Amylases

淀粉酶
  • 文章类型: Journal Article
    先前在一项无症状中年人的小型研究中已经描述了低血清淀粉酶与血浆胰岛素水平降低和肥胖增加之间的潜在关联。在本研究中,我们试图确定在非人类灵长类动物(NHP)从代谢正常到明显的2型糖尿病(T2DM)的纵向发展过程中这些变化的性质,一种在病理生理学和潜在机制上似乎与中年成年人中最常见的疾病相同的疾病。在157只无关的成年恒河猴(猕猴)中表征了淀粉酶和脂肪酶水平;38%的人在研究中发展为T2DM。在所有的猴子,多元线性回归分析显示,淀粉酶可以通过%体脂(β-0.29;p=0.002)负预测,年龄(β-0.27;p=0.005),和HbA1c(β-0.18;p=0.037)。在研究中包括的所有NHP中,淀粉酶水平由脂肪酶水平(β=0.19;p=-0.024)正预测。在代谢综合征的NHPs中,淀粉酶显著降低(p<0.001),糖尿病前期(PreDM)(p<0.001),与代谢正常的成人NHP相比,T2DM(p<0.001)。与正常NHP相比,具有PreDM(p=0.005)和T2DM(p=0.04)的NHP中的脂肪酶增加。这是对任何物种的第一次纵向研究,包括人类,为了显示从正常到代谢综合征的代谢过程中淀粉酶和脂肪酶的动力学,到PreDM,然后到明显的T2DM。人类和猴子在T2DM中的惊人相似性,在胰腺病理生理学和代谢功能中,这些发现具有很高的翻译价值。
    Latent associations between low serum amylase and reduced plasma insulin levels and increased adiposity have been described previously in a small study of asymptomatic middle-aged humans. In the present study, we sought to determine the nature of such changes during the longitudinal progression from metabolically normal to overt type 2 diabetes mellitus (T2DM) in nonhuman primates (NHPs), a disease that appears to be the same in both pathophysiology and underlying mechanisms as that which most commonly develops in middle-aged adult humans. Amylase and lipase levels were characterized in 157 unrelated adult rhesus monkeys (Macaca mulatta); 38% developed T2DM while under study. In all monkeys, multivariable linear regression analysis revealed that amylase could be negatively predicted by % body fat (β -0.29; p = 0.002), age (β -0.27; p = 0.005), and HbA1c (β -0.18; p = 0.037). Amylase levels were positively predicted by lipase levels (β = 0.19; p = -0.024) in all NHPs included in the study. Amylase was significantly lower in NHPs with metabolic syndrome (p < 0.001), prediabetes (PreDM) (p < 0.001), and T2DM (p < 0.001) compared to metabolically normal adult NHPs. Lipase increased in NHPs with PreDM (p = 0.005) and T2DM (p = 0.04) compared to normal NHPs. This is the first longitudinal study of any species, including humans, to show the dynamics of amylase and lipase during the metabolic progression from normal to metabolic syndrome, to PreDM and then to overt T2DM. The extraordinary similarity between humans and monkeys in T2DM, in pancreatic pathophysiology and in metabolic functions give these findings high translational value.
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  • 文章类型: Journal Article
    这项研究调查了饲喂淀粉酶启用的玉米青贮(ACS)对泌乳奶牛的性能和肠道气体排放的影响。在2周的协变量周期之后,在10周的随机完整区组设计实验中,将48只哺乳期中期的荷斯坦奶牛分配到3种处理中的1种。处理是含有相同比例的玉米青贮饲料(饮食DM的40%)的饮食如下:(1)常规杂交玉米青贮饲料对照(CON),(2)ACS取代控制青贮饲料(ADR),和(3)ADR日粮用磨碎的玉米粒代替大豆壳,以达到与CON(ASR)相同的日粮淀粉浓度。对照玉米青贮饲料和ACS在同一天收获,并含有40.3%和37.1%DM和(DM的%):37.2%和41.0%NDF以及37.1%和30.0%淀粉,分别。使用GreenFeed系统测量肠内气体排放。在协变量期间,由于与健康相关的问题,两头母牛被淘汰。使用口胃瘤胃取样技术从24头母牛(每个处理8头)收集瘤胃液。与CON相比,饲喂ADR的奶牛在第3、4和9周的实验期间增加了DMI,但处理不影响牛奶或ECM牛奶产量(平均39.0kg/d;SEM=0.89).与CON相比,饲料效率(每单位牛奶,但不是ECM)的ADR倾向于较低,而ASR的牛奶真蛋白浓度(趋势)和产量较低。相对于CON,ADR和ASR饮食均降低了牛奶尿素N。与CON相比,ADR增加了每日CH4排放量和排放强度,而ASR不增加。与对照玉米青贮相比,用ACS配制的两种饮食都倾向于增加原生动物总数。ASR的饮食NDF的总道消化率更高,ADR和ASR的ADF比CON更大。ADR增加了乙酸盐的摩尔比例(趋势)和乙酸盐与丙酸盐的比例,但不是ASR,与CON相比。在实验的最初几周,用ACS(具有较低的淀粉浓度)在奶牛的日粮中替换CON会增加STI。维护的ECM,倾向于降低饲料效率,增加了肠道CH4的排放,可能是由于易消化纤维的摄入量增加,与CON相比。
    This study investigated the effects of feeding an amylase-enabled corn silage (ACS) on the performance and enteric gas emissions in lactating dairy cows. Following a 2-wk covariate period, 48 mid-lactation Holstein cows were assigned to 1 of 3 treatments in a 10-wk randomized complete block design experiment. Treatments were diets containing the same proportion of corn silage (40% of dietary DM) as follows: (1) a conventional hybrid corn silage control (CON), (2) ACS replacing the control silage (ADR), and (3) the ADR diet replacing soybean hulls with ground corn grain to achieve the same dietary starch concentration as CON (ASR). Control corn silage and ACS were harvested on the same day and contained 40.3% and 37.1% DM and (% of DM): 37.2% and 41.0% NDF and 37.1% and 30.0% starch, respectively. Enteric gas emissions were measured using the GreenFeed system. Two cows were culled due to health-related issues during the covariate period. Ruminal fluid was collected from 24 cows (8 per treatment) using the orogastric ruminal sampling technique. When compared with CON, cows fed ADR had increased DMI during experimental wk 3, 4, and 9, but treatment did not affect milk or ECM milk yields (39.0 kg/d on average; SEM = 0.89). Compared with CON, feed efficiency (per unit of milk, but not ECM) tended to be lower for ADR, whereas milk true protein concentration (a tendency) and yield were lower for ASR. Milk urea N was decreased by both ADR and ASR diets relative to CON. Compared with CON, daily CH4 emission and emission intensity were increased by ADR but not ASR. Total protozoal count tended to be increased by both diets formulated with ACS when compared with control corn silage. Total-tract digestibility of dietary NDF was greater for ASR, and that of ADF was greater for both ADR and ASR versus CON. The molar proportion of acetate (a tendency) and acetate-to-propionate ratio were increased by ADR, but not ASR, when compared with CON. Replacement of CON with ACS (having lower starch concentration) in the diet of dairy cows increased DMI during the initial weeks of the experiment, maintained ECM, tended to decrease feed efficiency, and increased enteric CH4 emissions, likely due to increased intake of digestible fiber, compared with CON.
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  • 文章类型: Journal Article
    急性胰腺炎(AP)是一种复杂的炎症状态,可导致全身炎症反应和多器官功能障碍。这项研究调查了半乳糖凝集素-3(Gal-3)的作用,β-半乳糖苷结合凝集素,调节AP的获得性免疫反应。在野生型和半乳糖凝集素-3缺陷型C57BL/6小鼠中,通过胆管结扎诱发急性胰腺炎。我们确定了炎症细胞的表型和分子特征,血清淀粉酶浓度,胰腺胰蛋白酶活性,胰腺和肺部病理学。半乳糖凝集素-3缺乏减少CD3+CD49-T细胞和CD4+T辅助细胞的总数,下调炎性细胞因子和IFN-γ的产生,并增加了患病动物胰腺中产生IL-10的Foxp3T调节性细胞和调节性CD4T细胞的积累。半乳糖凝集素-3的缺失改善了以降低血清淀粉酶浓度和胰腺胰蛋白酶活性为特征的急性胰腺炎,和减弱肺的组织病理学。这些发现揭示了半乳糖凝集素-3在急性胰腺炎的获得性免疫应答中的作用,并确定半乳糖凝集素-3是研究疾病免疫发病机制的有吸引力的靶标,并被认为是患有急性炎症性疾病的患者的潜在治疗靶标。胰腺。
    Acute pancreatitis (AP) is a complex inflammatory condition that can lead to systemic inflammatory responses and multiple organ dysfunction. This study investigates the role of Galectin-3 (Gal-3), a β-galactoside-binding lectin, in modulating acquired immune responses in AP. Acute pancreatitis was induced by ligation of the bile-pancreatic duct in wild-type and Galectin-3-deficient C57BL/6 mice. We determined the phenotypic and molecular features of inflammatory cells, serum concentrations of amylase, pancreatic trypsin activity, and pancreatic and lung pathology. Galectin-3 deficiency decreased the total number of CD3+CD49- T cells and CD4+ T helper cells, downregulated the production of inflammatory cytokine and IFN-γ, and increased the accumulation of IL-10-producing Foxp3+ T regulatory cells and regulatory CD4+ T cells in the pancreata of diseased animals. The deletion of Galectin-3 ameliorates acute pancreatitis characterized by lowering serum amylase concentration and pancreatic trypsin activity, and attenuating of the histopathology of the lung. These findings shed light on the role of Galectin-3 in acquired immune response in acute pancreatitis and identify Galectin-3 as an attractive target for investigation of the immunopathogenesis of disease and for consideration as a potential therapeutic target for patients with acute inflammatory disease of the pancreas.
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  • 文章类型: Journal Article
    嗜热放线菌通常在极端环境中发现,可以茁壮成长并适应极端条件。这些生物体由于其显著的酶活性而表现出显著的变异并获得显著的兴趣。这项研究评估了灰链霉菌NBR14和诺卡氏菌NBRM9菌株通过使用小麦和豆秸秆的深层发酵产生热稳定淀粉酶的潜力。利用Box-Behnken设计来确定淀粉酶生物合成的最佳参数。随后,淀粉酶进行了部分纯化和表征。此外,将所得的水解物用于酿酒酵母的乙醇发酵。发现NBR14(7.72U/mL)和NBRM9(26.54U/mL)菌株获得最高淀粉酶活性的最佳参数为40和30°C,pH值7,孵育时间7天,和底物浓度(3和2g/100mL),分别。对NBR14和NBRM9淀粉酶进行了部分纯化,产生251.15和144.84U/mg的特定活性,以及3.91和2.69倍的纯化因子,分别。部分纯化后,从NBR14和NBRM9中提取的淀粉酶在9和7的pH值以及50和60°C的温度下显示出最高的活性水平,分别。结果还表明,NBR14和NBRM9淀粉酶的最大速度(Vmax)分别为57.80和59.88U/mL,分别,Km常数为1.39和1.479mM。48小时后,生物乙醇的生产浓度分别为5.95mg/mL和9.29mg/mL,分别,通过酿酒酵母发酵。嗜热放线菌及其α-淀粉酶产量显示出从农业副产品可持续生产生物乙醇的潜力。
    Thermophilic actinomycetes are commonly found in extreme environments and can thrive and adapt to extreme conditions. These organisms exhibit substantial variation and garnered significant interest due to their remarkable enzymatic activities. This study evaluated the potential of Streptomyces griseorubens NBR14 and Nocardiopsis synnemataformans NBRM9 strains to produce thermo-stable amylase via submerged fermentation using wheat and bean straw. The Box-Behnken design was utilized to determine the optimum parameters for amylase biosynthesis. Subsequently, amylase underwent partial purification and characterization. Furthermore, the obtained hydrolysate was applied for ethanol fermentation using Saccharomyces cerevisiae. The optimal parameters for obtaining the highest amylase activity by NBR14 (7.72 U/mL) and NBRM9 (26.54 U/mL) strains were found to be 40 and 30 °C, pH values of 7, incubation time of 7 days, and substrate concentration (3 and 2 g/100 mL), respectively. The NBR14 and NBRM9 amylase were partially purified, resulting in specific activities of 251.15 and 144.84 U/mg, as well as purification factors of 3.91 and 2.69-fold, respectively. After partial purification, the amylase extracted from NBR14 and NBRM9 showed the highest activity level at pH values of 9 and 7 and temperatures of 50 and 60 °C, respectively. The findings also indicated that the maximum velocity (Vmax) for NBR14 and NBRM9 amylase were 57.80 and 59.88 U/mL, respectively, with Km constants of 1.39 and 1.479 mM. After 48 h, bioethanol was produced at concentrations of 5.95 mg/mL and 9.29 mg/mL from hydrolyzed wheat and bean straw, respectively, through fermentation with S. cerevisiae. Thermophilic actinomycetes and their α-amylase yield demonstrated promising potential for sustainable bio-ethanol production from agro-byproducts.
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  • 文章类型: Journal Article
    真菌会破坏大多数烘焙产品。蛋糕在储存过程中的变质通常与真菌有关。因此,这项研究旨在评估市场上出售的不同类型蛋糕的质量。测试的蛋糕样品(14个样品)中最主要的真菌属是曲霉属。,和青霉菌属。马铃薯葡萄糖琼脂(PDA),培养基真菌总数为43.3菌落/g。曲霉属是最主要的属,是从六个蛋糕样品中分离出来的。曲霉由3种代表,即,A.黄花,A.尼日尔,和A.Nidulans,分别由13.32、19.99和3.33个菌落/g表示。在麦芽提取物琼脂(MEA)培养基上,真菌总数为123.24菌落/g。曲霉是11个蛋糕样品中最主要的分离属,以5个物种为代表,即,A.黄花,A.尼日尔,A.ochrapiary,A.Terreus,和A.versicolor(26。65、63.29、3.33、6.66和3.33个菌落/g,分别)。在测试的27种丝状真菌中,有24种分离株(88.88%)对淀粉酶产生呈阳性。10株(37.03%)受试丝状真菌产脂酶阳性,最后,在总真菌分离物中,有11个分离株(40.74%)显示出蛋白酶产生的阳性结果。在十四个收集的蛋糕样品中未检测到黄曲霉毒素B1,B2,G1,G2和曲霉毒素A。在这项研究中,当比较天然试剂时,丁香油是防止霉菌发展的最佳选择过薄荷油和橄榄油。这可能是由于丁香油中存在多种生物活性化合物,其主要生物活性成分是丁香酚,作为抗真菌试剂。因此,新鲜出炉的蛋糕应该在几天内食用,以避免个人患上食源性疾病。
    Fungi can spoil the majority of baked products. Spoilage of cake during storage is commonly associated with fungi. Therefore, this study aimed to assess the quality of different types of cakes sold in the market. The most predominant fungal genera in the tested cake samples (14 samples) were Aspergillus spp., and Penicillium spp. On Potato Dextrose Agar (PDA), the medium fungal total count was 43.3 colonies /g. Aspergillus was the most dominant genus and was isolated from six samples of cake. Aspergillus was represented by 3 species namely, A. flavus, A. niger, and A. nidulans, represented by 13.32, 19.99, and 3.33 colonies /g respectively. On Malt Extract Agar (MEA) Medium, the fungal total count was 123.24 colonies / g. Aspergillus was the most dominant isolated genus from 11 samples of cake and was represented by 5 species, namely, A. flavus, A. niger, A. ochraceous, A. terreus, and A. versicolor (26. 65, 63.29, 3.33, 6.66, and 3.33 colonies / g , respectively). Twenty-four isolates (88.88 %) of the total tested twenty-seven filamentous fungi showed positive results for amylase production. Ten isolates (37.03%) of the total tested filamentous fungi showed positive results for lipase production, and finally eleven isolates (40.74 %) of the total fungal isolates showed positive results for protease production. Aflatoxins B1, B2, G1, G2, and ochratoxin A were not detected in fourteen collected samples of cake. In this study, clove oil was the best choice overpeppermint oil and olive oil for preventing mold development when natural agents were compared. It might be due to the presence of a varietyof bioactive chemical compounds in clove oil, whose major bioactive component is eugenol, which acts as an antifungal reagent. Therefore, freshly baked cake should be consumed within afew days to avoid individuals experiencing foodborne illnesses.
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  • 文章类型: Journal Article
    人类胰腺的组织病理学异质性是有据可查的;然而,组织水平的功能证据很少。在这里,我们研究了通过胰头(PH)的原位葡萄糖刺激的胰岛和卡巴胆碱刺激的腺泡细胞分泌,主体(PB),和没有糖尿病的供体的尾部(PT)区域(ND;n=15),一种胰岛自身抗体阳性(1AAb+;n=7),和1型糖尿病(T1D;病程<14个月,n=5)。胰岛素,胰高血糖素,胰淀粉酶,脂肪酶,胰蛋白酶原分泌以及3D组织形态测量特征在ND的各个区域中具有可比性。在T1D中,所有区域的胰岛素分泌和β细胞体积都显著减少,而胰高血糖素和酶没有改变。β细胞体积较低,尽管1AAb+的胰岛素分泌正常,导致与ND相比,体积调节的胰岛素分泌增加。1AAb+中的胰岛和腺泡细胞分泌在整个PH中是一致的,PB,和PT。这项研究支持胰腺切片功能的低区域间变异,潜在的,1AAb+的代谢需求增加。
    Histopathological heterogeneity in the human pancreas is well documented; however, functional evidence at the tissue level is scarce. Herein, we investigate in situ glucose-stimulated islet and carbachol-stimulated acinar cell secretion across the pancreas head (PH), body (PB), and tail (PT) regions in donors without diabetes (ND; n = 15), positive for one islet autoantibody (1AAb+; n = 7), and with type 1 diabetes (T1D; <14 months duration, n = 5). Insulin, glucagon, pancreatic amylase, lipase, and trypsinogen secretion along with 3D tissue morphometrical features are comparable across regions in ND. In T1D, insulin secretion and beta-cell volume are significantly reduced within all regions, while glucagon and enzymes are unaltered. Beta-cell volume is lower despite normal insulin secretion in 1AAb+, resulting in increased volume-adjusted insulin secretion versus ND. Islet and acinar cell secretion in 1AAb+ are consistent across the PH, PB, and PT. This study supports low inter-regional variation in pancreas slice function and, potentially, increased metabolic demand in 1AAb+.
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  • 文章类型: Journal Article
    微生物产生的生物质降解酶在农业废弃物处理中具有巨大的潜力。为了产生合适的生物质降解酶,用于从烟草废料中释放糖和香气化合物,在这项研究中,研究了直接使用废料作为碳源进行酶生产的可行性。通过对从烟叶中分离出的十种真菌菌株的比较研究,发现布鲁氏曲霉Ab-10可产生用于糖化烟草残渣的有效酶混合物。蛋白质组学分析确定了酶混合物中一组植物生物质降解酶,包括淀粉酶,半纤维素酶,纤维素酶和果胶酶。在底物浓度为100g/L,酶用量为4mg/g时,使用由A.brunneuviolaceusAb-10产生的粗酶从烟草废料产生17.6g/L的葡萄糖。此外,23个挥发性分子的含量,包括芳香化合物4-酮异佛尔酮和苯甲醇,酶处理后显著增加。通过将生物质降解酶的生产整合到烟草废料加工系统中,该结果为烟草废料的增值提供了一种策略。
    Biomass-degrading enzymes produced by microorganisms have a great potential in the processing of agricultural wastes. In order to produce suitable biomass-degrading enzymes for releasing sugars and aroma compounds from tobacco scraps, the feasibility of directly using the scraps as a carbon source for enzyme production was investigated in this study. By comparative studies of ten fungal strains isolated from tobacco leaves, Aspergillus brunneoviolaceus Ab-10 was found to produce an efficient enzyme mixture for the saccharification of tobacco scraps. Proteomic analysis identified a set of plant biomass-degrading enzymes in the enzyme mixture, including amylases, hemicellulases, cellulases and pectinases. At a substrate concentration of 100 g/L and enzyme dosage of 4 mg/g, glucose of 17.6 g/L was produced from tobacco scraps using the crude enzyme produced by A. brunneoviolaceus Ab-10. In addition, the contents of 23 volatile molecules, including the aroma compounds 4-ketoisophorone and benzyl alcohol, were significantly increased after the enzymatic treatment. The results provide a strategy for valorization of tobacco waste by integrating the production of biomass-degrading enzymes into the tobacco scrap processing system.
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  • 文章类型: Journal Article
    枯草芽孢杆菌是一种革兰氏阳性细菌,经常用于生物工业中生产各种蛋白质,因为其优越的蛋白质分泌能力。为了确定枯草芽孢杆菌分泌蛋白质的最佳条件,快速灵敏的蛋白质分泌测量方法至关重要。快速和通用的测定对于以高通量方式检测不同的蛋白质是最有用的。在这项研究中,我们介绍了一种基于分裂荧光素酶的方法,用于测量枯草芽孢杆菌的蛋白质分泌。NanoBiT系统用于监测四种不同蛋白质的分泌:木聚糖酶A,淀粉酶M,蛋白谷氨酰胺酶A,和GFP纳米抗体。我们的发现强调了分裂荧光素酶系统作为一种快速的,敏感,和用户友好的方法。
    Bacillus subtilis is a Gram-positive bacterium that is frequently used in the bioindustry for the production of various proteins, because of its superior protein secretion capacities. To determine optimal conditions for protein secretion by B. subtilis, a quick and sensitive method for measuring protein secretion is crucial. A fast and universal assay is most useful for detecting diverse proteins in a high-throughput manner. In this study, we introduce a split-luciferase-based method for measuring protein secretion by B. subtilis. The NanoBiT system was used to monitor secretion of four different proteins: xylanase A, amylase M, protein glutaminase A, and GFP nanobody. Our findings underscore the split-luciferase system as a quick, sensitive, and user-friendly method.
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  • 文章类型: Journal Article
    同时胰肾(SPK)移植可提高生活质量并限制糖尿病并发症的进展。人们不愿接受血液检查异常的捐献者的胰腺,由于担心较差的结果。我们使用英国移植注册(2016-2021)调查了供体淀粉酶和肝脏血液测试(内脏缺血损伤的标志物)是否预测胰腺移植结果。包括857名SPK接受者(脑干死亡后619人,循环系统死亡后238)。供体淀粉酶的峰值范围为8至3300U/L(中位数=70),校正多重混杂因素后,这对胰腺移植物存活率没有影响(aHR=0.944,95%CI=0.754-1.81).在多变量模型中,峰值丙氨酸转氨酶也不影响胰腺移植物的存活(aHR=0.967,95%CI=0.848-1.102)。限制性三次样条用于评估供体血液测试和胰腺移植物存活之间的关联,而不假设线性关系;这些都没有证实淀粉酶,也不是转氨酶,显著影响胰腺移植结果。这是最大的,最具统计学意义的研究评估献血者血液检查和移植结果。如果其他因素是可以接受的,来自淀粉酶和转氨酶轻度或中度升高的供体的胰腺可以放心地接受。因此,使用来自此类供体的胰腺移植物是安全的,立即,和简单的方法来扩大捐助者池,以满足日益增长的需求。
    Simultaneous pancreas-kidney (SPK) transplantation improves quality of life and limits progression of diabetic complications. There is reluctance to accept pancreata from donors with abnormal blood tests, due to concern of inferior outcomes. We investigated whether donor amylase and liver blood tests (markers of visceral ischaemic injury) predict pancreas graft outcome using the UK Transplant Registry (2016-2021). 857 SPK recipients were included (619 following brainstem death, 238 following circulatory death). Peak donor amylase ranged from 8 to 3300 U/L (median = 70), and this had no impact on pancreas graft survival when adjusting for multiple confounders (aHR = 0.944, 95% CI = 0.754-1.81). Peak alanine transaminases also did not influence pancreas graft survival in multivariable models (aHR = 0.967, 95% CI = 0.848-1.102). Restricted cubic splines were used to assess associations between donor blood tests and pancreas graft survival without assuming linear relationships; these confirmed neither amylase, nor transaminases, significantly impact pancreas transplant outcome. This is the largest, most statistically robust study evaluating donor blood tests and transplant outcome. Provided other factors are acceptable, pancreata from donors with mild or moderately raised amylase and transaminases can be accepted with confidence. The use of pancreas grafts from such donors is therefore a safe, immediate, and simple approach to expand the donor pool to reach increasing demands.
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  • 文章类型: Journal Article
    脂肪储库或甘油三酯在鱼中通过脂肪酶的作用水解,以用于能量和/或用于生长和繁殖。在食草动物鱼类中,由非脂质底物(葡萄糖)从头合成脂质导致器官中的脂肪沉积和/或脂肪浸润,特别是在卵巢上,限制其正常功能。这项研究的目的是了解不同大小的成年猪的消化道(DT)的脂肪酶,他们的部分净化,表征及其同工酶。评估了碳水化合物与蛋白质相互作用的体外水解研究,以建立对抗过度葡萄糖释放的特定蛋白质选择。研究结果鉴定出4种脂肪酶同工酶~mol。wt19.88、24.29、32.86、54.56kDa,最佳pH为3.5和8,pH稳定性在5.5-10之间;最佳温度为35°C,热稳定性在35至45°C之间。表征研究表明,在其活性位点和Ca中存在巯基,Na和Zn离子激活脂肪酶活性。如从体外研究评估的,当与azolla(植物蛋白)和鱼粉(动物蛋白)一起使用时,米糠作为碳水化合物来源可以对抗过量葡萄糖的过度释放,所述过量葡萄糖导致在猪链球菌中形成内脏脂肪。
    Fat depots or triglycerides are hydrolysed by the action of lipases in fish to be used for energy and/or for growth and reproduction. In herbivores fishes, de novo synthesis of lipids from non- lipid substrates (glucose) leads to fat deposits and/or fatty infiltration in organs especially on ovaries limiting its normal functions. This study was aimed to understand lipases from the digestive tract (DT) of adult Hypselobarbus pulchellus of different sizes, their partial purification, characterisation and their isozymes. In-vitro hydrolysis study on interaction of carbohydrate with proteins was evaluated to establish specific protein selection that combat undue glucose release. Results of the study identified four lipase isoenzymes of ~ mol. wt 19.88, 24.29, 32.86, 54.56 kDa with optimal pH of 3.5 and 8, pH stability between pH 5.5-10; optimal temperature at 35 °C and heat stability between 35 and 45 °C. Characterisation studies indicated presence of thiol group in their active site and Ca, Na and Zn ions activated lipase activity. Rice bran as carbohydrate source when used along with azolla (plant protein) and fish meal (animal protein) may combat undue release of excess glucose that leads to visceral fat formation in H. pulchellus as assessed from in vitro studies.
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