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Alterações da formação hipocampal do Calidris pusilla associadas à migração outonal de longa distância
(Universidade Federal do Pará, 2017-08-31) MAGALHÃES, Nara Gyzely de Morais; DINIZ, Cristovam Guerreiro; http://lattes.cnpq.br/1025250990755299; DINIZ, Cristovam Wanderley Picanço; http://lattes.cnpq.br/2014918752636286
After breeding in the upper Arctic tundra, shorebirds affected by migratory restlessness trace an inherited preliminary route and use compasses, maps and visual landmarks, until they reach, in the northern hemisphere, stopover sites that have the necessary nutritional resources for fast and high gain of energy reserves for migratory journey, as in the Bay of Fundy-Canada. Following this stopover site that is used by 75% of the population of Calidris pusilla, the long-distance autumn migratory experience continues with uninterrupted 6-day non-stop flights over the Atlantic until these birds reach South America and then the island of Canela-Brazil. To test the hypothesis that the long-distance migratory process would influence neurogenesis, astrogenesis and activation of earlier-expression genes, we captured 12 individuals in full migratory activity in the Bay of Fundy and 9 individuals in the Island of Canela in Brazil. After selective immunostaining for mature neurons (NeuN), immature neurons (Dcx), astrocytes (GFAP), and neuronal activation by early genes (c-Fos), we quantified these markers in the hippocampal formation and compared the results of this quantification of the individuals in migration (Bay of Fundy) with those of wintering birds (Canela Island). We used quantitative stereological analyzes to estimate the total number of cells of hippocampal formation, number of active cells, total number of astrocytes and young and mature neurons. To verify if the differences found were statistically significant, we used the Student t test. Our results confirmed that autumnal migration alone, caused hippocampal changes in Calidris pusilla. After migration, we detected that the hippocampal formation has fewer activated cells and fewer astrocytes, more new neurons and greater relative volume in the quantified hemisphere (left hemisphere). However, this process did not influence the number of total cells and mature neurons. We suggest that the difference found between the volume and number of new neurons, of the full migration and wintering individuals, possibly occurred due to the migratory process in combination with local conditions found during the beginning of the wintering period. Taken together our findings demonstrate long-distance migratory shorebirds offer a unique opportunity to investigate many issues related to the cellular neurobiology of migration in general, and, on the neural plasticity associated with hippocampal neuronal and neurogenesis in adult birds.
Acesso aberto (Open Access)
Alterações histopatológicas dos rins de macacos prego, Cebusapella (Linnaeus 1758) após exposição crônica a baixas doses de metilmercúrio
(Universidade Federal do Pará, 2014-02-28) SOUSA, Andréa do Socorro Campos de Araújo; SILVEIRA, Luiz Carlos de Lima; http://lattes.cnpq.br/9383834641490219
Mercury has been a major environmental and occupational risk and it still remains a problem for human health in the Amazon region. Although studies have shown that mercury affects various tissues and organs, kidneys are the target organs to the metal toxicity. Thus, the aim of this study was to investigate the effects of chronic exposure to low doses of methylmercury on renal parenchyma of Cebusapella, adult males exposed during 120 consecutive days with daily oral doses of 1.5 μg in the diet. The concentrations of total mercury in the animals’ blood were monitored every 30 days using a cold vapor atomic spectrophotometer (201 Hg), compared to the control group. The method used for histopathological analysis was the immersion in paraffin for staining with hematoxylin and Eosin, Masson's CAB and PAS. The immunohistochemical investigations included reactions for detection of smooth muscle actin ( IA4 ), muscle actin ( HHF35 ) and cytokeratin (AE1 and AE2). The results showed that treatment with mercury caused significant differences (P < 0.001) between the exposed and control groups. As for total Hg levels, histopathologicalchanges just likehydrops in Proximal tubuleswere observed, a common finding in methylmercury exposure in other species, with no significant changes in creatinine and urea concentrations. The Person correlation test showed a strong negative relationship between mercury concentration and animal body weight loss (P < 0.0001). Another important finding was the decrease in mesangial cells number, which suggests that methylmercury executed its nephrotoxicity, affecting not only the renal tubular system, as well as the glomerular mesangium cells, making necessary a greater amount of experimental studies to clarify which mercury concentration alert level is capable of triggering aggression mechanisms and kidney injury in subjects exposed to methylmercury.
Acesso aberto (Open Access)
Análise morfológica e molecular dos filamentos das nadadeiras pélvicas do peixe pulmonado Lepidosiren paradoxa
(Universidade Federal do Pará, 2015-12-18) LIMA, Sérgio Queirós; SCHNEIDER, Igor; http://lattes.cnpq.br/4453558334183860
The Lepidosiren paradoxa species belongs to Dipnoi order, along with two genre and are considered true lungfishes. Adult males of L. paradoxa differ from females by the presence of filaments in the pelvic fin. These filaments resemble those found in gills of fish and neotenic salamanders. These filaments are grow and become vascularized during the reproduction period. In this work we tested the hypothesis that the pelvic filament of L. paradoxa share morphological and molecular characteristics with gill filaments. Thus, we performed morphological and molecular analyses of the filaments of the pelvic fins between the dry and rainy seasons. Morphological analysis was perfomed through hematoxylin and eosin (HE) and scanning electron microscopy (SEM). Finally, we quantified gene expression of gill markers by RT-PCR using pectoral fin as reference. The mean length and standard deviation of the filaments during rainy and dry season were 4,31mm of ± 0.186 and 1,63mm ± 0.104, respectively. In the SEM images, we observed with a few cell microvilli and/or microplicae and some smaller cells. In the HE analyzes we found that the filaments have uniform epithelium formed with four layers of cells being filled by connective tissue and finally become more vascularized the rainy season. Molecular analysis through RT-PCR did not show any change when compared to the pectoral fin and between the stations. In conclusion, despite morphological and molecular similarities between gill filaments and pelvic fin filaments, our findings do not support a role for L. paradoxa pelvic filaments in gas exchange.
Acesso aberto (Open Access)
Aspectos bioquímicos e morfológicos da matriz extracelular das cartilagens septal e alares do nariz suíno
(Universidade Estadual de Campinas, 1999-07-02) MESSIAS JÚNIOR, Nazário de Souza; GOMES, Laurecir; http://lattes.cnpq.br/0032653238117133
The objective of the present study was to characterize some aspects of the extracellular matrix components of the alar and septal cartilage of the nose of swine and to examine the histological and histochemical aspects of the two types of cartilage. The kaotropic agent Gu-HCl was used to extract the matrix components. The predominant glycosaminoglycan in the two caitilages was chondroitin sulfate. After ultracentrifugation, the D4 fraction of the septal and alar cartilages was fractioned on DEAE-Sephacel using na NaCl gradient. SDS-PAGE analysiys showed similarities between the two cartilages, with the presence of fíbromodulin, decorin and binding proteins. Only the septal cartilage presented the 115 kDa component, which possibly appears as 30 kDa subunits after the action of the reducing agent. The glycosaminoglycan populations were determined by fractionation on Sepharose CL-6B followed by analysis of the samples on the agarosepolyacrilamide gel. The two cartilages presented a strongly polydispersed population, which was less uniformly polydispersed in the septal than in the alar cartilage. AD2 fractionation on DEAE-Sephacel and sample analysis by SDS-PAGE and agarosepolyacrylamide electrophoresis showed the presence of the small proteoglycans biglycan and decorin, possible associated with the large proteoglycans. The septal cartilage presents chondrocytes roughly arranged in rows accompanyng the widest axis of the cartilage, whereas the alar cartilage presents chondrocytes characteristically arranged in isogenic groups. Histhochemistry was performed usisng Xylidine Ponceau for total proteins, Picrosirius-Hematoxylin for collagen, Toluidine Blue for total glycosaminoglycans. And alcian blues, pH 2.5 and pH 1.0, for sulfated and carboxylated glycosaminoglycans. Total glycosaminoglycans are more cleary visible in the territorial matrix, whereas total proteins are more cleary visible in the interterritorial matrix. The collagen of the septal cartilage is roughly organized along the longitudinal axis of the specimens between the chondrocytes rows.
Acesso aberto (Open Access)
Avaliação da atividade gonadal de machos e fêmeas de pescada branca, Plagioscion squamosissimus (Heckel, 1840), (Pisces, Teleostei, Sciaenidae) no rio Pará (PA)
(Universidade Federal do Pará, 2006) NEGRÃO, José Nazareno Cunha; OHASHI, Otávio Mitio; http://lattes.cnpq.br/5547874183666459
Male and female adults of P. squamosissimus (Pisces, Teleostei, Sciaenidae) were collected monthly in the Para River that surrounds Capim insland, state of Para (S 010 34. 971` ; W 0480 52.932`), during the month of february 2004 to february 2005, having 234 specimes. The gonads were collected, mounted and processed in accordance with the standard methods of paraffin and analysis using a light microscope. The species under analysis is one of the principal sources of animal protein for the local population, fishing being uninterrupted and intensive throughout the year based on information given by local fisherman, the collection process appears to be determined by an apparent qualiquantitative reduction in local stock. The species represents spawn divisions, the gonad development was characterized in state of repose, maturation, mature and empty or half-empty. Mature gonads were found on the months of december, january and july. Correlation analysis between the maturing state of gonads, seasonal variations in relation to gonadosomatic indexes (GSI) and also the occurrence of spawn in the respective months. Nevertheless, the information obtained in correlation to GSI and mature oocyte mediums and between spermatozoa percentages by seminiferous tubule, respectively, also indicate spawning in the respective months, and apparently that the species represents a more intensive spawning period or a primary spawning period between the months of december and january, and a second or less intensive spawning period in the months of july. Based on the tubular morphology method, eight of the seminiferous epithelium cycle stages (SEC) were determined, stage 1 being seminiferous tubule that are comprised of primary spermatogonia and cysts of the secondary spermatogonia; stage 2 is comprised of primary and seconday spermatogonia and spermatocytes cysts; stage 3 is characterized by primary and secondary spermatogonias, spermatocyte cysts and young spermatids; stage 4 with seminiferous tubule characterized by presence of spermatogonias primary and secondary, spermatocyte and by presence of young and later spermatids cysts; stage 5 represents all of the previously mentioned cells and is highlighted by presence of spermatozoa in the tubule lumen; stage 6 is characterized by reduction of cysts of germinative cells and a considerable increase in number of spermatozoa in the tubular lumen; stage 7 is characterized by seminiferous tubule that contain few cysts of germinative cells and is marked by the begining of a mass emptying of spermatozoa of the tubule lume; stage 8 is the last of the SEC and is characterized by the apparent disorganization of the remaining germinative cellular cysts in the seminiferous tubule.
Acesso aberto (Open Access)
Avaliação neuropatológica da lesão estriatal em ratos machos e fêmeas da linhagem lister hooded induzida experimentalmente por microinjeções de endotelina-1
(Universidade Federal do Pará, 2012-05-30) SANTOS, Ijair Rogerio Costa dos; LEAL, Walace Gomes; http://lattes.cnpq.br/2085871005197072
The inflammatory response may exacerbate the process harmful after acute neural disorders. Sexual dimorphism generated by different hormonal attendance between male and female have demonstrated neuroprotective abilities endogenous opposite, showing a better preservation of the integrity of the nervous tissue in female, putatively due to the presence of gynoid hormones. However, there is no research work comparing how this difference might affect the inflammatory response during stroke. In the present study, we investigated the differences in acute inflammatory processes of sexual dimorphism in adult rats of both sexes subjected to acute ischemic injury induced by endothelin (ET1) in the striatum. Six experimental groups were designed: male animals survival 24 hours (n = 8); male survival of 72 hours (n = 8); male survival of 7 days (n = 8) and female 24 hours survival (n = 8), females 72 hours of survival (n = 8); female seven days of survival. Histopathologic analysis was performed in the general sections stained with violet cresila. Macrophages, astrocytes, and neurons were identified by immunohistochemistry with antibodies specific for these inflammatory cells (ED1, anti-GFAP and Anti-NeuN, respectively). We conducted counts activated microglia / macrophages and neuron bodies mentioned in the experimental groups. No difference was observed quantitatively between different sexes, but there was an apparent decrease in the amount of macrophages / microglia at 3 days but in males and in females, presenting apparently some difference in the activation of astrocytes was stronger in males. The results suggest that sex differences, at least in lineage Lister hooded is not enough to cause significant differences in the preservation of nerve tissue and in some aspects of the inflammatory response after induction of cerebral ischemia by ET1.
Acesso aberto (Open Access)
Caracterização do genoma de tumores bem diferenciados da tireóide por hibridização genômica comparativa em matriz (aCGH)
(Universidade Federal do Pará, 2014-03-31) ALMEIDA, Deise Cibele Nunes de; OLIVEIRA, Edivaldo Herculano Correa de; http://lattes.cnpq.br/0094007714707651
More than 95% of thyroid gland malignancies are well-differentiated tumors. Preoperative diagnosis accuracy for papillary carcinomas, based on FNAC (Fine Needle Aspiration Cytology) and ultrasound screenings, has become nearly 100%. Nevertheless, when follicular neoplasm diagnosis is concerned, this reality is quite different. Using current preoperative screening tools, 60 – 80% of patients suspected to present follicular thyroid cancer, when operated turn out to suffer from benign disease. Therefore, in an effort to improve these results, genomic analysis of Copy Number Variations (CNV) that may more precisely differentiate follicular tumor from other thyroid conditions were carried out in 13 patients (3 bearing goiters, 2 hyperplasias, 4 follicular adenomas, 4 follicular carcinomas, 1 healthy individual). CNVs were tracked through aCGH method in order to detect follicular carcinoma most accurately. Results were compared with classic papillary carcinoma (4 subjects) and follicular variant (2 subjects). Samples demonstrated 725 CNVs, 703 of which belonging to patients with disease. Among these, 18 most frequent regions were selected. Patterns of amplification were more frequent in younger patients, while deletions were more frequent in the elder ones. Carcinoma patients presented very close CNV rates. Also, exclusive patterns of chromosomes 8 and 12 alterations were noticed in follicular carcinoma. Thus, it is possible to conclude that thyroid follicular carcinoma is a condition comprehending exclusive patterns of alterations, and progression of this disease is not related to follicular adenomas. Finally, regions -8p22 and 12p13.32-p12.33 were detected in 100% and 75% of samples respectively, therefore, may be deemed as possible tumoral markers.
Acesso aberto (Open Access)
Caracterização estrutural e ultraestrutural das células do epitélio gonadal de Hypophthalmus Marginatus (Valenciennes 1840) durante a espermatogênese
(Universidade Federal do Pará, 2014-09-05) GONÇALVES, Liziane Amaral Barbosa; ROCHA, Rossineide Martins da; http://lattes.cnpq.br/4371300451793081
Hypophthalmus marginatus is a siluriforme, belonging to the family Pimelodidae. This species is highly consumed in eastern Amazonia and has a wide socio-economic importance to the region. Studies on the histological characterization of the reproductive system are still unknown in this species. Therefore, this study serves as a tool for biotechnology techniques, necessary for the production in captivity, since this species is one of the fishing resource most intensively exploited by artisanal fisheries. The aim of this study was to characterize the structural and ultrastructural features of germinal epithelium cells of Hypophthalmus marginatus during spermatogenesis. 168 specimens were collected from December 2011 to December 2012. Macroscopically were identified 69 adult males. In field, ventral incision was held in fishes and fragments of testicles from the median region and semen were removed. Then the material was subjected to processing and analysis of light microscopy, transmission electron microscopy and scanning. The data revealed that the germ cells proliferate inside the spermatic cyst synchronously. Spermatids enter at the spermiogenesis process through three distinct stages of cell differentiation. The nucleus does not move relative to the axis flagellar and the centrioles don't migrate to the nucleus. The spermatozoids showed a spherical head with the highly condensed nucleus and no acrosome. The intermediary piece was short containing a single long flagellum. The axoneme presents the structure of microtubules within the pattern (9 + 2). The nuclear fossa housed the proximal centriole partially. The presence of spermatogenic cells in the same subphase of meiosis inside the cyst is characterized as cystic spermatogenesis, with synchronous growth of the cells. This suggests that spermiogenesis of H. marginatus is of type III and the presence of nuclear fossa is considered a distinctive feature for the species, which differs from other pimelodids.
Acesso aberto (Open Access)
Equinococose neotropicial: um estudo anatomopatológico para auxílio em uma sistematização diagnóstica correlacionada a morfologia
(Universidade Federal do Pará, 2012) ABREU JUNIOR, José Maria de Castro; SOARES, Manoel do Carmo Pereira; http://lattes.cnpq.br/1483308844442427; BATISTA, Evander de Jesus Oliveira; http://lattes.cnpq.br/2206444845201080
Equinococcosis is a disease caused by cestodes of the Echinococcus genus, that can be found at the small intestin of animals like bush-dog and domestic dog. Men behave as an accidental host by the ingestion of eggs of the parasite. The larval forms mainly develop at the liver, and sometimes at the lung, causing a cronic diesease that can be easily mistaken by neoplasias. At the State of Pará, North Brazil, there is a great number of cases of this rare diesease, related to Echinococcus vogeli in the Amazon.This study describes six new cases, five in the the liver, and one at the lung, making description a about the kind of inflammatory cells. Fibrosis, giant-cells, calcification and the laminated layer with the typical cerebroid pattern. Rostellar hooklets did not were found. The laminated layer, is not enough to describe the species of Echinococcus, but allows to consider that the worm´s gender is Echinococcus. Even though the number of cases is too small for a statistical analysis, were observed cells that can justify why the disease has a cronic evolution. More works like this, adding more cases are important, because equinococcosis is a neglected diesese.
Acesso aberto (Open Access)
Estrutura e ultraestrutura do epitélio epididimário de búfalos (Bubalus bubalis) em diferentes idades
(Universidade Estadual de Campinas, 1995-02-17) SANTOS, Jeannie Nascimento dos; DOLDER, Mary Anne Heidi; http://lattes.cnpq.br/7046643907575335
The zonation of the buffalo epididymis of Mediterranea (Me), Murrah (mu) breeds and croosbreed Me x Mu was studied in 4 to 36 month old animals. These buffalos were c1assified in two groups, the animals which have not reached puberty (sexually immature) and animals (mature animals) and their epididymis was divided in 7 subregions called A to G. each subregion was analysed with and transmission electron microscopy. Light microscopy inc1uded routine staining with Hematoxilin & Eosin (HE), histochemistry with P AS and morphometric analysis for tubular and luminal diameters and height of epithelium and stereocilia. Throughout the epididymary tubule it was observed with light microscopy that tubules of alI ages have Principal cell (P), basal cell (B), Apical ceeII (A) and intraepithelialleukocytes (LI). Mitotic figures in the apical region of the epithelium are also found and are more numerous in the B and C subregions, specially in buffalos between 5 and 9 months of age. Histochemical analysis revealed the existence of secretory activity from the 5th month of age, principally in the A subregion, where the activity increases with sexual maturation. Morphometric analysis supports the idea that the buffalos which have not reached puberty already havee weell determined diameters and height variations of the tubule, lumen, epithelium and stereocilia. These variations were similar to that observed in buffalos after puberty. Tubular and luminal diameters increase from the A to G subregions while the height of epithelium and stereocilia diminish. However, in both morphometric features a significant difference (p
Acesso aberto (Open Access)
Estudo morfofisiológico do epidídimo de caititus (Pecari tajacu) adultos
(Universidade Federal do Pará, 2015-05-28) OLIVEIRA, Anna Patrycia Martins de; GUIMARÃES, Diva Anelie de Araújo; http://lattes.cnpq.br/2891287458034896
This work aimed to study the histology of epididymis peccaries (Pecari tajacu) and sperm pathologies associated with different regions of the organ. A fact that will contribute to a better understanding of the reproductive biology of the species, and consequently for their conservation. Eight adult animals were used, aged two to four years, coming from breeding scientific research of Embrapa Amazônia Oriental, located in Belém, Pará, Brazil. Each of the four parts of the epididymis (proximal the head and distal the head, body and tail) was divided into two portions, and one of the portions was macerated, while the other remained intact. The epididymal tail underwent retrograde flow technique. The cell samples were stored in formol saline, and then used for the analysis of sperm morphology. Samples of the body were wrapped in gauze, identified and fixed in ALFAC. Later, they were submitted to routine histological processing. Diameters were measured up to 20 tubules and the heights of up to 30 tubules epithelium, the epididymis of each region for each animal. ANOVA (One Way), pointed out differences between the diameter of the proximal tubule of the head and distal head (p = 0.0008); between the proximal head and the body (p = 0.0009); between the proximal head and the tail (p = 0.0002). There were no significant differences (p> 0.05) among the other parts of the epididymis. It was found that the proximal head has a diameter of less tubules average compared to other portions of the epididymis. The Newman Keuls test showed similarities between the average body epithelium height and tail (p = 0.551). However, significant differences were observed between the distal head and the proximal head (p = 0.0009); the distal head and the tail (p = 0.0055); distal head and body (p = 0.0021); proximal head and body (p = 0.0008); proximal head and tail (p = 0.0055). In all regions of the epididymis of peccary, it observed the presence of pseudostratified columnar epithelium with stereocilia. 13 different types of morphological changes were found, and underdeveloped sperm and loose heads were the most frequent along the epididymis of peccaries. There were no differences in the number of pathologies in different regions of the epididymis (Man-Whitney test) (proximal head: U = 19.500, p = 0.189; distal head: U = 31.500, p = 0.958; body: U = 27,000; p = 0.599 and tail: U = 20.000, p = 0.207). In many species the histopathological analysis are often studied, however in wild animals, such as peccary, are still poorly researched. Thus, this study data are pioneers, and will serve as a basis for future comparative research on the reproductive biology of artiodactyls, contributing to their conservation and production.
Acesso aberto (Open Access)
Expressão gênica e viabilidade de folículos pré-antrais submetidos à vitrificação do córtex ovariano de Sapajus apella (macacas-prego)
(Universidade Federal do Pará, 2012-05-25) SANTANA, Luana de Nazaré da Silva; DOMINGUES, Sheyla Farhayldes Souza; http://lattes.cnpq.br/2794753357251149; SANTOS, Regiane Rodrigues dos; http://lattes.cnpq.br/0500967766886604
Vitrification is a biotech that has been increasingly showing promise in several species, among them domestic ruminants (such as sheep, goats and cows) and in nonhuman primates (NHP) (as cynomologus and rhesus), and consists in reducing ultra-rapid temperature by the presence of high concentrations of cryoprotective agents (PCA's) in liquid nitrogen. Thus the aim of this study was to develop a methodology for cryopreservation by vitrification of preantral follicles (PF's) in Sapajus apella. For this purpose, we used females (n = 9) adult Sapajus apella squad belonging to the National Primate Center (CENP). Samples were taken from the ovarian cortex by laparotomy so as not to destabilize the animals reproducibly. The fragments exposed to 8 different treatments: Ethylene glycol (EG) + 40% Sucrose (Sac) dissolved in 0.5 M TCM-199, added to Selenium (2,5, 5 or 10 ng / ml) or Trolox (25, 50 or 100 mM). Following exposure to cryoprotective agents was analyzed follicular viability before and after vitrification, from the follicular morphology, expression of Hsp70 genes, Erp29, Erp60 SOD1 and through the analysis by Polymerase Chain Reaction (PCR), in addition to performing measurement of oxidative stress thru the TEAC (English Total Equivalent Antioxidant Activity). The analyzes showed that vitrification allowed the maintenance of follicular viability by previous exposure to concentrations of ACP's alevadas, especially when supplemented with 50 mM trolox (which resulted in high follicular survival rates) and increased expression of the antioxidant enzyme SOD1. While in the absence of it was observed an increase in rates of follicles degenerate and vacuolated, and reduced expression of the antioxidant enzyme SOD1 and increased expression of chaperone Erp29.
Acesso aberto (Open Access)
Inibição nitrérgica local favorece o processo de regeneração do tendão de aquiles em ratos submetidos à tenotomia
(Universidade Federal do Pará, 2011-05-27) MORAES, Suellen Alessandra Soares de; SILVA, Anderson Manoel Herculano Oliveira da; http://lattes.cnpq.br/8407177208423247
The tendon injuries prevalence has been increased usually by repetitive movements in the occupational and recreational activities of society. The main features this injury involve healing dysfunction and loss of extracellular matrix. At this tissue, the extracellular matrix is basically made of type I collagen and the loss of this protein associated with problems in the remodeling have been considered the major responsible for the symptomatic and functional effects of injury. Additionally the literature reports the nitric oxide synthase expression was up-regulated throughout the repair process. At this context the propose of this work is investigated the nitrergic local inhibition in tissue and functional recovery after tendon injury. We used a experimental model of Achilles tendon rupture for establish pathophysiological changes similar to chronic tendinopathy. The animals were divided into experimental groups control, injury+vehicle (saline, 0,9%), injury+L- Nω-nitro-L-arginine methyl ester (L-NAME, 5mM) and injury+sodium nitroprusside (SNP, 3mM), which were followed until the 21º day, when were killed to collect samples. In order to access nitric oxide production Griess nitrite assay was used. Functional recovery was calculated by Achilles Functional Index (AFI) in 0, 7, 14 and 21 days. Tendons were also processed for histomorphological and autofluorescence analysis. The body weight gain was also evaluated. We characterized the effect of drugs about tissue without stronger systemic influences by maintenance of body weight and plasmatic levels of nitrite. The L-NAME led to significant decrease in the cellular density, vessel formation between collagen network, accelerated the tissue organization and restored early the functional pattern at 21 days after injury. On the other hand, the treatment with SNP and vehicle remained the tissue disorganization and low functional performance. Taken together our results suggest a positive effect of local suppression of NOS after rupture for tissue remodeling, contributing to tendon regeneration.
Acesso aberto (Open Access)
Regeneração tendínea em modelo murino: estudo da plasticidade central e investigação do efeito da modulação nitrérgica na plasticidade periférica
(Universidade Federal do Pará, 2015-08-10) MORAES, Suellen Alessandra Soares de; SILVA, Anderson Manoel Herculano Oliveira da; http://lattes.cnpq.br/8407177208423247
Tendon injuries cause strong impact on people due to pain and functional limitation resulting therefrom. After injury, the tissue starts to present a network of nerves. Furthermore, there is evidence for the occurrence of central plasticity after injury. Among the molecular factors involved in injury repair, nitric oxide (NO) is implicated in tissue remodeling, but its effects are not yet well understood. The purpose of this study is to ascertain the existence of central plasticity and the influence of NO in peripheral plasticity, functional limitation and tendon regeneration in murine model. To study the effects of NO in peripheral plasticity, we used control animals (CTRL, without injury) or animals treated with saline (SAL, 0.9% NaCl), L-nitro-arginine-methyl-ester (L-NAME, NO-synthesis inhibitor) and sodium nitroprusside (SNP, NO donor) every other day until the 21st day post injury (DPI). To evaluate central plasticity (L5 segment), an injury was performed alone and the spinal cord collected at 2 or 21 DPI. We analyzed the integrity and tissue organization in tendon samples by H&E, transmission electron microscopy and immunofluorescence, which was also used to evaluate peripheral plasticity. To assess tendon functional recovery, we determined the Achilles functional index, the joint angle and the open field. In spinal cord studies, we investigated glial reactivity and neuron involvement after injury by co-localizations with the cell activation indicator c-Fos. The findings of this research show that NO inhibition promotes tissue organization in association to an increase in collagen synthesis, secretion and deposition. Besides, L-NAME local administration seems to favor cell differentiation to tenocyte-like morphological types and improve the organization of nerve branches in between the collagen mesh in correlation with functional recovery at 21 DPI. On the other hand, increased levels of NO by SNP promoted worsening in almost all parameters analyzed. Our data also show tendon injury triggers a central plasticity process with an increase in glial reactivity at 2 DPI and ipsilateral cell activation at 2 and 21 DPI. Afterall, our findings point out occurrence of central plasticity after tendon injury and favoring of tissue repair and peripheral plasticity through nitrergic blockage, unraveling fundamental aspects of tissue recovery that may represent new targets for a therapeutical approach in tendon injuries.
Acesso aberto (Open Access)
O tratamento com ácido ascórbico acelera o processo de reparo do tendão calcâneo em modelo de lesão tendínea em ratos
(Universidade Federal do Pará, 2015-02-24) SOUZA, Martha Costa de; SILVA, Anderson Manoel Herculano Oliveira da; http://lattes.cnpq.br/8407177208423247
Tendon rupture affects a large part of the population in special seniors and athletes.The repair process requires more studies which can indicate the possibility of new treatments. The ascorbic acid (AA) is a well known substance by its requirement for proline and lysine hydroxylase activity during the collagen synthesisis and the efficiency of this vitamin for various treatments, because of its antioxidant properties. So, the aim of the present study is to evaluate the local treatments effects with this substance on tissue and functional repair the Achilles tendon from rats. The study was approved by the ethics committee of the institution (CEPAE-UFPA) according to the license 161-13. The animals were submitted to rupture of Achilles tendon, divided into three groups (n=27): control, injury+AA (30 mM), injury+vehicle (0.9% NaCl). All treatments were performed by local injection, from the second day after injury and every other day until day 14 or 21. The walk of the animals was assessed by functional index of Achilles (IFA) on days 7(n=6), 14(n=6) e 21(n=3), cells number was assessed by staining with DAPI and tissue organization by staining with HE and autofluorescence, at 14(n=9) and 21(n=9) days of injury. The animals did not differ in body mass gain. The injury+AA group (-39.51±15.3) showed functional improvement especially at day 14 when compared to the injury+vehicle (-89.22±16.57, p<0.01). The histological examination demonstrated in counting the number of cells that the injury+AA group (762±29.6) showed a smaller number of cells on day 21 (762±29.6) in relation to the groups injury+vehicle (916±57.0, p<0.01). The analysis of autofluorescence of collagen and HE showed that the injury+AA group achieved better ECM organization on day 14 and 21 in relation to the groups injury+vehicle, in turn, differs significantly from the control group. Our results suggest that AA accelerates the healing process of tendon injury, presenting tissue and functional improvements 21 days after injury.