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> 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) Thu, 17 Jan 2019 11:05:10 +0100 OJS 60 Relationship between lipid droplets size and integrated optical density <p>Lipid accumulation is largely investigated due to its role in many human diseases. The attention is mainly focused on the lipid droplets (LDs), spherical cytoplasmic organelles, which are devoted to the storage of the lipids. The amount of lipid content is often evaluated by measuring LDs size and/or the integrated optical density (IOD) in cultured cells. Both evaluations are directly associated to the lipid content and therefore they are correlated to each other, but a lack of theoretical relationship between size and IOD was observed in literature. Here we investigated the size-IOD relationship of LDs observed in microscopical images of cultured cells. The experimental data were obtained from immature and differentiated 3T3-L1 murine cells, which have been extensively used in studies on adipogenesis. A simple model based on the spherical shape of the LDs and the Lambert-Beer law, which describes the light absorption by an optical thick material, leads to a mathematical relationship. Despite only light rays’ absorption was considered in the model, neglecting their scattering, a very good agreement between the theoretical curve and the experimental data was found. Moreover, a computational simulation corroborates the model indicating the validity of the mathematically theoretical relationship between size and IOD. The theoretical model could be used to calculate the absorption coefficient in the LDs population and it could be applied to seek for morphologically and functionally LDs subpopulations. The identification of LDs dynamic by measuring size and IOD could be related to different pathophysiological conditions and useful for understand cellular lipid-associated diseases.</p> Federico Boschi, Vanni Rizzatti, Elena Zoico, Tommaso Montanari, Mauro Zamboni, Andrea Sbarbati, Monica Colitti ##submission.copyrightStatement## Mon, 25 Mar 2019 08:52:54 +0100 VEGF, VEGF165b and EG-VEGF expression is specifically related with hormone profile in pituitary adenomas <p>Vascular endothelial growth factor (VEGF), its inhibitory splice variant, VEGF<em>165b </em>and Endocrine Gland derived VEGF (EG-VEGF) have a controversial role in pituitary gland. We aim to study VEGF, VEGF<em>165b</em> and EG-VEGF expression in pituitary adenomas. A significant correlation was found between growth hormone (GH) and VEGF secretion (P=0.024). For prolactinomas, VEGF and prolactin expression, had a P-value of 0.02 for Kendall coefficient and a P-value of 0.043 for the Spearman coefficient. VEGF-mRNA amplification was detected in both tumor cells and folliculostellate cells. VEGF<em>165b</em> was positive in 16.66% of pituitary adenomas. EG-VEGF was significantly correlated with prolactin (P=0.025) and luteinizing hormone (P=0.028). Our data strongly support VEGF, VEGF<em>165b</em> and EG-VEGF as important players of pituitary adenomas tumorigenesis. Particular hormonal milieu heterogeneity, special vascular network with an unusual reactivity to tumor growth correlated with variability of VEGF, VEGF<em>165b</em> and EG-VEGF secretion may stratify pituitary adenomas in several molecular groups with a direct impact on therapy and prognosis.</p> Ana Silvia Corlan, Anca Maria Cîmpean, Eugen Melnic, Marius Raica, Simona Sarb ##submission.copyrightStatement## Tue, 05 Mar 2019 13:51:36 +0100 Immunohistochemical distribution of insulin-, glucagon- and somatostatin-containing cells in the pancreas of Lake Van fish (Alburnus tarichi Güldenstädt, 1814) (Cyprinidae) <p>The Lake Van fish (<em>Alburnus tarichi</em>) is a species that is endemic to Turkey’s Lake Van basin. In this study, the regional distribution, volume density, and relative frequency of some pancreatic endocrine cells in Lake Van fish were investigated via immunohistochemistry using specific mammalian antibodies. The pancreatic tissue was observed to be surrounded by adipose tissue, which was adjacent to the gall bladder or extrahepatic bile duct, or dispersed in the adipose tissue ranked among coils of post-esophageal swelling and intestine. The pancreatic endocrine cells were examined, including the islets, exocrine pancreas, and pancreatic ducts. According to the modified aldehyde fuchsin staining and immunohistochemistry, insulin-secreting beta cells were observed to localize throughout the islets. Glucagon immune-reactive (IR) cells were observed to be situated moderately on the islet periphery, and were rarely determined in the islet central region. A small number of somatostatin-IR cells were observed in the islet centers and peripheries. Similar distributions of those 3 endocrine cells were also determined in the secondary islets. Additionally, the endocrine cell percentages did not differ between the primary and secondary islets; insulin-, glucagon- and somatostatin-IR cells comprised approximately 54%, 29%, and 11% of the endocrine cells in the principal islets, whereas they comprised 52%, 27%, and 14% in the secondary islets, respectively. Insulin-, glucagon- and somatostatin-IR cells were also determined among the epithelium and subepithelial connective tissue in the pancreatic ducts or exocrine areas of the pancreas. With this study, the existence, regional distribution, and relative frequency of the insulin-, glucagon- and somatostatin-IR cells were first investigated in the pancreatic tissue of Lake Van fish and the results were discussed.</p> Burak Kaptaner ##submission.copyrightStatement## Mon, 25 Feb 2019 12:03:44 +0100 Deleted in Liver Cancer 2 (DLC2) protein expression in hepatocellular carcinoma <p>Deleted in Liver Cancer (DLC) proteins belong to the family of RhoGAPs and are believed to operate as negative regulators of the Rho family of small GTPases. So far, the role of the first identified member from the DLC family, DLC1, was established as a tumor suppressor in hepatocellular carcinoma. The function of its close family relative, DLC2 is unequivocal. In the present study we attempted to determine whether the loss of DLC2 is a common feature of hepatocellular carcinoma tissue. We examined two types of hepatocellular carcinoma- typical and fibrolamellar one. Our analysis revealed that DLC2 protein is not diminished in cancer tissue when compared to non-cancerous liver specimens. What is more, we observed DLC2 to be more abundantly expressed in cancer tissue, particularly in tumors with the inflammation background. In addition, we found that <em>DLC2</em> gene status was diploid in virtually all tumor samples examined. Our results indicate that DLC2 is not diminished in hepatocellular carcinoma cells. It appears that members of the DLC family, although structurally highly related, may function differently in cancer cells.</p> Dominika Wolosz, Agnieszka Walczak, Grzegorz Szparecki, Michal Dwojak, Magdalena Winiarska, Ewa Wolinska, Barbara Gornicka ##submission.copyrightStatement## Mon, 18 Feb 2019 08:32:28 +0100 Morphological and ultrastructural analysis of normal, injured and osteoarthritic human knee menisci <p>The human meniscus plays a crucial role for transmission and distribution of load across the knee, as well as shock absorption, joint stability, lubrication, and congruity. The aim of this study was to compare the complex geometry, and unique ultrastructure and tissue composition of the meniscus in healthy (control) and pathological conditions to provide understanding of structural changes that could be helpful in the future design of targetted therapies and improvement of treatment indications. We analyzed meniscus samples collected from 3 healthy multi-organ donors (median age, 66 years), 5 patients with traumatic meniscal tear (median age, 41 years) and 3 patients undergoing total knee replacement (TKR) for end-stage osteoarthritis (OA) (median age, 72 years). We evaluated the extracellular matrix (ECM) organization, the appearance and distribution of areas of calcification, and modifications of cellular organization and structure by electron microscopy and histology. The ECM structure was similar in specimens from traumatic meniscus tears compared to those from patients with late-stage OA, showing disorganization of collagen fibers and increased proteoglycan content. Cells of healthy menisci showed mainly diffuse chromatin and well preserved organelles. Both in traumatic and in OA menisci, we observed increased chromatin condensation, organelle degeneration, and cytoplasmic vacuolization, a portion of which contained markers of autophagic vacuoles. Areas of calcification were also observed in both traumatic and OA menisci, as well as apoptotic-like features that were particularly prominent in traumatic meniscal tear samples. We conclude that meniscal tissue from patients with traumatic meniscal injury demonstrate pathological alterations characteristic of tissue from older patients undergoing TKR, suggesting that they have high susceptibility to develop OA.</p> Michela Battistelli, Marta Favero, Debora Burini, Giovanni Trisolino, Dante Dallari, Lucia De Franceschi, Steven R. Goldring, Mary B. Goldring, Elisa Belluzzi, Giuseppe Filardo, Brunella Grigolo, Elisabetta Falcieri, Eleonora Olivotto ##submission.copyrightStatement## Mon, 11 Feb 2019 08:44:44 +0100 Antigen retrieval pre-treatment causes a different expression pattern of Cav3.2 in rat and mouse spinal dorsal horn <p>Ca<sub>v</sub>3 channels consist of three isoforms, Ca<sub>v</sub>3.1 (α1G), Ca<sub>v</sub>3.2 (α1H), and Ca<sub>v</sub>3.3 (α1I), which produce low-threshold spikes that trigger burst firings in nociceptive neurons of the spinal dorsal horn (SDH) and dorsal root ganglion (DRG). Although Ca<sub>v</sub>3.2 plays a crucial role in pathological pain, its distribution in SDH still remains controversial. One study showed that Ca<sub>v</sub>3.2 is ubiquitously expressed in neurons, but another study implied that Ca<sub>v</sub>3.2 is expressed restricted to astrocytes. To unravel these discrepancies, we used methods of immunohistochemistry either with or without antigen retrieval (AR) pre-treatment to detect Ca<sub>v</sub>3 in SDH and DRG from both rats and mice. Moreover, Ca<sub>v</sub>3.2 mRNA was detected in mice SDH using <em>in situ</em> hybridization. We found that the expression pattern of Ca<sub>v</sub>3.2 but not Ca<sub>v</sub>3.1 and Ca<sub>v</sub>3.3 in SDH were largely different with or without AR pre-treatment, which showed a neuron-like and an astrocyte-like appearance, respectively. Double staining further demonstrated that Ca<sub>v</sub>3.2 was mainly co-stained with the neuronal marker NeuN in the presence of AR but was with glial fibrillary acidic protein (GFAP, marker for astrocytes) in the absence of AR pre-treatment. Importantly, Ca<sub>v</sub>3.2 mRNA was mainly co-localized with Ca<sub>v</sub>3.2 but not GFAP. Together, our findings indicate that AR pre-treatment or not impacts the expression pattern of Ca<sub>v</sub>3.2, which may make a significant contribution to the future study of Ca<sub>v</sub>3.2 in SDH.</p> Xiao E Cheng, Long Xian Ma, Xiao Jin Feng, Meng Ye Zhu, Da Ying Zhang, Lin Lin Xu, Tao Liu ##submission.copyrightStatement## Wed, 23 Jan 2019 09:28:15 +0100 Seasonal expressions of prolactin, prolactin receptor and STAT5 in the scented glands of the male muskrats (Ondatra zibethicus) <p>Prolactin (PRL) production in mammals has been demonstrated in extrapituitary gland, which can activate autocrine/paracrine signaling pathways to regulate physiological activity. In the current study, we characterized the gene expression profiles of PRL, prolactin receptor (PRLR) and signal transducers and activators of transcription 5 (STAT5) in the scented glandular tissues of the muskrats, to further elucidate the relationship between PRL and the scented glandular functions of the muskrats. The weight and volume of the scented glands in the breeding season were significantly higher than those of the non-breeding season. Immunohistochemical data showed that PRL, PRLR and STAT5/phospho-STAT5 (pSTAT5) were found in the glandular and epithelial cells of the scented glands in both seasons. Furthermore, we found that PRL, PRLR and STAT5 had higher immunoreactivities in the scented glands during the breeding season when compared to those of the non-breeding season. In parallel, the gene expressions of PRL, PRLR and STAT5 were significantly higher in the scented glands during the breeding season than those of the non-breeding season. The concentrations of PRL in scented glandular tissues and sera were measured by enzyme-linked immunosorbent assay (ELISA), and their levels were both notably higher in the breeding season than those of the non-breeding season. These findings suggested that the scented glands of the muskrats were capable of extrapituitary synthesis of PRL, which might attribute PRL a specific function to an endocrine or autocrine/paracrine mediator.</p> Wenqian Xie, Hong Liu, Qian Liu, Qiong Gao, Fuli Gao, Yingying Han, Zhengrong Yuan, Haolin Zhang, Qiang Weng ##submission.copyrightStatement## Thu, 17 Jan 2019 10:59:27 +0100 Epithelial expression of the hormone leptin by bovine skin <p>Leptin (Lep) stimulates keratinocytes to proliferate, intervenes in the wound healing and participates to hair follicle morphogenesis and cycle. While it is secreted by skin structures including epidermis and hair follicles, intradermal adipose tissue also seems to have a role in Lep secretion and accordingly in the control of hair follicle growth in mice and humans. Lep was investigated in the skin of humans and laboratory animals but there are not data regarding bovine species. The aim of this work was to study the expression of Lep and its receptor (LepR) in the skin of bovine and, at the same time, to investigate the presence and extension of intradermal adipose tissue. A morphological evaluation of the skin was performed while the presence and localization of Lep and LepR were analyzed by RT-PCR and immunohistochemistry. A high and thick dermis without adipocytes was observed. Hair follicles and sebaceous and sweat glands were located in the proximal part of the skin while a thick layer of connective tissue, lacking adipose cells, separated these structures by subcutis. RT-PCR evidenced the transcripts for both molecules. By immunohistochemistry, Lep and LepR were observed in the epidermis and hair follicles. Based on the absence of intradermal adipose tissue and the presence of both Lep and LepR in the epidermis and in the hair follicle epithelium, it can be posited that in bovine skin Lep participates to the control of epidermis growth and hair follicle cycle through a paracrine and autocrine mechanisms.</p> Francesca Mercati, Cecilia Dall'Aglio, Ludovica Timperi, Paola Scocco, Elena De Felice, Margherita Maranesi ##submission.copyrightStatement## Thu, 17 Jan 2019 11:01:22 +0100 Embedding cell monolayers to investigate nanoparticle-plasmalemma interactions at transmission electron microscopy <p>Transmission electron microscopy is the technique of choice to visualize the spatial relationships between nanoconstructs and cells and especially to monitor the uptake process of nanomaterials. It is therefore crucial that the cell surface be preserved in its integrity, to obtain reliable ultrastructural evidence: the plasmalemma represents the biological barrier the nanomaterials have to cross, and the mode of membrane-nanoconstruct interaction is responsible for the intracellular fate of the nanomaterials. In this paper, we describe a simple and inexpensive method to process cell monolayers for ultrastructural morphology and immunocytochemistry, ensuring consistent preservation of the cell surface and of the occurring interactions with nanoparticles of different chemical composition.</p> Manuela Costanzo, Manuela Malatesta ##submission.copyrightStatement## Thu, 28 Mar 2019 11:30:11 +0100 Cell Migration - Methods and Protocols <p>As the title suggests, this book presents several techniques to study cells migration in vivo, in vitro, ex vivo and with different model systems to dissect many of the biochemical and biophysical properties (at both molecular and cellular levels) involved in the dynamics of migration and cell-to-cell communication...</p> Manuela Monti ##submission.copyrightStatement## Thu, 17 Jan 2019 11:03:09 +0100 Sertoli Cells - Methods and Protocols <p>The mammalian somatic cell providing the physiological milieu to germ cells growth and differentiation is the Sertoli cell, named after the Italian physiologist Enrico Sertoli. Sertoli described for the first time this functionally multitasking cell in 1865 (when he was 23 years-old!) in human testicular tissues while ending up his medicine studies at the University of Pavia...</p> CarloAlberto Redi ##submission.copyrightStatement## Thu, 17 Jan 2019 11:04:07 +0100