Tumour Budding in Pancreatic Cancer revisited: Validation of the ITBCC Scoring System

Abstract

Background

Pancreatic ductal adenocarcinoma (PDAC) is a highly lethal malignancy with rising incidence. Biomarkers that would help the prognostic stratification of patients are urgently needed. Although tumour budding (BD) is a strong and independent prognostic factor in PDAC, is not included in histopathology reports, partly due to lack of standardized scoring system.

Aim

Aim of the present work is to assess the reliability and reproducibility of the BD scoring system recently proposed by the International Tumour Budding Concensus Conference (ITBCC) 2016, in a well-characterized PDAC-cohort (n=120) with complete clinico-pathological and follow-up information.

Methods

BD was scored independently by two pathologists on H&E-stained PDAC-sections by assessing the densest budding area at 20x-magnification (one "hotspot", 0.785mm²) regardless of intra-or peritumoural localisation and assigned into four categories: BD0: 0 buds; BD1: 1-4 buds; BD2: 5-9 buds; and BD3: ≥10 buds. Findings were correlated to patient and tumour characteristics und inter-observer agreement was assessed.

Results

Weighted kappa value for BD-category was 0.62 (0.5-0.73) indicating strong agreement. Increasing BD-category (BD3 versus BD0-2) correlated with higher grade (p=0.002) and shorter overall (OS, p<0.0001; HR(95%CI)=3.234(1.95-5.37)) and disease-free survival (DFS, p=0.0135; HR(95%CI)=1.974(1.15-3.39)). BD (BD3 versus BD0-2) was an independent prognostic factor for OS and DFS, after adjusting for TNM-stage by using both the 8^th AJCC Edition (OS; p=0.0031;HR(95%CI)=2.298(1.32-.99)); (DFS; p=0.0458;HR(95%CI)=1.713(1.01-2.91)) and the 7^th AJCC Edition (OS; p<0.0001;HR(95%CI)=2.795(1.71-4.57)) and (DFS; p=0.00786;HR(95%CI)=1.643(0.95-2.86)).

Conclusions

ITBCC-scoring is a simple, reliable and reproducible method to evaluate BD in PDAC and facilitates its documentation in histopathology reports allowing the prognostic stratification of PDAC-patients.

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Assessment of HER2 status in breast cancer biopsies is not affected by accelerated tissue processing

Abstract

Aims

To establish whether core needle biopsy (CNB) specimens processed with an accelerated processing method with short fixation time can be used to accurately determine HER2 status of breast cancer.

Methods and results

A consecutive case series from two high volume breast clinics was created. We compared routine HER2 immunohistochemistry (IHC) assessment between accelerated processing CNB specimens and routinely processed postoperative excision specimens. Additional amplification-based testing was performed in cases with equivocal results. The formalin fixation time was less than 2 hours and between 6 and 72 hours, respectively. Fluorescent in situ hybridization and multiplex ligation-dependent probe amplification were used for amplification testing.

One hundred and forty-four cases were included, 15 of which were HER2 positive on the routinely processed excision specimens. On the CNB specimens, 44 were equivocal on IHC and required an amplification-based test. Correlation between the CNB specimens and the corresponding excision specimens was high for final HER2 status, with an accuracy of 97% and a kappa of 0.85.

Conclusions

HER2 status can be reliably determined on CNB specimens with accelerated processing time using standard clinical testing methods. Using this accelerated technology, the minimum six hours of formalin fixation which current guidelines consider necessary, can safely be decreased. This allows for a complete and expedited histology-based diagnosis of breast lesions in the setting of a one-stop-shop, same-day breast clinic.

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Antinociceptive potency of enkephalins and enkephalinase inhibitors in the mouse model of colorectal distension - proof of concept

Abstract

Irritable bowel syndrome (IBS) is a chronic disease characterized by abdominal pain and changes in bowel habits. Patients with IBS comprise a significant portion of attendants at the outpatient clinics. Targeting intestinal opioid receptors was found successful in alleviating pain and diarrhea – two major symptoms of IBS. In this study we aimed to evaluate a novel potential pharmacological option: the use of enkephalinase inhibitors in therapy of visceral pain occurring in the course of IBS. We thus assessed the antinociceptive efficacy of enkephalins: Leu-enkephalin and Met-enkephalin, and enkephalinase inhibitors: opiorphin and sialorphin in the mouse model of visceral pain induced by colorectal distension.Leu-enkephalin, Met-enkephalin and sialorphin, but not opiorphin, at the dose of 1 mg/kg injected subcutaneously potently decreased the visceromotor response to colon distension as compared to control. To conclude, enkephalinase inhibitors are worth being considered as potential therapeutics in patients with chronic abdominal pain and/or changed bowel habits, i.e. suffering from IBS.

This article is protected by copyright. All rights reserved.

The antinociceptive efficacy of enkephalins:Leu-enkephalin and Met-enkephalin,and enkephalinase inhibitors: opiorphin and sialorphinwas assessedin the mouse model of visceral pain induced by colorectal distension

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miR-340 alleviates chemoresistance of osteosarcoma cells by targeting ZEB1

Chemoresistance during treatment of osteosarcoma (OS) is attracting more and more attention as the main clinical obstacle. The purpose of this study was to elucidate the role of miR-340 in chemoresistance of OS. Plasmid construction and transfection, miRNA arrays, PCR analyses, and western blot analysis, as well as MTT, apoptosis, and luciferase assays were carried out in MG-63 cells and MG-63/cisplatin (DDP)-resistant cells. The results showed that miR-340 was downregulated in OS tissues and drug-resistant OS cells. Moreover, a negative correlation was observed between miR-340 and ZEB1 expression in OS tissues. Forced expression of miR-340 in drug-resistant OS cells significantly reduced multidrug resistance-1 and P-gp expression. Overexpression of miR-340 enhanced sensitivity to DDP by inhibiting viability and promoting apoptosis. The luciferase assay and western blot analysis identified ZEB1 as a direct target of miR-340, and miR-340 negatively regulated ZEB1 expression. Ectopic expression of ZEB1 reversed the effects of miR-340 on P-gp expression, cell viability, and apoptosis. miR-340 alleviated chemoresistance of OS cells by targeting ZEB1. Our results indicate that targeting miR-340 may be a potential therapeutic approach to treat drug-resistant OS. Correspondence to Liqiu Chen, BSc, Department of Orthopedics, the Affiliated Wenling Hospital of Wenzhou Medical University, Wenling 317500, China Tel/fax: +86 576 8620 6022; e-mail: lqchen66@163.com Received December 15, 2017 Accepted February 5, 2018 Copyright © 2018 Wolters Kluwer Health, Inc. All rights reserved.

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A novel penicillin derivative induces antitumor effect in melanoma cells

In this study, we explored the in-vitro and in-vivo mechanism of antitumor action of a novel synthetic nonantibiotic triazolylpeptidyl penicillin derivative, named TAP7f, on B16-F0 murine melanoma cells. In-vitro assays showed that TAP7f caused an inhibition of S phase progression and a concomitant decrease of the percentage of cells in G0/G1 phase. We also found that TAP7f treatment induced an apoptotic response characterized by an increase of the sub-G1 fraction of B16-F0 hypodiploid cells, the occurrence of cells with picnotic nuclei, and the detection of phosphatidylserine exposure on the outer side of the plasma membrane. Apoptotic cell death was further characterized by the activation of caspase-8, caspase-9, and caspase-3; the increase in the proapoptotic/antiapoptotic ratio of Bcl-2 family proteins; the higher expression levels of Fas receptor and TRAIL ligand; and the cleavage of poly(ADP-ribose) polymerase, a caspase-3 substrate. The in-vivo effect of TAP7f was studied in a syngeneic C57BL/6J mouse melanoma model. Results showed that TAP7f inhibited melanoma cell proliferation in vivo, as determined by a decreased expression of proliferating cell nuclear antigen, inducing a significant reduction of tumor growth. Apoptosis in vivo was assessed by detecting active caspase-3 in tumor slices from treated mice and the expression levels of Fas, TRAIL, and Bcl-2 proteins in tumor lysates. The administration of 80 mg/kg of TAP7f to non-tumor-bearing mice showed no histopathological effects on different organ tissues. Our results suggest that TAP7f might be considered as a potential therapeutic agent for cancer treatment. Correspondence to Leonor P. Roguin, PhD, Institute of Biochemistry and Biophysics (UBA-CONICET), School of Pharmacy and Biochemistry, University of Buenos Aires, Junín 956, C1113AAD Buenos Aires, Argentina Tel: +54 11 4964 8290; fax: +54 11 4962 5457; e-mail: rvroguin@qb.ffyb.uba.ar Received October 12, 2017 Accepted February 3, 2018 Copyright © 2018 Wolters Kluwer Health, Inc. All rights reserved.

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Functional Morphology of Thecal Glands in the Ovary of Japanese Quails (Coturnix japonica)

The role of thecal glands in the ovary of birds remains controversial. Using transmission electron microscopy and immunohistochemistry, immunohistochemical localisation of cyclooxygenase I and II (COX-1 and COX-2), oestrogen receptor α and β (ER-α and ER-β), androgen receptor (AR) and progesterone receptor (PR), a detailed analysis of the thecal glands was performed. Our ultrastructural studies revealed that the thecal glands of the quail ovary consist of 2 cell types, steroid-producing cells (SPCs) and enclosing cells (ENCs). The SPCs are large, light cells containing a varying number of lipid droplets. Their cytoplasm is characterised by a large amount of smooth endoplasmic reticulum. The ENCs are always located at the periphery of the gland. Some ENCs contain an abundant number of microfilaments, but lipid droplets and dense bodies were rare. Within 1 gland, SPCs with distinct COX-2 immunostaining were interspersed between usually larger numbers of moderately COX-2-positive cells. A completely different staining pattern was observed for COX-1, where the cytoplasm of the ENCs was distinctly immunopositive. The SPCs stained only weakly with antibodies to COX-1. The thecal glands showed distinct reactions for ER-β but only a weak to negative one for ER-α, PR, and AR. Our immunohistochemical and ultrastructural data support our hypothesis that the thecal glands of the quail are involved in steroid hormone and prostaglandin synthesis. The prostaglandins secreted by the thecal glands probably contribute to the ovulation of the follicle first in the hierarchy.
Cells Tissues Organs

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Living Up to the Hype: Protein Synthesis Promotes Hypertranscription in Embryonic Stem Cells

The rapid proliferation and unlimited self-renewal of embryonic stem cells depends upon a permissive chromatin landscape that enables hypertranscription. In this issue of Cell Stem Cell, Bulut-Karslioglu et al. report that euchromatin and transcriptional output are enhanced by protein synthesis in embryonic stem cells (Bulut-Karslioglu et al., 2018).

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