Publisher: Elsevier   (Total: 3206 journals)

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Showing 1 - 200 of 3206 Journals sorted alphabetically
Academic Pediatrics     Hybrid Journal   (Followers: 39, SJR: 1.655, CiteScore: 2)
Academic Radiology     Hybrid Journal   (Followers: 27, SJR: 1.015, CiteScore: 2)
Accident Analysis & Prevention     Partially Free   (Followers: 106, SJR: 1.462, CiteScore: 3)
Accounting Forum     Hybrid Journal   (Followers: 28, SJR: 0.932, CiteScore: 2)
Accounting, Organizations and Society     Hybrid Journal   (Followers: 44, SJR: 1.771, CiteScore: 3)
Achievements in the Life Sciences     Open Access   (Followers: 8)
Acta Anaesthesiologica Taiwanica     Open Access   (Followers: 6)
Acta Astronautica     Hybrid Journal   (Followers: 449, SJR: 0.758, CiteScore: 2)
Acta Automatica Sinica     Full-text available via subscription   (Followers: 2)
Acta Biomaterialia     Hybrid Journal   (Followers: 30, SJR: 1.967, CiteScore: 7)
Acta Colombiana de Cuidado Intensivo     Full-text available via subscription   (Followers: 3)
Acta de Investigación Psicológica     Open Access   (Followers: 2)
Acta Ecologica Sinica     Open Access   (Followers: 11, SJR: 0.18, CiteScore: 1)
Acta Histochemica     Hybrid Journal   (Followers: 5, SJR: 0.661, CiteScore: 2)
Acta Materialia     Hybrid Journal   (Followers: 340, SJR: 3.263, CiteScore: 6)
Acta Mathematica Scientia     Full-text available via subscription   (Followers: 5, SJR: 0.504, CiteScore: 1)
Acta Mechanica Solida Sinica     Full-text available via subscription   (Followers: 9, SJR: 0.542, CiteScore: 1)
Acta Oecologica     Hybrid Journal   (Followers: 12, SJR: 0.834, CiteScore: 2)
Acta Otorrinolaringologica (English Edition)     Full-text available via subscription  
Acta Otorrinolaringológica Española     Full-text available via subscription   (Followers: 2, SJR: 0.307, CiteScore: 0)
Acta Pharmaceutica Sinica B     Open Access   (Followers: 3, SJR: 1.793, CiteScore: 6)
Acta Psychologica     Hybrid Journal   (Followers: 26, SJR: 1.331, CiteScore: 2)
Acta Sociológica     Open Access   (Followers: 1)
Acta Tropica     Hybrid Journal   (Followers: 6, SJR: 1.052, CiteScore: 2)
Acta Urológica Portuguesa     Open Access   (Followers: 1)
Actas Dermo-Sifiliograficas     Full-text available via subscription   (Followers: 3, SJR: 0.374, CiteScore: 1)
Actas Dermo-Sifiliográficas (English Edition)     Full-text available via subscription   (Followers: 2)
Actas Urológicas Españolas     Full-text available via subscription   (Followers: 3, SJR: 0.344, CiteScore: 1)
Actas Urológicas Españolas (English Edition)     Full-text available via subscription   (Followers: 1)
Actualites Pharmaceutiques     Full-text available via subscription   (Followers: 7, SJR: 0.19, CiteScore: 0)
Actualites Pharmaceutiques Hospitalieres     Full-text available via subscription   (Followers: 3)
Acupuncture and Related Therapies     Hybrid Journal   (Followers: 8)
Acute Pain     Full-text available via subscription   (Followers: 15, SJR: 2.671, CiteScore: 5)
Ad Hoc Networks     Hybrid Journal   (Followers: 11, SJR: 0.53, CiteScore: 4)
Addictive Behaviors     Hybrid Journal   (Followers: 18, SJR: 1.29, CiteScore: 3)
Addictive Behaviors Reports     Open Access   (Followers: 9, SJR: 0.755, CiteScore: 2)
Additive Manufacturing     Hybrid Journal   (Followers: 14, SJR: 2.611, CiteScore: 8)
Additives for Polymers     Full-text available via subscription   (Followers: 22)
Advanced Drug Delivery Reviews     Hybrid Journal   (Followers: 198, SJR: 4.09, CiteScore: 13)
Advanced Engineering Informatics     Hybrid Journal   (Followers: 13, SJR: 1.167, CiteScore: 4)
Advanced Powder Technology     Hybrid Journal   (Followers: 17, SJR: 0.694, CiteScore: 3)
Advances in Accounting     Hybrid Journal   (Followers: 9, SJR: 0.277, CiteScore: 1)
Advances in Agronomy     Full-text available via subscription   (Followers: 20, SJR: 2.384, CiteScore: 5)
Advances in Anesthesia     Full-text available via subscription   (Followers: 30, SJR: 0.126, CiteScore: 0)
Advances in Antiviral Drug Design     Full-text available via subscription   (Followers: 2)
Advances in Applied Mathematics     Full-text available via subscription   (Followers: 12, SJR: 0.992, CiteScore: 1)
Advances in Applied Mechanics     Full-text available via subscription   (Followers: 12, SJR: 1.551, CiteScore: 4)
Advances in Applied Microbiology     Full-text available via subscription   (Followers: 24, SJR: 2.089, CiteScore: 5)
Advances In Atomic, Molecular, and Optical Physics     Full-text available via subscription   (Followers: 15, SJR: 0.572, CiteScore: 2)
Advances in Biological Regulation     Hybrid Journal   (Followers: 4, SJR: 2.61, CiteScore: 7)
Advances in Botanical Research     Full-text available via subscription   (Followers: 2, SJR: 0.686, CiteScore: 2)
Advances in Cancer Research     Full-text available via subscription   (Followers: 35, SJR: 3.043, CiteScore: 6)
Advances in Carbohydrate Chemistry and Biochemistry     Full-text available via subscription   (Followers: 9, SJR: 1.453, CiteScore: 2)
Advances in Catalysis     Full-text available via subscription   (Followers: 5, SJR: 1.992, CiteScore: 5)
Advances in Cell Aging and Gerontology     Full-text available via subscription   (Followers: 5)
Advances in Cellular and Molecular Biology of Membranes and Organelles     Full-text available via subscription   (Followers: 14)
Advances in Chemical Engineering     Full-text available via subscription   (Followers: 29, SJR: 0.156, CiteScore: 1)
Advances in Child Development and Behavior     Full-text available via subscription   (Followers: 11, SJR: 0.713, CiteScore: 1)
Advances in Chronic Kidney Disease     Full-text available via subscription   (Followers: 11, SJR: 1.316, CiteScore: 2)
Advances in Clinical Chemistry     Full-text available via subscription   (Followers: 27, SJR: 1.562, CiteScore: 3)
Advances in Clinical Radiology     Full-text available via subscription   (Followers: 1)
Advances in Colloid and Interface Science     Full-text available via subscription   (Followers: 21, SJR: 1.977, CiteScore: 8)
Advances in Computers     Full-text available via subscription   (Followers: 14, SJR: 0.205, CiteScore: 1)
Advances in Cosmetic Surgery     Full-text available via subscription   (Followers: 1)
Advances in Dermatology     Full-text available via subscription   (Followers: 16)
Advances in Developmental Biology     Full-text available via subscription   (Followers: 14)
Advances in Digestive Medicine     Open Access   (Followers: 14)
Advances in DNA Sequence-Specific Agents     Full-text available via subscription   (Followers: 7)
Advances in Drug Research     Full-text available via subscription   (Followers: 26)
Advances in Ecological Research     Full-text available via subscription   (Followers: 44, SJR: 2.524, CiteScore: 4)
Advances in Engineering Software     Hybrid Journal   (Followers: 30, SJR: 1.159, CiteScore: 4)
Advances in Experimental Biology     Full-text available via subscription   (Followers: 9)
Advances in Experimental Social Psychology     Full-text available via subscription   (Followers: 51, SJR: 5.39, CiteScore: 8)
Advances in Exploration Geophysics     Full-text available via subscription   (Followers: 2)
Advances in Family Practice Nursing     Full-text available via subscription   (Followers: 1)
Advances in Fluorine Science     Full-text available via subscription   (Followers: 9)
Advances in Food and Nutrition Research     Full-text available via subscription   (Followers: 69, SJR: 0.591, CiteScore: 2)
Advances in Fuel Cells     Full-text available via subscription   (Followers: 17)
Advances in Genetics     Full-text available via subscription   (Followers: 21, SJR: 1.354, CiteScore: 4)
Advances in Genome Biology     Full-text available via subscription   (Followers: 11, SJR: 12.74, CiteScore: 13)
Advances in Geophysics     Full-text available via subscription   (Followers: 8, SJR: 1.193, CiteScore: 3)
Advances in Heat Transfer     Full-text available via subscription   (Followers: 26, SJR: 0.368, CiteScore: 1)
Advances in Heterocyclic Chemistry     Full-text available via subscription   (Followers: 11, SJR: 0.749, CiteScore: 3)
Advances in Human Factors/Ergonomics     Full-text available via subscription   (Followers: 26)
Advances in Imaging and Electron Physics     Full-text available via subscription   (Followers: 4, SJR: 0.193, CiteScore: 0)
Advances in Immunology     Full-text available via subscription   (Followers: 37, SJR: 4.433, CiteScore: 6)
Advances in Inorganic Chemistry     Full-text available via subscription   (Followers: 10, SJR: 1.163, CiteScore: 2)
Advances in Insect Physiology     Full-text available via subscription   (Followers: 2, SJR: 1.938, CiteScore: 3)
Advances in Integrative Medicine     Hybrid Journal   (Followers: 6, SJR: 0.176, CiteScore: 0)
Advances in Intl. Accounting     Full-text available via subscription   (Followers: 3)
Advances in Life Course Research     Hybrid Journal   (Followers: 10, SJR: 0.682, CiteScore: 2)
Advances in Lipobiology     Full-text available via subscription   (Followers: 1)
Advances in Magnetic and Optical Resonance     Full-text available via subscription   (Followers: 8)
Advances in Marine Biology     Full-text available via subscription   (Followers: 21, SJR: 0.88, CiteScore: 2)
Advances in Mathematics     Full-text available via subscription   (Followers: 17, SJR: 3.027, CiteScore: 2)
Advances in Medical Sciences     Hybrid Journal   (Followers: 9, SJR: 0.694, CiteScore: 2)
Advances in Medicinal Chemistry     Full-text available via subscription   (Followers: 6)
Advances in Microbial Physiology     Full-text available via subscription   (Followers: 5, SJR: 1.158, CiteScore: 3)
Advances in Molecular and Cell Biology     Full-text available via subscription   (Followers: 26)
Advances in Molecular and Cellular Endocrinology     Full-text available via subscription   (Followers: 8)
Advances in Molecular Pathology     Hybrid Journal   (Followers: 1)
Advances in Molecular Toxicology     Full-text available via subscription   (Followers: 7, SJR: 0.182, CiteScore: 0)
Advances in Nanoporous Materials     Full-text available via subscription   (Followers: 5)
Advances in Oncobiology     Full-text available via subscription   (Followers: 2)
Advances in Ophthalmology and Optometry     Full-text available via subscription   (Followers: 1)
Advances in Organ Biology     Full-text available via subscription   (Followers: 2)
Advances in Organometallic Chemistry     Full-text available via subscription   (Followers: 18, SJR: 1.875, CiteScore: 4)
Advances in Parallel Computing     Full-text available via subscription   (Followers: 7, SJR: 0.174, CiteScore: 0)
Advances in Parasitology     Full-text available via subscription   (Followers: 6, SJR: 1.579, CiteScore: 4)
Advances in Pediatrics     Full-text available via subscription   (Followers: 27, SJR: 0.461, CiteScore: 1)
Advances in Pharmaceutical Sciences     Full-text available via subscription   (Followers: 19)
Advances in Pharmacology     Full-text available via subscription   (Followers: 17, SJR: 1.536, CiteScore: 3)
Advances in Physical Organic Chemistry     Full-text available via subscription   (Followers: 10, SJR: 0.574, CiteScore: 1)
Advances in Phytomedicine     Full-text available via subscription  
Advances in Planar Lipid Bilayers and Liposomes     Full-text available via subscription   (Followers: 3, SJR: 0.109, CiteScore: 1)
Advances in Plant Biochemistry and Molecular Biology     Full-text available via subscription   (Followers: 10)
Advances in Plant Pathology     Full-text available via subscription   (Followers: 6)
Advances in Porous Media     Full-text available via subscription   (Followers: 5)
Advances in Protein Chemistry     Full-text available via subscription   (Followers: 19)
Advances in Protein Chemistry and Structural Biology     Full-text available via subscription   (Followers: 20, SJR: 0.791, CiteScore: 2)
Advances in Psychology     Full-text available via subscription   (Followers: 69)
Advances in Quantum Chemistry     Full-text available via subscription   (Followers: 7, SJR: 0.371, CiteScore: 1)
Advances in Radiation Oncology     Open Access   (Followers: 3, SJR: 0.263, CiteScore: 1)
Advances in Small Animal Medicine and Surgery     Hybrid Journal   (Followers: 3, SJR: 0.101, CiteScore: 0)
Advances in Space Biology and Medicine     Full-text available via subscription   (Followers: 7)
Advances in Space Research     Full-text available via subscription   (Followers: 435, SJR: 0.569, CiteScore: 2)
Advances in Structural Biology     Full-text available via subscription   (Followers: 6)
Advances in Surgery     Full-text available via subscription   (Followers: 13, SJR: 0.555, CiteScore: 2)
Advances in the Study of Behavior     Full-text available via subscription   (Followers: 36, SJR: 2.208, CiteScore: 4)
Advances in Veterinary Medicine     Full-text available via subscription   (Followers: 20)
Advances in Veterinary Science and Comparative Medicine     Full-text available via subscription   (Followers: 15)
Advances in Virus Research     Full-text available via subscription   (Followers: 6, SJR: 2.262, CiteScore: 5)
Advances in Water Resources     Hybrid Journal   (Followers: 57, SJR: 1.551, CiteScore: 3)
Aeolian Research     Hybrid Journal   (Followers: 6, SJR: 1.117, CiteScore: 3)
Aerospace Science and Technology     Hybrid Journal   (Followers: 400, SJR: 0.796, CiteScore: 3)
AEU - Intl. J. of Electronics and Communications     Hybrid Journal   (Followers: 8, SJR: 0.42, CiteScore: 2)
African J. of Emergency Medicine     Open Access   (Followers: 6, SJR: 0.296, CiteScore: 0)
Ageing Research Reviews     Hybrid Journal   (Followers: 12, SJR: 3.671, CiteScore: 9)
Aggression and Violent Behavior     Hybrid Journal   (Followers: 483, SJR: 1.238, CiteScore: 3)
Agri Gene     Hybrid Journal   (Followers: 1, SJR: 0.13, CiteScore: 0)
Agricultural and Forest Meteorology     Hybrid Journal   (Followers: 18, SJR: 1.818, CiteScore: 5)
Agricultural Systems     Hybrid Journal   (Followers: 32, SJR: 1.156, CiteScore: 4)
Agricultural Water Management     Hybrid Journal   (Followers: 47, SJR: 1.272, CiteScore: 3)
Agriculture and Agricultural Science Procedia     Open Access   (Followers: 4)
Agriculture and Natural Resources     Open Access   (Followers: 3)
Agriculture, Ecosystems & Environment     Hybrid Journal   (Followers: 58, SJR: 1.747, CiteScore: 4)
Ain Shams Engineering J.     Open Access   (Followers: 5, SJR: 0.589, CiteScore: 3)
Air Medical J.     Hybrid Journal   (Followers: 8, SJR: 0.26, CiteScore: 0)
AKCE Intl. J. of Graphs and Combinatorics     Open Access   (SJR: 0.19, CiteScore: 0)
Alcohol     Hybrid Journal   (Followers: 12, SJR: 1.153, CiteScore: 3)
Alcoholism and Drug Addiction     Open Access   (Followers: 12)
Alergologia Polska : Polish J. of Allergology     Full-text available via subscription   (Followers: 1)
Alexandria Engineering J.     Open Access   (Followers: 2, SJR: 0.604, CiteScore: 3)
Alexandria J. of Medicine     Open Access   (Followers: 1, SJR: 0.191, CiteScore: 1)
Algal Research     Partially Free   (Followers: 11, SJR: 1.142, CiteScore: 4)
Alkaloids: Chemical and Biological Perspectives     Full-text available via subscription   (Followers: 2)
Allergologia et Immunopathologia     Full-text available via subscription   (Followers: 1, SJR: 0.504, CiteScore: 1)
Allergology Intl.     Open Access   (Followers: 5, SJR: 1.148, CiteScore: 2)
Alpha Omegan     Full-text available via subscription   (SJR: 3.521, CiteScore: 6)
ALTER - European J. of Disability Research / Revue Européenne de Recherche sur le Handicap     Full-text available via subscription   (Followers: 11, SJR: 0.201, CiteScore: 1)
Alzheimer's & Dementia     Hybrid Journal   (Followers: 56, SJR: 4.66, CiteScore: 10)
Alzheimer's & Dementia: Diagnosis, Assessment & Disease Monitoring     Open Access   (Followers: 6, SJR: 1.796, CiteScore: 4)
Alzheimer's & Dementia: Translational Research & Clinical Interventions     Open Access   (Followers: 6, SJR: 1.108, CiteScore: 3)
Ambulatory Pediatrics     Hybrid Journal   (Followers: 5)
American Heart J.     Hybrid Journal   (Followers: 58, SJR: 3.267, CiteScore: 4)
American J. of Cardiology     Hybrid Journal   (Followers: 67, SJR: 1.93, CiteScore: 3)
American J. of Emergency Medicine     Hybrid Journal   (Followers: 48, SJR: 0.604, CiteScore: 1)
American J. of Geriatric Pharmacotherapy     Full-text available via subscription   (Followers: 13)
American J. of Geriatric Psychiatry     Hybrid Journal   (Followers: 17, SJR: 1.524, CiteScore: 3)
American J. of Human Genetics     Hybrid Journal   (Followers: 40, SJR: 7.45, CiteScore: 8)
American J. of Infection Control     Hybrid Journal   (Followers: 35, SJR: 1.062, CiteScore: 2)
American J. of Kidney Diseases     Hybrid Journal   (Followers: 37, SJR: 2.973, CiteScore: 4)
American J. of Medicine     Hybrid Journal   (Followers: 51)
American J. of Medicine Supplements     Full-text available via subscription   (Followers: 3, SJR: 1.967, CiteScore: 2)
American J. of Obstetrics & Gynecology MFM     Hybrid Journal   (Followers: 1)
American J. of Obstetrics and Gynecology     Hybrid Journal   (Followers: 277, SJR: 2.7, CiteScore: 4)
American J. of Ophthalmology     Hybrid Journal   (Followers: 67, SJR: 3.184, CiteScore: 4)
American J. of Ophthalmology Case Reports     Open Access   (Followers: 5, SJR: 0.265, CiteScore: 0)
American J. of Orthodontics and Dentofacial Orthopedics     Full-text available via subscription   (Followers: 6, SJR: 1.289, CiteScore: 1)
American J. of Otolaryngology     Hybrid Journal   (Followers: 25, SJR: 0.59, CiteScore: 1)
American J. of Pathology     Hybrid Journal   (Followers: 32, SJR: 2.139, CiteScore: 4)
American J. of Preventive Medicine     Hybrid Journal   (Followers: 29, SJR: 2.164, CiteScore: 4)
American J. of Surgery     Hybrid Journal   (Followers: 39, SJR: 1.141, CiteScore: 2)
American J. of the Medical Sciences     Hybrid Journal   (Followers: 12, SJR: 0.767, CiteScore: 1)
Ampersand : An Intl. J. of General and Applied Linguistics     Open Access   (Followers: 7)
Anaerobe     Hybrid Journal   (Followers: 4, SJR: 1.144, CiteScore: 3)
Anaesthesia & Intensive Care Medicine     Full-text available via subscription   (Followers: 67, SJR: 0.138, CiteScore: 0)
Anaesthesia Critical Care & Pain Medicine     Full-text available via subscription   (Followers: 26, SJR: 0.411, CiteScore: 1)
Anales de Cirugia Vascular     Full-text available via subscription   (Followers: 1)
Anales de Pediatría     Full-text available via subscription   (Followers: 3, SJR: 0.277, CiteScore: 0)
Anales de Pediatría (English Edition)     Full-text available via subscription  
Anales de Pediatría Continuada     Full-text available via subscription  
Analytic Methods in Accident Research     Hybrid Journal   (Followers: 6, SJR: 4.849, CiteScore: 10)
Analytica Chimica Acta     Hybrid Journal   (Followers: 44, SJR: 1.512, CiteScore: 5)
Analytica Chimica Acta : X     Open Access  
Analytical Biochemistry     Hybrid Journal   (Followers: 224, SJR: 0.633, CiteScore: 2)
Analytical Chemistry Research     Open Access   (Followers: 13, SJR: 0.411, CiteScore: 2)
Analytical Spectroscopy Library     Full-text available via subscription   (Followers: 14)
Anesthésie & Réanimation     Full-text available via subscription   (Followers: 2)
Anesthesiology Clinics     Full-text available via subscription   (Followers: 25, SJR: 0.683, CiteScore: 2)

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Similar Journals
Journal Cover
Acta Histochemica
Journal Prestige (SJR): 0.661
Citation Impact (citeScore): 2
Number of Followers: 5  
 
  Hybrid Journal Hybrid journal (It can contain Open Access articles)
ISSN (Print) 0065-1281 - ISSN (Online) 1618-0372
Published by Elsevier Homepage  [3206 journals]
  • The meaning of non-classical estrogen receptors and peroxisome
           proliferator-activated receptor for boar Leydig cell of immature testis
    • Abstract: Publication date: Available online 22 February 2020Source: Acta HistochemicaAuthor(s): M. Kotula-Balak, M. Duliban, P. Pawlicki, R. Tuz, B. Bilinska, B.J. Plachno, Z.J. Arent, I. Krakowska, K. TarasiukAbstractCommunication in biological systems involves diverse-types of cell-cell interaction including cross-talk between receptors expressed by the target cells. Recently, novel sort of estrogen receptors (G protein - coupled estrogen receptor; GPER and estrogen-related receptor; ERR) that signal directly via estrogen binding and/or via mutual interaction-regulated estrogen signaling were reported in various organs including testis. Peroxisome proliferator - activated receptor (PPAR) is responsible for maintaining of lipid homeostasis that is critical for sex steroid production in the testis. Here, we investigated the role of interaction between GPER, ERRβ and PPARγ in steroidogenic Leydig cells of immature boar testis. Testicular fragments cultured ex vivo were treated with GPER or PPARγ antagonists. Then, cell ultrastructure, expression and localization of GPER, ERRβ, PPARγ together with the molecular receptor mechanism, through cyclic AMP and Raf/Ras/extracellular signal activated kinases (ERK), in the control of cholesterol concentration and estrogen production by Leydig cells were studied. In the ultrastructure of antagonist-treated Leydig cells, mitochondria were not branched and not bifurcated as they were found in control. Additionally, in PPARγ-blocked Leydig cells changes in the number of lipid droplets were revealed. Independent of used antagonist, western blot revealed decreased co-expression of GPER, ERRβ, PPARγ with exception of increased expression of ERRβ after PPARγ blockage. Immunohistochemistry confirmed presence of all receptors partially located in the nucleus or cytoplasm of Leydig cells of both control and treated testes. Changes in receptor expression, decreased cholesterol and increased estradiol tissue concentrations occurred through decreased cAMP level (with exception after GPER blockage) as well as Raf/Ras/ERK pathway expression. These all findings indicate that GPER-ERRβ-PPARγ interaction exists in immature boar testis and regulates Leydig cell function. Further detailed studies and considerations on GPER-ERRβ-PPARγ as possible diagnosis/therapy target in disturbances of testis steroidogenic function are needed.
       
  • Comparative histological studies on properties of polysaccharides secreted
           by vomeronasal glands of eight Laurasiatheria species
    • Abstract: Publication date: Available online 18 February 2020Source: Acta HistochemicaAuthor(s): Daisuke Kondoh, Jumpei Tomiyasu, Raito Itakura, Mizuho Sugahara, Masashi Yanagawa, Kenichi Watanabe, Phillip A. Alviola, Sheryl A. Yap, Edison A. Cosico, Florante A. Cruz, Ariel R. Larona, Allen J.F. Manalad, Joseph S. Masangkay, Yuki Sugiura, Shigeru Kyuwa, Shumpei Watanabe, Yumi Une, Tsutomu Omatsu, Hironori Bando, Kentaro KatoAbstractMost mammalian species have a vomeronasal organ that detects specific chemical substances, such as pheromones. Mucous fluid covering the vomeronasal sensory epithelium is secreted by vomeronasal glands, and the properties of these fluids have been suggested to be involved in chemical detection. Histological studies using periodic acid-Schiff (PAS) and Alcian blue pH 2.5 (AB) stains, which respectively detect natural and acidic polysaccharides, have suggested variations in the nature of the vomeronasal glands among species. Here, we investigated the responsivity of the vomeronasal glands to PAS and AB stains in eight Laurasiatheria species. All species studied herein possessed vomeronasal glands that stained positive for PAS, like other many reported species. The vomeronasal glands of dogs and minks – like rodents, were AB-negative, whereas those of cows, goats, sika deer, musk shrews and two bat species were positive. Considering the present findings and previous reports, the vomeronasal glands in most of Laurasiatheria species appear to be fundamentally abundant in acidic polysaccharides, whereas those in carnivores essentially contains neutral polysaccharides.
       
  • Discrete localization patterns of Arf6, and its activators EFA6A and
           BRAG2, and its effector PIP5kinaseγ on myofibrils of myotubes and plasma
           membranes of myoblasts in developing skeletal muscles of mice
    • Abstract: Publication date: Available online 12 February 2020Source: Acta HistochemicaAuthor(s): Surang Chomphoo, Hiroyuki Sakagami, Hisatake Kondo, Wiphawi HipkaeoAbstractArf6 (ADP ribosylation factor 6), activated by Arf-GEF (guanine nucleoside exchange factor), is involved in the membrane trafficking and actin-remodeling which are critical for maintenance of cell organization and activity and for fusion of myoblasts to form myotubes/myofibers. EFA6A (exchange factor for Arf6 type A) and BRAG2 (brefeldin A-resistant Arf-GEF 2) represent members of discrete subfamilies of Arf-GEF, while PIP5Kγ (phosphatidylinositol4-phosphate5-kinase γ) produces PI 4,5-bisphosphate (PIP2) and it is target for Arf6. In the present study, immunoreactive bands for Arf6, EFA6A, BRAG2 and PIP5Kγ were detected in immunoblots of skeletal muscle homogenates of mice at E18D (embryonic day 18), while the bands for Arf6, EFA6A and PIP5Kγ were reduced in density and no significant bands for BRAG2 were discerned at P1D (postnatal 1 day). No immunoblot bands for any of the molecules were eventually detected in skeletal fibers of adult mice. Immunoreactivities for endogenous Arf6, EFA6A and PIP5Kγ were visualized using immuno-light microscopy localized as periodic striations running perpendicular to the longitudinal axes of skeletal muscle fibers of mice at E18D and P1D. All the striations were co-immunoreactive for β-actin in double immunofluorescence microscopy, and the immunoreactivities were confined to thin myofilaments at sarcomeric I-domains in immuno-electron microscopy. On the other hand, immunoreactivities for Arf6, BRAG2 and PIP5Kγ were conspicuous on plasmalemma of myoblasts at E14D, while immunoreactivity for EFA6A was already distinct in striations perpendicular to myofibrils in myotubes at E14D. The present findings suggest three possibilities: involvement of EFA6A-activated Arf6 together with PIP5Kγ in maturation of myofibrils, movement of Arf6 and PIP5Kγ from the plasmalemma of myoblasts to myofibrils of myotubes, and that of BRAG2 to the cytoplasm of myotubes; and further a function of EFA6A independent of the activation of Arf6 in immature myofibrils. In addition, the involvement of Arf6, BRAG2 and PIP5Kγ in the fusion of myoblasts into myotubes was supported by the present finding.
       
  • In-vitro investigation of calcitonin associated effects on the
           trophoblastic cells
    • Abstract: Publication date: Available online 3 February 2020Source: Acta HistochemicaAuthor(s): Birsen Ozdemir, Serbay Ozkan, Elif Guzel, Meral KoyuturkAbstractCalcitonin is expressed in the epithelium of endometrium, and modulates zonula adherens junctions which are composed of cadherin-catenins complex during the implantation window. Trophoblastic cells which have complex interaction with the epithelial cells of endometrium during implantation were demonstrated to have calcitonin receptors. Mechanism of action of calcitonin on trophoblastic cells has not yet been elucidated. Therefore, it was aimed to determine the effects of calcitonin on the expressions of β-catenin and phospho-β-catenin in a dose depended manner under the influence of progesterone and estrogen hormones (P + E) by using JAR cell line through the immunocytochemical and Western blot analyses. Moreover, adherens junctions (AJs) were ultrastructurally investigated to assess the involvement of cadherin-catenin complex in accordance with the changes in the specified parameters. Immunocytochemical analysis showed that only 10 nM calcitonin treated group had increased expression of membranous β-catenin compared to the control group, while there was decreased expression of β-catenin in the nucleus of all the experimental groups. Cytoplasmic expressions of the phospho-β-catenin decreased in all experimental groups compared to the control group while the decrease in the nuclear expression was remarkable in the groups treated with P + E, and P + E + 250 nM calcitonin. Western blot analysis showed that total β-catenin and phospho-β-catenin expressions were not significantly different. Ultrastructural analysis showed that increase in the number of AJs was noticeable in the group treated with 10 nM calcitonin. Overall, the localization and expression levels of β-catenin and phospho-β-catenin suggest that calcitonin could show its effects through the non-canonical pathway in the trophoblastic cells.
       
  • Neuroprotective effects of andrographolide on chronic cerebral
           hypoperfusion-induced hippocampal neuronal damage in rats possibly via
           PTEN/AKT signaling pathway
    • Abstract: Publication date: Available online 1 February 2020Source: Acta HistochemicaAuthor(s): Da-Peng Wang, Shu-Hui Chen, Di Wang, Kai Kang, Yi-Fang Wu, Shao-Hua Su, Ying-Ying Zhang, Jian HaiAbstractTo explore the potential effects of andrographolide on chronic cerebral hypoperfusion (CCH)-induced neuronal damage as well as the underlying mechanisms. Rat CCH model was established by 2-vessel occlusion (2VO). The CCH rats received andrographolide treatment for 4 weeks. The neuron loss was detected by using neuronal nuclei (NeuN) immunofluorescent staining. The expression levels of phospho-phosphatase and tensin homolog deleted on chromosome ten (p-PTEN), protein kinase B (AKT), p-AKT, and cysteinyl aspartate specific proteinase-3 (Caspase-3) proteins were accessed by Western blotting. Moreover, the neuronal apoptosis of hippocampus tissues was detected via terminal deoxynucleotidyl transferase- mediated dUTP nick end labeling (TUNEL) staining. CCH reduced the number of NeuN-positive cells, while the number was significant increased after andrographolide treatment. CCH increased the proteins expression level of p-PTEN, Caspase-3, and decreased the p-AKT, which were reversed by andrographolide treatment. Furthermore, andrographolide treatment also down-regulated CCH-induced TUNEL-apoptosis rate. Our results suggest that the PTEN/AKT pathway may be modulated by andrographolide and the damaging effects of CCH on hippocampus may be ameliorated by andrographolide treatment. Andrographolide may act as a potential therapeutic approach for chronic ischemic insults.
       
  • Motivating students to enter science research careers
    • Abstract: Publication date: Available online 31 January 2020Source: Acta HistochemicaAuthor(s): Steve Oppenheimer
       
  • The roles of cirRNA in the development of germ cells
    • Abstract: Publication date: Available online 31 January 2020Source: Acta HistochemicaAuthor(s): Yaqin Cai, Xiaocan Lei, Zhuo Chen, Zhongcheng MoAbstractCircular RNA (CircRNA), a type of endogenous non-coding RNAs (ncRNAs), is generally generated from precursor mRNA (pre-mRNA) by canonical splicing and head-to-tail back splicing. The structure without a polyA tail renders circRNA highly insensitive to ribonuclease. Simultaneously, the distribution of circRNAs is tissue and developmental stage-specific. There are five potential biological functions of circRNAs: 1) promote transcription of their parental genes; 2) function as a miRNA sponge; 3) RNA binding protein (RBP) sponge; 4) encode protein; 5) act as an mRNA trap. Recently, circRNA has attracted attention because studies have shown that circRNAs are associated with follicular development, ovarian senescence, spermatogenesis, and germ cell development process, suggesting that circRNAs may function in germ cells regulation. The investigation of circRNAs in germ cells will provide an excellent opportunity to understand its potential molecular basis, and potentially improving reproduction status in human. In this article, the relationship between circRNA and germ cell development will be discussed.
       
  • Inhibition of the migration of MCP-1 positive cells by icing applied soon
           after crush injury to rat skeletal muscle
    • Abstract: Publication date: Available online 25 January 2020Source: Acta HistochemicaAuthor(s): Motoi Miyakawa, Masato Kawashima, Daijiro Haba, Megumi Sugiyama, Kaho Taniguchi, Takamitsu ArakawaAbstractMigration of the macrophages to the injured site soon after the skeletal muscle injury is crucial for subsequent regeneration of the muscle fibers. The Monocyte chemoattractant protein-1 (MCP-1) is important chemokine for regulating migration of the monocytes/macrophages. Earlier reports have discussed that icing applied soon after muscle crush injury retards muscle regeneration through retardation of macrophage migration. The MCP-1+ cells and neutrophils might promote the migration of the macrophages. To test the hypothesis that icing soon after the skeletal muscle injury affects MCP-1+ cells and neutrophils, we examined the effect of icing on MCP-1+ cells and neutrophils after crush injury to skeletal muscle in rats. Owing to the icing application for 20 min soon after the injury, accumulation of the macrophages was inhibited until 12 h after injury. Numbers of the neutrophils at 3 h after the injury and the MCP-1+ cells at 6 h and later after the injury in the icing group were significantly lower than those in the non-icing group, suggesting that these phenomena contribute to the retardation of macrophage migration.
       
  • miRNA-183∼96∼182 regulates melanogenesis, cell proliferation
           and migration in B16 cells
    • Abstract: Publication date: Available online 21 January 2020Source: Acta HistochemicaAuthor(s): Bin Du, Xuexian Liu, Ajab Khan, Shuangxiu Wan, Xiang Guo, Jixuan Xue, Ruiwen FanAbstractMelanoma is a highly invasive malignant skin tumor having high metastatic rate and poor prognosis. The biology of melanoma is controled by miRNAs. The miRNA-183 cluster, which is composed of miRNA-183∼96∼182 genes, plays an important roles in tumor development. In order to investigate the role and action of miRNA-183 cluster in B16 cells, we overexpressed and knocked down miRNA-183 cluster in B16 cells. Using bioinformatics analysis, we predicted that the key framscript factor of melangenic genes. Microphthalmia-associated transcription factor (MITF) is one of the targets of miRNA-183 cluster. The results of Luciferase activity assays confirmed that MITF was targeted by miRNA-183 cluster. Overexpression and knockdown of miRNA-183 cluster in B16 cells resulted in down and up regulation of MITF expression, respectively at both mRNA and protein levels. Furthmore, overexpression and knockdown of the miRNA-183 cluster in B16 cells decreased and increased the expression of mRNA and protein of melangenic genes tyrosinase (TYR), and tyrosinase-related protein 1 (TYRP1), dopachrome-tautomerase (DCT), as well as the production of melanins and eumelanin production, respectively. On the proliferation and migration pathway, overexpression and knockdown of miRNA-183 cluster increased and decreased, respectively the expression of mRNA and protein of mitogen-activated protein kinase 1 (MEK1), extracellular regulated protein kinases1/2 (ERK1/2) and cAMP-responsive-element binding protein (CREB). These results indicated that miRNA-183 cluster regulated melanogenesis in B16 cells as well as cell proliferation and migration by directly targeting MITF through migration pathway.
       
  • Expression and localization of meiosis-associated protein in gonads of
           female rats at different stages
    • Abstract: Publication date: Available online 18 January 2020Source: Acta HistochemicaAuthor(s): Yihui Liu, Xiaorui Fan, Meishan Yue, Weidong Yue, Xinrong Zhang, Jingwen Zhang, Gaoya Ren, Junping HeAbstractIt was well known that a critical process of oogenesis in the female mammalian was the entry of mitotic oogonia into meiosis. Early studies from model animal mice suggested that the retinoic acid (RA) response signal protein STRA8 (stimulated by retinoic acid gene 8) and the meiosis-specific chromosomal behavior marker protein SCP3 (Synaptonemal Complex Protein 3) were two crucial molecular markers during meiosis. The expression of STRA8 and SCP3 at different stages in rat ovaries was investigated by immunohistochemistry, qRT-PCR and Western Blot. Immunohistochemistry results showed that STRA8 and SCP3 were mainly expressed in embryonic stage. And STRA8 was expressed in the cytoplasm and nucleus of the ovaries after birth. qRT-PCR and Western Blot results showed that the mRNA and protein levels of STRA8 and SCP3 were expressed in embryonic stage. The expression of STRA8 and SCP3 indicated germ cells enter meiosis in rats embryo, and STRA8 and SCP3 could serve as molecular markers for the meiosis in rats. The localization of STRA8 in the nucleus increased the possibility that STRA8 might act as transcription factor or activate transcription to function after birth.
       
  • Eicosapentaenoic and docosapentaenoic acids lessen the expression of
           PPARγ/Cidec affecting adipogenesis in cultured 3T3-L1 adipocytes
    • Abstract: Publication date: Available online 17 January 2020Source: Acta HistochemicaAuthor(s): Fabiane Ferreira Martins, Marcia Barbosa Aguila, Carlos Alberto Mandarim-de-LacerdaAbstractEicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have benefits in the metabolism of adipose tissue. However, its contribution to the adipogenesis is not entirely elucidated. The study aimed to evaluate the effects of EPA and DHA on adipogenesis, especially in the PPARγ (peroxisome proliferator-activated receptor-gamma) and Cidec (cell death-inducing DFFA-like effector c) pathway. Twenty-four hours after confluence, 3T3-L1 adipocytes were treated with EPA (100 μM), DHA (50μM) and EPA (100μM) + DHA (50μM) and at the end of differentiation (day 11) the cells were collected for analysis. Cell viability analysis indicated that the concentrations used for EPA and DHA did not cause cytotoxicity in cultured 3T3l1 adipocytes. The treatments have lessened the triacylglycerol accumulation in the adipocyte cytoplasm that, compared to the control group, were EPA-32%, DHA-38%, EPA + DHA −24%. The double-labeling immunofluorescence showed a signal attenuation of protein expressions of PPARγ, CIDEC, and SREBP-1c (sterol regulatory element-binding protein). EPA and DHA had a significant impact on the expression of cleaved CASPASE 3, which increases cell apoptosis and gene expressions of Pparγ and Cidec in the treated groups. Also, there was a reduction of C/ebpα (CCAAT/enhancer-binding protein alpha), Cd36 (cluster differentiation 36), and Foxo1 (forkhead box O). In conclusion, the study determined the ability of both EPA and DHA, alone or combined, in the adipogenesis modulation in cultured 3T3-L1 adipocytes, affecting the cell differentiation, maturation, and consequently, reducing adipogenesis via PPARγ-CIDEC suppression.
       
  • Utility of the immunohistochemical analysis of DNA mismatch-repair
           proteins in endometrial hyperplasia
    • Abstract: Publication date: Available online 17 January 2020Source: Acta HistochemicaAuthor(s): Nabiha Missaoui, Nesrine Boukhari, Sarra Limam, Sihem Hmissa, Moncef MokniAbstractThe utility of the expression lack of DNA mismatch-repair (MMR) proteins in the detection of Lynch syndrome in endometrial hyperplasia as precursor lesion of endometrial carcinoma has not been well-established. The study investigated the immunoexpression pattern of MMR proteins in endometrial hyperplasia from Tunisian patients. We carried out a retrospective study of 60 endometrial hyperplasias diagnosed among Tunisian patients. Expression of MLH1, MSH2, MSH6, and PMS2 proteins was performed by immunohistochemistry on whole-slide sections of archival tissues. Analysis of MLH1 promoter methylation and microsatellite alterations was conducted in appropriate cases. Microsatellite instability screening was assessed using the Bethesda panel, including BAT25, BAT26, D17S250, D2S123, and D5S346 markers. Expression of MMR proteins was observed in all hyperplasias without atypia as well as in 27 out of 29 atypical hyperplasias. Only two atypical hyperplasias exhibited expression loss of MMR proteins. A single case revealed MSH6 expression lack. Expression loss of MLH1 and PMS2 was identified in another atypical hyperplasia and was associated with hypermethylation of MLH1 promoter. This patient had no familial history of endometrial cancer at the diagnostic time. The two deficient MMR cases showed microsatellite stable pattern. In conclusion, only two endometrial hyperplasias displayed an altered pattern of MMR expression. Our results suggest the limited utility of the immunohistochemical analysis of MMR protein in the early detection of Lynch syndrome in Tunisian patients diagnosed with endometrial hyperplasias. Multicenter studies with larger sample size are needed to more explore these findings.
       
  • Anatomical features of the tongue of two chiropterans endemic in the
           Egyptian fauna; the Egyptian fruit bat (Rousettus aegyptiacus) and
           insectivorous bat (Pipistrellus kuhlii)
    • Abstract: Publication date: Available online 16 January 2020Source: Acta HistochemicaAuthor(s): Diaa Massoud, Mohamed M.A. AbumandourAbstractThe current study aimed to investigate the tongue (lingual) morphometry, histology, and histochemistry of two chiropterans endemic in the Egyptian fauna, and having different feeding preferences. The tongues of nine adult individuals of each species were utilized in our investigation. The tongue of fruit-eating bat was observed relatively longer than the one of insect-eating bat. Grossly, the insect-eating bat had a lingual prominence on the dorsum of lingual body, while the fruit-eating bat had a concave midline over the lingual body. Histologically, numerous forms of lingual papillae were scattered along the dorsal epithelium of the tongue. The lingual papillae of the fruit-eating bat seem to be well adapted for piercing the skin of a fruit and liquid sap retention. The lingual glands of both species were lodged in the muscular layer. Two main sets were identified; the serous von Ebner's gland usually seen accompanied by the circumvallate papillae and Weber's gland with mixed mucoserous secretions. Von Ebner's gland showed more prominent acidic mucins, while Weber's gland expressed neutral mucins. The lingual epithelium of the fruit-eating bat had an outer covering of cornified non-nucleated epithelium. On the other hand, the insect-eating bat had an outer covering of nucleated epithelium. It is for the first time to record the existence of the entoglossal plates of both species which consisted of a bony core in the fruit-eating bat and a cartilaginous element in the insect-eating bat. The current study represents an attempt to shed more light on the tongue evolution among mammalian vertebrates.Graphical abstractGraphical abstract for this article
       
  • Differences in the expression of catecholamine-synthesizing enzymes
           between vesicular monoamine transporter 1- and 2-immunoreactive glomus
           cells in the rat carotid body
    • Abstract: Publication date: Available online 16 January 2020Source: Acta HistochemicaAuthor(s): Kouki Kato, Takuya Yokoyama, Tatsumi Kusakabe, Katsuhiko Hata, Seigo Fushuku, Nobuaki Nakamuta, Yoshio YamamotoAbstractVesicular monoamine transporters (VMAT) 1 and 2 are responsible for monoamine transportation into secretary vesicles and are tissue-specifically expressed in central and peripheral monoaminergic tissues, including the carotid body (CB). The aim of the present study was to examine the expression of catecholamine-synthesizing enzymes in VMAT1- and VMAT2-immunoreactive glomus cells in the rat CB using multiple immunolabeling. The expression of VMAT1 and VMAT2 mRNA in the CB was confirmed by RT-PCR. Immunohistochemistry revealed that VMAT1 immunoreactivity was predominant in glomus cells rather than VMAT2 immunoreactivity. Glomus cells with VMAT1 immunoreactivity exhibited weak/negative VMAT2 immunoreactivity, and vice versa. Immunoreactivities for VMAT1 and tyrosine hydroxylase, the rate-limiting enzyme for catecholamine biosynthesis, were co-localized in the same glomus cells and a positive correlation was confirmed between the two immunoreactivities (Spearman’s coefficient = 0.82; p 
       
  • Ultrastructural characterization of vitamin D receptors and metabolizing
           enzymes in the lipid droplets of the fatty liver in rat
    • Abstract: Publication date: Available online 10 January 2020Source: Acta HistochemicaAuthor(s): Filipović Natalija, Bočina Ivana, Restović Ivana, Grobe Maximilian, Kretzschmar Genia, Kević Nives, Mašek Tomislav, Vitlov Uljević Marija, Jurić Marija, Vukojević Katarina, Saraga-Babić Mirna, Vuica AnaAbstractVitamin D is a steroid hormone with numerous actions in the organism. There are strong evidences that relate vitamin D deficiency with liver lipid metabolism disturbances, but the mechanism of this action is still unknown. In our previous work we postulated the localization and accumulation of vitamin D receptor (VDR) in membrane of the lipid droplets (LDs) in hepatocytes. In this study, we applied the transmission electron microscopy (TEM) to confirm this hypothesis by using a long-term (6 months) high sucrose intake rat model that was previously found to be appropriate for research of the hepatic lipid accumulation. In addition to the VDR, we also found key vitamin D metabolizing enzymes, 1α-hydroxylase and CYP 24 associated with the membrane of the LDs. A light-microscopy data revealed significant increase in expression of VDR and CYP 24 in liver of high-sucrose treated rats, in comparison to controlones.According to the best of our knowledge, this is a first study confirming the presence of the VDR in the membrane of the LDs in general and also in particular in LDs of the hepatocytes that were accumulated as a consequence of the prolonged high sucrose intake.Moreover, we found association of main vitamin D metabolizing enzymes with LD membrane. These results provide a new insight in the possible relation of vitamin D signalling system with LD morphology and function and with the lipid metabolism in general.
       
  • Paeonol antagonizes oncogenesis of osteosarcoma by inhibiting the function
           of TLR4/MAPK/NF-κB pathway
    • Abstract: Publication date: January 2020Source: Acta Histochemica, Volume 122, Issue 1Author(s): Jianguo Zhou, Qinglin Liu, Rui Qian, Shiwei Liu, Weiquan Hu, Zhenyu LiuAbstractAs the the major functional component of Paeonia suffruticosa, paeonol (PAE) has shown its potential to inhibit the progression of multiple cancer types. In the current study, the mechanism driving the effect of PAE on osteosarcoma (OS) was investigated by focusing on its influence on TLR4-mediated MAPK/NF-κB pathway. Human OS cells were firstly administrated with PAE of different concentrations to assess its effect on the proliferation, apoptosis, metastasis, and TLR4/MAPK/NF-κB pathway in OS cells. Thereafter, the level of TLR4 was induced in OS cells before PAE administration to explore the role of the molecule in the anti-OS function of PAE. The results of in vitro assays were further validated with xenograft mice models. The administration of PAE of two doses both suppressed the proliferation and induced apoptosis in OS cells in a dose-dependent manner. Regarding the effect on the metastasis potential of OS cells, PAE inhibited the migration and invasion potential of the cells, but the effect did not change with concentrations. The administration of PAE also inhibited the expression of TLR4 and deactivated MAPK/NF-κB pathway. Moreover, the induced expression of TLR4 counteracted the anti-OS function of PAE. Further validation with xenograft models also showed that PAE inhibited solid tumor growth and TLR4 expression in OS mice. In conclusion, it was inferred that the anti-OS function of PAE depended on the inhibition of TLR4 and its downstream MAPK/NF-κB pathway.
       
  • Effect of gonadotropin releasing hormone on the expression of luteinizing
           hormone and estrogen in the nerve ganglia and ovary of a tropical abalone,
           Haliotis asinina Linnaeus
    • Abstract: Publication date: January 2020Source: Acta Histochemica, Volume 122, Issue 1Author(s): Parinyaporn Nuurai, Chaitip Wanichanon, Ratanasate WanichanonGonadotropin releasing hormone (GnRH) is a peptide brain hormone that is involved in the regulation of reproduction in vertebrates via stimulation of the secretion of the pituitary hormones, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), which in their turn stimulate sexual development and sex steroid hormone secretion by the gonads. The tropical abalone, Haliotis asinina, in common with many other invertebrates contains a peptide with a similar structure to GnRH. This study looks at its possible involvement in reproduction by injecting groups of one-year-old female abalone at the mature phase by injecting them with synthetic H. asinina (Has) GnRH at doses of 0, 250 and 500 ng/g and then measuring the amount of material in nerve ganglia, ovary and hemolymph that cross-reacted with enzyme-linked immunosorbent assays (ELISA) for vertebrate LH and steroid, estradiol. Immunohistochemistry, using antibodies for the same two compounds, was also carried out to examine the location of immunoactivity in the tissues of the animals. There were slight (in some cases statistically significant) increases in LH-immunoactivity and estradiol in the hemolymph and tissues. However, this applied to the lower dose only (i.e the dose-response relationship was non-monotonic). Using immunohistochemistry, LH-immunoreactive cells were observed in types 1 and 2 neurosecretory (NS1 and NS2) cells within the cerebral and pleuropedal ganglia of H. asinina. In addition, LH-immunoreactive nerve fiber bundles were strongly detected in both ganglia. The immunoactivity against the estrogen appeared to be localized in the granulated cells within the connective tissue and trabeculae of the mature ovary. There was no positive staining in the cytoplasm of any stage of the germ cells. The interpretation of these findings is presently hindered by the fact that the homologous gene for vertebrate LH has not yet been identified in the genomes of any mollusks (so the cause of the immunostaining is as yet unknown) and also by the fact that mollusks are known to readily absorb steroids from the environment and store them long-term in the form of fatty acid esters. More work, involving identification of the protein that cross-reacts with the LH antiserum and also exclusion of the possibility that the estradiol is of exogenous origin, will have to be carried out before these findings can be used to manipulate reproduction in this species.Graphical abstractGraphical abstract for this article
       
  • Actinic cheilitis: Morphometric parameters and its relationship with the
           degree of epithelial dysplasia
    • Abstract: Publication date: January 2020Source: Acta Histochemica, Volume 122, Issue 1Author(s): Amanda Katarinny Goes Gonzaga, Rodrigo Porpino Mafra, Leorik Pereira da Silva, Roseana de Almeida Freitas, Lélia Batista de Souza, Leão Pereira PintoAbstractActinic cheilitis (AC) is a potentially malignant lesion caused by chronic sun exposure. This study aimed to evaluate the relationship between the degree of epithelial dysplasia and morphometric findings in AC. Sixty-eight slides of AC cases were selected and classified according to the grade of epithelial dysplasia, following morphologic criteria of World Health Organization. For morphometric analysis, the slides were scanned and images were analyzed using Pannoramic Viewer software. We obtained vertical measurements of the parameters: thicknesses of the keratin layer, lamina propria and zone of solar elastosis in three selected fields. Thirty-seven (54.4%) of the analyzed cases were classified as none/mild dysplasia and 31 (45.6%) as moderate/severe epithelial dysplasia. Cases with a moderate/severe dysplasia exhibited a thicker layer of keratin (median = 0.055 mm) than none/mild dysplasia (median = 0.045 mm) (p = 0.033). No significant differences in the thicknesses of lamina propria and zone of solar elastosis were observed according to the grade of epithelial dysplasia. A positive significant correlation between keratin layer and lamina propria thicknesses was found (p = 0.019). Based on our findings, rigorous clinical follow-up should be recommended for patients whose histopathological examination shows a greater thickness of the keratin layer.
       
  • Histochemical and immunohistochemical localization of nitrergic structures
           in the carotid body of spontaneously hypertensive rats
    • Abstract: Publication date: Available online 7 January 2020Source: Acta HistochemicaAuthor(s): Dimitrinka Y. Atanasova, Angel D. Dandov, Nikolay D. Dimitrov, Nikolai E. LazarovAbstractThe carotid body (CB) is a multipurpose metabolic sensor that acts to initiate cardiorespiratory reflex adjustments to maintain homeostasis of blood-borne chemicals. Emerging evidence suggests that nitric oxide increases the CB chemosensory activity and this enhanced peripheral chemoreflex sensitivity contributes to sympathoexcitation and consequent pathology. The aim of this study was to examine by means of NADPH-diaphorase histochemistry and nitric oxide synthase (NOS) immunohistochemistry the presence and distribution of nitrergic structures in the CB of spontaneously hypertensive rats (SHRs) and to compare their expression patterns to that of age-matched normotensive Wistar rats (NWRs). Histochemistry revealed that the chemosensory glomus cells were NADPH-d-negative but were encircled by fine positive varicosities, which were also dispersed in the stroma around the glomeruli. The NADPH-d-reactive fibers showed the same distributional pattern in the CB of SHRs, however their staining activity was weaker when compared with NWRs. Thin periglomerular, intraglomerular and perivascular varicose fibers, but not glomus or sustentacular cells in the hypertensive CB, constitutively expressed two isoforms of NOS, nNOS and eNOS. In addition, clusters of glomus cells and blood vessels in the CB of SHRs exhibited moderate immunoreactivity for the third known NOS isoenzyme, iNOS. The present study demonstrates that in the hypertensive CB nNOS and eNOS protein expression shows statistically significant down-regulation whereas iNOS expression is up-regulated in the glomic tissue compared to normotensive controls. Our results suggest that impaired NO synthesis could contribute to elevated blood pressure in rats via an increase in chemoexcitation and sympathetic nerve activity in the CB.
       
  • Use of synthetic polymers improves the quality of vitrified caprine
           preantral follicles in the ovarian tissue
    • Abstract: Publication date: Available online 2 January 2020Source: Acta HistochemicaAuthor(s): Diego Alberto Montano Vizcarra, Yago Pinto Silva, Jamily Bezerra Bruno, Danielle Cristina Calado Brito, Deysi Dipaz Berrocal, Luciana Mascena Silva, Maria Luana Gaudencia dos Santos Morais, Benner Gerardo Alves, Kele Amaral Alves, Francielli Weber Santos Cibin, José Ricardo Figueiredo, Mary B. Zelinski, Ana Paula Ribeiro RodriguesAbstractThe aim of this study was to evaluate whether the addition of synthetic polymers to the vitrification solution affected follicular morphology and development and the expression of Ki-67, Aquaporin 3 (AQP3) and cleaved Caspase-3 proteins in ovarian tissue of the caprine species. Caprine ovaries were fragmented and two fragments were immediately fixed (Fresh Control) for morphological evaluation, while other two were in vitro cultured for 7 days (Cultured Control) and fixed as well. The remaining fragments were distributed in two different vitrification groups: Vitrified and Vitrified/Cultured. Each group was composed of 4 different treatments: 1) Sucrose (SUC); 2) SuperCool X-1000 0.2 % (X-1000); 3) SuperCool Z-1000 0.4 % (Z-1000) or 4) with polyvinylpyrrolidone K-12 0.2 % (PVP). Also, Fresh Control, Cultured Control, SUC and X-1000 were destined to immunohistochemical detection of Ki-67, AQP3 and cleaved Caspase-3 proteins. Morphologically, the treatment with X-1000 showed no significant difference with the Fresh Control group and was superior to the other treatments. After the cleaved caspase-3 analysis, X-1000 showed the lowest percentages of strong immunostaining while Cultured Control showed the highest. Also, a positive correlation was found between the percentages of degenerated follicles and the percentages of strong staining intensity follicles. Regarding the AQP3 analysis, the highest percentages of strong AQP3 staining intensity were found in X-1000. In conclusion, we have demonstrated that the addition of the synthetic polymer SuperCool X-1000 to the vitrification solution improved the current vitrification protocol of caprine ovarian tissue.
       
  • Expression and localization of endogenous phospholipase Cβ3 confined to
           basal cells in situ of immature ducts and adult excretory ducts of
           submandibular gland of mice
    • Abstract: Publication date: Available online 30 December 2019Source: Acta HistochemicaAuthor(s): Atsara Rawangwong, Masahiko Watanabe, Hisatake Kondo, Wiphawi HipkaeoAbstractOur previous study demonstrated that, different from the parotid and sublingual glands, the submandibular glands of adult mice did not show an immunoblot band for PLCβ3 which is critical in the secretion mechanism by muscarinic cholinergic signaling. Therefore, the submandibular glands of mice at various stages of postnatal development were examined for this enzyme molecule in immunoblot and immunohistochemistry. In immunoblot, a weak band for PLCβ3-expression was detected only at early postnatal stages. In immunohistochemistry, PLCβ3-immunoreactivity was distinctly found in most basally located cells of immature ducts, while the immunoreactivity was weakly seen in terminal tubule cells without significant immunoreactivity in adjacent acinar cells. In contrast, the immunoreactivity was distinctly found in some basal cells of adult excretory ducts, and it was ultrastructurally localized densely in close association with bundles of tonofilaments in the cells. The present finding suggests the possibility that Ca2+ signaling governed by phospholipase Cβ3 is involved in the differentiation of ductal basal cells into apical cells through control of keratin molecule(s) in the cells.
       
  • Age-related changes in the articular cartilage of the mandible of rats
    • Abstract: Publication date: Available online 28 December 2019Source: Acta HistochemicaAuthor(s): M.F. Calderon, R.A.B. Nucci, R.R. de SouzaAbstractThis study investigated the effects of aging on the articular cartilage of the mandible. Wistar rats were divided in two groups (n = 10/per group): 3-months-old group (young group); and 13-months-old group (aged group). After euthanasia, the head of the mandible was collected and stained with hematoxylin and eosin (HE) to evaluate the thickness of the articular layer and cartilage. Sections stained with Picrosirius red and Safranin O were used to evaluate the collagen and proteoglycans deposition, respectively. First, aging has decreased the articular layer thickness. Second, the results suggest a decrease of chondrocytes followed by an increase of the matrix to maintain the mandible homeostasis. Finally, both collagen and proteoglycans increased with aging. Aging displayed important effects to the mandible of aged rats.
       
  • Impact of vildagliptin on vascular and fibrotic remodeling of myocardium
           in experimental diabetic cardiomyopathy
    • Abstract: Publication date: Available online 28 December 2019Source: Acta HistochemicaAuthor(s): Ahmed A.M. Abdel-Hamid, Alaa El-Din L. FirganyAbstractThe effect of dipeptidyl peptidase-4 inhibitors (DPP-4is) on myocardium in diabetic cardiomyopathy (DCM) remains a matter of debate. In the current study we investigated the effect of vildagliptin (VILDA, 3 mg/kg/d) on myocardium of DCM focusing on coronary microcirculation as well as on endothelial stress markers (ICAM and VCAM). We divided animals equally into 4 groups; nondiabetic (ND), VILDA per se, DCM and DCM + VILDA and their myocardium was evaluated for the fibro-vascular remodeling immunohistochemically as well as for molecular changes. VILDA had reversed the histological changes occurred in DCM including the disintegration, degeneration, and steatosis of cardiomyocytes with disappearance of the edema fluid. In addition VILDA significantly increased (p 
       
  • Could cathepsin-k be a driver of the myofibroblastic differentiation
           observed in dermatofibroma, atypical fibroxanthoma and pleomorphic dermal
           sarcoma'
    • Abstract: Publication date: Available online 27 December 2019Source: Acta HistochemicaAuthor(s): Costantino Ricci, Antonio De Leo, Emi Dika, Martina Lambertini, Giulia Veronesi, Barbara CortiAbstractDermatofibroma (BFH), atypical fibroxanthoma (AFX) and dermal pleomorphic sarcoma (DPS) are skin-based soft-tissue neoplasms of uncertain lineage. They are classified as “fibrohistiocytic” neoplasms, even if the World Health Organization stated that this term connotes a polymorphic group of lesions that histologically resemble fibroblasts and histiocytes. It is well-known that this group of lesions shows a “fibro-histiocytic-dendritic” and/or a “myofibroblastic” phenotype, even within the same lesion. We studied the expression of cathepsin-k in 34 cases (25 BFH, 5 AFX, 4 DPS) with a broad panel of antibodies. 20 cases (5 dermatofibrosarcoma protuberans, 5 melanomas, 5 basal cell carcinomas, 5 squamous cell carcinomas) were chosen as controls. Although our results need to be validated, they support a myofibroblastic and/or partial myofibroblastic (“proto-myofibroblastic”) phenotype and the lineage-plasticity of these neoplasms, highlighting the potential role of cathepsin-k in myofibroblastic trans-differentiation. Cathepsin-k proved to be an additional immunoistochemical marker potentially useful in the diagnostic algorithm.
       
  • Effect of 1,25(OH)2-vitamin D3 on expression and phosphorylation of
           progesterone receptor in cultured endometrial stromal cells of patients
           with repeated implantation failure
    • Abstract: Publication date: Available online 24 December 2019Source: Acta HistochemicaAuthor(s): Hossein Hosseinirad, Marefat Ghaffari Novin, Sedighe Hosseini, Hamid Nazarian, Fardin Amidi, Shahrokh Paktinat, Elham Azizi, Zahra Shams MofaraheAbstractRepeated implantation failure (RIF) occurs in a condition when good quality embryos fail to implant in the endometrium following several in vitro fertilization (IVF) cycles. Suboptimal endometrial receptivity is one of the main underlying factors that causes this failure. Progesterone is the key regulator of endometrial receptivity which regulates gene expression through binding to its receptors in the endometrial stromal cells (eSC). The aim of this study was to evaluate the effect of 1,25(OH)2-vitamin D3 on progesterone receptor (PR) expression level and its phosphorylation on Ser294 residues in eSC of RIF patients and healthy fertile women. After isolation of the eSC from biopsy samples of RIF patients and healthy fertile women and their characterization, the cells were incubated with vitamin D3 and the expression level of PR mRNA, PR protein and phospho-Ser294 PR protein were evaluated after treatment. The results showed that vitamin D3 treatment increases PR mRNA and protein level and phospho-Ser294 PR protein level in the isolated eSC of both RIF patients and the control group. These results suggest that vitamin D3 may possibly play a key role during the embryo implantation process by affecting the expression pattern and regulatory modifications of the PR in the eSC.
       
  • Prognostic value of connective tissue growth factor and c-Myb expression
           in IgA nephropathy and Henoch-Schönlein purpura—A pilot
           immunohistochemical study
    • Abstract: Publication date: Available online 20 December 2019Source: Acta HistochemicaAuthor(s): Maja Mizdrak, Natalija Filipović, Katarina Vukojević, Vesna Čapkun, Ivan Mizdrak, Merica Glavina DurdovAbstractAimAdverse and advanced prognostic signs in IgA nephropathy (IgAN) are interstitial fibrosis and tubular atrophy, but early predictors of bad outcome are still lacking. We investigated expression of connective tissue growth factor (CTGF) and c-Myb in renal biopsies of IgAN and Henoch-Schönlein purpura (HSP), because these gene products are indirectly included in fibrosis and epithelial-mesenchymal transition (EMT).MethodsThe sample included 23 patients and 8 controls who underwent nephrectomy due to renal cancer. The slides cut from the paraffin blocks were prepared for standard indirect immunoflourescence, using antibodies to CTGF and c-Myb. Ten high-power non-overlapping fields were photographed on Olympus IX51 microscope. Average percent of positive tubular cells, as well as number of positive cells per glomerulus were calculated.ResultsThe cytoplasmic tubular CTGF expression was higher in IgAN/HSP than in controls (P 
       
  • High mobility group box 1 antibody represses autophagy and alleviates
           hippocampus damage in pilocarpine-induced mouse epilepsy model
    • Abstract: Publication date: Available online 20 December 2019Source: Acta HistochemicaAuthor(s): Cui Ying, Liang Ying, Liu Yanxia, Wang Le, Cao LiliAbstractAs a neurological disorder, epilepsy has affected over 65 million people all over the world because of the unforeseeable seizures it might cause. However, in-depth understandings of the pathogenesis of epilepsy and effective treatments for the disease are still lacked. Recent discoveries suggest that autophagy, as an endogenous self-cleansing pathway in mammals, might be involved in the onset of epilepsy. Our study assumes that a non-histone DNA binding protein, high mobility group box-1 (HMGB1), formerly considered as a crucial inflammatory factor, may mediate the autophagy of neurons in epileptic mouse brain. To verify this hypothesis, pilocarpine induced epilepsy mouse model was constructed. The mice were treated with HMGB1 antibody for 4 weeks after the initial epileptic seizure. Behavioral test results suggested a recovery of learning ability and memory in epileptic mice when treated with HMGB1 antibody. Pathological changes in hippocampus were inspected under microscopes and hippocampus damages caused by seizures in mouse with epilepsy such as increased intracellular space were alleviated by HMGB1 antibody treatment. Moreover, the expressions of the proteins involved in autophagy pathways were detected by immunofluorescence staining and western blot. microtubule-associated protein 1A/1B-light chain 3 (LC3), Beclin 1, autophagy protein-5 (ATG5), and ATG7 levels were significantly decreased by HMGB1 antibody while the level of p62 was increased. TdT-mediated dUTP Nick-End Labeling (TUNEL) illustrated that cell apoptosis induced by seizures in hippocampus was mitigated by HMGB1 antibody. In conclusion, we propose that HMGB1 may induce increased autophagy in epilepsy mouse model.
       
  • Differential mucins secretion by intestinal mucous cells of Chelon ramada
           in response to an enteric helminth Neoechinorhynchus agilis
           (Acanthocephala)
    • Abstract: Publication date: Available online 18 December 2019Source: Acta HistochemicaAuthor(s): Giampaolo Bosi, Joseph A. DePasquale, Emanuele Rossetti, Bahram Sayyaf DezfuliAbstractIntestinal mucous cells produce and secrete mucins which hydrate, lubricate and protect the intestinal epithelium from mechanical injuries due to the transition of digesta or action of pathogens. Intestinal mucous cells are considered elements of the innate immune system as they secrete lectins, toxins, immunoglobulins, and anti-microbial peptides. Acid mucins can surround and eliminate many pathogenic microorganisms. We performed a quantitative analysis of the density and mucus composition of different intestinal mucous cell types from mullet (Chelon ramada) that were infected solely with Neoechinorhynchus agilis. Most N. agilis were encountered in the middle region of the intestine. Mucous cell types were identified with Alcian Blue (pH2.5) and Periodic acid-Schiff (PAS) histochemistry, and by staining with a panel of seven lectins. Mucus enriched for high viscosity acid mucins was accumulated near points of worm attachment. Parasites were surrounded by an adherent mucus layer or blanket. Ultrastructural examination showed intestinal mucous cells typically possessed an elongated, basally positioned nucleus and numerous electron dense and lucent vesicles in the cytoplasm. The results show both an increase in mucus production and changes in mucin composition in infected mullet in comparison with uninfected conspecifics.
       
  • The protective effect of curcumin on the diabetic rat kidney: A
           stereological, electron microscopic and immunohistochemical study
    • Abstract: Publication date: Available online 18 December 2019Source: Acta HistochemicaAuthor(s): Mahmut Ulubay, Işınsu Alkan, Kıymet Kübra Yurt, Süleyman KaplanAbstractThe prevalence of diabetes in the world is increasing rapidly. Kidney diseases are among the most common medical disorders. The aim of this study was to investigate the effect of curcumin on the diabetic kidney. Thirty-five female Wistar albino rats were divided into seven groups. No procedure was performed on the Cont group. The Sham group received corn oil via gavage for 14 days. The curcumin (Curc) group received 30-mg/kg curcumin for 14 days, while the diabetes mellitus (DM) group received 50-mg/kg streptozotocin (STZ) in a single dose intraperitoneally. The DM + curcumin 1 (DC1) group received 30 mg/kg curcumin for 14 days, seven days after the application of STZ, while the DM + curcumin 2 (DC2) received 30 mg/kg curcumin for 14 days, 21 days after the application of STZ, and the DM + curcumin 3 (DC3) group received single-dose STZ at the same time as the application of 30 mg/kg curcumin for 14 days. Medulla, cortex, tubule, and glomerulus volume ratios were calculated using stereological techniques. Cortex volumes in the Sham and DM groups were significantly lower than in the Cont group (p 
       
  • Diaphragm muscle structure in the elderly: Findings from an autopsy study
    • Abstract: Publication date: Available online 14 December 2019Source: Acta HistochemicaAuthor(s): Ricardo Aparecido Baptista Nucci, Romeu Rodrigues de Souza, Claudia Kimie Suemoto, Alexandre Leopold Busse, Laura Beatriz Mesiano Maifrino, Carlos Alberto Anaruma, Carlos Augusto Pasqualucci, Wilson Jacob-FilhoAbstractSarcopenia is characterized as aging-related weakness and atrophy, which decreases the diaphragm force generating capacity leading to a decrease of expulsive non-ventilatory motor behaviors, which are critical for airway clearance. Thus, this study aimed to analyze the histopathology of the diaphragm in postmortem samples. Thirty individuals were included. Diaphragm samples were stained with hematoxylin and eosin for histopathological analysis. Picrosirius stain was used to highlight the collagen fibers. We observed a positive association between advancing age and histopathological findings in the diaphragm structure. We suggest that age is a key factor in increasing diaphragm muscle histopathology. However, further clinicopathological studies are needed to confirm our findings.
       
  • The effects of magnesium sulfate on cyclophosphamide-induced ovarian
           damage: Folliculogenesis
    • Abstract: Publication date: Available online 4 December 2019Source: Acta HistochemicaAuthor(s): Tuğba Ekiz Yılmaz, Müge Taşdemir, Mehmet Kaya, Nadir Arıcan, Bülent AhıshalıAbstractCyclophosphamide (CYP) is one of the alkylating chemotherapeutic agents and its adverse effects on folliculogenesis in the ovary are well-known due to the previous scientific research on this topic. Magnesium has various effects in organisms, including catalytic functions on the activation and inhibition of many enzymes, and regulatory functions on cell proliferation, cell cycle, and differentiation. In this study, the effects of magnesium sulfate (MgSO4) on CYP induced ovarian damage were investigated. Immature Wistar-Albino female rats of 28-days were treated with pregnant mare serum gonadotrophin (PMSG) to develop the first generation of preovulatory follicles. Rats of the experimental groups were then treated with either CYP (100 mg/kg, i.p) and MgSO4 (270 mg/kg loading dose; 27 mg/kg maintenance doseX12, i.p) solely or in combination. Following in-vivo 5-bromo-2-deoxyuridine (BrdU) labeling, animals were sacrificed and ovaries were embedded in paraffin and Epon. In the ovaries, added to the evaluation of general morphology and follicle count; BrdU and TUNEL-labeling, cleaved caspase-3 and p27 (cyclin-dependent kinase inhibitor) staining was also performed immunohistochemically and an ultrastructural evaluation was performed by transmission electron microscopy (TEM). The number of primordial follicles were decreased and multilaminar primary and atretic follicles were increased in CYP group. After MgSO4 treatment, while primordial follicle pool were elevated, the number of atretic follicles were decreased. Additionally, decreased BrdU-labeling, increased cleaved caspase 3 immunoreactivity and increased TUNEL labeling were observed in CYP group. In CYP treated animals, observations showed that while MgSO4 administration caused no alterations in BrdU proliferation index and caspase-3 immunoreactivity, it significantly reduced the TUNEL labeling. It was also observed that, while p27 immunoreactivity significantly increased in the nuclei of granulosa and theca cells in the CYP group; MgSO4 treatment significantly reduced these immunoreactivities. The ultrastructural observations showed frequent apoptotic profiles in granulosa and theca cells in both early and advanced stages of follicles in the CYP group and the MgSO4 treatment before the CYP application led to ultrastructural alleviation of the apoptotic process. In conclusion, our data suggest that MgSO4 may provide an option of pharmacologic treatment for fertility preservation owing to the beneficial effects of on chemotherapy-induced accelerated follicular apoptotic process, and the protection of the primordial follicle pool.
       
  • Involvement of proliferative and apoptotic factors in the development of
           hindgut in rat fetuses with ethylenethiourea-induced anorectal
           malformations
    • Abstract: Publication date: Available online 28 November 2019Source: Acta HistochemicaAuthor(s): Caiyun Long, Yunxia Xiao, Siying Li, Xiaobing Tang, Zhengwei Yuan, Yuzuo BaiAbstractBackgroundAnorectal malformations (ARMs) are common congenital malformations of the terminal digestive tract, but little is known regarding their pathogenesis. Aberrant cell proliferation/apoptosis are believed to be involved in ARMs. However, there are no studies on proliferation/apoptosis-related genes.PurposeWe aimed to investigate the spatiotemporal expression patterns of two proliferation/apoptosis-related genes (MYC proto-oncogene and tumor protein p53) and explore their potential functions in the hindguts of ethylene thiourea-induced ARMs rat fetuses.MethodsMYC and p53 expression was evaluated using immunohistochemical staining, western blotting, and quantitative real-time polymerase chain reaction (RT-qPCR). Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and p53 costaining were performed to assay the colocalization of apoptotic and p53-expressing cells.ResultsRat fetuses with ARMs displayed fusion failure of the urogenital septum and cloacal membrane. In the control group, MYC was persistently expressed from gestational day (GD)14 to GD16 and distributed throughout the hindgut, while p53 was weakly detected in the terminal segment of the urethra and hindgut; in the ARMs group, MYC expression was obviously reduced, while p53 was widely and highly expressed in the urethra and hindgut. Western blotting and RT-qPCR confirmed the decrease in MYC and increase in p53 expression in ARMs. TUNEL and p53 co-staining revealed considerable overlap between apoptotic and p53-expressing cells.ConclusionThe expression patterns of c-myc and p53 were disrupted in ARMs rat embryos, and the downregulation of c-myc and upregulation of p53 might be related to the development of ARMs at the key time points of ARMs morphogenesis.
       
  • Expression of Langerin/CD207 in airways, lung and associated lymph nodes
           of a stranded striped dolphin (Stenella coeruleoalba)
    • Abstract: Publication date: Available online 27 November 2019Source: Acta HistochemicaAuthor(s): E.R. Lauriano, S. Pergolizzi, P. Lo Cascio, M. Kuciel, N. Zizzo, M.C. Guerrera, M. Aragona, G CapilloAbstractThe airways and lungs of vertebrates are an entrance way for several microbial pathogens. Cetaceans present an upper and lower respiratory anatomy that allows the rapid flow of large air volumes, which may lead to high susceptibility to respiratory infections. Mortality and stranding rate of Cetaceans increased dramatically, so wide the knowledge about the immune system and specific antibodies identifying immune cells populations, is of fundamental importance to monitor and document cetacean health.The aim of this study was to identify the localization of dendritic cells marked by Langerin/CD207 in airways, lungs and associated lymph nodes, of the striped dolphin Stenella coeruleoalba. Samples of trachea, bronchi, lungs and lung-associated lymph nodes were obtained from a stranded adult male of Stenella coeruleoalba. Our results showed abundant lymphoid aggregates (LAs) in the lung of S. ceruleoalba. Langerhans-like dendritic cells were well distributed along the epithelium and interstitium of respiratory tract and in associated lymph nodes.The present study deepens the knowledge about the cetacean’s immune system and report about the exploitability of a commercial antibody (Langerin/CD207) for cetacean species.
       
  • Inferring lanosterol functions in the female rabbit reproductive tract
           based on the immunolocalization of lanosterol 14-demethylase and farnesoid
           beta-receptor
    • Abstract: Publication date: Available online 26 November 2019Source: Acta HistochemicaAuthor(s): Maribel Méndez-Tepepa, Dafne Zepeda-Pérez, Leticia Nicolás-Toledo, Lourdes Arteaga-Casteñeda, Gabriel Gutiérrez-Ospina, Estela Cuevas-RomeroAbstractFemale reproductive organs have de novo synthesis of cholesterol. Some sterol molecules, intermediaries in the cholesterol synthesis, have important paracrine/autocrine actions. Lanosterol binds to the farnesoid beta-receptor (FXRβ), a molecule widely expressed in the ovaries, suggesting that it may play a role in reproduction. Up to date, we know little about lanosterol functions across female reproductive organs. We described immunolocalized lanosterol 14-demethylase (LDM or CYP51A1), responsible for catalyzing the conversion of lanosterol in cholesterol, and FXRβ in the ovary, oviduct, uterus, and vagina of virgin and pregnant rabbits. In virgin rats, we found CYP51A1 and FXRβ immunoreactivity was found in all ovarian follicles, epithelial cells, stroma, and Graafian follicles. Also, the epithelium and stroma, as well as the smooth muscle of the oviduct, vagina, and uterus showed CYP51A1 and FXRβ immunoreactivity. In pregnant dams, we observed the presence of CYP51A1 and FXRβ immunoreactivity in the corpora lutea, giant uterine cells, and trophoblastic cells. The presence of CYP51A1 and FXRβ support that lanosterol participates in diverse reproductive processes, including follicular maturation, transport of gametes and zygote, implantation of blastocyst, lubrication, and contraction of the vagina, secretion of female prostate, and control of delivery mediated by pelvic muscles contraction.
       
  • Vesicular glutamate transporter 2-immunoreactive afferent nerve terminals
           in rat carotid sinus baroreceptors
    • Abstract: Publication date: Available online 26 November 2019Source: Acta HistochemicaAuthor(s): Takuya Yokoyama, Kazuya Settai, Nobuaki Nakamuta, Yoshio YamamotoAbstractSensory nerve endings respond to various stimuli and subsequently transmit afferent informations to central nervous system, but their responsibility has been suggested to be modulated by glutamate. In the present study, we examined the immunohistochemical localization of vesicular glutamate transporter 1 (vGLUT1) and vGLUT2 in baroreceptor nerve endings immunoreactive for P2X2 and P2X3 purinoceptors in the rat carotid sinus by immunohistochemistry of whole-mount preparations with confocal scanning laser microscopy. P2X3-immunoreactive flat leaf-like axon terminals were immunoreactive to vGLUT2, but not to vGLUT1. Among members of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex, immunoreactivities for synaptosomal-associated protein, 25 kDa, Syntaxin1, and vesicle-associated membrane protein 2 (VAMP2) were localized in P2X2- and P2X3-immunoreactive axon terminals. Punctate immunoreactive products for VAMP2 and vGLUT2 were co-localized in axon terminals. These results suggest that vGLUT2 is localized in P2X3-immunoreactive baroreceptor terminals in the carotid sinus, and these terminals may release glutamate by exocytosis in order to modulate baroreceptor function in the carotid sinus.
       
  • Fibroblast growth factor-2 signaling modulates matrix reorganization and
           cell cycle turnover rate in the regenerating tail of Hemidactylus
           flaviviridis
    • Abstract: Publication date: Available online 26 November 2019Source: Acta HistochemicaAuthor(s): Anusree Pillai, Sonam Patel, Isha Ranadive, Isha Desai, Suresh BalakrishnanAbstractLizards restore their lost tail by the recruitment of multipotent cells which are selectively differentiated into varied cell types so as to sculpt a new tail. The precise coordination of the events involved in this complex process requires crosstalk between many signaling molecules and differential regulation of several mediators that facilitate the achievements of various milestones of regeneration. Fibroblast growth factor-2 is one such signaling molecule which activates a number of intracellular signaling pathways. Herein, the regulatory role of FGF2 during tail regeneration in Hemidactylus flaviviridis was investigated. Upon inhibition of FGFR using SU5402, the FGF2 levels were found to be significantly reduced at both transcript and protein level. Further, the compromised levels of the gelatinases, namely MMP2 and MMP9 in the tail tissues of treated lizards indicate that FGF2 regulates the activity of these enzymes perhaps to facilitate the recruitment of multipotent mesenchymal cells (blastema). The in vivo 5BrdU incorporation assay showed a lower cell proliferation rate in FGF2 signal inhibited animals during all the proliferative stages of regeneration studied. This observation was substantiated by decreased levels of PCNA in treated group. Moreover, from the combined results of Caspase-3 localization and its expression levels in the regenerates of control and SU5402 treated lizards it can be deduced that FGF2 signal regulates apoptosis as well during early stages of regeneration. Overall, the current study indicates beyond doubt that FGF2 signaling plays a pivotal role in orchestrating the matrix reorganization and cell cycle turnover during lizard tail regeneration.
       
  • Biochemical, immunohistochemical and morphometrical investigation of the
           effect of thymoquinone on the rat testis following exposure to a 900-MHz
           electromagnetic field
    • Abstract: Publication date: Available online 26 November 2019Source: Acta HistochemicaAuthor(s): Ahmad Yahyazadeh, Berrin Zuhal Altunkaynak, Suleyman KaplanLong-term use of cell phones emitting electromagnetic fields (EMFs) have raised concerns regarding public health in recent year. We aimed to investigate the possible effects of 900 MHz EMF exposure (60 min/day for 28 days) on the rat testis. Another objective was to determine whether the deleterious effect of EMF radiation would be reduced by the administration of thymoquinone (TQ) (10 mg/kg/day). Twenty-four male adult Wistar albino rats were randomly selected, then assigned into four groups as followControl, EMF, TQ and EMF + TQ. Testicular samples were analyzed using histological, stereological, biochemical and immunohistochemical techniques. Total numbers of primary spermatocytes and spermatids as well as Leydig cells were significantly decreased in the EMF group compared to the Control group (p 
       
  • Effects of uterus derived mesenchymal stem cells and their exosomes on
           asherman’s syndrome
    • Abstract: Publication date: Available online 24 November 2019Source: Acta HistochemicaAuthor(s): Gulistan Sanem Saribas, Candan Ozogul, Meral Tiryaki, Ferda Alpaslan Pinarli, Sevtap Hamdemir KilicAbstractAsherman’s syndrome has become a growing problem with the incidence of cesarean and endometrial surgical procedures. A surgical procedure that can damage to the basal layer of the endometrium is formed as intrauterine adhesion and can cause asherman’s syndrome. Mesenchymal stem cells (MSCs) are characterized by some characteristics such as non-immunogenic, angiogenic, antifibrotic, antiapoptotic and antiinflammatory properties, also they support tissue repair by secretion of various factors and chemokines in cellular therapy. Exosomes are active paracrine components with a great potential for repairing damaged tissue. Exosomes include many paracrine factors responsible for regeneration and angiogenesis. In this study, 10 newborn Wistar rats were used to obtain MSCs. A total of 24 adult Wistar rats were also used. The rats were divided into 4 groups: untreated control group; asherman control group; asherman + uterine-derived MSCs group; asherman + uterine-derived MSCs-exosomes group. At the end of the experiment, uterine tissues were evaluated by histochemical and immunohistochemical. As a result of MSCs and exosomes treatments, proliferation and vascularization in uterine tissue was increased. It was also shown to reduce fibrosis with masson’s trichrome staining. MMP-2 and MMP-9 expression was enhanced by MSC and exosomal therapy; in addition, TIMP-2 expression was decreased. In our study, it was shown that proliferation and vascularization increased and fibrosis decreased in uterus as a result of MSC and exosome treatments. Our results indicate that the exosomal treatment restored the damage of asherman’s syndrome at tissue at a shorter time than the MSCs group.
       
  • Pd-ligand 1 is expressed in inflammatory cells but not in neoplastic cells
           in hepatocellular carcinoma: An immunohistochemical study
    • Abstract: Publication date: Available online 22 November 2019Source: Acta HistochemicaAuthor(s): Francesco Vasuri, Azzurra Nerpiti, Stefano Zagnoni, Matteo Ravaioli, Antonia D’Errico, Michelangelo FiorentinoAbstractNowadays, the major limit to the immunohistochemical (IHC) analysis of tissue PD-L1 is the high variability of the monoclonal antibodies commercially available. Aims of the present paper are to assess the best clone and the most suitable scoring for PD-L1 IHC determination on human hepatocellular carcinoma (HCC) among three commercially available clones, and to evaluate which PD-L1 clone is the best in predicting HCC aggressiveness in vivo. We built a tissue microarray (TMA) with 60 retrospective HCC cases, including the correspondent non-tumoral tissue. IHC was automatically performed using the following anti-PD-L1 clones: 28.8, SP142, and SP263.As results, we did not find any immunoreactivity for PD-L1 in both neoplastic and normal hepatocytes included in the TMA using the three antibodies. Positivity for PD-L1 was exclusively seen in inflammatory cells within the HCC tissue and in cirrhotic parenchyma. When a gold standard was assessed, the sensitivity of SP142, 28.8 and SP263 was 46 %, 54 % and 85 % respectively. Using the SP263 clone, the absolute number of PD-L1-positive inflammatory cells in the HCC cores was paired with the number of PD-L1-positive inflammatory cells in the corresponding non-tumoral tissue (P = 0.001). Finally, using SP263, the mean number of PD-L1-positive cells was 11.3 ± 12.6 in HCC from deceased patients, versus 4.7 ± 5.2 in alive patients (p = 0.039). SP263 is the most sensitive clone for PD-L1 IHC tissue determination in HCC, as well as the best antibody for the assessment of its biological behavior.
       
  • Clenbuterol accelerates recovery after immobilization-induced atrophy of
           rat hindlimb muscle
    • Abstract: Publication date: Available online 21 November 2019Source: Acta HistochemicaAuthor(s): Hideki Suzuki, Yuki Yoshikawa, Hisaya Tsujimoto, Takashi Kitaura, Isao MuraokaAbstractUsing immunohistochemistry, we investigated whether daily administration of clenbuterol (CLE; 1 mg/kg body weight per day) accelerates recovery after casted immobilization(IMM)-induced atrophy of fast-twitch plantaris and slow-twitch soleus muscles. Adult male Sprague-Dawley rats were assigned to the control (CON), casted immobilization (IMM), casted immobilization following recovery control (RCON), and casted immobilization following recovery with CLE administration (RCLE) groups. Casted immobilization and recovery periods were 9 and 14days, respectively. Rats of the CON group were subjected to the experiment simultaneously with the IMM group.Nine days of immobilization induced muscle fiber atrophy, which was greater in the soleus muscle than in the plantaris muscle. After the 2-week recovery period, the cross-sectional areas of each fiber type in both muscles were higher in the RCON group than in the IMM group. The cross-sectional areas of each fiber type in both muscles in the RCLE group were larger than those in the RCON group. The myonuclear number of each fiber type of the plantaris muscle in the RCON and RCLE groups was higher than that in the CON group. In contrast, the myonuclear number per fiber of the soleus muscle was not affected by hindlimb immobilization, reloading, and clenbuterol administration regardless of muscle fiber type.These results suggest that CLE accelerates the recovery of atrophied plantaris and soleus muscles fibers and that their mechanisms of responses to CLE in both muscles may be different during recovery period after muscle atrophy.
       
  • Comparative analysis of fresh chondrocytes, cultured chondrocytes and
           chondroprogenitors derived from human articular cartilage
    • Abstract: Publication date: Available online 13 November 2019Source: Acta HistochemicaAuthor(s): Elizabeth Vinod, Upasana Kachroo, Soosai Manickam Amirtham, Boopalan Ramasamy, Solomon SathishkumarIntroductionInterest in chondroprogenitors arose due to their inherent stem cell like properties, and their initial characterization was based on identification of a small percentage of CD49e positive cells in cultured chondrocytes (CC). It was further noted that when fresh chondrocytes (FC; reported to express low CD49e) were subjected to fibronectin adhesion assay, an isolate of chondroprogenitors was obtained, which was highly positive for CD49e, thus making it a distinguishing marker for this cell population. However, this notion was challenged when reports demonstrated high CD49e expression in CC as well. Therefore, our aim was to compare CD49e expression in FC, CC and chondroprogenitors.MethodsChondrocytes and chondroprogenitors were isolated from articular cartilage of osteoarthritic joints from three patients. Assessment of classic fibronectin receptor (CD49e, CD29), positive (CD105, CD73, CD90) and negative (CD45, CD34) mesenchymal stem cell marker expression in all groups was performed, as chondroprogenitors fulfill the minimal criteria laid down by International Society for Cellular Therapy. Following this, adipogenic, osteogenic and chondrogenic differentiation was assessed by Oil red O, Alizarin Red and Alcian Blue staining respectively.Results and conclusionOur observations indicate that FC show significantly low surface marker expression as compared to CC and chondroprogenitors, whereas no significant difference was seen in values when CC and chondroprogenitors were compared. Moreover, comparable results were exhibited when trilineage differentiation potential was compared across groups. Since CC and chondroprogenitors show similar characteristics, there is a pressing need for a specific differentiating marker to isolate a pure population of chondroprogenitors.Graphical abstractGraphical abstract for this article
       
  • In vitro effects of PNP and PNMC on apoptosis and proliferation in the hen
           ovarian stroma and prehierarchal follicles
    • Abstract: Publication date: Available online 8 November 2019Source: Acta HistochemicaAuthor(s): A.K. Grzegorzewska, A. Hrabia, K. Kowalik, D. Katarzyńska-Banasik, A. Kozubek, A. SechmanAbstractThis study aimed to examine the mRNA expression, activity, and immunolocalisation of apoptosis/proliferation regulating factors following in vitro exposure of the stroma, white (WFs), and yellowish (YFs) follicles of the chicken ovary to 4-nitrophenol (PNP) or 3-methyl-4-nitrophenol (PNMC). PNMC increased the mRNA expression of caspase-3, -8, Apaf-1, and cytochrome c in the ovarian stroma. The activity of caspase-3, -8, and -9 decreased in WFs in both nitrophenol-treated groups. PNP reduced the number of caspase-3-positive cells in the stromal connective tissue (CT) and the theca interna and externa layers of WFs. In the stroma, the proliferating index decreased in the wall of primary follicles in both nitrophenol-treated groups, however, in the CT, the effect of PNMC was opposite. In the theca interna of WFs, PNP diminished the proliferating index. These results suggest that nitrophenols might impact the development of chicken ovarian follicles by affecting cell death and proliferation.
       
  • Presence of serotonin and its receptor in the central nervous system and
           ovary and molecular cloning of the novel crab serotonin receptor of the
           blue swimming crab, Portunus pelagicus
    • Abstract: Publication date: Available online 7 November 2019Source: Acta HistochemicaAuthor(s): Jirasuda Nakeim, Napamanee Kornthong, Jirawat Saetan, Supawadee Duangprom, Prasert Sobhon, Prapee SretarugsaAbstractSerotonin (5-HT) plays pivotal roles in many physiological processes including reproduction of crustaceans, which are mediated 5-HT receptors. The distributions of 5-HT and its receptor have never been explored in Portunus pelagicus. To validate the targets which indirectly indicate the roles of 5-HT in this crab, we have investigated the distribution of 5-HT in the central nervous system (CNS) and ovary using immunohistochemistry and tissue expression of its receptor by RT-PCR. In the brain, 5-HT immunoreactivity (-ir) was detected in clusters 6, 7, 8, 11, 14, 15 and the fibers. In the ventral nerve cord (VNC), 5-HT-ir was detected in pairs of neurons and the fibers connected to the neurons. In the ovary, 5-HT-ir was intense in the oocyte step 1 (Oc1) and Oc2, and its intensity was slightly decreased in Oc3 and Oc4. The 5-HT receptor was molecularly characterized to be type 7, and it was strongly expressed in the eyestalk, brain, VNC, mature ovary and muscle. Due to the presence of 5-HT receptor we suggest that 5-HT acts primarily at the CNS and ovary, thus implicating its role in reproduction especially in the development of oocytes though its exact function in this crab needed to be explored further.
       
  • Prophylactic supplementation of 20-HETE ameliorates hypoxia/reoxygenation
           injury in pulmonary vascular endothelial cells by inhibiting apoptosis
    • Abstract: Publication date: Available online 6 November 2019Source: Acta HistochemicaAuthor(s): Praveenkumar Sugumaran, Vishnusekar Narayanan, Daling Zhu, Meetha Medhora, Elizabeth R. Jacobs, Yamini Chandramohan, Vimalraj Selvaraj, Anuradha DhanasekaranAbstractHypoxia reoxygenation (HR) injury perturbs structural and functional syncytium in lung tissues. It is commonly implicated in conditions such as stroke, lung transplant or severe pneumonia. In the present study, we investigated the cytoprotective action of 20-hydroxyeicosatetraenoic acid (20-HETE) on pulmonary vascular endothelial cells (PMVECs) under normoxic and hypoxic niche followed by HR. 20-HETE pretreatment showed a protective effect at a concentration of 1μM as there was a marked increase (20%) in the cell viability compared to control and HR groups. Pretreatment of 20-HETE in HR induced injury decreased ROS production dictated its antioxidant property. Similarly, SOD and ATP levels were also downregulated by 20-HETE pretreatment. Cell apoptosis was detected by TUNEL assay, Acridine orange, and procaspase-3 cleavage, caspase-3 activity assay, respectively. JC-1 mitochondrial membrane potential assay and protein expression pattern of BCL-2, and BAD phosphorylation status were examined. The results showed that HR induced significant increase of apoptotic PMVECs, while 20-HETE pretreatment attenuated the effects. Further, 20-HETE pretreatment activated PI3K/Akt and HIF-1α signaling pathway to exhibit its protective effects against HR-induced oxidative stress and apoptosis. Overall, the results concluded the potent antioxidant role of 20-HETE in aiding cytoprotection upon HR injury.
       
  • Presence of metalloproteinases 2 and 9 and 8-OHdG in the fibrotic process
           in skeletal muscle of Mdx mice
    • Abstract: Publication date: Available online 4 November 2019Source: Acta HistochemicaAuthor(s): Lidiane Begalli de Souza, Carla Maziero, Mariana Cruz Lazzarin, Hananiah Tardivo Quintana, Tabata de Carvalho Tomé, Vivianne Izabelle de Araújo Baptista, Flavia de OliveiraAbstractInflammation and oxidative stress occurs in muscle of Duchenne muscular dystrophy (DMD). The relationship between a panel of biomarkers and the DMD outcome is necessary to indicate of disease progression and response to rehabilitation programs. The aim was to analyze the connective tissue of muscle of Mdx mice and immunoexpression of MMP-2, MMP-9, and 8-OHdG, which signalizes oxidative stress related to DNA damage. Biceps brachii of male C57BL/10 and C57BL/10-Dmdmdx mice was submitted to Hematoxylin-Eosin, Sirius red and immunohistochemistry (MMP-2, MMP-9 and 8-OHdG) analysis. Mdx showed focal lesions with intense inflammation and fibrosis related to immunoexpression of MMP-2 and MMP-9, proving the hypothesis that these MMPs are linked to muscular tissue degeneration, which can be regenerated by their inhibition, improving the treatment of DMD carriers. Histopathological findings related to centralized nuclei increase were related to higher 8-OHdG immunomarked nuclei in Mdx, which signalizes oxidative stress associated with DNA damage provoked by DMD. Such result shows that the evaluation of 8-OHdG during the evolution of the disease could be a method to evaluate DMD disease progression.
       
  • Changes in snail and SRF expression in the kidneys of diabetic rats during
           ageing
    • Abstract: Publication date: Available online 24 October 2019Source: Acta HistochemicaAuthor(s): Sandra Kostic, Brandon Williams, Samy Ksouri, Leon Hardung, Natalija Filipovic, Lejla Ferhatovic Hamzic, Livia Puljak, Nasrollah Ghahramani, Katarina VukojevicAbstractBackgroundDiabetic nephropathy is a progressive condition which develops for many years. We analyzed expression of Snail and serum response factor (SRF), epithelial-mesenchymal transition (EMT) regulatory transcription factors with a key role in renal fibrosis, in different renal areas of diabetic rats during ageing.MethodsMale Sprague–Dawley rats were treated with 55 mg/kg streptozotocin (model of type 1 diabetes mellitus; DM group) or citrate buffer (control). DM group received insulin weekly to prevent ketoacidosis. After 2 weeks, 2, 6 and 12 months kidney samples were collected and analysed in different renal areas.ResultsSnail expression was located within cortex in proximal convoluted tubules, in control and DM groups, in the cytoplasm. Percentage of Snail-positive cells in control groups was high and decreased with time, whereas in DM groups the highest percentage was after 2 weeks. In all time points, smaller percentage of Snail expression was seen in DM groups compared to controls.SRF expression was mostly located in the proximal convoluted tubules, always in the cytoplasm. In control groups SRF was expressed in all time periods in proximal convoluted tubules, with decrement after 12 months. Percentage of SRF-positive cells was higher in control groups compared to DM in all time points, with the exception of 12 months. To a smaller degree, SRF expression was seen in the glomeruli and distal convoluted tubules, with more SRF positive cells in DM compared to their control groups.ConclusionsWhile Snail expression remained lower in diabetic tissues, compared to controls, expression of SRF increased in diabetic tissues in the second part of the year. These changes may need long time to develop, and, in line with earlier reports, it is possible that insulin treatment of DM rats once a week reduces possibility of EMT and development of renal fibrosis even in the long term.
       
  • Comparative expression of cell adhesion molecule1 (CADM1) in the testes of
           experimental mice and some farm animals
    • Abstract: Publication date: Available online 18 October 2019Source: Acta HistochemicaAuthor(s): Mahmoud S. Gewaily, Mohamed Kassab, Foad A. Farrag, Essam A. Almadaly, Mustafa S. Atta, Ahmed Abd-Elmaksoud, Tomohiko WakayamaAbstractCell adhesion molecule1 (CADM1) is a member of the immunoglobulin superfamily (IGSF) that has been found in mammalian testis and plays a substantial role in cell-to-cell interaction via either hemophilic (between spermatogenic cells) or heterophilic (between spermatogenic and somatic Sertoli cells) binding. The present study investigated the immunohistochemical localization of CADM1 in the testes of adult mice (Mus musculus), as well as sexually mature bull (Bos taurus), camel (Camelus dromedarius), and donkey (Equus asinus), using immunohistochemical techniques. The results revealed that CADM1 expression was observed in the spermatogonia and early spermatocytes as well as elongated spermatids in the mice testes; however, in the bull testis, its expression was restricted to the elongated spermatids. This expression was found in some of the early spermatocytes and elongated spermatids of the rutting camel testis but only found in the elongated spermatids of the non-rutting camel testis. Interestingly, CADM1 expression was detected in the spermatogonia, early spermatocytes, and elongated spermatids of the donkey testis. On the other hand, there was no expression of CADM1 observed in the Sertoli or interstitial cells. In conclusion, the expression of CADM1 during spermatogenesis differed among species and between rutting and non-rutting camel. Accordingly, this study emphasized the crucial role of CADM1 in the process of spermatogenesis and how it is related to sexual activity in both experimental and farm animals.
       
  • Chronological changes in rat heel skin following depressurization of
           pressure ulcer-like dermal lesions
    • Abstract: Publication date: Available online 16 October 2019Source: Acta HistochemicaAuthor(s): Daijiro Haba, Takamitsu ArakawaAbstractIn our previous study, we proposed an animal model in which pressure ulcer-like dermal lesions can be produced by denervation of the sciatic nerve and application of a pressure load to rat heel skin. In the present study, we divided these animals into non-treated and pressure loading groups, and initiated hindlimb unloading (depressurization) by tail suspension at 1, 3, 5, 7, and 14 days after inflicting lesions (1–14d pressurization groups). Chronological changes in heel lesions were examined morphologically in all treatment groups after 1, 3, 7, 14, 28, and 40 days. Open dermal lesions were formed by 14 days in the loading group and numerous macrophages were present. In the 14d pressurization group, numerous macrophages were still distributed in and around lesions and Vascular endothelial cell growth factor (VEGF) expression was strongly detected by 3 days, but a thin germinal layer began to appear and CD68-positive macrophages and VEGF immunoreactions decreased gradually by 7 days later. By 14 days after depressurization, the germinal layer was repaired, and macrophages and immunoreactions of VEGF were similar to those of non-treated skin. These chronological changes were similar to those in human pressure ulcers, but from 5d after depressurization, different chronological changes were observed. Specifically, epidermis was thickened and macrophages were hardly detected at 5 days in the loading group, but the epidermis disappeared by 1 day in the 5d pressurization group. Subsequently, numerous macrophages aggregated and VEGF expression was increased by 3 days, and the remaining healing process was similar to that in the 14d pressurization group. Even when unloading was performed during the early stages (5d pressurization group), the epidermis disappeared and macrophages were then distributed before repair of the lesion was observed. These results suggest that earlier migration of macrophages to skin lesions might be associated with rapid wound healing.
       
 
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