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The terminology applied to canine prostatic epithelial lesions, especially carcinomas, is currently not standardized and this hampers the ability of pathologists to study the biological and clinical significance of these lesions. The aim of this review is to present the essential histomorphological diagnostic attributes of a wide spectrum of prostatic epithelial lesions in dogs. In addition to the traditionally recognized prostatic hyperplasia, hormonal atrophy, prostatitis, squamous metaplasia, adenocarcinoma and transitional cell (urothelial) carcinoma, new entities are described and discussed in order to provide veterinary pathologists with a basic atlas of common histological lesions of the canine prostate that is comprehensive and easy to use.

Additional file 5. pathway_components. Abstract of the WNT and BCR signaling pathway and the components.

Additional file 3. supplement figures. Heatmaps of the top-100 up- and downregulated genes of the biological triplicates.

Additional file 1. FACS plots. Plots of apoptotic (Annexin V FITC+ and Propidium iodide-) and necrotic cells (Annexin V FITC+ and Propidium iodide+) detected by flow cytometry analysis at 72h of pro-B ALL cell lines RS4;11 and SEM exposed with AraC, DEX and IDEL and two drugs combined (AraC+IDEL, DEX+IDEL).

Additional file 2. supplement tables. Comparison of the differential expression analysis of RS4;11 and SEM exposed with AraC, DEX and IDEL and two drugs combined (AraC+IDEL, DEX+IDEL) for 72 h.

Additional file 6. pw_ranking_Idel. Pathway ranking of pathways with components mTOR, BTK, AKT, and PI3K.

Background The introduction of combined conventional cytostatics and pathway-specific inhibitors has opened new treatment options for several cancer types including hematologic neoplasia such as leukaemias. As the detailed understanding of the combination-induced molecular effects is often lacking, the identification of combination-induced molecular mechanisms bears significant value for the further development of interventional approaches. Methods Combined application of conventional cytostatic agents (cytarabine and dexamethasone) with the PI3K-inhibitor Idelalisib was analysed on cell-biologic parameters in two acute pro-B lymphoblastic leukaemia (B-ALL) cell lines. In particular, for comparative characterisation of the molecular signatures induced by the combined and mono application, whole transcriptome sequencing was performed. Emphasis was placed on pathways and genes exclusively regulated by drug combinations. Results Idelalisib + cytostatics combinations changed pathway activation for, e.g., “Retinoblastoma in cancer”, “TGF-b signalling”, “Cell cycle” and “DNA-damage response” to a greater extent than the two cytostatics alone. Analyses of the top-20 regulated genes revealed that both combinations induce characteristic gene expression changes. Conclusion A specific set of genes was exclusively deregulated by the drug combinations, matching the combination-specific anti-proliferative cell-biologic effects. The addition of Idelalisib suggests minor synergistic effects which are rather to be classified as additive.

Ultrasound-guided fine-needle aspiration (US-FNA) biopsy is a widely used minimally invasive sampling procedure for cytological diagnosis. This study investigates the feasibility of using US-FNA samples for both cytological diagnosis and whole transcriptome RNA-sequencing analysis (RNA-Seq), with the ultimate aim of improving canine prostate cancer management. The feasibility of the US-FNA procedure was evaluated intra vitam on 43 dogs. Additionally, aspirates from 31 euthanised dogs were collected for standardising the procedure. Each aspirate was separated into two subsamples: for cytology and RNA extraction. Additional prostate tissue samples served as control for RNA quantity and quality evaluation, and differential expression analysis. The US-FNA sampling procedure was feasible in 95% of dogs. RNA isolation of US-FNA samples was successfully performed using phenol-chloroform extraction. The extracted RNA of 56% of a subset of US-FNA samples met the quality requirements for RNA-Seq. Expression analysis revealed that only 153 genes were exclusively differentially expressed between non-malignant US-FNAs and tissues. Moreover, only 36 differentially expressed genes were associated with the US-FNA sampling technique and unrelated to the diagnosis. Furthermore, the gene expression profiles clearly distinguished between non-malignant and malignant samples. This proves US-FNA to be useful for molecular profiling. peerReviewed

Ultrasound-guided fine-needle aspiration (US-FNA) biopsy is a widely used minimally invasive sampling procedure for cytological diagnosis. This study investigates the feasibility of using US-FNA samples for both cytological diagnosis and whole transcriptome RNA-sequencing analysis (RNA-Seq), with the ultimate aim of improving canine prostate cancer management. The feasibility of the US-FNA procedure was evaluated intra vitam on 43 dogs. Additionally, aspirates from 31 euthanised dogs were collected for standardising the procedure. Each aspirate was separated into two subsamples: for cytology and RNA extraction. Additional prostate tissue samples served as control for RNA quantity and quality evaluation, and differential expression analysis. The US-FNA sampling procedure was feasible in 95% of dogs. RNA isolation of US-FNA samples was successfully performed using phenol-chloroform extraction. The extracted RNA of 56% of a subset of US-FNA samples met the quality requirements for RNA-Seq. Expression analysis revealed that only 153 genes were exclusively differentially expressed between non-malignant US-FNAs and tissues. Moreover, only 36 differentially expressed genes were associated with the US-FNA sampling technique and unrelated to the diagnosis. Furthermore, the gene expression profiles clearly distinguished between non-malignant and malignant samples. This proves US-FNA to be useful for molecular profiling.

Background Promotor hypermethylation of CpG islands is common in B cell precursor acute lymphoblastic leukemia (BCP-ALL) with mixed lineage leukemia (MLL) gene rearrangements. Hypomethylating agents (HMA) such as azacitidine (AZA) and decitabine (DEC) reduce DNA hypermethylation by incorporation into DNA and were successfully introduced into the clinic for the treatment of myeloid neoplasias. Methods Here, we investigated whether HMA induce comparable biological effects in MLL-positive BCP-ALL. Further, efficacy of HMA and concomitant application of cytostatic drugs (cytarabine and doxorubicin) were evaluated on established SEM and RS4;11 cell lines. In addition, promising approaches were studied on BCP-ALL cell line- and patient-derived xenograft models. Results In general, DEC effects were stronger compared to AZA on MLL-positive BCP-ALL cells. DEC significantly reduced proliferation by induction of cell cycle arrest in G0/G1 phase and apoptosis. Most sensitive to HMA were SEM cells which are characterized by a fast cell doubling time. The combination of low-dose HMA and conventional cytostatic agents revealed a heterogeneous response pattern. The strongest antiproliferative effects were observed when ALL cells were simultaneously exposed to HMA and cytostatic drugs. Most potent synergistic effects of HMA were induced with cytarabine. Finally, the therapeutic potential of DEC was evaluated on BCP-ALL xenograft models. DEC significantly delayed leukemic proliferation in xenograft models as demonstrated longitudinally by non-invasive bioluminescence as well as 18F-FDG-PET/CT imaging. Unexpectedly, in vivo concomitant application of DEC and cytarabine did not enhance the antiproliferative effect compared to DEC monotherapy. Conclusions Our data reveal that DEC is active in MLL-positive BCP-ALL and warrant clinical evaluation. Electronic supplementary material The online version of this article (10.1186/s13045-018-0607-3) contains supplementary material, which is available to authorized users.

Patient characteristics. (DOCX 17Â kb)

18F-FDG uptake parameter. (DOCX 43Â kb)

List of RS4;11-ffluc xenograft mice. (DOCX 14Â kb)

Effects of HMA on proliferation. (DOCX 114Â kb)

Effects of HMA and cytostatic drug combinations on proliferation. (DOCX 231Â kb)

DEC reduces methylation of LINE-1 and CDH13. (DOCX 49Â kb)

List of SEM-ffluc xenograft mice used for imaging studies. (DOCX 16Â kb)

Methylation-specific quantitative PCR. (MSqPCR) (DOCX 15Â kb)

Special issue Deep-Sea Biodiversity and Life History Processes.-- 10 pages, 7 figures, 5 tables, Nezumia aequalis and Coelorinchus mediterraneus are abundant species on the upper and lower continental slopes, respectively, in the Mediterranean Sea. A study on the reproductive strategy of the two species was conducted on the Catalan margin (NW Mediterranean). The reproductive cycle of both species was investigated using visual analyses of gonads and histological screening. The shallower species, N. aequalis, showed continuous reproduction with a peak of spawning females in winter months. In contrast, the deeper-living species, C. mediterraneus, showed semi-continuous reproduction with a regression period during the spring. Juveniles of N. aequalis were present in all seasons, but most abundant in the spring. Only two juveniles of C. mediterraneus were found. Both species had asynchronous oocyte development. The average fecundity of N. aequalis was 10,630 oocytes per individual, lower than known for the same species in the Atlantic Ocean. The fecundity of C. mediterraneus was measured for the first time in this study, with an average of 7693 oocytes per individual. Males of both species appear to have semi-cystic spermatogenesis. © 2013 Elsevier Ltd., This study was conducted in the framework of the RECS II, PROMETEO and DOS MARES projects funded by the Spanish Science and Innovation Ministry (CICYT: REN02/04556/C02/MAR; CTM2007-66316-C02/MAR; CTM2010- 21810-C03-03/MAR respectively, to JBC) and the European Community’s Seventh Framework Program under the HERMIONE project (Grant agreement 226354, to ERLL). UF-A is funded by a Spanish doctoral fellowship FPI-2008. ERLL is funded by a Spanish JAE-CSIC postdoctoral grant with co-funding of the European Social Fund. This paper is contribution no. 619 from AZTI- Tecnalia (Marine Research Division)

Summary Reasons for performing study: CD14 positive (CD14+) cells are the precursor cells of monocyte-derived dendritic cells (DCs). In horses their potent antigen-presenting capacity and ability to induce an effective immune response classify these cells suitable for several therapeutic approaches such as for equine sarcoid. However, in horses, the generation efficiency of DCs from adherent peripheral blood mononuclear cells (PBMCs) is currently still poor. Objectives: Establishment of a simple short protocol to enhance DC generation in horses by using a human CD14 monoclonal antibody (mAb) and an automated magnetic activated cell sorting (MACS) system. Methods: Peripheral blood mononuclear cells were isolated from fresh heparinised blood samples of 3 horses and primarily stained for flow cytometric analysis (FACS) with a mAb against human CD14 as well as a secondary phycoerythrin (PE) conjugated antibody to determine the initial percentage of CD14 cells in the sample. Peripheral blood mononuclear cells were used for automated MACS using the same primary and secondary antibodies and analysed by FACS. CD14+ selected cells were cultured for 4 days adding granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) to the culture media. Dendritic cell generation was assessed analysing cell morphology and surface marker expression (hCD83, hCD86, eqMHCII). Results: Prior to selection, the mean percentage of CD14+ cells in the total cell population was 5.5%, further gaiting of this cell population resulted in 78.46% CD14+ monocytes. After our positive selection the mean percentage of CD14+ cells in the population was 98% without affecting viability. After culture, DC yield was 2-fold higher than in previous published outcomes. Conclusions: The additional CD14 cell separation step after PBMC isolation significantly amplified the number of CD14+ cells, increasing the number of generated DCs. Potential relevance: The number of DCs available is critical for further use of these cells and the herein described protocol will therefore help to improved DC generation for therapeutic approaches in horses.