European Journal of Histochemistry <p>The <strong>European Journal of Histochemistry&nbsp;</strong>has been an influential cytology journal for over 60 years, publishing research articles on functional cytology and histology in animals and plants. The&nbsp;<strong>European Journal of Histochemistry&nbsp;</strong>offers original research articles investigating on structural and molecular components performed by histochemical and immunohistochemical methods, at light and electron microscopy, cytometry and imaging techniques.</p> <p>Areas of particular interest include cell differentiation, senescence and death, and cell-cell interactions in normal and pathological tissues; attention is also given to articles on newly developed or originally applied histochemical and microscopical techniques.</p> <p>Since its foundation in 1954,&nbsp;the <strong>European Journal of Histochemistry&nbsp;</strong>is the official organ of the Italian Society of Histochemistry.</p> <p>&nbsp;</p> <h4>The Proceedings of the Conference&nbsp;<strong><em>Therapeutic nanoproducts: from biology to innovative technology -&nbsp;</em>Rome, 19-20 June 2019<br></strong>are now <a href="" target="_blank" rel="noopener">available</a></h4> <h4>The Proceedings of the&nbsp;<strong><em>65th Congress GEI-SIBSC and the 38th Congress Italian Society of Histochemistry (SII) </em></strong><strong><em>-&nbsp;</em>Ancona, 24-27 June 2019</strong><strong><em><strong><em><br></em> </strong></em></strong>are now <a href="" target="_blank" rel="noopener">available</a></h4> <p>&nbsp;</p> en-US <p><strong>PAGEPress</strong> has chosen to apply the&nbsp;<a href="" target="_blank" rel="noopener"><strong>Creative Commons Attribution NonCommercial 4.0 International License</strong></a>&nbsp;(CC BY-NC 4.0) to all manuscripts to be published.<br><br> An Open Access Publication is one that meets the following two conditions:</p> <ol> <li>the author(s) and copyright holder(s) grant(s) to all users a free, irrevocable, worldwide, perpetual right of access to, and a license to copy, use, distribute, transmit and display the work publicly and to make and distribute derivative works, in any digital medium for any responsible purpose, subject to proper attribution of authorship, as well as the right to make small numbers of printed copies for their personal use.</li> <li>a complete version of the work and all supplemental materials, including a copy of the permission as stated above, in a suitable standard electronic format is deposited immediately upon initial publication in at least one online repository that is supported by an academic institution, scholarly society, government agency, or other well-established organization that seeks to enable open access, unrestricted distribution, interoperability, and long-term archiving.</li> </ol> <p>Authors who publish with this journal agree to the following terms:</p> <ol> <li>Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.</li> <li>Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.</li> <li>Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.</li> </ol> (Nadia Moscato) (Tiziano Taccini) Fri, 03 May 2019 00:00:00 +0200 OJS 60 miR-132 regulates the expression of synaptic proteins in APP/PS1 transgenic mice through C1q <p>Cognitive impairment in Alzheimer’s disease (AD) is usually accompanied by synaptic loss in both the hippocampus and neocortex. In the early stage of AD, amyloid β-induced synapse changes is the main reason, while in the later stage, the accumulation of Tau protein promotes synapse degeneration as the key factor leading to dementia. MicroRNA (miRNA) is closely related to the expression changes of many AD-related genes. One of the most abundant brain-enriched miRNAs is miR-132, which has been shown to regulate both neuron morphogenesis and plasticity. It has been reported that miR-132 is significantly reduced in the brains of Alzheimer’s patients. Genetic deletion of miR-132 in mice promotes Aβ deposition, leading to impaired memory and enhanced Tau pathology, but how the miRNA-mediated gene expression dysregulation contributes to AD pathology remains unclear. Here we found the possible downstream target of miR-132 by <em>in silico</em> analysis, namely C1q. C1q is the primary protein of classical complement cascade, which is highly expressed in the synaptic regions of the central nervous system in Alzheimer’s patients. However, it is not clear whether miR-132 plays a role in AD through regulating C1q. To address this question, the APP/PS1 transgenic mice were transfected with miR-132 and given C1 inhibitors. Behavior tests were conducted to assess memory and cognitive abilities seven days after administration. In addition, we analyzed the expression of PSD95, Synapsin-1 and phosphorylated (p)-Synapsin. We found that the expression levels of the synaptic proteins treated with miR-132 or C1INH were significantly increased compared with the AD group. Further RT-qPCR result suggested that miR-132 might regulate C1q expression in AD.</p> Nan Xu, Ang-Di Li, Li-Li Ji, Yao Ye, Zhen-Yu Wang, Lei Tong Copyright (c) 2019 Nan Xu, Ang-Di Li, Li-Li Ji, Yao Ye, Zhen-Yu Wang, Lei Tong Fri, 03 May 2019 00:00:00 +0200 Peribiliary gland damage due to liver transplantation involves peribiliary vascular plexus and vascular endothelial growth factor <p>Extrahepatic bile ducts are characterized by the presence of peribiliary glands (PBGs), which represent stem cell niches implicated in biliary regeneration. Orthotopic liver transplantation may be complicated by non-anastomotic strictures (NAS) of the bile ducts, which have been associated with ischemic injury of PBGs and occur more frequently in livers obtained from donors after circulatory death than in those from brain-dead donors. The aims of the present study were to investigate the PBG phenotype in bile ducts after transplantation, the integrity of the peribiliary vascular plexus (PVP) around PBGs, and the expression of vascular endothelial growth factor-A (VEGF-A) by PBGs. Transplanted ducts obtained from patients who underwent liver transplantation were studied (N=62). Controls included explanted bile duct samples not used for transplantation (N=10) with normal histology. Samples were processed for histology, immunohistochemistry and immunofluorescence. Surface epithelium is severely injured in transplanted ducts; PBGs are diffusely damaged, particularly in ducts obtained from circulatory-dead compared to brain-dead donors. PVP is reduced in transplanted compared to controls. PBGs in transplanted ducts contain more numerous progenitor and proliferating cells compared to controls, show higher positivity for VEGF-A compared to controls, and express VEGF receptor-2. In conclusion, PBGs and associated PVP are damaged in transplanted extrahepatic bile ducts; however, an activation of the PBG niche takes place and is characterized by proliferation and VEGF-A expression. This response could have a relevant role in reconstituting biliary epithelium and vascular plexus and could be implicated in the genesis of non-anastomotic strictures.</p> Antonio Franchitto, Diletta Overi, Romina Mancinelli, Anna Paola Mitterhofer, Paolo Muiesan, Francesca Tinti, Ilaria Umbro, Stefan G. Hubscher, Paolo Onori, Eugenio Gaudio, Guido Carpino Copyright (c) 2019 Antonio Franchitto, Diletta Overi, Romina Mancinelli, Anna Paola Mitterhofer, Paolo Muiesan, Francesca Tinti, Ilaria Umbro, Stefan G. Hubscher, Paolo Onori, Eugenio Gaudio, Guido Carpino Fri, 10 May 2019 13:58:16 +0200 A method for semi-automated image analysis of HLA class I tumour epithelium expression in rectal cancer <p>Biomarkers may hold the key towards development and improvement of personalized cancer treatment. For instance, tumour expression of immune system-related proteins may reveal the tumour immune status and, accordingly, determine choice for type of immunotherapy. Therefore, objective evaluation of tumour biomarker expression is needed but often challenging. For instance, human leukocyte antigen (HLA) class I tumour epithelium expression is cumbersome to quantify by eye due to its presence on both tumour epithelial cells and tumour stromal cells, as well as tumour-infiltrating immune cells. In this study, we solved this problem by setting up an immunohistochemical (IHC) double staining using a tissue microarray (TMA) of rectal tumours wherein HLA class I expression was coloured with a blue chromogen, whereas non-epithelial tissue was visualized with a brown chromogen. We subsequently developed a semi-automated image analysis method that identified tumour epithelium as well as the percentage of HLA class I-positive tumour epithelium. Using this technique, we compared HCA2/HC10 and EMR8-5 antibodies for the assessment of HLA class I tumour expression and concluded that EMR8-5 is the superior antibody for this purpose. This IHC double staining can in principle be used for scoring of any biomarker expressed by tumour epithelium.</p> Daniëlle Krijgsman, Ronald L.P. van Vlierberghe, Vaios Evangelou, Alexander L. Vahrmeijer, Cornelis J.H. van de Velde, Cornelis F.M. Sier, Peter J.K. Kuppen Copyright (c) 2019 Daniëlle Krijgsman, Ronald L.P. van Vlierberghe, Vaios Evangelou, Alexander L. Vahrmeijer, Cornelis J.H. van de Velde, Cornelis F.M. Sier, Peter J.K. Kuppen Mon, 20 May 2019 16:00:06 +0200 Ultrastructure of telocytes, a new type of interstitial cells in the myocardium of the Chinese giant salamander (Andrias davidianus) <p>Telocytes (TCs) are new interstitial cells, and they are involved in tissue regeneration, particularly in heart. Therefore, TCs are suggested to be a promising cell in regenerative medicine. However, the information of location structural characteristics and functions of TCs is still limited. In this study, cardiac TCs of the Chinese giant salamanders (<em>Andrias davidianus</em>) were identified by transmission electron microscopy. TCs were located in the interstitium between cardiomyocytes (CM). TCs possessed distinctive ultrastructural characteristics, including one to two very long and thin moniliform telopodes (Tps), emerging points from the cell body, caveolae, dichotomous branchings, labyrinthic systems, neighbouring exosomes and homo-cellular contacts between Tps. TCs/Tps were frequently observed in close proximity to cardiomyocytes. Moreover, Tps established hetero-cellular contacts with cardiomyocytes. Our results confirm the presence of TCs in the myocardium of the <em>A. davidianus</em>. This will help us to better understand roles of TCs in amphibian hearts.</p> Tingting Ge, Yaqiong Ye, Hui Zhang Copyright (c) 2019 Tingting Ge, Yaqiong Ye, Hui Zhang Thu, 23 May 2019 10:34:41 +0200 Autophagy precedes apoptosis during degeneration of the Kölliker’s organ in the development of rat cochlea <p>The Kölliker’s organ is a transient epithelial structure during cochlea development that gradually degenerates and disappears at postnatal 12-14 days (P12-14). While apoptosis has been shown to play an essential role in the degeneration of the Kölliker’s organ, the role of another programmed cell death, autophagy, remains unclear. In our study, autophagy markers including microtubule associated protein light chain 3-II (LC3-II), sequestosome 1 (SQSTM1/p62) and Beclin1 were detected in the supporting cells of the Kölliker’s organ through immunohistochemistry staining. In addition, Western blot and real-time PCR revealed a gradually decreased expression of LC3-II and an increased expression of p62 during early postnatal development. Compared to apoptosis markers that peaks between P7 and P10, autophagy flux peaked earlier at P1 and decreased from P1 to P14. By transmission electron microscopy, we observed representative autophagosome and autolysosome that packaged various organelles in the supporting cells of the Kölliker’s organ. During the degeneration, these organelles were digested <em>via</em> autophagy well ahead of the cellular apoptosis. These results suggest that autophagy plays an important role in transition and degeneration of the Kölliker’s organ prior to apoptosis during the early postnatal development.</p> Shule Hou, Jiarui Chen, Jun Yang Copyright (c) 2019 Shule Hou, Jiarui Chen, Jun Yang Wed, 05 Jun 2019 09:47:31 +0200 Mucosubstances in the porcine gastrointestinal tract: Fixation, staining and quantification <p>Mucins are of great interest in intestinal research and histochemical methods are often employed to identify them. Since it is in the nature of mucins that they are “hard to hold onto” once they come into contact with water, a frequently used medium in histochemistry, there are a number of challenges that may decrease diagnostic accuracy. As the outcome of methods published for microscopic detection of mucosubstances proved to be unsatisfactory in our hands, the aim was the establishment of a reliable and reproducible protocol. Tissue samples were available from pig feeding experiments. In the present study, we focus on a fixation / staining procedure without making comparisons between differently fed pigs. Several fixation and staining procedures were evaluated for their use in semiautomatic quantification and quality assessment of different mucus fractions simultaneous on one tissue section. Cryostat sectioning, subsequent fixation steps with heat, ethanol and modified Bouin’s solution, followed by triple staining with high iron diamine, alcian blue and periodic acid-Schiff turned out to be the best method to identify sulfomucin, sialomucin and neutral mucin simultaneous on one tissue section. This methodology resulted in very good morphology of goblet cells with intact mucin containing vesicles within the cells, which was comparable to ultrastructural electron microscopical observations. Semiautomatic quantification of different mucins was possible. In conclusion, reliable mucus quantification and assessment of mucus quality requires strictly tested procedures. According to our experience, the most important aim after cryosectioning is fast fixation of the mucosubstances, which requires a combination of different fixation steps.</p> Juliane Rieger, Barbara Drewes, Hana Hünigen, Johanna Plendl Copyright (c) 2019 Juliane Rieger, Barbara Drewes, Hana Hünigen, Johanna Plendl Mon, 24 Jun 2019 10:29:26 +0200 Immunohistochemical identification of resistin in the uterus of ewes subjected to different diets: Preliminary results <p>Resistin is a polypeptide hormone of the adipokine-family, primarily, but not exclusively, produced by the adipose tissue. Recent studies suggested that resistin may affect the male and female reproductive activity. The study aim was to immunohistochemically evaluate the presence and distribution of resistin in the ovine uterus. Uterine samples were collected from two groups of ewes at the end of an experimental trial during which the animals of the first group (CTRL) were fed only by grazing while those of the second one (EXP) were supplemented with barley and corn. Using a monoclonal antibody against resistin, tested by Western Blot, the immunopositive reaction was identified in the cytoplasm of epithelial lining cells and uterine glands. The endogenous production of resistin seemed to be affected by different diet, as evidenced by staining differences between the CTRL and EXP groups. Our findings support the existence of a peripheral resistin system in the sheep uterus. It is possible that this system is involved in the functionality of the uterus, which is also affected by the animal’s nutritional status.&nbsp;</p> Cecilia Dall'Aglio, Paola Scocco, Margherita Maranesi, Linda Petrucci, Gabriele Acuti, Elena De Felice, Francesca Mercati Copyright (c) 2019 Cecilia Dall'Aglio, Paola Scocco, Margherita Maranesi, Linda Petrucci, Gabriele Acuti, Elena De Felice, Francesca Mercati Fri, 03 May 2019 15:25:10 +0200