The development of angiotensin I-converting enzyme inhibitor derived from chicken bone protein

In order to produce angiotensin I‐converting enzyme (ACE) inhibitor for application in functional food, chicken bones were gathered from a meat processing factory and then hydrolyzed with Alcalase, pepsin and trypsin for 12 h. The hydrolysates were lyophilized, stored at ?80°C and tested experimentally every 2 h for pH value, peptide content, degree of hydrolysis (DH), electrophoresis and activity of ACE inhibitor. The hydrolysates of Alcalase had the highest peptide content and DH. The components of more than 66 kDa had disappeared in hydrolysates of Alcalase and trypsin after 2 h of hydrolysis. The hydrolysates of Alcalase were more active in inhibiting ACE, especially when hydrolyzed at 4 and 8 h, and also had low IC50 values of 1.960 and 0.945 mg/mL. According to the results of DH and electrophoresis, the higher activity of ACE inhibitor is assumed to be derived from the low molecular peptides in hydrolysates of Alcalase. Chicken leg bone has a high potential to be utilized to develop ACE inhibitory peptides as a potential ingredient of functional food intended to alleviate hypertension.     

Effects of acute exercise on angiotensin I-converting enzyme (ACE) activity in horses

Angiotensin I-converting enzyme (ACE) level measurement in blood samples is an important tool in human medicine for the detection, treatment and control of diseases such as sarcoidosis and hypertension. Recently ACE has been advocated as being correlated to athletic aptitude in human athletes and a genetic polymorphism has been shown to be responsible for the enzymatic levels in the circulation. The objective of this research was to evaluate the effects of acute exercise in horses in order to increase the understanding of a possible correlation between ACE levels in plasma and performance in equine athletes. A standardised exercise test (SET) to fatigue was conducted on 8 horses and repeated venous blood collections carried out for ACE activity measurements before, during and after the SET. Our results show an increase in ACE activity up to fatigue and a return to baseline values at 30 min post exercise.     

Effects of two training protocols on Angiotensin I-converting enzyme (ACE) activity in horses

Reasons for performing study: Studies in man have shown a correlation between Angiotensin I‐converting enzyme (ACE) genetic polymorphisms, ACE activity in the blood and superior athletic performance in sports requiring endurance. It has been hypothesised that the same correlation occurs in horses. There is no information in the literature concerning the effects of training on ACE activity in equine plasma. Hypothesis: Exercise training influences the activity of circulating ACE and the response observed is dependent on the exercise protocol. Methods: Thirteen horses of mixed breeds were randomly allocated 2 different training protocols to be carried out for a period of 15 weeks. Blood samples were collected from each horse before the beginning of training to determine baseline values. Subsequent sampling took place every 15 days throughout the training phase and for 8 weeks of paddock rest. Angiotensin I‐converting enzyme activity was determined by automated spectrophotometry. Results: Training for 15 weeks significantly increased plasma ACE activity, irrespective of training protocol. Differences observed in ACE activity pattern between the 2 training protocols were not statistically significant. Increase in ACE activity peaked with maximum workload. As soon as training was interrupted, ACE levels significantly decreased. Conclusions and discussion: Exercise training affects levels of ACE activity in equine plasma. The mechanism for this is not yet elucidated, but cardiovascular adaptation to exercise and blood pressure changes might be involved in this regulation. Potential relevance: Exercise training produced a gradual increase in enzymatic activity and might warrant the use of ACE as a tool for fitness monitoring. Angiotensin I‐converting enzyme enzymatic activity in the plasma might be directly correlated to a change in genetic expression and that variability must be taken into account when evaluating results from horses undergoing a physical training programme.     

In vitro analysis of antiangiogenic activity of fungi isolated from clinical cases of equine keratomycosis

OBJECTIVE: The goal of this project was to explore the possibility that fungal organisms produce metabolites that inhibit angiogenesis. Procedures Fungal cultures were obtained from cases of keratomycosis, grown in Sabouraud's dextrose broth, and sterile filtered for use in experiments. The Matrigel assay was used to screen the filtrate samples for antiangiogenic activity. Matrigel is a basement membrane matrix that supports the differentiation of human umbilical vein endothelial (HUVE) cells into a capillary-like network of tubules. HUVE cells were cultured using standard techniques and passaged at confluence, with all cells being used at passage 3-6. HUVE cells (40 000 cells) were pipetted into each well of a 24-well tissue-culture plate coated with Matrigel. An aliquot of fungal media filtrate was added to each well and the plates allowed to incubate for 18 h, at which time they were evaluated for tubule formation. RESULTS: Two fungal isolates showed inhibition of tubule formation. The addition of 100, 200 and 400 &mgr;L of the fungal media filtrate from the first isolate (Fusarium sp. 99A34574) produced a consistent and dose-dependent inhibition of tubule formation. The second isolate (Aspergillus sp. 271599) did not show inhibition of tubule formation with 100 or 200 &mgr;L added to the wells, however, it did show inhibition at 400 &mgr;L/well. The remaining three isolates did not cause inhibition at any concentration. CONCLUSIONS: Our findings suggest that certain fungal organisms produce metabolites that inhibit tubule formation in vitro, and that these metabolites may play a significant role in altering the host vascular response to fungal infections of the cornea.     

Stability of sorbitol dehydrogenase activity in bovine and equine sera

Serum sorbitol dehydrogenase (SDH) activities in 10 cows and nine horses were measured using an automated clinical analyzer. The serum samples were divided into aliquots that were stored at room temperature (21 degrees C), refrigerated (0-5 degrees C), or frozen (-30 degrees C). The stability of the SDH activity was monitored at various intervals. SDH activity in bovine sera remained stable for at least 5 hours at room temperature, 24 hours refrigerated, and 72 hours frozen without any significant (p < 0.05) differences from the initial serum values. In equine sera, SDH activity remained stable for at least 5 hours at room temperature and 48 hours frozen. The activity of the refrigerated equine sera was stable for at least 5 hours but less than 24 hours. An evaluation of fresh bovine serum and heparinized plasma samples indicated that there was no significant difference (p < 0.05) between the two sampling methods and that either may be employed for automated measurement of SDH activity following the established protocol. Sample type comparison indicated that there was a small but statistically significant (p < 0.05) difference between the results obtained comparing fresh serum and heparinized plasma samples for the horse. A reference range for Holstein cows was established using sera from 71 clinically healthy cattle (mean -/+ 2 SD = 32 -/+ 26 U/L).     

高效液相色谱法测定珠蛋白肽对血管紧张素转化酶的抑制活性

建立了测定珠蛋白肽对血管紧张素转化酶抑制活性的高效液相色谱分析方法。以马尿酰-组胺酰-亮氨酸为底物,血管紧张素转化酶为催化剂,反应产物马尿酸为检测指标,未加珠蛋白肽的反应为空白对照。最佳色谱条件为:ZorbaxSB-C18(4.6mm×260mm,粒径5μm),柱温25℃,流动相为乙腈/水=25/75[体积比,各含0.05%三氟乙酸(体积分数)],流速1.0mL/min,紫外检测器,检测波长为228nm,进样量10μL。马尿酸浓度为0.005～1mmol/L,线性相关系数为0.9998,最低检测浓度为1μmol/L,回收率在96.66%～101.34%之间,相对标准偏差为1.34%之间。该方法操作简单、分析时间短、准确性和精密度高,利于对珠蛋白肽生产进行质量控制。     

香菇木屑菌糖中5种饲用添加酶活性的测定与分析

对香菇木屑菌糠中的纤维素酶、木聚糖酶、α-半乳糖苷酶、果胶酶、蛋白酶的活性进行研究.结果表明,纤维素酶和木聚糖酶的活性分别为16.56 IU和13.86 IU,α-半乳糖苷酶和果胶酶的活性较低,蛋白酶则没有活性.     

Comparison of 4 blood storage methods in a protocol for equine pre-operative autologous donation

OBJECTIVE: To compare viability of equine whole blood stored by 4 different methods, and to establish optimal storage protocols for an equine autologous blood donation program. STUDY DESIGN: In vitro study of stored equine whole blood. Animals- Six healthy adult horses. METHODS: Blood from each horse was collected into 4 different containers: glass bottles containing acid-citrate-dextrose solution (ACD), plastic bags containing ACD, citrate-phosphate-dextrose (CPD), and CPD with supplemental adenine (CPDA-1). Blood was stored for 5 weeks and sampled at 2-day intervals. Standard hematologic and biochemical variables were evaluated, and adenosine-5-triphosphate (ATP) and 2,3-diphosphoglycerate (2,3-DPG) concentrations were measured and normalized to total hemoglobin content. RESULTS: Plasma hemoglobin, % hemolysis, lactate, potassium, ammonia, and lactate dehydrogenase (LDH) increased, whereas glucose concentration and pH decreased in all stored blood over 5 weeks. There was a temporal increase in hemolysis with all storage methods, but the increase was greatest in glass bottles. Lactate and ammonia were highest in CPD and CPDA-1 samples, indicating more active red blood cell (RBC) metabolism. 2,3-DPG concentrations decreased during storage, but were optimally preserved with CPDA-1. ATP concentrations were significantly higher for blood stored in CPDA-1, and were lowest in glass bottles. CONCLUSIONS: Hematologic and biochemical values measured for blood stored in CPDA-1 are suggestive of improved RBC viability compared with other storage methods. With the exception of ATP, results from stored equine blood were similar to those reported for other species. CLINICAL RELEVANCE: Commercial CPDA-1 bags appear to be the optimal storage method for equine whole blood.     

Effect of benazepril and pimobendan on serum angiotensin‐converting enzyme activity in dogs

To support their combined use, the objective of the study was to evaluate the effects of benazepril and pimobendan on serum angiotensin‐converting enzyme (ACE) activity in dogs. A total of 48 healthy beagle dogs were randomized into four groups (n = 12 per group) in a parallel‐group design study: A (control, placebo twice daily (BID)); B (0.5–1.0 mg/kg benazepril once daily (SID) in the morning, placebo in the evening); C (0.25–0.5 mg/kg benazepril BID); D (0.25–0.5 mg/kg benazepril and 0.125–0.25 mg/kg pimobendan, both BID). The test items were administered orally for 15 days. Serum ACE activity was measured on days 1 and 15. Groups B, C and D had significantly lower average serum ACE activity compared to baseline and to the control group, on both days 1 and 15. There were no significant differences in average ACE activity between groups B, C and D. Noninferiority of group C to B was demonstrated. In conclusion, 0.25–0.5 mg/kg benazepril administered BID produced noninferior inhibition of serum ACE activity compared to 0.5–1.0 mg/kg benazepril dosed SID. Pimobendan had no significant effect on benazepril's action on serum ACE activity. The results support the use of benazepril BID in dogs and in combination with pimobendan.     

肾素血管紧张素系统(RAS)两条轴相互负向调节与大鼠非酒精性脂肪肝病(NAFLD)肝损伤的关系研究

探讨了肾素血管紧张素系统(renin angiotensin system,RAS)两条轴对大鼠非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)肝脏损伤的相互负向调节作用。30只雄性SD大鼠随机分为正常对照组、模型组和用药组。除正常对照组外,其余2组饲喂高脂饲料,用药组另外给伐他汀50 mg/kg·d。3周后,宰杀大鼠,测定血清中甘油三酯(TG)、谷丙转氨酶(ALT)、谷草转氨酶(AST)的含量;测定肝组织匀浆羟自由基(·OH)、一氧化氮合酶(NOS)、超氧化物歧化酶(SOD)、总抗氧化能力(T-AOC)酶活性;ELISA法测定组织匀浆中血管紧张素Ⅱ(AngⅡ)、血管紧张素1-7(Ang1-7)及白细胞介素1β(IL-1β)、肿瘤坏死因子α(TNF-α)含量;Western blot分析肝组织血管紧张素转化酶(ACE)、血管紧张素转化酶2(ACE2)、血管紧张素Ⅱ受体1亚型(AT1R)、Mas受体(MasR)蛋白水平。结果:与正常对照组相比,模型组大鼠血清TG、AST和ALT含量以及·OH和NOS活性均显著升高(P<0.05), T-AOC、SOD活性显著降低(P<0.05);IL-1β和TNF-α含量极显著升高(P<0.01);ACE、ACE2的蛋白表达水平与ACE/ACE2的比值均显著升高(P<0.05),AngⅡ、Ang1-7含量与AT1R表达升高(P<0.05),MasR表达有升高趋势(P>0.05)。结论:高脂饲料连续饲喂3周可诱导大鼠发生NAFLD,肝脏局部RAS两条轴均处于激活状态,ACE介导AngⅡ-AT1R经典轴促进肝损伤,而ACE2介导Ang1-7-MasR轴抵抗肝损伤;ACE2负性调节RAS作用,对大鼠NAFLD时肝脏氧化应激及炎性损伤有一定保护作用。     

Potential effect of the microbial fermented feed utilization on physicochemical traits,antioxidant enzyme and trace mineral analysis in rabbit meat

The study investigated the potential effect of the microbial fermented feed utilization on physicochemical traits, antioxidant enzyme and trace mineral analysis in rabbit meat. A total of 72 six-week-old male rabbits were weighed and randomly divided into four groups (1) (SRKC) control; (2) (SRKP) Lactobacillus plantarum 1 × 10⁶ cfu/g fresh weight (FW); (3) (SRKG) Pediococcus acidilactici 1 × 10⁶cfu/g FW and (4) (SRKPG) P. acidilactici + L. plantarum 1 × 10⁶ cfu/g FW. Performance characteristic, weekly body weight, was positively (p < .05) enhanced, while daily feed intake (DFI) and feed convention ratio (FCR) were not influenced in treatments group as compared to untreated. The water, protein, water holding capacity (WHC) and dry matter (DM) concentration were positively (p < .05) influenced, while ash, pH, lightness, redness and yellowness were not influenced in treated group as compared to untreated. The concentration of glutathione peroxidase (Gpx), superoxide dismutase (SOD) and aspartate aminotransferase (AST) was positively (p < .05) influenced in treatments group as compared to control. Regarding trace minerals, copper (Cu), iron (Fe), manganese (Mn) and zinc (Zn) were positively (p > .05) reduced in treated group as compared to untreated. It is concluded that the addition of lactic acid bacteria (L. plantarum and P. acidilactici) in Hybrid pennisetum silage had a constructive influence on rabbit health performance and meat biochemistry.     

中药复方制剂对球虫感染鸡血液免疫指标和酶活性影响

为了研究中草药复方制剂对人工感染柔嫩艾美耳球虫的鸡血液免疫指标和酶活性影响,选取80只罗曼蛋公雏饲喂至24日龄,随机分为4组,分别为非感染给药组、非感染非给药组、感染非给药组和感染给药组。27日龄时,感染组每只鸡接种1×105个孢子化球虫卵囊。感染球虫8d后,心脏采血,收集血清,采用半自动生化分析仪测定血清免疫指标:总蛋白(TP),白蛋白(ALB),球蛋白,免疫球蛋白IgA、IgG、IgM,血清酶指标:谷丙转氨酶(GPT)、谷草转氨酶(GOT)、碱性磷酸酶(ALP)。结果表明非感染非给药组的总蛋白、白蛋白、球蛋白血清浓度明显高于感染组,而感染给药组与非感染非给药组之间差异不显著(P0.05);感染组IgA、IgM含量显著高于非感染非给药组(P0.01),而IgG含量则显著低于非感染非给药组(P0.01),IgA、IgM含量感染给药组显著低于感染非给药组(P0.01),IgG含量感染给药组显著高于感染非给药组(P0.01);与非感染非给药组相比,感染组3种血清酶的活性均明显下降,感染给药组各种酶的活性均显著高于感染非给药组(P0.01)。表明该中草药复方制剂可降低柔嫩艾美耳球虫对鸡血清蛋白浓度的影响,使血清酶活性升高。     

Comparison of the Accu-Chek Aviva point-of-care glucometer with blood gas and laboratory methods of analysis of glucose measurement in equine emergency patients

Background: More information is needed regarding accuracy of commonly used methods of glucose measurement in the critically ill horse.
Hypothesis: Glucometry will have good agreement with a laboratory standard. Glucometry with plasma will have better agreement than when performed with whole blood.
Animals: Fifty sequentially admitted equine emergency patients, aged >1year.
Methods: Venous blood was collected at admission and immediately analyzed by point-of-care glucometry on both whole blood (POC/WB) and plasma (POC/PL), a multielectrode blood gas analyzer with whole blood (BLG), and a standard laboratory method with plasma (CHEM). Paired data were compared using Lin's concordance correlation, Pearson's correlation, and robust regression. Bias and limits of agreement were tested by the Bland-Altman technique. Bivariate regression analysis was used to explore confounding factors.
Results: Concordance was significant for all comparisons, and was strongest for CHEM-POC/PL (0.977) and weakest for POC/WB-POC/PL (0.668). Pearson's correlation was excellent for all comparisons except those with POC/WB. All comparisons had excellent robust regression coefficients except those with POC/WB.
Conclusions and Clinical Importance: POC glucometry with plasma had excellent agreement with a laboratory standard, as did blood gas analysis. POC glucometry with whole blood correlated poorly with a laboratory standard. These differences may be clinically important, and could affect decisions based on glucose concentrations.     

Comparison of 2 glucose analytical methodologies in immature Kemp’s ridley sea turtles: dry chemistry of plasma versus point-of-care glucometer analysis of whole blood

Blood glucose measurements provide important diagnostic information regarding stress, disease, and nutritional status. Glucose analytical methodologies include dry chemistry analysis (DCA) of plasma and point-of-care (POC) glucometer analysis of whole blood; however, these 2 methods differ in cost, required sample volume, and processing time. Because POC glucometers use built-in equations based on features of mammalian blood to convert whole blood measurements to plasma equivalent units, obtained glucose data must be compared and validated using gold-standard chemistry analytical methodology in reptiles. For in-water, trawl-captured, immature Kemp’s ridley sea turtles (Lepidochelys kempii) from Georgia, USA, we observed significant, positive agreement between the 2 glucose determination methods; however, the glucometer overestimated glucose concentrations by 1.4 mmol/L on average in comparison to DCA and produced a wider range of results. The discordance of these results suggests that POC glucometer glucose data should be interpreted in the context of methodology- and brand-specific reference intervals along with concurrent packed cell volume data.