980 resultados para tissue nonspecific alkaline phosphatase


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Twenty-four horses were distributed into four different groups, instrumented control (GI), duodenum obstruction (GII), ileum obstruction (GIII) and large colon obstruction (GIV). Serum and peritoneal fluid analysis of aspartate aminotransferase, creatine kinase, lactate dehydrogenase, alkaline phosphatase, inorganic phosphorus and lactate were measured. Samples were collected one hour before the surgical procedure (T0); 3 hours after the obstruction (T3ob), 1, 3, 12, 24, 120 and 168 hours after the beginning of reperfusion/deobstruction. Duodenal (GII) and ileum (GIII) obstructions changed serum and peritoneal fluid biochemical analysis. However, only lactate, lactate dehydrogenase, creatine kinase and inorganic phosphorus concentrations were abnormal in peritoneal fluid three hours after the obstruction. The biochemical analysis of peritoneal fluid allowed a faster diagnostic of intestinal alterations than the serum analysis; hence it should be prioritized when pre-operatory colic assessment is carried out.

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The newborn goat kids suffer several adjustments to life outside the uterus that affect different body functions that they need to produce heat, muscle activity and start searching for food. All these events lead to changes in several blood constituents such as proteins and serum biochemical parameters. Other studies have shown these variations, but not extend beyond the neonatal period to phase in young animals. The aim was to test the hypothesis that there is variation of the protein profile, and biochemical components of blood glucose in goat kids from birth to 75 days of life in terms of adaptation to extra uterine life. To achieve these objectives have been collected blood samples from 25 goats born by normal delivery, regardless of their sex. The variables serum total protein (TP), albumin, α-globulin, β-globulin, γ-globulin, which includes the immunoglobulin G (IgG), aspartate (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), creatinine and urea, and glucose were determined at times zero (immediately after birth), two, seven, 15, 30 and 75 days old. We observed significant differences in all variables between times, but only the creatinine concentration was higher than those of other moments in time zero and two days old, probably due to immaturity of renal function in newborn animals. The blood constituents of the kids had variations in the study period, related to physiological and nutritional causes.

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The objective was to test the hypothesis that there is variation of some protein components, biochemical and glucose from the blood of goats in the postpartum period. ElevenBoer goats were used to evaluate the serum following variables: total protein (TP), albumin, α-globulin, β-globulin, γ-globulin, immunoglobulin G (IgG), aspartate (AST), alkaline phosphatase (ALP), gamma-glutamyltransferase (GGT), creatinine, urea and glucose. These components were determined in moments zero (immediately after delivery), two, seven, 15, 30 and 75 days postpartum. The β-globulin, IgG, creatinine and urea from the goats did not show significant variations, the blood glucose at time zero was greater than the baseline values due to physiological stimulation of glycogenesis determined by the increase of cortisol in delivery. The blood constituents of goats showed variations in the period to 75 days postpartum as a result physiological causes related to delivery and postpartum.

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In vertebrate species, testosterone seems to inhibit spermatogonial differentiation and proliferation. However, this androgen can also be converted, via aromatase, into estrogen which stimulates spermatogonial differentiation and mitotic activity. During seasonal spermatogenesis of adult bullfrogs Lithobates catesbeianus, primordial germ cells (PGCs) show enhanced testosterone cytoplasm immunoexpression in winter; however, in summer, weak or no testosterone immunolabelling was observed. The aim of this study was to confirm if PGCs express stem cell markers-alkaline phosphatase (AP) activity and GFRα1 (glial-cell-line-derived neurotrophic factor)-and verify whether testosterone is maintained in these cells by androgen receptors (ARs) and/or sex hormone-binding globulin (SHBG) in winter. Furthermore, regarding the possibility that testosterone is converted into estrogen by PGCs in summer, the immunoexpression of estrogen receptor (ER)β was investigated. Bullfrog testes were collected in winter and in summer and were embedded in glycol methacrylate for morphological analyses or in paraffin for the histochemical detection of AP activity. GFRα1, AR, SHBG and ERβ expression were detected by Western blot and immunohistochemical analyses. The expression of AP activity and GFRα1 in the PGCs suggest that these cells are spermatogonial stem cells. In winter, the cytoplasmic immunoexpression of ARs and SHBG in the PGCs indicates that testosterone is maintained by these proteins in these cells. The cytoplasmic immunoexpression of ERβ, in summer, also points to an ER-mediated action of estrogen in PGCs. The results indicate a participation of testosterone and estrogen in the control of the primordial spermatogonia during the seasonal spermatogenesis of L. catesbeianus. © 2012 S. Karger AG, Basel.

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Objectives: This study aimed to comparatively evaluate the in vitro osteogenic potential of cells obtained from the mandibular ramus (MR, autogenous bone donor site) and from the maxillary sinus (MS) bone grafted with a mixture of anorganic bovine bone (ABB) and MR prior to titanium implant placement (MS, grafted implant site). Material and methods: Cells were obtained from three patients subjected to MS floor augmentation with a 1: 1 mixture of ABB (GenOx Inorg®) and MR. At the time of the sinus lift procedure and after 8 months, prior to implant placement, bone fragments were taken from MR and MS, respectively, and subjected to trypsin-collagenase digestion for primary cell culturing. Subcultured cells were grown under osteogenic condition for up to 21 days and assayed for proliferation/viability, osteoblast marker mRNA levels, alkaline phosphatase (ALP) activity and calcium content/Alizarin red staining. ALP activity was also determined in primary explant cultures exposed to GenOx Inorg® (1: 1 with MR) for 7 days. Data were compared using either the Mann-Whitney U-test or the Kruskal-Wallis test. Results: MS cultures exhibited a significantly lower osteogenic potential compared with MR cultures, with a progressive increase in cell proliferation together with a decrease in osteoblast markers, reduced ALP activity and calcium content. Exposure of MR-derived primary cultures to GenOx Inorg® inhibited ALP activity. Conclusion: These results suggest that the use of GenOx Inorg® in combination with MR fragments for MS floor augmentation inhibits the osteoblast cell differentiation at the implant site in the long term. © 2013 John Wiley & Sons A/S.

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The aim of this study was to evaluate the possibility of fluoride solutions applied to enamel to protect pulp cells against the trans-enamel and transdentinal cytotoxicity of a 16% carbamide peroxide (CP) bleaching gel. The CP gel was applied to enamel/ dentin discs adapted to artificial pulp chambers (8 h/day) during 1, 7 or 14 days, followed by fluoride (0.05% or 0.2%) application for 1 min. The extracts (culture medium in contact with dentin) were applied to MDPC-23 cells for 1 h, and cell metabolism (MTT assay), alkaline phosphatase (ALP) activity and cell membrane damage (flow cytometry) were analyzed. Knoop microhardness of enamel was also evaluated. Data were analyzed statistically by ANOVA and Kruskal-Wallis tests (a=0.05). For the MTT assay and ALP activity, significant reductions between the control and the bleached groups were observed (p<0.05). No statistically significant difference occurred among bleached groups (p>0.05), regardless of fluoride application or treatment days. Flow cytometry analysis demonstrated 30% of cell membrane damage in all bleached groups. After 14 days of treatment, the fluoride-treated enamel presented significantly higher microhardness values than the bleached-only group (p<0.05). It was concluded that, regardless of the increase in enamel hardness due to the application of fluoride solutions, the treated enamel surface did not prevent the toxic effects caused by the 16% CP gel to odontoblast-like cells.

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An uncommon disseminated Mycobacterium tuberculosis infection is described in a 12-year-old female dog presenting with fever, dyspnea, cough, weight loss, lymphadenopathy, melena, epistaxis, and emesis. The dog had a history of close contact with its owner, who died of pulmonary tuberculosis. Radiographic examination revealed diffuse radio-opaque images in both lung lobes, diffuse visible masses in abdominal organs, and hilar and mesenteric lymphadenopathy. Bronchial washing samples and feces were negative for acid-fast organisms. Polymerase chain reaction (PCR)-based species identification of bronchial washing samples, feces, and urine revealed M. tuberculosis using PCR-restriction enzyme pattern analysis-PRA. Because of public health concerns, which were worsened by the physical condition of the dog, euthanasia of the animal was recommended. Rough and tough colonies suggestive of M. tuberculosis were observed after microbiological culture of lung, liver, spleen, heart, and lymph node fragments in Löwenstein-Jensen and Stonebrink media. The PRA analysis enabled diagnosis of M. tuberculosis strains isolated from organs. Copyright © 2013 by The American Society of Tropical Medicine and Hygiene.

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Purposes: The purposes of this study were to evaluate the influence of chronic stress (CS) on implant osseointegration and also to analyze whether alendronate (ALN) therapy could prevent these eventual stress-negative effects. Materials and Methods: Adult male Holtzmann rats were assigned to one of the four experimental groups: AL (ALN; 1mg/kg/week; n=12), ALS (ALN+CS; 1mg/kg/week; n=12), CTL (sterile physiological saline; n=12), or CTLS (sterile physiological saline+CS; n=12). After 58 days of drug therapy, the ALS and CTLS groups were exposed to CS, and 2 days later all animals underwent tibial implant installation. The animals were euthanized 28 days following the operative surgical procedure. Results: It was observed that the CTLS group presented an impairment of bone metabolism represented by lowest levels of bone-specific alkaline phosphatase and bone area fraction occupancy values. Furthermore, these animals presented a higher proportion of empty osteocytic lacunae. In contrast, the ALN therapy showed increased osseointegration and torque value parameters, regardless of stress exposition. Conclusions: Analysis of the data presented suggests that CS partially impairs the osseointegration of tibial implants and that ALN therapy is able to prevent these negative effects. © 2013 Wiley Periodicals, Inc.

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Inappropriate reference limits increase the risks of unnecessary investigations and diagnostic failures. Species, breed, environment, handling, and physiologic stage can influence serum biochemical variables. This study aimed to establish serum biochemical reference intervals of Pêga donkeys and the influence of age and sex on these variables. Samples were taken from 110 donkeys (79 females and 31 males; 8 under 1 year, 33 between 1 and 3 years, and 69 over 3 years old). There were no differences for creatine kinase (CK), albumin, urea, and magnesium among age groups. Animals under 1 year old had the lowest aspartate aminotransferase (AST), creatinine, triglycerides, total calcium, and chloride means; lower indirect bilirubin, gamma-glutamyl transferase (GGT), alanine aminotransferase (ALT), and sodium levels than over 3-year-old group and the highest phosphorus, glucose, cholesterol, ionized calcium, and alkaline phosphatase (AP) values. Total protein increased with age. The group from 1 to 3 years had higher potassium than up to 1-year-old group. Animals over 3 years old had the highest means of direct and total bilirubin. ALT, CK, GGT, direct and total bilirubin, urea, triglycerides, sodium, potassium, and chloride levels did not differ between genders. Females had higher AST, total protein, total calcium, and indirect bilirubin levels. Males showed greater AP, albumin, phosphorus, glucose, creatinine, cholesterol, magnesium, and ionized calcium values. Discrepancies among present results and previous studies show influencing factors on biochemical profile of donkeys and reinforce the importance of establishment of specific reference intervals. This study is useful in clinical routine and as basis to other scientific researches with Pêga donkeys. © 2013 Springer-Verlag London.

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The aim of the present study was to evaluate the effect of low-level laser therapy (LLLT) on odontoblast-like MDPC-23 cells exposed to carbamide peroxide (CP 0.01 %-2.21 μg/mL of H2O2). The cells were seeded in sterile 24-well plates for 72 h. Eight groups were established according to the exposure or not to the bleaching agents and the laser energy doses tested (0, 4, 10, and 15 J/cm2). After exposing the cells to 0.01 % CP for 1 h, this bleaching solution was replaced by fresh culture medium. The cells were then irradiated (three sections) with a near-infrared diode laser (InGaAsP-780 ± 3 nm, 40 mW), with intervals of 24 h. The 0.01 % CP solution caused statistically significant reductions in cell metabolism and alkaline phosphate (ALP) activity when compared with those of the groups not exposed to the bleaching agent. The LLLT did not modulate cell metabolism; however, the dose of 4 J/cm2 increased the ALP activity. It was concluded that 0.01 % CP reduces the MDPC-23 cell metabolism and ALP activity. The LLLT in the parameters tested did not influence the cell metabolism of the cultured cells; nevertheless, the laser dose of 4 J/cm2 increases the ALP activity in groups both with and without exposure to the bleaching agent. © 2013 Springer-Verlag London.

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The aim of the study was to evaluate the effects of a highly potent bisphosphonate, zoledronic acid (ZOL), on cultured odontoblast-like cells MDPC-23. The cells (1.5 × 104 cells/cm2) were seeded for 48 h in wells of 24-well dished. Then, the plain culture medium (DMEM) was replaced by fresh medium without fetal bovine serum. After 24 h, ZOL (1 or 5 μM) was added to the medium and maintained in contact with the cells for 24 h. After this period, the succinic dehydrogenase (SDH) enzyme production (cell viability-MTT assay), total protein (TP) production, alkaline phosphatase (ALP) activity, and gene expression (qPCR) of collagen type I (Col-I) and ALP were evaluated. Cell morphology was assessed by SEM. Five μM ZOL caused a significant decrease in SDH production. Both ZOL concentrations caused a dose-dependent significant decrease in TP production and ALP activity. ZOL also produced discret morphological alterations in the MDPC-23 cells. Regarding gene expression, 1 μM ZOL caused a significant increase in Col-I expression. Although 5 μM ZOL did not affect Col-I expression, it caused a significant alteration in ALP expression (ANOVA and Tukey's test, p < 0.05). ZOL presented a dose-dependent cytotoxic effect on the odontoblast-like cells, suggesting that under clinical conditions the release of this drug from dentin could cause damage to the pulpo-dentin complex. © 2012 Elsevier Ltd.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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This study evaluated the cytotoxicity of experimental adhesive systems (EASs) on odontoblast-like cells. Paper discs (n=132) were impregnated with 10 μL of each EAS-R1, R2, R3, R4, and R5 (in an ascending order of hydrophilicity), followed by photoactivation. R1 and R2 are nonsolvated hydrophobic blends, R3 represents a simplified etch-and-rinse adhesive system, and R4 and R5 represent simplified self-etch adhesive systems. Discs were immersed in Dulbecco's modified Eagle's medium for 24 h to obtain eluates applied on MDPC-23 cell cultures. No material was applied on discs used as control (R0). Cell viability [3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], total protein (TP) production, alkaline phosphatase (ALP) activity, type of cell death, and degree of monomer conversion Fourier transform infrared (%DC-FTIR) were evaluated. Data were analyzed by Kruskal-Wallis and Mann-Whitney tests (α=0.05). Considering R0 (control) as having 100% of cell viability, R1, R2, R3, R4, and R5 reduced the metabolic activity of cells by 36.4, 3.1, 0.2, 21.5, and 65.7%, respectively, but only R1 and R5 differed from R0. Comparing with R0, lower TP production was observed for R1, R4, and R5, while ALP activity decreased for R1 and R5. Necrotic cell death was predominant for all EASs, but only R1, R4, and R5 differed from R0. Only R5 presented a different apoptotic cell death ratio from R0. R1 presented the lowest %DC (ca. 37%), whereas R4 and R5 presented the highest (ca. 56%). In conclusion, R2 and R3 were not toxic to the MDPC-23 cells, suggesting that the degree of hydrophilicity or %DC of the EASs alone were not responsible for their cytopathic effects. © 2013 Wiley Periodicals, Inc.

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The yolk protein precursor, vitellogenin (Vg), in bees is synthesized in the fat body trophocytes, delivered to the hemolymph and ultimately absorbed from there during the vitellogenic phase of oocytes in the active ovary. The routes tracing the material exchange that occurs between the trophocytes and the hemolymph, in addition to the transportation from the hemolymph to the ovarian follicles, were marked by alkaline phosphatase and lanthanum nitrate (LN). Active ovaries from nurse workers and physogastric queens, as well as inactive ovaries of virgin queens, were examined by transmission electron microscopy. The LN permitted better visualization of the routes of exchanges between the organs and the hemolymph. Both methods demonstrate the apparent differences between the phases of the ovary and the bee caste. In inactive ovaries of the virgin queens, the routes from the follicular epithelium to the oocyte remain closed; conversely, they are open in active ovaries of the nurse workers and physogastric queens. The differences between the methods and classes of bees are discussed. © The Author 2013. Published by Oxford University Press [on behalf of The Japanese Society of Microscopy]. All rights reserved.