figure 3. Molecular findings in ERG-positive and ERG-negative tumors. The figure describes previously collected molecular findings obtained by FISH and IHC using various methods according to specific properties to the analyzed gene/protein. Overall the data show that for the majority of analyzed parameters, the difference between ERG-positive and ERG-negative tumors is rather low in absolute numbers but statistically significant. The color code varies between the analyzed parameter: AMACR, CD166, chromogranin A (CHA), EGFR, mTOR, p53, SSTR2, synaptophysin (SYN): weak (dark grey), moderate (light grey), strong (medium grey). Bcl2: low (dark grey), high (light grey). CD10: apical (dark grey), cytoplasmatic (light grey), mixed staining (medium grey). C-Myc: gene polysomy (dark grey), gene gain (light grey), gene amplification (medium grey). HER2 immunostaining: 1þ, 2þ, and 3þ combined. Lipoprotein lipase (LPL): gene loss. Annexin A3 (ANX3): weak (dark grey), moderate (light grey), and strong staining (medium grey).

Article Snippet: Additional IHC data have previously been generated on a different TMA containing different samples from the identical tumor blocks of the same tumor set by using the following antibodies (pretreatments and dilutions): AMACR (clone 13H4, pH 9, 1:200; Dako), Annexin A3 (ANXA3, clone tgc7, pH 2.5, 1:8,100; TgcBIOMICS; ref. 23), androgen receptor (AR, clone 2F12, pH 9.0, 1:10; Novocastra), Bcl2 (clone 124, pH 9.0, 1:250; Dako), CD10 (MME, clone 56C6, pH 9, 1:50; Novocastra; ref. 24), CD166 (ALCAM, clone MOG/07, pH 7.8, 1:2,700; Novocastra), chromogranin A (CHGA, clone LK2H19, 1:500; Biocare), epidermal growth factor receptor (EGFR; clone 31G7, Pronase type XIV, 1:100; Zymed), FOXP1 (polyclonal, pH 9, 1:1,350; Abcam), HER2 (HercepTest; Dako; ref. 25), mTOR (polyclonal, pH 2.0, 1:75; Cell Signaling Technology), p53 (clone D01, pH 7.8, 1:3,600; Oncogene; ref. 26), PSMA (clone 3E6, pH 6.0, 1:150; Dako; ref. 27), SSTR2 (somatostatin receptor 2, polyclonal, pH 6.0, 1:150; Atlas Antibodies), synaptophysin (clone SY38, pH 9.0, 1:20; Dako).With the exception of some previously published IHC data (23–26), the following scoring system was used: The staining intensity (0, 1þ, 2þ, and 3þ) and the fraction of positive tumor cells were recorded for each tissue spot.

Techniques: Staining, Amplification, Immunostaining

Journal: Cancers

Article Title: Enhancer of Zeste Homolog 2 (EZH2) Is a Marker of High-Grade Neuroendocrine Neoplasia in Gastroenteropancreatic and Pulmonary Tract and Predicts Poor Prognosis

doi: 10.3390/cancers14122828

Figure Lengend Snippet: H3K27me3 expression in NEN. H3K27me3 staining was performed in all specimens. ( A ) shows an example of very low H3K27me3 expression, while ( B ) shows very high expression. H3K27me3 expression was quantified using the H-score and correlated with grading and entity ( C ). We performed a correlation analysis of H3K27me3 and EZH2 ( D ). Overall survival (OS) in correlation with H3K27me3 expression ( E ). GEP: gastroenteropancreatic, NEN: neuroendocrine neoplasia, OS: overall survival, P: pulmonary.

Article Snippet: After this procedure, the probes were incubated with primary antibodies against EZH2 (Leica, Wetzlar, Germany, EZH2-L-CE, 1:100, high), EZH1 (Merck, Darmstadt, Germany, #ABE281, 1:1000, low), SUZ12 (Sigma, St. Louis, MO, USA, HPA057436, 1:100, low), EED (Sigma, HPA061140, 1:100, high), H3K27me3 (Cell signaling, Danvers, MA, USA, 9733S, 1:500, high), Ki-67 (Dako/Agilent, Santa Clara, CA, USA, clone MIB-1, 1:200, low), DAXX (Sigma, HPA008736, 1:200, low), ATRX (ThermoFisher, Waltham, MA, USA, clone CLO 537, 1:100, low), TP53 (Agilent, Santa Clara, CA, USA, GAG16, RTU, high), RB (Sigma, rabbit polyclonal, 1:100, high), SSTR2A (Zytomed Systems, Berlin, Germany, rabbit polyclonal, 1:100, high) or Chromogranin A (Cell Marque, Rocklin, CA, USA, LKH210, RTU, high) at room temperature for 20 min. Polymeric secondary antibodies coupled to HRPO peroxidase (EnVision Flex+, Dako) and DAB (Dako) were applied for visualization of the sites of immunoprecipitations.

Techniques: Expressing, Staining