RESUMO
Soil enzymes mediate key processes and functions of the soils, such as organic matter decomposition and nutrient cycling in both natural and agricultural ecosystems. Here, we studied the activity of five extracellular soil enzymes involved in the C, N, and P-mineralizing process in both litter and surface soil layer of rainforest in the northwest region of the Colombian Amazon and the response of those soil enzymes to land use change. The experimental study design included six study sites for comparing long-term pasture systems to native forest and regeneration practices after pasture, within the main landscapes of the region, mountain and hill landscapes separately. Results showed considerable enzymatic activity in the litter layer of the forest, highlighting the vital role of this compartment in the nutrient cycling of low fertility soils from tropical regions. With the land use transition to pastures, changes in soil enzymatic activities were driven by the management of pastures, with SOC and N losses and reduced absolute activity of soil enzymes in long-term pastures under continuous grazing (25 years). However, the enzyme activities expressed per unit of SOC did not show changes in C and N-acquiring enzymes, suggesting a higher mineralization potential in pastures. Enzymatic stoichiometry analysis indicated a microbial P limitation that could lead to a high catabolic activity with a potential increase in the use of SOC by microbial communities in the search for P, thus affecting soil C sequestration, soil quality and the provision of soil-related ecosystem services.
Assuntos
Acetilglucosaminidase/análise , Fosfatase Ácida/análise , Agricultura/métodos , Celulose 1,4-beta-Celobiosidase/análise , Glucosidases/análise , Floresta Úmida , Solo/química , Xilosidases/análise , Carbono/análise , Colômbia , Conservação dos Recursos Naturais , Microbiota , Nitrogênio/análise , Fósforo/análise , Microbiologia do Solo , Clima TropicalRESUMO
The diagnosis of acute kidney injury by evaluating the activity of urinary enzymes is an important tool especially for its precocity in relation to methods of assessment of renal function considered late as the installation of injury precedes the function change. This study was performed to determine the reference interval for urinary NAG activity (N-Acetyl-β-D-glucosaminidase) in urine of healthy dogs of different breeds, ages and sexes. It was utilized an automated method for the determination of NAG activity standardized for dog urine samples. Through statistical analysis it was observed that NAG is positively correlated with the age of the animal and urine specific gravity but is not interfered by sex or weight. Based on the determination of urinary activity and after correcting the values for a 1,025 urine specific gravity was obtained an reference interval of X '= 3.62U/L±0.66U/L.(AU)
O diagnóstico da injúria renal aguda pela avaliação da atividade de enzimas urinárias é uma importante ferramenta para o médico veterinário, especialmente por sua precocidade em relação aos métodos de avaliação da função renal, considerados tardios, visto que a instalação da injúria precede a alteração da função. Neste estudo foi realizada a determinação do intervalo de referência médio para a atividade urinária de NAG (N-Acetyl- β-D-Glucosaminidase), em urinas de cães hígidos, de diferentes raças, sexos e faixas etárias. Para isso, trabalhou-se com um método automatizado para determinação da atividade de NAG, padronizado para amostras de urina de cães. Por meio de análise estatística, observou-se que a NAG apresenta correlação positiva com a idade do animal e com a densidade urinária, mas não sofre interferência quanto ao sexo ou peso. Com base na determinação da atividade urinária e após corrigir os valores para uma densidade de 1,025, obteve-se um intervalo de referência médio de X´ = 3,62U/L±0,66U/L.(AU)
Assuntos
Animais , Cães , Acetilglucosaminidase/análise , Urina/química , Cães/fisiologiaRESUMO
The diagnosis of acute kidney injury by evaluating the activity of urinary enzymes is an important tool especially for its precocity in relation to methods of assessment of renal function considered late as the installation of injury precedes the function change. This study was performed to determine the reference interval for urinary NAG activity (N-Acetyl-β-D-glucosaminidase) in urine of healthy dogs of different breeds, ages and sexes. It was utilized an automated method for the determination of NAG activity standardized for dog urine samples. Through statistical analysis it was observed that NAG is positively correlated with the age of the animal and urine specific gravity but is not interfered by sex or weight. Based on the determination of urinary activity and after correcting the values for a 1,025 urine specific gravity was obtained an reference interval of X '= 3.62U/L±0.66U/L.(AU)
O diagnóstico da injúria renal aguda pela avaliação da atividade de enzimas urinárias é uma importante ferramenta para o médico veterinário, especialmente por sua precocidade em relação aos métodos de avaliação da função renal, considerados tardios, visto que a instalação da injúria precede a alteração da função. Neste estudo foi realizada a determinação do intervalo de referência médio para a atividade urinária de NAG (N-Acetyl- β-D-Glucosaminidase), em urinas de cães hígidos, de diferentes raças, sexos e faixas etárias. Para isso, trabalhou-se com um método automatizado para determinação da atividade de NAG, padronizado para amostras de urina de cães. Por meio de análise estatística, observou-se que a NAG apresenta correlação positiva com a idade do animal e com a densidade urinária, mas não sofre interferência quanto ao sexo ou peso. Com base na determinação da atividade urinária e após corrigir os valores para uma densidade de 1,025, obteve-se um intervalo de referência médio de X´ = 3,62U/L±0,66U/L.(AU)
Assuntos
Animais , Cães , Acetilglucosaminidase/análise , Urina/química , Cães/fisiologiaRESUMO
OBJETIVO: Este estudo teve como objetivo averiguar a atividade enzimática da N-acetil-β-D-glicosaminidase (NAG) como possível biomarcador precoce de disfunção renal para a exposição ocupacional ao chumbo inorgânico. MATERIAIS E MÉTODOS: Foi selecionado um grupo de 30 pessoas do sexo masculino expostas ao chumbo inorgânico em uma fábrica de baterias localizada no estado do Paraná. Fizeram parte do grupo os funcionários que mostraram valores de chumbo sanguíneo inferiores a 40 mg/dl. O grupo controle foi representado por 15 adultos saudáveis com similaridade em relação à idade e ao gênero do grupo exposto. Foram determinados os níveis de plumbemia, do ácido d-aminolevulínico urinário e a atividade da NAG urinária. RESULTADOS E DISCUSSÃO: Foi evidenciado que a atividade urinária da NAG foi significativamente maior (p < 0,05; teste U de Mann-Whitney) no grupo exposto ao chumbo inorgânico quando comparado ao grupo controle, e houve uma correlação negativa com significância (p < 0,05; correlação de Spearman Rank Order) entre o indicador biológico de exposição plúmbica e a atividade urinária da NAG. CONCLUSÃO: Os resultados demonstraram que o aumento da atividade urinária da NAG pode ser utilizado como um biomarcador precoce da exposição ao chumbo inorgânico.
OBJECTIVE: This study aimed to verify the enzymatic activity of N-acetyl-β-D-glucosaminidase (NAG) as a possible early biomarker of renal dysfunction due to occupational exposure to inorganic lead. MATERIALS AND METHODS: We selected a group of 30 males that had been exposed to inorganic lead in a battery factory in the state of Paraná. This group comprised those employees whose blood lead levels were below 40 mg/dl. The control group consisted of 15 healthy adults of similar age and gender compared with the exposed group. Blood lead concentrations, d-aminolevulinic acid levels and urinary NAG activity were measured. RESULTS AND DISCUSSION: It was shown that urinary NAG activity was significantly higher (p < 0.05, U test of Mann-Whitney) in the exposed group in comparison to the control group, and there was a significant negative correlation (p < 0.05, Spearman Rank Order correlation) between the biological indicator of lead exposure and urinary NAG activity. CONCLUSION: The results showed that the increase of urinary NAG activity may be used as an early biomarker of the exposure to inorganic lead.
Assuntos
Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Acetilglucosaminidase/análise , Acetilglucosaminidase/urina , Acetilglucosaminidase , Doenças Profissionais/diagnóstico , Diagnóstico Precoce , Biomarcadores/análise , Biomarcadores/urina , Nefropatias/diagnóstico , Nefropatias/enzimologia , Ácido Aminolevulínico/análise , Ácido Aminolevulínico , Creatinina/análise , Intoxicação por Chumbo/diagnóstico , Intoxicação por Chumbo/sangue , Intoxicação por Chumbo/urinaRESUMO
Schistosoma mansoni is a parasitic trematode of the portal-mesenteric veins with a closed-end intestine. Adult worms regurgitate their intestinal content after digestion, together with constituents of the lining gut. Some of these molecules circulate in the blood and are antigenic. We obtain a "vomit" preparation and preliminary evaluate its biochemical composition and antigenic capacity. The "vomit" preparation was obtained after changes in temperature and solutions of incubation of adult worms between 4 and 37 degrees C. Supernatant was assayed for protein, carbohydrate concentration and enzymatic activities associated to the intestine and to the worm tegument. The antigenicity of the product was evaluated using Western blot (WB) analysis against sera of experimentally infected mice, before and after drug cure, sera from people infected with S. mansoni and from individuals infected with other parasitoses. More carbohydrate than protein was detected in the preparations. Cysteine proteinase (CP), N-acetyl-beta-D: -glucosaminidase and alkaline phosphatase activities were detected. The latter enzyme activity is a marker of the tegument, suggesting that in spite of careful conditions used to avoid the presence of tegumental material, manipulation of the worms always resulted in the release of tegumental molecules. Cationic exchange chromatography was useful to separate various components of this "vomit" preparation, particularly enzymes responsible for CP activity. Two highly immunogenic and specific duplets were observed in the WB analysis, 31/32- and 38/40-kDa components, the former probably referring to the intestinal CPs Sm31/Sm32. None of the two duplets disappeared after successful chemotherapy during the time of evaluation in mice or humans.
Assuntos
Antígenos de Helmintos/isolamento & purificação , Schistosoma mansoni/imunologia , Esquistossomose mansoni/diagnóstico , Acetilglucosaminidase/análise , Fosfatase Alcalina/análise , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/química , Western Blotting , Cromatografia por Troca Iônica , Cricetinae , Cisteína Endopeptidases/análise , Humanos , Camundongos , Peso Molecular , Schistosoma mansoni/química , Esquistossomose mansoni/tratamento farmacológico , Esquistossomose mansoni/imunologiaRESUMO
One of the most fascinating aspects of the Entamoeba histolytica trophozoite ultrastructure is the lack of a typical secretory pathway, particularly of rough endoplasmic reticulum and Golgi system, in a cell with such a high secretory activity. Here, we describe the isolation of amoeba cell structures containing ER-typical activities. Following isopycnic centrifugation of plasma membrane-free extracts, microsomes enriched in enzymatic activities such as dolichol-P-mannose synthase (DPMS; EC 2.4.1.83), UDP-GlcNAc:dolichol-P GlcNAc-1-P transferase (NAGPT; EC 2.7.8.15), and UDP-D-GlcNAc:dolichol-PP GlcNAc (NAGT; EC 2.4.1.141) were resolved from phagolysosomal fractions. Sec61alpha-subunit, an ER-marker involved in the translocation of nascent proteins to the ER, was found to co-fractionate with DPMS activity indicating that they are contained in microsomes with a similar density. Further, we optimized conditions for trophozoite homogenization and differential centrifugation that resulted in the separation of a 57,000 g-sedimenting microsomal fraction containing EhSec61alpha-subunit, EhDPMS, and EhPDI (protein disulfide isomerase, a soluble marker of the lumen of the ER). A relevant observation was the lack of ER markers associated to the nuclear fraction. Large macromolecular structures such as Ehproteasome were sedimented at a higher speed. Our knowledge of the molecular machinery involved in the biosynthesis of dolichol-linked oligosaccharide was enriched with the identification of putative genes related to the stepwise assembly of the dolichol-PP-GlcNAc(2)Man(5) core. No evidence of genes supporting further assembly steps was obtained at this time.
Assuntos
Entamoeba histolytica/ultraestrutura , Microssomos/enzimologia , Proteínas de Protozoários/metabolismo , Acetilglucosaminidase/análise , Fosfatase Ácida/análise , Animais , Western Blotting , Centrifugação com Gradiente de Concentração , Dolicóis/metabolismo , Retículo Endoplasmático/enzimologia , Retículo Endoplasmático/fisiologia , Entamoeba histolytica/enzimologia , Entamoeba histolytica/genética , Entamoeba histolytica/fisiologia , Glucosiltransferases/análise , Glicosilação , Manosiltransferases/análise , Manosiltransferases/genética , Proteínas de Membrana/análise , Microssomos/fisiologia , Microssomos/ultraestrutura , Oligossacarídeos/biossíntese , Complexo de Endopeptidases do Proteassoma/análise , Isomerases de Dissulfetos de Proteínas/análise , Canais de Translocação SECRESUMO
The majority of biological responses classically attributed to tumor necrosis factor alpha (TNF-alpha) is mediated by p55 receptor (TNFR1). Here, we aimed to clarify the biological role of TNFR1-mediated signals in an in vivo inflammatory angiogenesis model. Polyester-polyurethane sponges, used as a framework for tissue growth, were implanted in C57Bl/6 mice. These implants were collected at days 1, 7, and 14 post-implant for enzyme-linked immunosorbent assay or at days 7 and 14 for hemoglobin, myeloperoxidase, and N-acetylglucosaminidase measurements, used as indexes for angiogenesis, neutrophil, and macrophage accumulation, respectively. In TNFR1-deficient C57Bl/6 mice, there was a significant decrease in sponge vascularization but not in late inflammatory cell influx. It is interesting that levels of vascular endothelial growth factor were significantly lower in TNFR1-deficient than in wild-type mice at days 1 and 7. Levels of angiogenic chemokines, CC chemokine ligand 2/murine homologue of monocyte chemoattractant protein-1 and CXC chemokine ligand 1-3/keratinocyte-derived chemokine, were significantly lower in TNFR1-deficient mice at days 1 and 7 after implantation, respectively. These observations suggest that TNFR1-mediated signals have a critical role in sponge-induced angiogenesis, possibly by influencing the effector state of inflammatory cells and hence, modulating the angiogenic molecular network.
Assuntos
Neovascularização Patológica/metabolismo , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Transdução de Sinais , Acetilglucosaminidase/análise , Animais , Quimiocinas/metabolismo , Feminino , Hemoglobinas/análise , Inflamação/genética , Inflamação/metabolismo , Inflamação/patologia , Macrófagos/metabolismo , Camundongos , Camundongos Knockout , Infiltração de Neutrófilos/genética , Peroxidase/análise , Poliésteres , Poliuretanos , Próteses e Implantes , Receptores Tipo I de Fatores de Necrose Tumoral/genética , Transdução de Sinais/genética , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Creatinine plays a key role in the function and maturation of fetal kidneys throughout pregnancy. It is important to identify other markers that may help in the diagnosis of renal dysfunction. Our aim was to determine the profile of and the correlation between biochemical markers to be used to assess renal function and maturation of the fetus in the amniotic fluid during pregnancy and to determine the distribution of normal values for creatinine, N-acetyl-beta-D-glucosaminidase (NAG), beta2-microglobulin, glucose, urea, sodium, potassium, phosphorus, calcium, uric acid, albumin, and osmolality in three gestational age groups. This was a cross-section study that assessed 115 samples of amniotic fluid during three different periods of pregnancy, i.e., 13 to 20, 27 to 34, and 36 to 42 weeks. Concentrations of creatinine, NAG, urea, potassium and uric acid increased during pregnancy (P<0.05). Beta2-microglobulin, glucose, sodium, phosphorus, calcium, and albumin concentration and osmolality decreased (P<0.05), whereas beta2-microglobulin, glucose and uric acid presented significant correlations with gestational age and creatinine, respectively (r>0.6, P<0.05). Urea, potassium and phosphorus showed mild correlations with both (r>0.5, P<0.05). NAG, sodium, albumin and osmolality did not show significant correlations (r<0.5, P<0.05). These tests confirmed the important role of creatinine in terms of correlation with gestational age. beta2-Microglobulin, glucose and uric acid were significant as markers of function and maturation of fetal kidneys, whereas NAG did not demonstrate a useful role for the assessment of renal maturation.
Assuntos
Líquido Amniótico/química , Creatinina/análise , Desenvolvimento Embrionário e Fetal , Rim/embriologia , Acetilglucosaminidase/análise , Acetilglucosaminidase/fisiologia , Adulto , Biomarcadores , Estudos Transversais , Desenvolvimento Embrionário e Fetal/fisiologia , Feminino , Idade Gestacional , Glucose/análise , Glucose/fisiologia , Humanos , Rim/metabolismo , Rim/fisiologia , Gravidez , Sódio/análise , Sódio/fisiologia , Ácido Úrico/análise , Microglobulina beta-2/análise , Microglobulina beta-2/fisiologiaRESUMO
Increased activity of several lysosomal hydrolases was demonstrated in amniotic fluid from a fifteenth week pregnancy in which the fetus had I-cell disease. Cultured cells from amniotic fluid had a decreased activity of the same enzymes. The diagnosis of I-cell disease was later confirmed by enzyme assays in cell cultures of fetal skin and by morphologic studies of several tissues from the aborted fetus. Electron microscopic studies of the fetal tissues and cultured fibroblasts had large numbers of typical inclusions of I-cell disease, thus substantiating the diagnosis and intrauterine manifestation of the disease. The results indicate that prenatal diagnosis of I-cell disease is possible with enzyme assays of amniotic fluid and in cultures of fetal cells from the fluid. Enzyme studies of amniotic fluid can provide a preliminary diagnosis within a few hours, but it is suggested that the definitive diagnosis should be based on assays in cultured cells from amniotic fluid.