Effect of pH on hemoglobin-catalyzed lipid oxidation in cod muscle membranes in vitro and in situ

The effect of pH and hemoglobin on oxidation of the microsomal lipids of cod was determined in isolated microsomes and in washed cod muscle. An increase of hemoglobin concentration from 0.5 to 15 microM accelerated lipid oxidation in both systems. In cod microsomes the rate of lipid oxidation increased in the order pH 6.8 > pH 7.6 > pH 8.4 > pH 6.0 > pH 3.5. However, in washed cod muscle a decrease of pH from 7.8 to 6.8 greatly increased the lag phase and decreased the rate of lipid oxidation. A further decrease in pH to 3.5 decreased the lag phase and increased the rate of lipid oxidation further. A decrease of pH from 7.6 to 6.4 greatly reduced the affinity of hemoglobin for oxygen. Formation of methemoglobin due to autoxidation occurred more rapidly at pH 6.0 than at pH 7.5. Structural changes of the isolated microsomal membranes could be the reason for the unexpected slow lipid oxidation in microsomes at pH 6.0 and below.     

Separation of muscle membrane from alkali-solubilized fish muscle proteins

Treatment with Ca2+ and citric acid improved membrane removal from muscle homogenates solubilized at pH 10.5 by centrifugation at 4000 g for 15 min. The percentage of phospholipid removed from muscle homogenates increased with increasing Ca2+ concentrations at 1 mM citric acid. More than 85% phospholipid and 45% protein in the muscle homogenates were removed at Ca2+ concentrations of >20 mM in the presence of 1 mM citric acid. At 8 mM Ca2+, addition of citric acid at 5 mM improved phospholipid removal to approximately 78% from 58% in its absence. Because treatment with 8 mM Ca2+ alone can remove significant amounts of phospholipid, it is likely that Ca2+ played the major role in membrane removal in muscle homogenates solubilized at pH 10.5.     

Assessment of the catabolic effects of interleukin-1beta on proteoglycan metabolism in equine cartilage cocultured with synoviocytes

OBJECTIVE: To evaluate the effects of interleukin (IL)-1beta on proteoglycan metabolism in equine cartilage explants when cultured in the presence of synoviocytes. SAMPLE POPULATION: Samples of cartilage and synovium collected from the femoropatellar joints of three 2- to 3-year-old horses. PROCEDURES: 3 experimental groups were established: cartilage explants only, synoviocytes only, and cartilage explants-synoviocytes in coculture. In each group, samples were cultured with or without IL-1beta (10 ng/mL) for 96 hours. Glycosaminoglycan (GAG) content of cartilage and medium samples was measured by use of a spectrophotometric assay; RNA was isolated from synoviocytes and cartilage and analyzed for expression of matrix metalloproteinases (MMP)-3 and -13 (cartilage and synoviocytes), aggrecan (cartilage), collagen type IIB (cartilage), and 18S as a control (cartilage and synoviocytes) by use of quantitative PCR assays. Cartilage matrix metachromasia was assessed histochemically. RESULTS: IL-1beta-induced GAG loss from cartilage was significantly less in cocultures than in cartilage-only cultures. Cartilage aggrecan gene expression was also significantly less downregulated and synoviocyte MMP-3 expression was less upregulated by IL-1beta in cocultures, compared with cartilage- and synoviocyte only cultures. Histochemical findings supported the molecular and biochemical results and revealed maintenance of matrix metachromasia in cocultured cartilage treated with IL-1beta. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that synoviocytes secrete 1 or more mediators that preferentially protect matrix GAG metabolism from the degradative effects of IL-1beta. Further studies involving proteomic and microarray approaches in similar coculture systems may elucidate novel therapeutic targets for the treatment of osteoarthritis.     

Safety issues of methylglyoxal and potential scavengers

The health safety of methylglyoxal (MGO) has been recognized as a key issue owing to its ultra-high reactivity toward some key biomolecules such as amino acids, proteins, DNA, sulfhydryl- and basic nitrogen-containing compounds, including amino-bearing neurotransmitters. In this review, we have summarized the endo- and exogenous sources of MGO and its accumulation inside the body due to high intake, abnormal glucose metabolism and or malfunctioning glyoxalases, and review the debate concerning the adverse functionality of MGO ingested from foods. Higher than normal concentrations of MGO in the circulatory system and tissues have been found to be closely associated with the production of advanced glycation end products (AGEs), increased oxidative stress, elevated inflammation and RAGE (AGE receptors) activity, which subsequently progresses to a pathological stage of human health, such as diabetes complications, cancer, cardiovascular and degenerative diseases. Having illustrated the mechanisms of MGO trapping in vivo, we advocate the development of efficient and efficacious MGO scavengers, either assisting or enhancing the activity of endogenous glyoxalases to facilitate MGO removal, or providing phytochemicals and functional foods containing them, or pharmaceuticals to irreversibly bind MGO and thus form MGO-complexes that are cleared from the body.     

泵装置动力特性随转速变化关系的研究

通过对不同叶片角下不同转速时各性能特性的试验研究,获得了泵装置各性能特性随转速的变化规律,研究了泵装置最高效率及其最高效率点位置和转速的关系,探讨了不同转速下最高效率点处各参数之间的关系及各性能曲线换算到额定转速时的变化,其结论可为泵装置模型试验及泵站的高效运行提供理论的实际指导。     

Effect of the antiparasitic drugs fenbendazole and ivermectin on the soil nematode Pristionchus maupasi

Pristionchus maupasi, a soil nematode, was used to elucidate the potential ecotoxic effect of the two anthelmintics fenbendazole and ivermectin in cattle dung. The population growth of P. maupasi was greater in faeces from cattle harbouring active Panacur- or Ivomec-boli, which are releasing fenbendazole and ivermectin to the rumen, respectively, compared to the growth in control faeces. In dose-response experiments it could be shown that the pure chemical compound of fenbendazole was increasingly nematocidal to P. maupasi in concentrations from 10 to 20 microg/g faeces (ww, i.e. wet weight) and the pure compound of ivermectin was effective above 3 microg/g faeces (ww). The results indicate that neither fenbendazole nor ivermectin have any acute toxic effect on P. maupasi in naturally excreted concentrations of the pure drugs, together with their metabolites in faeces from bolus-treated cattle. Both drugs are excreted in concentrations that are non-toxic to P. maupasi.     

Production of a transgenic pig expressing human albumin and enhanced green fluorescent protein

We introduced a fusion gene of human albumin and enhanced green fluorescent protein (EGFP) into porcine oocytes using the sperm vector method, and produced a piglet that showed clear expression of GFP in the hooves and skin. PCR and Southern blotting analysis of genomic DNA extracted from the piglet's tissues, including the liver, showed that the tissues carried the transgene. RT-PCR analysis demonstrated that both the human albumin and EGFP genes were expressed in the tissues. The fact that human albumin gene was integrated and expressed in the liver of the transgenic pig opened a way for us to achieve our goal, which was the use of transgenic pigs for the bioartificial liver support system.     

Effects of platelet rich plasma and acellular bone marrow on gene expression patterns and DNA content of equine suspensory ligament explant cultures

REASONS FOR PERFORMING STUDY: Suspensory ligament (SL) desmitis is a common source of lameness. The results of this study will determine if blood-derived products stimulate SL matrix synthesis and have potential as regenerative therapies for SL desmitis OBJECTIVES: To determine if various blood-based biological products including plasma, blood, PRP, platelet poor plasma (PPP) and ABM aspirate stimulates anabolic and/or catabolic pathways in suspensory ligaments (SL). METHODS: The body of the SL was harvested from 6 horses and used to establish explant cultures. Explants were cultured in plasma, blood, PRP, PPP or ABM at concentrations of 10, 50 or 100%. Anabolic responses were assessed by use of quantitative PCR for collagens type I and III, cartilage oligomeric matrix protein (COMP) and decorin. Total DNA and collagen protein content were also measured. Catabolic reactions were measured by quantitative PCR for matrix metalloproteinases 3 and 13 (MMP-3, MMP-13). Results: Acellular bone marrow aspirate at 100% stimulated decorin and COMP mRNA synthesis more than all other treatments at all concentrations. No treatment at any concentration stimulated the catabolic gene MMP-13; only 50% ABM stimulated MMP-13 mRNA expression. CONCLUSIONS: Acellular bone marrow is indicated, and might be preferred to plasma, blood or PPP, as a blood-based biological source for SL tissue regenerative therapy. Long-term, placebo controlled case studies are indicated to determine if ABM aids in recovery from SL desmitis. POTENTIAL RELEVANCE: Bone marrow aspirate is an autogenous, readily available biological source for SL regenerative therapy where the aim is to stimulate matrix synthesis.