Reference intervals for the activity of lactate dehydrogenase and its isoenzymes in the serum and cerebrospinal fluid of healthy canines

Background

Lactate dehydrogenase (LD) exists as 5 isoenzymes (LD‐1 through LD‐5) that are expressed throughout the body and can be detected in both serum and cerebrospinal fluid (CSF). LD and its isoenzymes have been relatively unstudied in veterinary medicine, although studies in human medicine have demonstrated that changes in total LD activity and atypical isoenzyme patterns can indicate disease processes, including neurologic abnormalities.

Objectives

The purpose of this study was to establish RIs for LD and its isoenzymes in the serum and CSF of clinically healthy dogs. By establishing a definitive RI for this enzyme in healthy canines, further study of the clinical and diagnostic usefulness of LD can be undertaken.

Methods

Serum and atlantoaxial CSF were collected from clinically healthy dogs. Total LD activity was measured spectrophotometrically immediately after collection. Isoenzyme distributions were also determined within 8 hours of collection using the QuickGel LD Isoenzyme technique and a densitometric scanner.

Results

The median serum total LD in healthy canines was 69.0 U/L (n = 41; range: 21.0‐217.0 U/L), while the median CSF total LD was 10.0 U/L (n = 40; range: 6.0‐19.3 U/L). LD‐5 is the predominant isoenzyme in canine serum (n = 40), contributing over half of the total enzyme activity. Conversely, in canine CSF (n = 42), LD‐1 is the predominant isoenzyme, followed by LD‐2 and LD‐3.

Conclusions

Knowledge of the distribution and concentration of LD in the serum and CSF of healthy dogs will set the foundation for future studies of canine LD as a potentially clinically useful biomarker.     

Effects of post-anthesis fertilizer on the protein composition of the gluten polymer in a US bread wheat

Both genetic and environmental factors influence the types and amounts of wheat proteins that link together to form polymers essential for flour quality. To understand how plant growth conditions might influence gluten polymer formation, protein fractions containing small and large polymers were separated from flour from the US wheat Butte 86 grown in the absence or presence of post-anthesis fertilizer. Proteins in the polymer fractions were analyzed by quantitative two-dimensional gel electrophoresis (2-DE). The ratio of high molecular weight glutenin subunits (HMW-GS) to low molecular weight glutenin subunits (LMW-GS) increased in both fractions in response to fertilizer, due in part to small increases in the proportions of individual HMW-GS. There were also changes within the LMW-GS. In particular, omega and alpha chain terminators increased in proportion in both polymer fractions, but changes were more pronounced in the large polymer fractions. Serpins also increased in both polymer fractions. Additionally, the study revealed differences in the proportions of traditional LMW-GS in small and large polymer fractions. LMW-s type proteins were more abundant in the large polymers while LMW-i type proteins were more prevalent in the small polymers, suggesting that these proteins may play different roles in the gluten polymer.     

Genotyping of isolates ofTaylorella equigenitalis from thoroughbred brood mares in Japan

Profiles of the genomic DNA of 104 strains ofT. equigenitalis isolated from brood mares with contagious equine metritis in Hokkaido during the breeding seasons from 1980 to 1993, as well as those of five strains (SS28, EQ56, EQ59, EQ70 and HH139) previously isolated in Japan were examined after restriction digestion and crossed-field gel electrophoresis. These profiles were essentially identical to each other and the various isolates and strains appeared to have a common genotype, designated genotype J, with respect to two restriction enzymes,ApaI andNotI. These results suggest a common source for all these isolates obtained over the course of more than 10 years in Japan.Abbreviations CEM contagious equine metritis - CFGE crossed-field gel electrophoresis - PFGE pulsed-field gel electrophoresis     

Purification and characterization of a flavor-related enzyme, γ-glutamyl-transpeptidase,from Toona sinensis leaves

A flavor-related enzyme from Toona sinensis (TS), ?-glutamyl-transpeptidase (GGT), was purified 40.67-fold with 13.40% recovery using ammonium sulfate precipitation, dialysis and gel filtration chromatography. Gel filtration chromatography and sodium dodecyl sulfate acryl amide gel electrophoresis (SDS-PAGE) revealed that GGT was a heterodimer with a molecular weight of 90 kDa, and the molecular size of its two subunits was 26 kDa and 64 kDa, respectively. The apparent Km and Vmax of GGT were 3.623 μmol/mL and 2.831 μmol/mL/min of protein, respectively. The optimum reaction temperature was 60°C, while the optimum pH was 7.0. GGT can tolerate temperatures of 70°C for a short time and can keep stable activity at 60°C for more than 2 hours. The effect of metal ions on GGT activity showed that Cu²⁺ and Zn²⁺ had strong inhibition, while Fe³⁺ had strong activation, and other metal ions had only a slight effect. LC-MS analysis indicated that GGT catalyzed (S, S)-γ-glutamyl-(cis-S-1-propenyl)-thioglycine, a flavor precursor found in TS, to release glutamic acid moiety.     

Comparative cardiovascular and pulmonary effects of sedatives and anesthetic agents and anesthetic drug selection for the trauma patient

Objective: To integrate and compare the effects of tranquilizer/sedatives and anesthetic drugs on various parameters of cardiovascular function in normal dogs and in dogs stressed by hypovolemia, anemia, and endotoxemia, and to discuss the relative merits and appropriate precautions of anesthetic drugs with respect to specific patient physiologic complications. Data sources: Personal data and experiences in conjunction with veterinary and human clinical and research studies. Human and veterinary data synthesis: Drugs that produce calming, sedation, muscle relaxation, analgesia, and loss of consciousness have the potential to produce marked cardiorespiratory effects particularly in hemorrhaged, hypovolemic‐traumatized animals. Acute but key cardiovascular components that are affected by sedative and anesthetic drugs include heart rate and rhythm, venous return (preload), systemic vascular resistance (afterload), and myocardial contractile (inotropic) and relaxation (lusitropic) properties. In addition, all sedative and anesthetic drugs alter or depress normal baroreceptor reflex activity, thereby inhibiting or eliminating the animal's normal physiologic response to decreases in arterial blood pressure and predisposing to tissue hypoperfusion, decreased oxygen delivery, and oxygenation. Oxygen delivery needs to be adequate to meet the metabolic (oxygen) requirements of the patient. Decreases in oxygen delivery to tissues increases oxygen extraction, thereby maintaining tissue oxygenation (supply‐independent oxygen consumption phase) until compensatory processes reach their limit and any further decrease in oxygen delivery causes a decrease in oxygen consumption (supply‐dependent oxygen consumption phase). The critical oxygen delivery that defines the transition between these 2 phases is generally higher in the anesthetized state than in the awake state. The effect of anesthetics on critical oxygen delivery at comparable anesthetic dosages is pentobarbital=ketamine>alfentanil>etomidate=propofol>inhalational anesthetics. Anesthetics generally decrease oxygen consumption from the awake, baseline state; exceptions are ketamine and ether. Ketamine, however, increases oxygen delivery and oxygen extraction. Conclusions: The transition from the awake to the anesthetized state is a huge imposition on the physiology of animals and, therefore, should be accomplished with great care and proper vigilance. Rapid, ‘crash’ induction of anesthesia should be avoided in hypotension‐prone animals and slow, prolonged induction should be avoided in animals with respiratory disorders. It is not recommended to implement an unfamiliar protocol in critical patients, even if it might be pharmacologically preferable. Familiarity with an anesthetic drug is a very important reason for its selection.     

猪卵泡液中抑制素α亚基的鉴定

利用SephadexG 2 0 0和SephadexG 1 0 0 2次凝胶过滤层析的方法 ,从猪卵泡液中初步分离得到抑制素及其α亚基的粗提物。通过免疫印迹的方法检测 ,表明粗提物中含有抑制素α亚基     

冷冻干燥对丝素蛋白凝胶结构的影响

为了探讨冷冻干燥对丝素蛋白凝胶结构的影响,利用傅里叶变换红外光谱(FTIR)研究了丝素凝胶在冷冻干燥前后二级结构的变化。发现在冷冻干燥过程中,丝素蛋白中分子链内β-折叠结构含量增加,分子链间β-折叠结构含量减少,并有部分无规卷曲结构转变为β-折叠结构;但冷冻干燥前后总的β-折叠结构含量变化不大。     

不同相对湿度对间歇性26℃环境下肉鸡盲肠菌群多样性的影响

本试验旨在研究不同相对湿度(RH)对间歇性26℃环境下肉鸡盲肠菌群多样性的影响。选取29日龄爱拔益加(AA)肉鸡180只转入环境控制舱,随机分成3个组(RH分别为30%、60%和85%),每组6个重复,每个重复10只鸡(公母各5只)。从29日龄开始,每天10:00—16:00(6 h)温度维持26℃,RH分别为30%、60%和85%,剩余时间温度为21℃,RH为60%。试验共14 d。采用16S rDNA的变性梯度凝胶电泳(DGGE)技术,结合特异性和共性条带割胶回收DNA进行克隆和测序,分析RH在间歇性26℃偏热处理第7天和第14天时对盲肠内容物菌群结构和多样性的影响。结果表明:1)试验第7天,30%RH组肉鸡盲肠DGGE条带数(菌群丰富程度)高于60%RH组,而85%RH组低于60%RH组;试验第14天,30%、85%RH组肉鸡盲肠DGGE条带数均高于60%RH组。2)聚类分析显示,试验第7天,85%RH对盲肠菌群影响明显;试验第14天,30%RH对盲肠菌群影响明显。但随着处理时间推移,30%RH对肉鸡盲肠菌群影响越大。3)间歇性26℃环境下不同RH组肉鸡盲肠内共性菌群是Faecalibacterium prausnitzii;在试验第7天,30%、85%RH组肉鸡盲肠中特异性菌群是Stomatobaculum longum。结果提示:间歇性26℃环境下,低湿(30%RH)和高湿(85%RH)影响肉鸡盲肠菌群的结构和多样性,且不同处理时间RH的影响不同     

Identification of serum protein markers for early diagnosis of pregnancy in buffalo

Improper or delayed pregnancy diagnosis has significant impact over animal production, particularly in buffaloes which inherently suffer from several reproductive inefficiencies. Thus the present study has undertaken to identify serum protein markers pertaining to early pregnancy diagnosis in buffaloes. Serum samples were collected from 10 pregnant Murrah Buffalo heifers at weekly intervals from days 0‐35 post‐artificial insemination and from 12 inseminated non‐pregnant cyclic buffalo heifers on days 0, 7, 14 and 21. Two‐dimensional gel electrophoresis and densitometric analysis revealed the presence of five protein spots showing average density fold change of ≥4 during early pregnancy. Mass spectrometry analysis identified these up‐regulated proteins as anti‐testosterone antibody light chain, apolipoprotein A‐II precursor, serum amyloid A, cytokeratin type II, component IV isoform 1, which are have established roles in embryogenesis, but over‐expression of the fifth identified protein immunoglobulin lambda light chain in pregnancy has been elucidated as a novel finding in the current study. Further, with bioinformatics analysis, potential antigenic B‐cell epitopes were predicted for all these five proteins. An antibody cocktail‐based approach involving antibodies against all these five up‐regulated entire proteins or their epitopes could be developed for early detection of pregnancy in buffaloes. © 2016 Japanese Society of Animal Science     

Effects of coated and non-coated ZnO nano particles on cucumber seedlings grown in gel chamber

Zinc oxide-engineered nanoparticles (ZnO ENPs) have received the most attention in recent years. This increasing interest has been directed towards studying the environmental fate and effects of ZnO ENPs on ecological terrestrial species. In this study, ZnO NPs were synthesized by atmospheric pressure solution evaporation method and were coated or uncoated with humic acid (HA). The root uptakes of uncoated and HA-coated ZnO NPs and zinc (Zn) were investigated by gel-grown cucumber. Two ZnO levels (1 and 200 µM) were applied in the form of coated (T₃) and non-coated (T₂) NPs or bulk particles (T₁). The results showed that coating NPs by HA increases zeta potential of NPs and decreases their aggregation size due to the increase in the repulsion forces among the particles. Addition of 1 mgL^?1 ZnO into gel chamber enhanced root and shoot biomass; however, the shoot growth was higher in the presence of NPs compared to its bulk counterpart. Moreover, greater phytotoxicity of ZnO from the source of NPs than bulk particles in shoot was observed. Scanning electron microscopy results showed a clear evidence of the penetration of NPs into root cells.