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European Journal of Histochemistry: EJH

Manuela Costanzo, Federico Boschi, Flavia Carton, Giamaica Conti, Viviana Covi, Gabriele Tabaracci, Andrea Sbarbati, Manuela Malatesta
Ozone is a strong oxidant, highly unstable atmospheric gas. Its medical use at low concentrations has been progressively increasing as an alternative/adjuvant treatment for several diseases. In this study, we investigated the effects of mild ozonisation on human adipose-derived adult stem (hADAS) cells i.e., mesenchymal stem cells occurring in the stromal-vascular fraction of the fat tissue and involved in the tissue regeneration processes. hADAS cells were induced to differentiate into the adipoblastic lineage, and the effect of low ozone concentrations on the adipogenic process was studied by combining histochemical, morphometric and ultrastructural analyses...
September 3, 2018: European Journal of Histochemistry: EJH
Beatrice Paradiso, Michele Simonato, Gaetano Thiene, Anna Lavezzi
Formalin-fixed, paraffinembedded (FFPE) human brain tissues are very often stored in formalin for long time. Formalin fixation reduces immunostaining, and the DNA/RNA extraction from FFPE brain tissue becomes suboptimal. At present, there are different protocols of fixation and several procedures and kits to extract DNA/RNA from paraffin embedding tissue, but a gold standard protocol remains distant. In this study, we analyzed four types of fixation systems and compared histo and immuno-staining. Based on our results, we propose a modified method of combined fixation in formalin and formic acid for the autoptic adult brain to obtain easy, fast, safe and efficient immunolabelling of long-stored FFPE tissue...
August 27, 2018: European Journal of Histochemistry: EJH
Mariko Kawai, Yohei Kataoka, Junya Sonobe, Hiromitsu Yamamoto, Hiroki Maruyama, Toshio Yamamoto, Kazuhisa Bessho, Kiyoshi Ohura
Alveolar bone is not spontaneously regenerated following trauma or periodontitis. We previously proposed an animal model for new alveolar bone regeneration therapy based on the non-viral BMP-2/7 gene expression vector and in vivo electroporation, which induced the formation of new alveolar bone over the course of a week. Here, we analysed alveolar bone during a period of three weeks following gene transfer to periodontal tissue. Non-viral plasmid vector pCAGGS-BMP-2/7 or pCAGGS control was injected into palatal periodontal tissue of the first molar of the rat maxilla and immediately electroporated with 32 pulses of 50 V for 50 msec...
August 9, 2018: European Journal of Histochemistry: EJH
Wenjing Liu, Chuanxi Wang, Hao Yu, Shaofeng Liu, Jun Yang
Acetylation tubulin is one of the major post-translational modifications of microtubules. Stable microtubules are well known to contain acetylated tubulin. Here, we examined the spatiotemporal expression of acetylated tubulin in the mouse cochlea during postnatal development. At postnatal day 1 (P1), acetylated tubulin was localized primarily to the auditory nerve inside the cochlea and their synaptic contacts with the inner and outer hair cells (IHCs and OHCs). In the organ of Corti, acetylated tubulin occurred first at the apex of pillar cells...
August 8, 2018: European Journal of Histochemistry: EJH
Roberta Sferra, Simona Pompili, Luca Ventura, Caroline Dubuquoy, Silvia Speca, Eugenio Gaudio, Giovanni Latella, Antonella Vetuschi
A concomitant action of multiple profibrotic mediators appears crucial in the development and progression of fibrosis. Sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways are both involved in pathogenesis of fibrosis in several organs by controlling differentiation of fibroblasts to myofibroblasts and the epithelial to-mesenchymal transition. However, their direct involvement in chronic colitis-associated fibrosis it is not yet known. In this study we evaluated the immunohistochemical expression of some proteins implicated in sphingosine kinase/sphingosine 1 phosphate and transforming growth factor-β/Smads pathways in Dextrane Sodium Sulphate (DSS)-induced colorectal fibrosis in mice...
July 31, 2018: European Journal of Histochemistry: EJH
Vika Smerdu, Martina Perše
Skeletal muscle fibre types, whose characteristics are determined by myosin heavy chain (MyHC) isoforms, can adapt to changed physiological demands with changed MyHC isoform expression resulting in the fibre type transitions. The endurance training is known to induce fast-to-slow transitions and has beneficial effect in carcinogenesis, whereas the effect of an excessive fat intake and its interaction with the effect of swimming are less conclusive. Therefore, we studied the effect of high-fat mixed lipid (HFML) diet and long-term (21-week) swimming on fibre type transitions and their average diameters by immunohistochemical demonstration of MyHC isoforms in slow soleus (SOL), fast extensor digitorum longus (EDL), and mixed gastrocnemius medialis and lateralis (GM, GL) muscles, divided to deep and superficial portions (GMd, GMs, GLd, GLs), of sedentary and swimming Wistar rats with experimentally (dimethylhydrazine) induced colon tumours and fed either with HFML or low-fat corn oil (LFCO) diet...
July 25, 2018: European Journal of Histochemistry: EJH
Kotaro Yamagishi, Ichiro Tsukamoto, Fumihisa Nakamura, Kazuhiko Hashimoto, Kazuhiro Ohtani, Masao Akagi
Epidemiological studies have shown an association between hypertension and knee osteoarthritis (OA). The purpose of this study was to investigate whether activation of the renin-angiotensin system (RAS) can aggravate mechanical loading-induced knee OA in mice. Eight-week-old male Tsukuba hypertensive mice (THM) and C57BL/6 mice were divided into running and non-running groups. Mice in the running group were forced to run (25 m/min, 30 min/day, 5 days/week) on a treadmill. All mice in the four groups (n=10 in each group) were euthanized after 0, 2, 4, 6, or 8 weeks of running or natural breeding...
July 24, 2018: European Journal of Histochemistry: EJH
Arianna Casini, Rosa Vaccaro, Mattia Toni, Carla Cioni
Cholinergic systems play a role in basic cerebral functions and its dysfunction is associated with deficit in neurodegenerative disease. Mechanisms involved in human brain diseases, are often approached by using fish models, especially cyprinids, given basic similarities of the fish brain to that of mammals. In the present paper, the organization of central cholinergic systems have been described in the cyprinid Cyprinus carpio, the common carp, by using specific polyclonal antibodies against ChAT, the synthetic enzyme of acetylcholine, that is currently used as a specific marker for cholinergic neurons in all vertebrates...
July 24, 2018: European Journal of Histochemistry: EJH
Morium Begam, Joseph A Roche
We report that, labeling mouse muscle tissue, with mouse monoclonal antibodies specific to slow or fast myosin heavy chain (sMyHC and fMyHC, respectively), can lead to artefactual labeling of damaged muscle fibers, as hybrid fibers (sMyHC+ and fMyHC+).  We demonstrate that such erroneous immunophenotyping of muscle may be avoided, by performing colabeling or serial-section-labeling, to identify damaged fibers. The quadriceps femoris muscle group (QF) in 7-month-old, male, C57BL/6J mice had: 1.21 ± 0.21%, 98...
July 24, 2018: European Journal of Histochemistry: EJH
Mário S De Souza Albuquerque, Antônio F Da Silva-Filho, Marina Ferraz Cordeiro, Maria de Fátima Deodato de Souza, Michael W L Quirino, Luiza R Amorim Lima, Maira G Da Rocha Pitta, Mario R Martins, Moacyr J B De Melo Rêgo
Gastric cancer (GC) is the second most common cause of cancer-related deaths in the world. This study aims to investigate the differential tissue expression of ppGalNAc-T15 and to evaluate its possible association with clinical-pathological parameters and outcome of gastric adenocarcinoma patients. For these 70 patients were evaluated the expression by immunohistochemistry to ppGalNAc-T15. Our results showed that 33 (47.1%) patients were ppGalNAC-T15+ positive and 37 (52.9%) negative. Positive staining for ppGalNAc-T15 was significantly present in patients older than 60 years (P=0...
June 14, 2018: European Journal of Histochemistry: EJH
Mihály Kálmán, Erzsébet Oszwald, István Adorján
Dystroglycan has an important role in binding of perivascular glial end-feet tothe basal lamina. Its β-subunit is localized in the glial end-feet. The investigation period lasted from E(embryonic day)12 to E20. Laminin and β-dystroglycan were detected by immunohistochemistry, the glial localization of the latter one was supported  by electron microscopy. The immatureglial structures were visualized by the immunostaining of nestin. The β-dystroglycan immunoreactivity appeared at E16 following the laminin of basal lamina but preceding the perivascular processes of radial glia (E18) and astrocyte-like cells (E20)...
May 18, 2018: European Journal of Histochemistry: EJH
Roman M Kassa, Roberta Bonafede, Federico Boschi, Manuela Malatesta, Raffaella Mariotti
Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by motoneuron death. Several cellular pathways have been described to be involved in ALS pathogenesis; however, the involvement of presynaptic stripping and the related MHC class I molecules in mutant SOD1 motoneurons remains to be clarified. To this purpose, we here investigated, for the first time, the motoneurons behavior, di per seand after facial axonal injury, in terms of synaptic stripping and MHC class I expression in wild-type (Wt) mice and in a murine model of ALS, the SOD1(G93A) mice, at the presymptomatic and symptomatic stage of the disease...
May 17, 2018: European Journal of Histochemistry: EJH
Sara Salucci, Sabrina Burattini, Francesca Buontempo, Ester Orsini, Lucia Furiassi, Michele Mari, Simone Lucarini, Alberto M Martelli, Elisabetta Falcieri
Marine organisms such as corals, sponges and tunicates produce active molecules which could represent a valid starting point for new drug development processes. Among the various structural classes, the attention has been focused on 2,2-bis(6-bromo-3-indolyl) ethylamine, a marine alkaloid which showed a good anticancer activity against several tumor cell lines. Here, for the first time, the mechanisms of action of 2,2-bis(6-bromo-3-indolyl) ethylamine have been evaluated in a U937 tumor cell model. Morpho-functional and molecular analyses, highlighting its preferred signaling pathway, demonstrated that apoptosis is the major death response induced by this marine compund...
April 10, 2018: European Journal of Histochemistry: EJH
Kentaro Nishida, Yuka Nomura, Kanako Kawamori, Akihiro Ohishi, Kazuki Nagasawa
In dorsal root ganglion (DRG) neurons, ATP is an important neurotransmitter in nociceptive signaling through P2 receptors (P2Rs) such as P2X2/3R, and adenosine is also involved in anti-nociceptive signaling through adenosine A1R. Thus, the clearance system for adenine nucleotide/nucleoside plays a critical role in regulation of nociceptive signaling, but there is little information on it, especially ectoenzyme expression profiles in DRG. In this study, we examined expression and localization of ecto-nucleotide pyrophosphatase/phosphodiesterases (ENPPs), by which ATP is metabolized to AMP, in rat DRG...
April 9, 2018: European Journal of Histochemistry: EJH
Bin Ma, Changfu Yin, Dailun Hu, Mark Newman, Philip K Nicholls, Zhanjun Wu, Wayne K Greene, Zhongli Shi
The nervous system and the immune system communicate extensively with each other in order to maintain homeostasis and to regulate the immune response. The peripheral nervous system (PNS) communicates specifically with the immune system according to local interactions, including the "hardwiring" of sympathetic/parasympathetic (efferent) and sensory nerves (afferent) to lymphoid tissue and organs. To reveal this type of bidirectional neuroimmune interaction at the microscopic level, we used immunofluorescent staining of glial fibrillary acidic protein (GFAP) coupled with confocal microscopy/3D reconstruction to reveal the distribution of non-myelinating Schwann cells (NMSCs) and their interactions with immune cells inside mouse spleen...
April 3, 2018: European Journal of Histochemistry: EJH
Gabriele Bobek, Tim Stait-Gardner, William Price, Angela Makris, Annemarie Hennessy
Abnormal development of the placenta is postulated to be central to the aetiology of preeclampsia. This study investigates changes in placental histopathology in mouse models of preeclampsia compared to the morphology using magnetic resonance microscopy (MRM) (11.7 T) of intact ex vivo tissue followed by 3D analysis of the image data. Here, C57BL/6JArc pregnant mice were subject to either normal pregnancy (n=3), or to one of two experimental models of preeclampsia; TNF-α infusion  (n=3) or reduced uterine perfusion pressure(RUPP) (n=3)...
April 3, 2018: European Journal of Histochemistry: EJH
Vittorio Farina, Gianluca Lepore, Francesca Biagi, Marcella Carcupino, Marco Zedda
A possible response to aging is autophagy, a self-digestion process in which portions of cytoplasm are encapsulated by double-membrane-bound structures and delivered to lysosome for degradation. A previous work of our group showed that astrocytes under starving conditions are characterized by a higher upregulation of the marker of autophagy LC3 II than neurons. Aim of the present work was to evaluate LC3 II expression in an aging model consisting in fetal sheep neurons and astrocytes at 10, 20 and 30 days of culture...
April 3, 2018: European Journal of Histochemistry: EJH
Silvia Damiana Visonà, Donatella Benati, Maria Cristina Monti, Mirco Galiè, Luisa Andrello, Andrea Frontini, Antonio Osculati
The aim of this post-mortem ultrastructural and immunohistochemical study is to explore the characteristics of acute myocardial ischemia in the context of sudden death, using the combination of two different methods, both more insightful than ordinary histology. Transmission electron microscope and immunohistochemistry, in addition to the traditional histology, were applied to study human heart specimens collected during forensic autopsies. The whole series was sub-grouped into cases (n=17) and controls (n=10)...
April 3, 2018: European Journal of Histochemistry: EJH
Valeriia Gusel'nikova, Olga Antimonova, Elena Fedorova, Mikhail Shavlovsky, Aleksandr Krutikov, Ekaterina Mikhailova, Aleksandra Gudkova, Vyacheslav Mikhailov, Dmitry Korzhevskii
Amyloidosis is a group of diseases that occurs when amyloid proteins are deposited in tissues and organs. The traditional way of identifying amyloid in tissue sections is staining with Congo red. However, this method has a number of limitations including background staining (background fluorescence), low fluorescence intensity and false-positive staining. Therefore, a complex of fluorescence-based methods should be applied to characterize tissue localization of amyloid deposits. The aim of this study was to identify amyloid deposits in the kidneys of dystrophin-deficient mdx mice using different fluorescent dyes...
April 3, 2018: European Journal of Histochemistry: EJH
Manuel Scimeca, Simone Bischetti, Harpreet Kaur Lamsira, Rita Bonfiglio, Elena Bonanno
The Energy Dispersive X-ray (EDX) microanalysis is a technique of elemental analysis associated to electron microscopy based on the generation of characteristic Xrays that reveals the presence of elements present in the specimens. The EDX microanalysis is used in different biomedical fields by many researchers and clinicians. Nevertheless, most of the scientific community is not fully aware of its possible applications. The spectrum of EDX microanalysis contains both semi-qualitative and semi-quantitative information...
March 15, 2018: European Journal of Histochemistry: EJH
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