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      Lipocalina associada à gelatinase de neutrófilos (NGAL) e calprotectina no tecido laminar de equinos após obstrução jejunal, tratados ou não com hidrocortisona Translated title: Neutrophil gelatinase associated lipocalin (NGAL) and calprotectin in equine laminar tissue after jejunal obstruction, treated or not with hidrocortisone

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          Abstract

          A laminite é uma doença podal grave que acomete os equídeos, sendo responsável por intenso sofrimento. Neste estudo foram pesquisadas a presença de calprotectina por meio da imunoistoquímica, e de lipocalina associada à gelatinase de neutrófilos (NGAL), por zimografia, no tecido laminar do casco de equinos após obstrução intestinal. Os animais foram divididos em quatro grupos: Grupo controle (Gc), contendo sete animais normais, sem procedimento cirúrgico; Grupo Instrumentado (Gi), contendo cinco animais, os quais passaram por todo o procedimento cirúrgico sem sofrerem obstrução intestinal; Grupo Não Tratado (Gnt), contendo quatro equinos submetidos a obstrução intestinal do jejuno por distensão de balão intraluminal, sem tratamento; e Grupo Tratado (Gt), contendo quatro equinos submetidos a obstrução intestinal, e tratados preventivamente com hidrocortisona. Houve imunomarcação de calprotectina em todos os grupos experimentais, com aumento nos equinos do grupo distendido em relação ao Gc. Com relação ao NGAL, houve aumento também do Gnt e do Gi em relação ao Gc. O Gt não diferiu dos demais. Conclui-se que a distensão do intestino delgado pode promover acúmulos de leucócitos nos cascos de equinos e que o NGAL é um método viável para se detectar infiltração neutrofílica em equinos. Novos estudos deverão ser realizados para se verificar possível benefício anti-inflamatório da hidrocortisona no casco de equinos com obstrução intestinal.

          Translated abstract

          Laminitis is a severe hoof condition in horses that may cause intense suffering. In this study, leukocyte infiltration in hoof laminar tissue was investigated in horses subject to intestinal obstruction using immunohistochemistry to detect calprotectin, and zymography to detect neutrophil gelatinase associated lipocalin (NGAL). There were four groups: the Control Group (Gc), with seven horses, without surgical procedures; the Sham-operated Group (Gi), with five horses that were subjected to surgical procedure without intestinal obstruction; the No Treat group (Gnt), with four horses subjected to intestinal obstruction (jejunal distention using an intraluminal balloon) without treatment; and Treated group (Gt), with four horses subjected to intestinal obstruction and treated with hydrocortisone. Positive calprotectin imunostaining was detected in all experimental groups, with increase cell counts in horses of the distended group compared with the control group. NGAL expression was increased in Gd compared with Gc e Gi. The Gt did not differ from the others. In conclusion, small intestine distension can promote leukocyte infiltration in equine hoof laminar tissue, and NGAL zymography was considered a useful method for leukocyte tissue detection in horses. New studies will be conducted to verify the possible beneficial anti-inflammatory effects of hydrocortisone in hoof of horses with intestinal obstruction.

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          Most cited references68

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          The lipocalin protein family: structure and function.

          The lipocalin protein family is a large group of small extracellular proteins. The family demonstrates great diversity at the sequence level; however, most lipocalins share three characteristic conserved sequence motifs, the kernel lipocalins, while a group of more divergent family members, the outlier lipocalins, share only one. Belying this sequence dissimilarity, lipocalin crystal structures are highly conserved and comprise a single eight-stranded continuously hydrogen-bonded antiparallel beta-barrel, which encloses an internal ligand-binding site. Together with two other families of ligand-binding proteins, the fatty-acid-binding proteins (FABPs) and the avidins, the lipocalins form part of an overall structural superfamily: the calycins. Members of the lipocalin family are characterized by several common molecular-recognition properties: the ability to bind a range of small hydrophobic molecules, binding to specific cell-surface receptors and the formation of complexes with soluble macromolecules. The varied biological functions of the lipocalins are mediated by one or more of these properties. In the past, the lipocalins have been classified as transport proteins; however, it is now clear that the lipocalins exhibit great functional diversity, with roles in retinol transport, invertebrate cryptic coloration, olfaction and pheromone transport, and prostaglandin synthesis. The lipocalins have also been implicated in the regulation of cell homoeostasis and the modulation of the immune response, and, as carrier proteins, to act in the general clearance of endogenous and exogenous compounds.
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            Granules of the human neutrophilic polymorphonuclear leukocyte.

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              Basement membrane pathology: a feature of acute equine laminitis.

              Thirty-two dorsal, mid-hoof wall, lamellar sections from 8 Standardbred horses, humanely killed 48 h after the administration of an alimentary carbohydrate overload, were sectioned and examined by light microscopy. Sections were stained with the connective tissue and basement membrane stains periodic acid-Schiff (PAS), Azan and periodic acid silver methanamine (PASM) and with routine haematoxylin and eosin (H&E). Lesions of the epidermal lamellae, attributable to laminitis, were graded in order of increasing severity from Grade N (normal), Grade 1 (mild), Grade 2 (moderate) to Grade 3 (severe and extensive). The grading system was based principally on changes to lamellar basement membrane (BM) which were clearly visible when the connective tissue stains PAS and PASM were used. Earliest changes were rounding of the basal cell nuclei and elongation of secondary epidermal lamellae (SELs). Secondary epidermal lamellae tips were pointed instead of round and the basement membrane had separated from the lamellae. In early Grade 1 lesions, this was obvious at the tips of the SELs where the BM had lifted to form teat-shaped bubbles. The absence of BM at the tips of secondary dermal lamellae, along with varying amounts of connective tissue, was considered a progression in severity and classified as Grade 2. Eventually, even the primary epidermal and primary dermal lamellae separated from each other and the empty shells of isolated BM, in what was once the tip of the primary epidermal lamella, signified that a global separation of the epidermal and dermal lamellae had occurred (Grade 3 lesion). The histopathological grading system correlated well with the degree of lameness at the time of euthanasia, (r2 = 0.94) and apparently described the severity of laminitis accurately. Disintegration of the BM and failure of its attachment to the basal cells of the epidermis appears to be one of the earliest pathological events to occur in acute laminitis and could be the change that initiates the collapse of the lamellar architecture. Histopathological diagnoses of laminitis are strengthened when based on sections stained with at least PAS, in addition to routine H&E and should exhibit evidence of the BM pathology described here.
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                Author and article information

                Contributors
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Role: ND
                Journal
                pvb
                Pesquisa Veterinária Brasileira
                Pesq. Vet. Bras.
                Colégio Brasileiro de Patologia Animal - CBPA (Rio de Janeiro )
                1678-5150
                September 2012
                : 32
                : 9
                : 817-823
                Affiliations
                [1 ] Estadual Paulista Brasil
                [2 ] Universidade Estadual Paulista Brazil
                [3 ] Universidade Federal de Minas Gerais Brazil
                [4 ] Universidade Estadual de Campinas Brazil
                Article
                S0100-736X2012000900001
                10.1590/S0100-736X2012000900001
                a707501d-4f0c-4e77-b6dc-25cab1f15c67

                http://creativecommons.org/licenses/by/4.0/

                History
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                SciELO Brazil

                Self URI (journal page): http://www.scielo.br/scielo.php?script=sci_serial&pid=0100-736X&lng=en
                Categories
                VETERINARY SCIENCES

                General veterinary medicine
                Horse,laminitis,calprotectin,NGAL,jejunal obstruction,Equinos,laminite,calprotectina,obstrução jejunal

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