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) Tue, 06 Aug 2019 09:55:21 +0200 OJS 60 Effects of physical training on sarcomere lengths and muscle-tendon interface of the cervical region in an experimental model of menopause <p>The aim of this study was to describe the structural and ultrastructural aspects of the myotendinous junction (MTJ) and the proximal and distal sarcomeres of the sternomastoid of aged Wistar rats subjected to an experimental model of menopause and swimming training. A total of 20 female elderly rats were divided into the following four groups (n=5 in each group): sedentary/no-menopausal (SNM), trained/no-menopausal (TNM), sedentary/menopausal (SM), and trained/menopausal (TM). The MTJ samples were dissected and analyzed using transmission electron microscopy. We showed that the TNM Group rats exhibited changes in morphological characteristics as a consequence of physical exercise, which included an increase of 36.60% (P&lt;0.001) in the evagination length of the MTJ and a reduction in the length of the distal (77.38%) (P&lt;0.0001) and proximal (68.15%) (P&lt;0.0001) sarcomeres. The SM Group exhibited a reduction of about 275.93% (P&lt;0.001) in the muscle-tendon interface and in the lengths of distal sarcomeres (55.87%) (P&lt;0.0001) compared with SNM Group. Our results suggest that the swimming training under experimental model of menopause promoted tissue reorganization and increased muscle-tendon interaction with a drastic development in the length and thickness of the sarcoplasmatic invaginations and evaginations. In addition, the sarcomeres exhibited different lengths and a reduction in both groups subjected to swimming training.</p> Carolina dos Santos Jacob, Lara Caetano Rocha, Jurandyr Pimentel Neto, Ii-sei Watanabe, Adriano Polican Ciena Copyright (c) 2019 The Author(s) Tue, 06 Aug 2019 00:00:00 +0200 Ferritin expression in the periodontal tissues of primates <p>Ferritin, an iron-binding protein, is composed of two subunits, ferritin heavy chain and ferritin light chain. It regulates many biological functions, such as proliferation, angiogenesis, and immunosuppression. The objective of this study was to determine the expression and distribution of ferritin in the periodontal tissues of primates. First, we assessed the expression of ferritin in primary cultured cells isolated from human periodontal tissues using the polymerase chain reaction and immunofluorescent staining <em>in vitro</em>. Second, we investigated the expression and distribution of ferritin in the periodontal tissues of <em>Macaca fascicularis</em>, human gingival tissues, and human gingival carcinoma tissues using immunohistochemistry <em>in</em> <em>vivo</em>. Both protein and mRNA of ferritin were constitutively present in human primary cultured cells, including those from the dental apical papilla, periodontal ligament, dental pulp, and gingival epithelium, as well as gingival fibroblasts. In <em>M. fascicularis</em> tissues, the immunohistochemical staining was particularly strong in blood vessel and mineralizing areas of the dental pulp and periodontal ligament. Ferritin heavy chain exhibited specific immunopositivity in the stratum basal of the epithelium in human gingival tissue, and strong immunostaining was found in peripheral regions of gingival carcinoma sites. Ferritin is constitutively present and widely distributed in the periodontal tissues of primates. Ferritin may play roles in epithelial proliferation, vascular angiogenesis, and mineralization in these tissues.</p> Wenxue Huang, Wei Li, Juan Liu, Jianxia Hou, Huanxin Meng Copyright (c) 2019 The Author(s) Tue, 03 Sep 2019 10:09:27 +0200 Presence of N-acetylgalactosamine/galactose residues on bronchioloalveolar cells during rat postnatal development <p>In mammals, the alveolarization process develops predominantly after birth. Airway cells display a complex assemblage of glycans on their surface. These glycans, particularly terminal glycan extensions, are important effective carriers of information that change during the differentiation process. Nevertheless, few systematic data are reported about the cell surface sugar residue content during post-natal lung development. In the present work, we aimed to identify and semi-quantify N-acetylgalactosamine (GalNAc)/galactose (Gal) residues on the bronchioloalveolar cell surface in rat lung sections from 1-, 4-, 8- day old and adult animals and link these data with the lung glycocalyx composition. Horseradish peroxidase-conjugated lectin from <em>Glycine max</em> (soybean agglutinin, SBA) was used, and light microscopy methodologies were performed. SBA labelling intensity was studied before and after sialidase pre-treatment, at one-, four- and eight-day-old animals and adult animals. For semi-quantitative evaluation of SBA binding intensity, two investigators performed the analysis independently, blinded to the type of experiment. Reactivity of the lectin was assessed in bronchiolar and respiratory portion/alveolar epithelial cell surfaces. We evidenced a stronger positive reaction when lung sections were pre-treated with neuraminidase before incubation with the lectin in one- and four-day-old animals and adult animals. These results were not so manifest in eight-day-old animals. This binding pattern, generally points towards the presence of terminal but mainly sub-terminal GalNAc/Gal residues probably capped by sialic acids on the rat bronchiolar/respiratory tract epithelial cells. As this glycan extension is common in O- and N-glycans, our results suggest that these glycan classes can be present in bronchioloalveolar cells immediately after birth and exist during the postnatal period. The results observed in eight-day-old rat lung sections may be due to the dramatic lung morphologic changes and the possible underlying biological mechanisms that occur during this age-moment.</p> Maria de Fátima Martins, Paula Martins, Carlos Alberto Gonçalves Copyright (c) 2019 The Author(s) Wed, 11 Sep 2019 00:00:00 +0200 Molecular characterization of nephron progenitors and their early epithelial derivative structures in the nephrogenic zone of the canine fetal kidney <p>Nephron progenitors (NPs) and nephrogenesis have been extensively studied in mice and humans and have provided insights into the mechanisms of renal development, disease and possibility of NP-based therapies. However, molecular features of NPs and their derivatives in the canine fetal kidney (CFK) remain unknown. This study was focused to characterize the expression of potential markers of canine NPs and their derivatives by immuno-fluorescence and western blot analysis. Transcription factors (TFs) SIX1 and SIX2, well-characterized human NP markers, were expressed in NPs surrounding the ureteric bud in the CFK. Canine NPs also expressed ITGA8 and NCAM1, surface markers previously used to isolate NPs from the mouse and human fetal kidneys. TF, PAX2 was detected in the ureteric bud, NPs and their derivative structures such as renal vesicle and S-shaped body. This study highlights the similarities in dog, mouse and human renal development and characterizes markers to identify canine NPs and their derivatives. These results will facilitate the isolation of canine NPs and their functional characterization to develop NP-based therapies for canine renal diseases.</p> Rawah Faraj, Angela Irizarry-Alfonzo, Pawan Puri Copyright (c) 2019 The Author(s) Mon, 23 Sep 2019 11:09:36 +0200 Expressions of TLR4, MyD88, IRAK4 and NF-κΒ in the oviduct of Chinese brown frog (Rana dybowskii) <p>One special physiological phenomenon of the Chinese brown frog (<em>Rana dybowskii</em>) is that its oviduct goes through expansion prior to pre-hibernation instead of expanding during the breeding period. Our previous study discovered that some cytokines such as interleukin-1 beta (IL-1β) and its receptor type I (IL1R1) proteins were expressed in the oviduct of <em>Rana dybowskii</em>. In this study, we continued to investigate the localizations and expression levels of cytokines including toll-like receptor 4 (TLR4) and its related factors, myeloid differentiation factor 88 (MyD88), interleukin-1 receptor-associated kinase (IRAK) and nuclear factor kappa B (NF-κΒ) in the oviduct of <em>Rana dybowskii </em>during the breeding period and pre-hibernation. Morphological results showed that there were significant differences in oviductal weight and pipe diameter with significantly higher values in pre-hibernation than those of the breeding period. Strong immunostaining of TLR4, MyD88, IRAK4 and NF-κΒ were observed in the pre-hibernation. These proteins levels had no significant difference between these two periods except MyD88 which was significantly higher in the pre-hibernation. The mRNA expression levels of MyD88 and NF-κΒ were obviously higher in pre-hibernation than those of the breeding period. These findings suggested that TLR4/MyD88 signal pathway might participate in the oviductal functions of <em>Rana dybowskii </em>during the breeding period and pre-hibernation.</p> Yue Zhang, Ao Zhang, Yue Zhao, Xiaohang Feng, Yuyao Sheng, Haolin Zhang, Qiang Weng, Meiyu Xu Copyright (c) 2019 The Author(s) Wed, 25 Sep 2019 08:05:23 +0200 Bovine pericardium membrane, gingival stem cells, and ascorbic acid: a novel team in regenerative medicine <p>Recently, the development and the application of 3D scaffold able to promote stem cell differentiation represented an essential field of interest in regenerative medicine. In particular, functionalized scaffolds improve bone tissue formation and promote bone defects repair. This research aims to evaluate the role of ascorbic acid (AS) supplementation in an <em>in vitro</em> model, in which a novel 3D-scaffold, bovine pericardium collagen membrane called BioRipar (BioR) was functionalized with human Gingival Mesenchymal Stem Cells (hGMSCs). As extensively reported in the literature, AS is an essential antioxidant molecule involved in the extracellular matrix secretion and in the osteogenic induction. Specifically, hGMSCs were seeded on BioR and treated with 60 and 90 μg/mL of AS in order to assess their growth behavior, the expression of bone specific markers involved in osteogenesis (runt-related transcription factor 2, RUNX2; collagen1A1, COL1A1; osteopontin, OPN; bone morphogenetic protein2/4, BMP2/4), and <em>de novo</em> deposition of calcium. The expression of COL1A1, RUNX2, BMP2/4 and OPN was evaluated by RT-PCR, Western blotting and immunocytochemistry, and proved to be upregulated. Our results demonstrate that after three weeks of treatment AS at 60 and 90 μg/mL operates as an osteogenic inductor in hGMSCs. These data indicate that the AS supplementation produces an enhancement of osteogenic phenotype commitment in an <em>in vitro</em> environment. For this reason, AS could represent a valid support for basic and translational research in tissue engineering and regenerative medicine.</p> Jacopo Pizzicannella, Guya D. Marconi, Sante D. Pierdomenico, Marcos F.X.B. Cavalcanti, Francesca Diomede, Oriana Trubiani Copyright (c) 2019 The Author(s) Wed, 25 Sep 2019 09:18:56 +0200 Hematopoietic stem cells debut in embryonic lymphomyeloid tissues of elasmobranchs <p>The evolutionary initiation of the appearance in lymphomyeloid tissue of the hemopoietic stem cell in the earliest (most primitive) vertebrate model, <em>i.e.</em> the elasmobranch (chondroichthyan) <em>Torpedo marmorata</em> Risso, has been studied. The three consecutive developmental stages of torpedo embryos were obtained by cesarean section from a total of six pregnant torpedoes. Lymphomyeloid tissue was identified in the Leydig organ and epigonal tissue. The sections were treated with monoclonal anti-CD34 and anti-CD38 antibodies to detect hematopoietic stem cells. At stage I (2-cm-long embryos with external gills) and at stage II (3-4 cm-long embryos with a discoidal shape and internal gills), some lymphoid-like cells that do not demonstrate any immunolabeling for these antibodies are present. Neither CD34+ nor CD38+ cells are identifiable in lymphomyeloid tissue of stage I and stage II embryos, while a CD34+CD38- cell was identified in the external yolk sac of stage II embryo. The stage III (10-11-cm-long embryos), the lymphomyeloid tissue contained four cell populations, respectively CD34+CD38-, CD34+CD38+, CD34-CD38+, and CD34-CD38- cells. The spleen and lymphomyeloid tissue are the principal sites for the development of hematopoietic progenitors in embryonic <em>Torpedo marmorata</em> Risso. The results demonstrated that the CD34 expression on hematopoietic progenitor cells and its extraembryonic origin is conserved throughout the&nbsp;vertebrate&nbsp;evolutionary scale.</p> Rosa Manca, Chester Glomski , Alessandra Pica Copyright (c) 2019 The Author(s) Mon, 30 Sep 2019 00:00:00 +0200 Comparison between Lipofectamine RNAiMAX and GenMute transfection agents in two cellular models of human hepatoma <p>RNA interference is a powerful approach to understand gene function both for therapeutic and experimental purposes. Since the lack of knowledge in the gene silencing of various hepatic cell lines, this work was aimed to compare two transfection agents, the liposome-based Lipofectamine™ RNAiMAX and the HepG2-specific, polymer-based GenMute™, in two cellular models of human hepatoma, HepG2 and Huh7.5. In the first part, we assessed transfection efficiency of a fluorescent Cy3-labeled negative control siRNA by cell imaging analysis; we found that cells treated with GenMute present a higher uptake of the fluorescent negative control siRNA when compared to Lipofectamine RNAiMAX-transfected cells, both in HepG2 and in Huh7.5 cells. In the second part, we evaluated GAPDH silencing with the two transfection reagents by RT-PCR similar GAPDH mRNA expression after each transfection treatment. Finally, we measured cell viability by the MTT assay, observing that cells transfected with GenMute have higher viability with respect to Lipofectamine RNAiMAX-administered cells. These results suggest that GenMute reagent might be considered the most suitable transfection agent for hepatic gene silencing.</p> Clarissa Berardo, Veronica Siciliano, Laura G. Di Pasqua, Plinio Richelmi, Mariapia Vairetti, Andrea Ferrigno Copyright (c) 2019 The Author(s) Tue, 06 Aug 2019 00:00:00 +0200 Clathrin mediated endocytosis - Methods and Protocols <p>Clathrin is one of the interesting “<em>moonlighting proteins</em>” which perform multiple functions relevant to biochemical or biophysical aspects. It can be considered the master regulator of vesicular trafficking being the main player of the Clathrin-Mediated Endocytosis (CME).....</p> Manuela Monti Copyright (c) 2019 The Author(s) Mon, 16 Sep 2019 14:16:07 +0200