N-acetylglucosaminidase Research Articles

Effects of yak excreta on soil organic carbon mineralization and microbial communities in alpine wetlands of southwest of China

Improving knowledge of how soil organic carbon (SOC) mineralization responds to excreta application is essential to better understand whether wetland carbon (C) pools will react to grazing. We investigated microbial activity and community structure in the different treatments of excreta addition experiments to examine how soil C mineralization responds to the excreta input in terms of microbial activities and compositions in wetland soils. The microcosms of mineralization incubation of excreta addition were established. The structure of the microbial community was described by the fatty acid composition of the phospholipids (PLFA). The methylumbelliferyl-linked substrates (MUB) and l-dihydroxyphenylalanine (L-DOPA) substrates were used to investigate the activities of β-glucosidase (BG), N-acetyl-glucosaminidase (NAG), acid phosphatase (AP), cellobiohydrolase (CBH), and phenol oxidase (PO). Excreta addition altered the cumulative C mineralization in swamp meadow (SM) and peatland (PL) soils, but SM was lower than PL. Excreta addition increased the biomass of individual PLFA and the fungi/bacteria ratio, suggesting that microbes are stimulated by nutrients and that the soil microbial community composition is modified by excreta inputs. The hydrolytic enzyme activities were higher in the PL soils than in the SM soils, but the trend was opposite for PO activity. The changes in pH, fungi, actinomycetes (ACT), AP, and CBH after yak fecal input significantly influenced the soil CO2 efflux. Our findings suggest that yak grazing could influence the rate of C cycling in wetland soils by influencing microbial communities, enzyme activities, and soil pH. This study suggest that the yak excreta addition increased cumulative C mineralization in SM and PL soils, and the effect of dung addition was more significant than urine addition. The effect of yak excreta addition on SOC mineralization was related with the soil pH, microorganism structure, and enzyme activity which modified by the excreta addition. Soil pH, fungi, AP, and CBH were positively correlated with SOC mineralization, but ACT was negatively correlated with SOC mineralization. In addition, the changes in C and N sources with yak excreta addition play an important role in altering microbial enzyme activities. The input of yak feces into wetlands because of grazing could increase SOC mineralization and thereby promote C emission.

Beta-2 Adrenergic and Glucocorticoid Receptor Agonists Modulate Ozone-Induced Pulmonary Protein Leakage and Inflammation in Healthy and Adrenalectomized Rats.

We have shown that acute ozone inhalation activates sympathetic-adrenal-medullary and hypothalamus-pituitary-adrenal stress axes, and adrenalectomy (AD) inhibits ozone-induced lung injury and inflammation. Therefore, we hypothesized that stress hormone receptor agonists (β2 adrenergic-β2AR and glucocorticoid-GR) will restore the ozone injury phenotype in AD, while exacerbating effects in sham-surgery (SH) rats. Male Wistar Kyoto rats that underwent SH or AD were treated with vehicles (saline + corn oil) or β2AR agonist clenbuterol (CLEN, 0.2 mg/kg, i.p.) + GR agonist dexamethasone (DEX, 2 mg/kg, s.c.) for 1 day and immediately prior to each day of exposure to filtered air or ozone (0.8 ppm, 4 h/day for 1 or 2 days). Ozone-induced increases in PenH and peak-expiratory flow were exacerbated in CLEN+DEX-treated SH and AD rats. CLEN+DEX affected breath waveform in all rats. Ozone exposure in vehicle-treated SH rats increased bronchoalveolar lavage fluid (BALF) protein, N-acetyl glucosaminidase activity (macrophage activation), neutrophils, and lung cytokine expression while reducing circulating lymphocyte subpopulations. AD reduced these ozone effects in vehicle-treated rats. At the doses used herein, CLEN+DEX treatment reversed the protection offered by AD and exacerbated most ozone-induced lung effects while diminishing circulating lymphocytes. CLEN+DEX in air-exposed SH rats also induced marked protein leakage and reduced circulating lymphocytes but did not increase BALF neutrophils. In conclusion, circulating stress hormones and their receptors mediate ozone-induced vascular leakage and inflammatory cell trafficking to the lung. Those receiving β2AR and GR agonists for chronic pulmonary diseases, or with increased circulating stress hormones due to psychosocial stresses, might have altered sensitivity to air pollution.

Protective effects of breviscapine adjuvant therapy in renal injury caused by extracorporeal shock wave therapy for renal calculi

Objective To investigate the protective effects of breviscapine in renal injury caused by extracorporeal shock wave therapy for renal calculi. Methods Totally 120 patients with renal calculi treated in our hospital from January 2015 to January 2017 were selected and randomly divided into the control group and the study group with 60 cases in each group. The patients in the control group were treated with extracorporeal shock wave, and patients in the study group were treated with breviscapine assisted extracorporeal shock wave. Serum related factors [serum endothelin 1 (ET-1), malondialdehyde (MDA), kidney injury molecule-1 (Kim-1)], biochemical parameters of renal injury [blood urea nitrogen (BUN), β2 microglobulin (β2-MG), urinary N-acetyl β-D glucosaminidase (NAG)], clinical curative effect and prognosis [the World Health Organization Quality of Life Scale (WHOQOL-BRFE) score] were observed before and after treatment. Results There were no significant differences in ET-1, MDA, Kim-1, BUN, β2-MG, NAG and WHOQOL-BRFE scores between the two groups before treatment (P>0.05). After treatment, ET-1, MDA, BUN and WHOQOL-BRFE scores were significantly higher, and Kim-1, β2-MG and NAG were significantly lower than those before treatment, and the changes in the study group were more obvious than those in the control group (P<0.05). After treatment, the total effective rate of the study group was [90.00%(54/60)] was higher than that of the control group [75.00%(45/60)] (P<0.05). Conclusions Breviscapine adjuvant therapy is significantly effective in the treatment of patients with renal injury caused by extracorporeal shock wave therapy for renal calculi. It can improve the clinical curative effect, the prognosis and quality of life through effective regulation of related factors in serum biochemical indexes of renal injury. Key words: Kidney Calculi; ERIGERON BREVISCAPUS; Kidney; High-Energy Shock Waves

Changes of soil microbial and enzyme activities are linked to soil C, N and P stoichiometry in afforested ecosystems

Afforestation has been shown to strongly affect substrate stoichiometry and profoundly influence the microbial community. However, the degree to which microbial and activity are linked to soil carbon (C), nitrogen (N) and phosphorus (P) stoichiometry in afforested ecosystems remains unclear. In this study, soil samples were collected from Robinia pseudoacacia L. (RP42, RP27, and RP17) and farmland (FL) sites with a chronosequence of 42, 27, and 17 years. We determined the microbial biomass C (Cmic), N (Nmic), Cmic to organic C (Csoil) ratio (Cmic:Csoil), Nmic to total nitrogen (Nsoil) ratio (Nmic:Nsoil), and metabolic quotient (Rmic:Cmic) to investigate microbial. We also measured soil b-glucosidase (BG), N-acetylglucosaminidase (NAG), acid phosphatase (AP), as well as Csoil, Nsoil, and total phosphorus (Psoil). The results showed that, compared with FL, Csoil:Psoil and Nsoil:Psoil were increased during aggradation, whereas Csoil:Nsoil increased in RP27 and RP17 but decreased in RP42. Cmic:Csoil and Nmic:Nsoil were 94% and 182% higher in RP42 than FL, respectively. However, Rmic:Cmic was lower in RP42, RP27, and RP17 than FL by 64%, 36% and 25%, respectively. Moreover, we found that BG, NAG, AP, BG:NAG, and BG:AP in RP42, RP27, and RP17 were higher than in FL. Compared with global soil (0.62 and 0.13), the lower BG:AP (0.42) and BG:NAG (0.03) ratios in present study indicated that Psoil may be limited. Additionally, redundancy analysis (RDA) revealed that the Csoil: Nsoil was positively correlated with Rmic: Cmic but negatively with Cmic: Csoil and Nmic: Nsoil, while Csoil:Psoil and Nsoil:Psoil were also significantly and negatively correlated with BG:AP, BG:NAG, and NAG. Therefore, our results indicated that afforested ecosystem are highly susceptible to changes in soil microbial and enzyme stoichiometry during aggradation and may become P-limited. Such changes were modulated by soil nutrient stoichiometry.

Influence of mangrove roots on microbial abundance and ecoenzyme activity in sediments of a subtropical coastal mangrove ecosystem

Although the rhizosphere effect has been widely studied, the relevant information about how mangrove roots affect microbial and enzymatic activity was still lacking due to the particularity of mangroves and intertidal zone. In order to understand the effects of mangrove roots on microbial growth and the indicators of nutrient cycling, microbial abundance as well as extracellular enzyme activities were investigated in both rhizosphere and non-rhizosphere sediments of a coastal mangrove wetland ecosystem. Comparing to non-rhizosphere sediments, bacterial abundance was slightly lower, but fungal abundance was notably higher in the rhizosphere sediments. Moreover, β-glucosidase (GLU), N-acetyl-glucosaminidase (NAG) and acid phosphatase (ACP) activities were enhanced clearly, whereas phenol oxidase (PHO) activity was significantly reduced in the rhizosphere sediments. Interestingly, it was found that soluble phenolics were closely related to fungal abundance and PHO activity (p < 0.05). Both fungal abundance and PHO activity were also significantly correlated with each other (p < 0.05). Therefore, the strong correlation between fungal abundance and both phenolics and PHO activity might partially suggest that fungi might contribute to the reduction of soluble phenolics.

Tissue Indices of Telomere Length and p53 in Patients with Different Gastrointestinal Tumors: Correlation with Clinicopathological Status.

Telomere length dysfunction is involved in the generation of genomic rearrangements that drive progression to malignancy. A set of serological markers for telomere dysfunction, namely chitinase and N-acetylglucosaminidase (NAG), DNA damage, and tissue alteration of p53 have been identified. The probability that genomic damage, accumulation of reactive oxygen species, and shorter telomeres may be related to the onset and advancement of gastrointestinal (GI) tumors. A total of 40 patients with GI tumors and 20 healthy controls with matched age and sex were included. Estimation of serum chitinase, NAG, lipid peroxide (LPER), glutathione peroxidase (GPx), superoxide dismutase (SOD), catalase by colorimetric methods, and p53 by ELISA were assessed. Related clinicopathological features were determined. Serological chitinase, NO, LPER, and p53 were significantly increased, SOD was significantly decreased (p˂0.001 for each) in GI tumor patients compared with controls and correlated significantly with age. There was a significant correlation between telomere dysfunction indices, p53, oxidative stress indices, and malignant stages of GI cancer patients. Moreover, a significant difference in the mean serum levels of indices between control, malignant, and benign subjects was found. Accordingly, these biomarkers play an important role in the pathogenesis of GI cancer and their estimation may predict the GI tumor behavior.

Depth rather than microrelief controls microbial biomass and kinetics of C-, N-, P- and S-cycle enzymes in peatland

The formation of microrelief forms in peatlands - elevated and dry hummocks, depressed wet hollows and intermediate lawns - is controlled by the interaction of water table, nutrient availability and dominant plant communities. This affects the composition and activity of various functional groups of microorganisms. With depth, the change in peat quality from less to more highly processed organic material additionally regulates microbial activity. We hypothesized that microbial biomass and enzyme activities are driven by aeration and by peat quality and therefore (i) they increase from hollows (water saturated/anaerobic) through lawns (intermediate) to hummocks (aerobic) in the top peat and ii) they decrease with depth due to increasing distance from fresh plant-derived inputs and lower oxygen availability. These hypotheses were tested for enzymes catalysing the decomposition of C-, N-, P- and S-containing organic compounds in peat of the three microform types at three depths (15, 50 and 200 cm). Microbial biomass and peat chemical characteristics were compared with enzyme kinetic parameters, i.e. maximal potential activity (Vmax) and the Michaelis constant (Km).Microbial biomass carbon (MBC) and Vmax of β-glucosidase and N-acetyl glucosaminidase increased by 30–70% from hummocks and lawns to hollows in the top 15 cm, contradicting the hypothesis. Similarly, Km and the catalytic efficiency of enzymes (Ka = Vmax/Km) were best related to MBC distribution and not to the aeration gradient. With depth, Vmax of β-glucosidase, xylosidase and leucine aminopeptidase followed the hypothesized pattern in hollows. In contrast, MBC was 1.3–4 times higher at 50 cm, followed by successively lower contents at 15 and 200 cm in all microforms. The same depth pattern characterized the Vmax distribution of 6 out of 8 enzymes. Phosphatase activity decreased from drier hummock to wetter hollows and the higher activity throughout the peat profile suggested a high microbial demand for P. Enzyme activities and catalytic efficiency in peat were closely linked to the distribution of microbial biomass with depth, which in turn was best explained by P content. From the ecological perspective, these results clearly show that peat decomposition will be accelerated when microbial activity is stimulated e.g. by increased P availability.

Divergence of dominant factors in soil microbial communities and functions in forest ecosystems along a climatic gradient

Abstract. Soil microorganisms play an important role in regulating nutrient cycling in terrestrial ecosystems. Most of the studies conducted thus far have been confined to a single forest biome or have focused on one or two controlling factors, and few have dealt with the integrated effects of climate, vegetation, and soil substrate availability on soil microbial communities and functions among different forests. In this study, we used phospholipid-derived fatty acid (PLFA) analysis to investigate soil microbial community structure and extracellular enzymatic activities to evaluate the functional potential of soil microbes of different types of forests in three different climatic zones along the north–south transect in eastern China (NSTEC). Both climate and forest type had significant effects on soil enzyme activities and microbial communities with considerable interactive effects. Except for soil acid phosphatase (AP), the other three enzyme activities were much higher in the warm temperate zone than in the temperate and the subtropical climate zones. The soil total PLFAs and bacteria were much higher in the temperate zone than in the warm temperate and the subtropical zones. The soil β-glucosidase (BG) and N-acetylglucosaminidase (NAG) activities were highest in the coniferous forest. Except for the soil fungi and fungi–bacteria (F/B), the different groups of microbial PLFAs were much higher in the conifer broad-leaved mixed forests than in the coniferous forests and the broad-leaved forests. In general, soil enzyme activities and microbial PLFAs were higher in primary forests than in secondary forests in temperate and warm temperate regions. In the subtropical region, soil enzyme activities were lower in the primary forests than in the secondary forests and microbial PLFAs did not differ significantly between primary and secondary forests. Different compositions of the tree species may cause variations in soil microbial communities and enzyme activities. Our results showed that the main controls on soil microbes and functions vary in different climatic zones and that the effects of soil moisture content, soil temperature, clay content, and the soil N ∕ P ratio were considerable. This information will add value to the modeling of microbial processes and will contribute to carbon cycling in large-scale carbon models.

Costunolide suppresses an inflammatory angiogenic response in a subcutaneous murine sponge model.

Costunolide is known to possess anti-inflammatory and antitumor activity, but its role in tumor angiogenesis, the key step involved in tumor growth and metastasis, and the involved molecular mechanism is still unknown. We aimed to investigate the effects of costunolide on key components of inflammatory angiogenesis in the murine cannulated sponge implant angiogenesis model. Polyester-polyurethane sponges, used as a framework for fibrovascular tissue growth, were implanted in Swiss albino mice and costunolide (5, 10 and 20mg/kg/day) was administered for 14days through installed cannula. The implants collected at day 14 post-implantation were processed for the assessment of hemoglobin (Hb), myeloperoxidase (MPO), N-acetylglucosaminidase (NAG) and collagen, which were used as indices for angiogenesis, neutrophil and macrophage accumulation, and extracellular matrix deposition, respectively. Relevant inflammatory, angiogenic and fibrogenic cytokines were also determined. Costunolide treatment attenuated the main components of the fibrovascular tissue, wet weight, vascularization (Hb content), macrophage recruitment (NAG activity), collagen deposition, and the levels of vascular endothelial growth factor (VEGF), interleukin (IL)-1β, IL-6, IL-17, tumor necrosis factor (TNF)-α and transforming growth factor (TGF-β). Regulatory function of costunolide on multiple parameters of the main components of inflammatory angiogenesis has been revealed giving insight into the potential therapeutic benefit underlying the anti-angiogenic actions of costunolide.

氮磷添加对杉木林土壤碳氮矿化速率及酶动力学特征的影响

PDF HTML阅读 XML下载 导出引用 引用提醒 氮磷添加对杉木林土壤碳氮矿化速率及酶动力学特征的影响 DOI: 10.5846/stxb201608231718 作者: 作者单位: 作者简介: 通讯作者: 中图分类号: 基金项目: 国家自然科学基金项目（41571251，41571130043）；中科院技术创新项目（201604） Effects of nitrogen and phosphorus additions on soil organic carbon and nitrogen mineralization and hydrolase kinetics in Chinese fir plantations Author: Affiliation: Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:以江西杉木林红壤为研究对象，开展野外长期氮（N）、磷（P）添加控制试验，设置对照（CK）、N （50kg N hm-2 a-1）、P （50kg P hm-2 a-1）、NP （50kg N hm-2 a-1+50kg P hm-2 a-1）处理，分析N、P添加对土壤碳矿化速率（Cmin）、氮矿化速率（Nmin）和相关的β-1，4-葡萄糖苷酶（βG）和β-1，4-N-乙酰葡糖氨糖苷酶（NAG）动力学参数的影响。结果表明：（1） N添加明显降低了Cmin和Nmin，比CK分别减少了25%和18%；N添加减小了NAG的潜在最大酶活性（Vmax）、半饱和常数（Km）、催化效率（Vmax/Km），但差异不显著（P>0.05）；N添加显著增加了βG的Vmax、Km，但对Vmax/Km有抑制作用。（2） P输入（P、NP）较CK使NAG的Vmax、Km减小26%-60%；NP同时添加明显提高βG和NAG的Vmax/Km（P<0.05），但P输入（P、NP）对Cmin和Nmin影响不显著（P>0.05）。（3）Cmin与土壤溶解性有机碳正相关，Nmin与pH显著正相关，与土壤NH4+-N、NO3--N显著负相关；βG和NAG的Vmax/Km均与NH4+-N、NO3--N负相关（P<0.05），Km均与NH4+-N、NO3--N正相关（P<0.05）。βG的Vmax与NH4+-N、NO3--N正相关（P<0.05），NAG的Vmax与有机碳、全氮、全磷、有效磷负相关（P<0.05）。研究结果表明，在亚热带杉木人工林中，N添加降低土壤pH，增加土壤有效氮含量，抑制βG和NAG的Vmax/Km，对土壤Cmin和Nmin产生抑制作用；而NP添加增加土壤有效磷含量，增加土壤βG和NAG的Vmax/Km。 Abstract:A long-term field experiment of nitrogen (N) and phosphorus (P) additions was established in Chinese fir plantations in subtropical China. Four treatments, i.e. control (CK), N (50kg N hm-2 a-1), P (50kg P hm-2 a-1), and NP (50kg N hm-2 a-1+ 50kg P hm-2 a-1),were selected in this study. We studied the effects of N and P additions on the soil mineralization rate of carbon (Cmin) and nitrogen (Nmin) and the kinetics of β-glucosidase(βG)and N-acetyl-glucosaminidase(NAG). The results showed that (1) N additions had negative effects on Cmin and Nmin, which were lower by 25% and 18%, respectively, compared to CK. (2) P (P, NP) additions decreased the Vmax and Km of NAG by 26%-60% compared to CK, whereas NP combined additions greatly enhanced the catalytic efficiency (Vmax/Km) of βG and NAG (P<0.05). (3) Cmin was positively correlated with dissolved organic carbon (DOC) contents, whereasNmin was positively correlated with pH and negatively correlated with NH4+-N and NO3--N contents. The catalytic efficiencies of βG and NAG were negatively correlated with NH4+-N and NO3--N contents (P<0.05). The Km of βG and NAG was positively correlated with NH4+-N and NO3--N (P<0.05), The Vmax of βG was positively correlated with NH4+-N and NO3--N (P<0.05), and the Vmax of NAG was negatively correlated with soil organic carbon, total nitrogen, total phosphorus, and available contents (P<0.05). Our results suggest that N additions had negative effects on Cmin and Nmin by decreasing soil pH, increasing soil available N contents, and inhibiting the Vmax/Km of βG and NAG, whereas NP additions increased soil available P and the Vmax/Km of βG and NAG in southern subtropical Chinese fir plantations. 参考文献 相似文献 引证文献

Renoprotective Effect of Platelet-Rich Plasma on Cisplatin-Induced Nephrotoxicity in Rats.

Platelet-rich plasma (PRP) has grown as an attractive biologic instrument in regenerative medicine for its powerful healing properties. It is considered as a source of growth factors that may induce tissue repairing and improve fibrosis. This product has proven its efficacy in multiple studies, but its effect on cisplatin-induced nephrotoxicity has not yet been elucidated. The present investigation was performed to estimate the protective impact of platelet-rich plasma against cisplatin- (CP-) evoked nephrotoxicity in male rats. Nephrotoxicity was induced in male Wistar rats by right uninephrectomy followed by CP administration. Uninephrectomized rats were assigned into four groups: (1) control group, (2) PRP group, (3) CP group, and (4) CP + PRP group. PRP was administered by subcapsular renal injection. Renal function, inflammatory cytokines, and growth factor level as well as histopathological investigation were carried out. Treatment with PRP attenuated the severity of CP-induced nephrotoxicity as evidenced by suppressed creatinine, blood urea nitrogen (BUN), and N-acetyl glucosaminidase (NAG) levels. Moreover, PRP depressed intercellular adhesion molecule-1 (ICAM-1), kidney injury molecule-1 (KIM-1), caspase-3, and transforming growth factor-beta 1 (TGF-β1) levels, while enhanced the epidermal growth factor (EGF) level. These biochemical results were reinforced by the histopathological investigation, which revealed restoration of normal renal tissue architectures. These findings highlight evidence for the possible protective effects of PRP in a rat model of CP-induced nephrotoxicity, suggesting a new avenue for using PRP to improve the therapeutic index of cisplatin.

Variations in eco-enzymatic stoichiometric and microbial characteristics in paddy soil as affected by long-term integrated organic-inorganic fertilization.

To investigate the effects of different nutrient management regimes on the soil chemical, eco-enzymatic stoichiometric and microbial characteristics, soil samples were collected from a 30-year, long-term field experiment with six plots growing rice. The results showed that as integrated fertilization increased, so did the concentrations of soil total or available nutrients and microbial biomass carbon (MBC). Our results also found enhanced soil basal respiration and cumulative carbon mineralization compared to chemical fertilization alone at the same nutrient doses. The activities of soil protease (Pro), β-glucosidase (βG), N-acetyl-glucosaminidase (NAG) and acid phosphatase (AP) from the integrated fertilization treatments were significantly higher than those of the treatments without organic manure, so did the activities of soil leucyl aminopeptidase (LAP) and urease (Ure) from the treatment with organic manure in addition to farmer practise fertilization (NPKM2). The stoichiometric ratios, expressed as lnβG/ln(NAG+LAP)/lnPro/lnUre/lnAP, ranged from 1:0.94:1.04:0.67:1.01 to 1:0.98:1.10:0.78:1.25, indicating that the acquisition of C, N and P changed consistently and synchronously under different nutrient management strategies. Integrated fertilization was more beneficial to the acquisition and utilization of soil organic carbon compared to low-molecular-weight organic nitrogen. We concluded that protease and urease should be considered in eco-enzymatic stoichiometric assessments for the hydrolysis of proteins, amino acids, carbohydrates and phosphomonoesters in soil, and integrated fertilization with chemical fertilizers and organic manure should be recommended as a preferable nutrient management system for intensive rice cultivation.

Adrenergic and glucocorticoid receptor antagonists reduce ozone-induced lung injury and inflammation

Recent studies showed that the circulating stress hormones, epinephrine and corticosterone/cortisol, are involved in mediating ozone-induced pulmonary effects through the activation of the sympathetic-adrenal-medullary (SAM) and hypothalamus-pituitary-adrenal (HPA) axes. Hence, we examined the role of adrenergic and glucocorticoid receptor inhibition in ozone-induced pulmonary injury and inflammation. Male 12-week old Wistar-Kyoto rats were pretreated daily for 7days with propranolol (PROP; a non-selective β adrenergic receptor [AR] antagonist, 10mg/kg, i.p.), mifepristone (MIFE; a glucocorticoid receptor [GR] antagonist, 30mg/kg, s.c.), both drugs (PROP+MIFE), or respective vehicles, and then exposed to air or ozone (0.8ppm), 4h/d for 1 or 2 consecutive days while continuing drug treatment. Ozone exposure alone led to increased peak expiratory flow rates and enhanced pause (Penh); with greater increases by day 2. Receptors blockade minimally affected ventilation in either air- or ozone-exposed rats. Ozone exposure alone was also associated with marked increases in pulmonary vascular leakage, macrophage activation, neutrophilic inflammation and lymphopenia. Notably, PROP, MIFE and PROP+MIFE pretreatments significantly reduced ozone-induced pulmonary vascular leakage; whereas PROP or PROP+MIFE reduced neutrophilic inflammation. PROP also reduced ozone-induced increases in bronchoalveolar lavage fluid (BALF) IL-6 and TNF-α proteins and/or lung Il6 and Tnfα mRNA. MIFE and PROP+MIFE pretreatments reduced ozone-induced increases in BALF N-acetyl glucosaminidase activity, and lymphopenia. We conclude that stress hormones released after ozone exposure modulate pulmonary injury and inflammatory effects through AR and GR in a receptor-specific manner. Individuals with pulmonary diseases receiving AR and GR-related therapy might experience changed sensitivity to air pollution.

Relevance of cystatin-C, N-acetylglucosaminidase, and Interleukin-18 with the diagnosis of acute kidney injury induced by cadmium in rats.

The diagnosis of acute kidney injury (AKI) is frequently established on modifications in serum creatinine (SCr). The discriminative and prognostic aptitudes of serum cystatin-C as well as N-acetylglucosaminidase (NAG) and Interleukin-18 (IL-18) were inspected for the estimation of AKI. In this study twelve rats were alienated into two groups: control group received saline, second group received cadmium chloride at a dose (2.4mg Cd/kg/day, i.p) for 30 days. Blood urea nitrogen (BUN), SCr, and IL-18 serum level were measured in addition to serum and tissue content of cystatin-C and NAG. AKI model showed significant increase in BUN, creatinine, and IL-18. RT-PCR showed upregulation of cystatin-C gene besides significant increase of its level in serum. Additionally, tissue content of NAG was significantly increased. Our findings may provide that grouping of several biomarkers for diagnosis of AKI is a more valuable diagnostic tool than single-marker measurement.

Ensemble variant interpretation methods to predict enzyme activity and assign pathogenicity in the CAGI4 NAGLU (Human N-acetyl-glucosaminidase) and UBE2I (Human SUMO-ligase) challenges.

CAGI (Critical Assessment of Genome Interpretation) conducts community experiments to determine the state of the art in relating genotype to phenotype. Here, we report results obtained using newly developed ensemble methods to address two CAGI4 challenges: enzyme activity for population missense variants found in NAGLU (Human N-acetyl-glucosaminidase) and random missense mutations in Human UBE2I (Human SUMO E2 ligase), assayed in a high-throughput competitive yeast complementation procedure. The ensemble methods are effective, ranked second for SUMO-ligase and third for NAGLU, according to the CAGI independent assessors. However, in common with other methods used in CAGI, there are large discrepancies between predicted and experimental activities for a subset of variants. Analysis of the structural context provides some insight into these. Post-challenge analysis shows that the ensemble methods are also effective at assigning pathogenicity for the NAGLU variants. In the clinic, providing an estimate of the reliability of pathogenic assignments is the key. We have also used the NAGLU dataset to show that ensemble methods have considerable potential for this task, and are already reliable enough for use with a subset of mutations.

Correlation analysis of renal ultrasound elastography and clinical and pathological changes in patients with chronic kidney disease .

To analyze the correlations of renal tissue elastography with clinical biochemical indicators and pathological changes in patients with chronic kidney disease (CKD) as well as to explore the potential for renal tissue elastography as a new, noninvasive method for the dynamic monitoring of renal disease progression, efficacy assessment, and prognosis evaluation. Patients admitted to the Department of Nephrology of the First Affiliated Hospital of China Medical University from August 2014 to January 2015 who had undergone renal biopsies were selected. A total of 113 patients with CKD and 16 healthy controls were enrolled in this study, including 61 males and 52 females. In total, 23 cases of IgA nephropathy, 39 cases of membranous nephropathy, 15 cases of minimal-change nephropathy (MCN), and 7 cases of focal segmental glomerulosclerosis were included. The Young's moduli (YM) of the renal cortex and medulla were measured using an AixPlorer Doppler ultrasound with full digital color from Supersonic Imagine. The correlations between the YM of renal tissue and clinical biochemical indicators of blood and urine and the differences in Young's moduli among the different pathological changes in the patients with CKD were analyzed. The YM of the CKD patients was significantly higher than that of the control group (p<0.05), and the YM of the renal cortex and medulla gradually increased with the progression of CKD. The YM of the renal cortex in the stage-G5 CKD patients was significantly higher than that of the CKD patients in stages G1 - G3 (p<0.05). The YM of the renal medulla of the CKD patients in stages G3-5 was significantly higher than that of the CKD patients in stages G1-G2. On univariate analysis, the YM of the renal cortex was correlated with systolic blood pressure, serum creatinine, cystatin C, serum albumin, serum phosphorus, calcium and phosphorus products, uric acid, iPTH, urinary N-acetyl-glucosaminidase (NAG), eGFR, and hemoglobin levels. And the YM of the renal medulla was correlated with systolic blood pressure, serum creatinine, serum albumin, uric acid, iPTH, urinary microalbumin (MA), urinary NAG, and hemoglobin levels. On multivariate analysis, serum cystatin C (β=0.485, p=0.018) and uric acid (β=0.418, p=0.039) levels were independently correlated with the YM of the renal cortex, while serum creatinine (β=0.380, p=0.019) and uric acid (β=0.482, p=0.004) levels, as well as smoking (β=0.337, p=0.009), were independently correlated with the YM of the renal medulla. The YMs of the renal cortex in patients with membranous nephropathy and IgA nephropathy were significantly higher than those in the patients with CN (p<0.05). The YM of the renal cortices of the patients in phases IV and V of IgA nephropathy based on the Lee grading system were significantly higher than those of the patients in phases II and III (p<0.05). According to the Oxford classification for IgA nephropathy, the Young's moduli of the renal cortex and medulla in T1 and T2 patients were significantly higher than those in T0 patients (p<0.05). The YM of the renal cortex and medulla showed no statistically-significant differences among the different stages of membranous nephropathy. The YM of the renal cortex and medulla are associated with the progression of renal insufficiency, and renal ultrasound elastography shows promise as a new means of assessing the stage of CKD. Renal ultrasound elastography is expected to become a new, noninvasive method for the early diagnosis of CKD and the dynamic monitoring of disease progression as well as the assessment of efficacy and prognosis. .

Relationship of Urinary Biomarkers NAG, NGAL and KIM-1 to Kidney Transplantation Complications

Introduction: Kidney transplantation (KT) is the best choice for patients that suffer End Stage Renal Disease (ESRD), because of the better survival rate compared to patients under dialysis. Our objective should be to improve allograft survival by acting in a preemptive way. Methods: We analysed the urinary biomarkers NAG (N-acetyl glucosaminidase), NGAL (Neutrophil Gelatinase associated Lipocaine) and KIM-1 (Kidney Injury Molecule-1) in urine samples from 70 kidney or kidney and pancreas transplant patients. The samples were taken at post-transplant days +1, +3, +5, +7, and the day of renal stabilisation in the first 3 months. The biomarker levels were related to the different complications after KT, such as acute rejection, delayed graft function (DGF), cold ischemia time, toxic drugs for kidney and the renal function on the day of stabilisation. The statistical test used was Pearson c2. The 70th percentile was taken and a significant value was established if p < 0.05. The statistical program used was IBM SPSS version 20. Results: During the first week after KT, in theory renal function improves as measured by creatinine levels. However, increasing levels of the urinary biomarkers NAG, NGAL and KIM-1 indicate possible tubular damage or other pathology of the kidney. NGAL has shown to be a more sensitive marker than NAG and KIM-1 because it increases more than the other markers from the first day after KT. Urinary values of NGAL at post-transplant day 7 predict statistically significant probability of suffering DGF. Low NAG values at post-transplant day 3 have been related to a cold ischemia time of between 120 and 900 minutes, while high values in the first week post-transplant predict more renal function stabilisation time; the same occurs with NGAL. In this study KIM-1 did not show any relationship with the events described above. Conclusions: The use of the urinary biomarkers NAG and NGAL can help us act in a preemptive way to avoid the complications of KT. This could improve the global results of renal transplantation.Figure

Chronic Nitrogen Enrichment at the Watershed Scale Does Not Enhance Microbial Phosphorus Limitation

Increased N inputs through chronic atmospheric deposition has enriched temperate forest ecosystems, altering critical ecosystem functions such as decomposition and potentially resulting in a shift to P limitation. We used a combination of microbial biomass stoichiometry and enzymatic activity analyses to evaluate the potential for microbial nutrient limitation over the course of a growing season in response to multi-decadal, whole-watershed N enrichments and a one time, plot-scale P addition that occurred in the 22nd year of whole-watershed treatments. The one-time P addition increased microbial biomass threefold and reduced N-acetyl-glucosaminidase (NAG) and acid phosphatase (AP) activity 1 week after application, but there was no interaction with long-term experimental N enrichment to indicate a shift to P limitation. However, both N and P treatments increased C limitation independently of each other over the duration of the study based on measured increases in β-1,4-glucosidase (BG) activity relative to NAG and AP. Microbial biomass stoichiometry and enzyme activity indicated that BBWM is P limited regardless of N status. Our findings highlight the complex interactions between C, N, and P use and limitation in a forested ecosystem subjected to long-term N enrichment.

Angiogenenic effects of BpLec, a C-type lectin isolated from Bothrops pauloensis snake venom

The present work reports the effects of a C-type lectin (BpLec) isolated from Bothrops pauloensis snake venom upon in vitro and in vivo angiogenesis models. Initially, we noted that BpLec was not cytotoxic to endothelial cells (tEnd) in doses up to 40μg/mL, but lower doses (2.5μg/mL, 5μg/mL, 10μg/mL and 20μg/mL) reduced tEnd cells adhesion to some extracellular matrix proteins and inhibited the in vitro vessel formation in Matrigel assay stimulated by bFGF. β-galactosides (d-lactose, N-acetyl-d-galactosamine and d-galactose) at 400mM reversed the effect of BpLec on tEnd cells adhesion, whereas d-galactose (400mM) partially reversed BpLec property of inhibiting vessel formation by tEnd cells in Matrigel. In vivo assays showed that BpLec increased hemoglobin content and capillary vessels number in polyether-polyurethane sponge discs subcutaneously implanted into dorsal skin mice. Additionally, BpLec also reduced collagen deposition and did not induce a pro-inflammatory response, as demonstrated by the decreased the secretion of some inflammatory cytokines, whereas myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activities were not altered by BpLec. Taken together, our results indicate that BpLec might represent an interesting angiogenesis and inflammatory modulator that could also be used for searching possible therapeutic targets involved in these processes.

Identification of Glycoproteins on the Surface of Bovine Milk Exosomes and Intestinal Cells that Facilitate Exosome Uptake in Human Colon Carcinoma Caco‐2 Cells

BackgroundExosomes are small, cargo‐containing vesicles that are secreted by donor cells to elicit changes in gene expression and metabolism in recipient cells. In previous studies, we provided evidence that, in addition to endogenous synthesis, exosomes may also be obtained from dietary sources such as bovine milk, and intestinal transport is facilitated by endocytosis.HypothesisGlycoproteins on the surface of bovine milk exosomes and human intestinal cells play essential roles in exosome transport.MethodsExosomes were isolated from bovine milk by differential centrifugation and authenticated as recommended by International Society for Extracellular Vesicles (Fig. 1). Surface proteins or glycan modifications in exosomes or Caco‐2 cells were removed using proteases (Glu‐C, trypsin, Arg‐C, Asp‐N, or proteinase K) or glycosidases [N‐glycosidase F (PNGase F), galactosidase, O‐glycosidase, neuraminidase, N‐acetyl glucosaminidase, or a combination of all]. Controls were incubated with solvent. Exosomes were labeled with FM4–64, and unlabeled fluorophore was removed. Surface peptides released after the treatment were identified using LC/MS‐MS, Mascot and Sequest databases. Glycoproteins on the external surface were identified using a series of tools based on neuronal networks including NetNglyc, NetOglyc, NetCglyc 1.0 and GlycoEP. Transport kinetics were modelled using the Michaelis Menten equation. One‐way ANOVA and Bonferroni's multiple comparison were used for statistical analyses.ResultsWhen exosomes or cells were treated with individual proteases (Fig. 2) or glycosidases (Fig. 3), exosome uptake decreased up to 80% when compared to controls even at prolonged incubation times. Treatment with galactosidase, PNGase F and O‐glycosidase had a stronger effect than other glycosidases. When exosomes or cells were treated with the glycosidase mixture, no uptake was detected or uptake was decreased by 80%, respectively, in comparison to controls (Fig. 4). 149 total proteins were identified in bovine exosome, including 4 (N), 2 (O) and 2 (C) glycan binding sites; 298 proteins were identified in Caco‐2 cells, including 46 membrane proteins that included 29 (N), 33 (O) and 6 (C) glycan binding sites.ConclusionsGlycoproteins on the surface of bovine milk exosomes and intestinal cells facilitate the uptake of exosomes in human intestinal cells. The β‐galactoside, Core 1 &amp; Core 3‐O‐linked disaccharide and N‐acetylglucosamine modifications appear to be of greater importance than other glycan features present on complex glycoproteins.Future DirectionsThe transport of bovine exosomes will be studied in conditional glycan knockout mice.Support or Funding InformationNIFA 2015‐67017‐23181, NIFA 2016‐67001‐25301/NIH DK107264, NIH 1P20GM104320, the Gerber Foundation, and USDA Hatch Act and W3002.