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1.
Kazuma Takahashi Takashi Kondo Yasunaga Yoshikawa Kiyotaka Watanabe Koichi Orino 《Acta veterinaria Scandinavica》2013,55(1):70
Background
Horse fibrinogen has been identified as a plasma specific ferritin-binding protein. There are two ways in the binding of ferritin-binding protein with ferritin: one is direct binding and the other is indirect binding which is heme-mediated. The aim of this study was to analyze the binding between horse fibrinogen and ferritin.Findings
Although fibrinogen in horse plasma did not show the binding to ferritin coated on the plate wells, after following heat-treatment (60°C, 30 min) of horse plasma, plasma fibrinogen as well as purified horse fibrinogen bound to plates coated with horse spleen ferritin, but not with its apoferritin which lost heme as well as iron after the treatment of reducing reagent. Binding of purified or plasma fibrinogen to ferritin was inhibited by hemin and Sn-protoporphyrin IX (Sn-PPIX), but not by PPIX or Zn-PPIX.Conclusions
Heat-treatment of horse plasma enabled plasma fibrinogen to bind to plate well coated with holo-ferritin. From the binding analysis of fibrinogen and ferritin, it is suggested that horse fibrinogen recognized iron or tin in complexed with the heme- or the hemin-ring, and also suggest that some fibrinogens circulate in the form of a complex with ferritin and/or heat-labile factors which inhibit the binding of fibrinogen with ferritin. 相似文献2.
The ferritins were purified from liver homogenates of buffalo, camel, cattle, sheep and shark by thermal denaturation, ammonium sulphate fractionation, Sephacryl S-300 gel filtration and DEAE-blue gel affinity chromatography. The yield and iron content of affinity-purified liver ferritins ranged from 0.008 to 0.052 mg/g and 3.17% to 11.4% respectively. As they are glycoproteins, the ferritins contained variable amounts of neutral carbohydrates. Except for shark ferritin, the ferritins all exhibited immunological cross-reactivity with anti-buffalo liver ferritin and anti-horse spleen ferritin by immunodiffusion and immunoelectrophoresis. Gel electrophoresis, gel filtration and ultracentrifugal analysis indicated the presence of a monomeric ferritin in all cases. SDS-gel electrophoresis of shark ferritin gave a protein band of 18 kDa. Ovine, buffalo and bovine ferritin comprised two protein subunits, the H (20 and 21 kDa) and the L types (18 and 19 kDa). Oligomeric ferritin subunits with molecular weights of 27, 37 and 55 kDa were also found for bovine and buffalo ferritin. SDS-PAGE of camel ferritin revealed a complex pattern with four prominent bands of 61, 51, 44 and 39 kDa. Two fast-migrating components of 15 and 16 kDa were also found in the purified liver ferritins, including reference preparations. The PO4
3–/Fe ratios of purified shark (0.10) and bovine ferritin (0.12) were similar to that of standard equine spleen ferritin (0.11). However, the ratio was higher in ovine (0.17), camel (0.22) and bovine (0.26) ferritins. The amino acid compositions, molecular weights and sedimentation coefficients of the different liver ferritins were similar. 相似文献
3.
Masami Numata Takashi Kondo Yasuo Nambo Yasunaga Yoshikawa Kiyotaka Watanabe Koichi Orino 《Animal Science Journal》2013,84(12):782-789
Antibody (immunoglobulin G (IgG), IgM or IgA) levels relative to ferritin in six foal sera (three male and three female) after birth (day 0 and 2, 6, 10, 20, 28, 36, 40, 52 and 56 weeks of age) were semi‐quantitatively measured with normalization with antibody activity to ferritin in one adult horse serum. After addition of horse spleen ferritin to the serum sample, the complex formed between antibodies to ferritin in the serum and ferritin was co‐immunoprecipitated using antibody to horse spleen ferritin. Antibody classes of the co‐immnoprecipitate were detected with antibodies specific for horse IgG, IgM or IgA heavy chain. Six adult horse serum samples were found to have ferritin‐binding activities in all immunoglobulin classes examined. Although ferritin antibody activities (IgG, IgM and IgA) were scant in the foal sera before sucking colostrum (day 0), their activities increased at 2 weeks of age. IgG antibodies showed a biphasic response and IgM antibody activity increased up to 40 weeks of age. Antibody (IgG, IgM and IgA) activities to ferritin in three colostrum samples were significantly higher than in adult horse serum samples. These results demonstrate that antibody to ferritin in foal serum is derived from colostrum after birth and is produced thereafter. 相似文献
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5.
为寻找免疫失败的原因,有效防治犬瘟热的流行,对临床一水貂犬瘟热疑似病例,取其肝、脾、脑等组织研磨后接种Vero和BHK细胞分离病毒,并对分离毒株进行一系列鉴定。通过电镜观察,发现了大小约150 nm的副黏病毒样粒子。结果表明,分离株对鹅、鸡、小鼠、家兔、山羊、猪红细胞均无凝集性,该分离株的毒价TCID50为10-4.87;分离株病毒对乙醚、氯仿敏感,病毒的核酸型为RNA。经间接免疫荧光试验,接种病毒的BHK细胞出现特异性的亮绿色荧光。对分离株和对照毒株分别提取RNA进行RT-PCR扩增,最后得到与预期扩增片段(294 bp)相符的核酸电泳带,充分证明分离病毒为犬瘟热病毒。 相似文献
6.
本研究利用不连续聚丙烯酰胺凝胶电泳法对绵羊的８种组织器官ＬＤＨ同工酶的分布及相对活性进行了分析测定。比较ＬＤＨ同工酶的酶谱特征。结果表明，绵羊的ＬＤＨ同工酶分布特征不同于其它动物，其相对活性有显著差异。骨骼肌中ＬＤＨ５活性比ＬＤＨ１强，Ｍ亚基比例大于Ｈ亚基；肝脏和心脏中ＬＤＨ１的活性最强。 相似文献
7.
The binding ability of staphylococcal protein A (SpA) and streptococcal protein G (SpG) to egg yolk antibodies of four fowl species (turkey, duck, moskovy duck and goose) was studied and compared with the binding ability to three serum antibodies from chicken, horse and cattle. SpA and SpG were not able to bind to any of the avian immunoglobulins. 相似文献
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9.
Masafumi HASHIMOTO Yasuo NAMBO Takashi KONDO Kiyotaka WATANABE Koichi ORINO 《Journal of Equine Science》2011,22(1):1-7
In mammal circulation, ferritin-binding proteins (FBPs) are thought to be involved in
clearance of circulating ferritin after complex formation with it through
receptor-mediated uptake. However, there is no report on fetal FBP in fetal circulation.
Although iron concentrations of fetal horse plasma were higher than those of adult horse
plasma, plasma ferritin concentrations and ferritin-binding activities were found to be
significantly lower in fetus than in adult. FBPs were purified from fetal or adult horse
plasma on horse spleen ferritin-Sepharose 4B affinity column. Partially affinity-purified
fetal horse plasma FBPs were mainly separated into 65 and 41 kDa bands in addition to
minor bands with higher molecular masses ranged from 102 to 140 kDa on SDS-PAGE under
reducing condition. The adult horse plasma FBPs were separated into 74, 54 and 28 kDa
bands, and the 74 and 54 kDa bands reacted with antibodies specific for horse IgM and IgG
heavy chains, respectively, by immunoblotting analyses. On the other hand, no antibodies
to horse immunoglobulin classes detected any bands in fetal horse plasma FBPs. The
affinity-purified adult and fetal horse plasma FBPs did not contain fibrinogen as a plasma
specific FBP, probably due to its lower affinity to the ligand ferritin. These results
demonstrate the presence of FBPs which are different from adult horse plasma FBPs
including anti-ferritin autoantibodies in fetal plasma. 相似文献
10.
Molecular cloning and tissue expression of chicken AdipoR1 and AdipoR2 complementary deoxyribonucleic acids 总被引：3，自引：0，他引：3
AdipoR1 and AdipoR2 belong to a novel class of transmembrane receptors that mediate the effects of adiponectin. We have cloned the chicken AdipoR1 and AdipoR2 complementary deoxyribonucleic acids (cDNA) and determined their expression in various tissues. We also investigated the effect of feed deprivation on the expression of AdipoR1 or AdipoR2 mRNA in the chicken diencephalon, liver, anterior pituitary gland, and adipose tissue. The chicken AdipoR1 and AdipoR2 cDNA sequences were 76-83% identical to the respective mammalian sequences. A hydrophobicity analysis of the deduced amino acid sequences of chicken AdipoR1/AdipoR2 revealed seven distinct hydrophobic regions representing seven transmembrane domains. By RT-PCR, we detected AdipoR1 and AdipoR2 mRNA in adipose tissue, liver, anterior pituitary gland, diencephalon, skeletal muscle, kidney, spleen, ovary, and blood. AdipoR1 or AdipoR2 mRNA expression in various tissues was quantified by real-time quantitative PCR, and AdipoR1 mRNA expression was the highest in skeletal muscle, adipose tissue and diencephalon, followed by kidney, ovary, liver, anterior pituitary gland, and spleen. AdipoR2 mRNA expression was the highest in adipose tissue followed by skeletal muscle, liver, ovary, diencephalon, anterior pituitary gland, kidney, and spleen. We also found that a 48 h feed deprivation significantly decreased AdipoR1 mRNA quantity in the chicken pituitary gland, while AdipoR2 mRNA quantity was significantly increased in adipose tissue (P<0.05). We conclude that the AdipoR1 and AdipoR2 genes are ubiquitously expressed in chicken tissues and that their expression is altered by feed deprivation in the anterior pituitary gland and adipose tissue. 相似文献
11.
Yoshiya Kanno Hiromichi Ohtsuka Yasunaga Yoshikawa Kiyotaka Watanabe Koichi Orino 《Animal Science Journal》2013,84(7):556-561
Anti‐ferritin autoantibody is a ferritin‐binding protein commonly found in mammals; it is thought to form an immune complex with ferritin and thereby mediate the rapid clearance of circulating ferritin. The aim of this study is to determine concentrations of ferritin and anti‐ferritin autoantibodies (immunoglobulin (Ig)M, IgG and IgA) in serum and colostrum of Holstein (H) and Japanese Black (JB) cows within 24 h of normal calving. Blood and colostrum samples were collected from cows of various ages (2–11 years) and calving number (1–8 live births). Mean ferritin concentrations were higher in colostrum than in serum for both breeds, and higher colostrum ferritin concentrations were found in H than JB cows. IgA antibodies in serum and colostrum from both breeds had negligible ferritin‐binding activity. For both breeds, IgM and IgG antibodies had higher ferritin‐binding activity in colostrum than in serum. There was a significant correlation between IgM and IgG ferritin‐binding activities in serum and colostrum of H and JB cows. These results suggest that ferritin and IgM and IgG autoantibodies are actively transferred from the blood stream to the colostrum at prepartum or early lactation. 相似文献
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13.
Rayalam S Eizenstat LD Davis RR Hoenig M Ferguson DC 《Domestic animal endocrinology》2006,30(3):185-202
The goal of this study was to express and purify recombinant feline TSH as a possible immunoassay standard or pharmaceutical agent. Previously cloned feline common glycoprotein alpha (CGA) and beta subunits were ligated into the mammalian expression vector pEAK10. The feline CGA-FLAG and beta subunits were cloned separately into the pEAK10 expression vector, and transiently co-transfected into PEAK cells. Similarly, previously cloned and sequenced yoked (single chain) fTSH (yfTSH) and the CGA-FLAG sequences were ligated into the same vector, and stable cell lines selected by puromycin resistance. Expression levels of at least 1 microg/ml were achieved for both heterodimeric and yoked fTSH forms. The glycoproteins were purified in one step using anti-FLAG immunoaffinity column chromatography to high purity. The molecular weights of feline CGA-FLAG subunit, beta subunit and yfTSH were 20.4, 17, and 45 kDa, respectively. Both heterodimeric and yoked glycoproteins were recognized with approximately 40% detection by both a commercial canine TSH immunoassay and an in-house canine TSH ELISA. The yoked glycoprotein exhibited parallelism with the heterodimeric form in the in-house ELISA, supporting their possible use as immunoassay standards. In bioactivity assays, the heterodimeric and yoked forms of fTSH were 12.5 and 3.4% as potent as pituitary source bovine TSH at displacing (125)I-bTSH and 45 and 24% as potent in stimulating adenylate cyclase activity in human TSH receptor-expressing JP09 cells. However, in addition to reduced receptor binding affinity, the recombinant glycohormones produced a reduced maximal effect at maximal concentration (E(max)) suggesting the possibility of the recombinant glycohormone constructs acting as partial agonists at the human TSH receptor. 相似文献
14.
Chicken macrophage activation by interferon: do birds lack the molecular homologue of mammalian interferon-gamma? 总被引：2，自引：0，他引：2
In mammalian species, interferon-gamma (IFN-gamma) is a lymphokine with a wide range of biological effects, of which the antiviral and macrophage-activating capacities are those best characterized. In birds, no equivalent with a similar range of actions has as yet been isolated. Chicken splenocytes were stimulated by mitogens in conditions that were similar to those used for the induction of mammalian IFN-gamma. Culture fluids were assayed for antiviral and macrophage-activating capacities. As much as 1000 units/ml of an interferon-like antiviral activity was found in the culture fluid of Staphylococcus aureus lysate-induced spleen cells. Seroneutralization assays with a polyclonal antiserum against purified interferon and physicochemical studies revealed that the antiviral activity is identical to or closely related to type I interferon (interferon-alpha/beta). The presence of macrophage activating factors (MAF) in the splenocyte medium was demonstrated by measuring increased production of H2O2 by chicken peritoneal macrophage cultures and a chicken macrophage cell line (HD11). The heat stability of this MAF activity was similar to that of the antiviral factor, and was completely neutralized by the anti-IFN-alpha/beta antiserum. These results show that when the classical procedure used for the production of mammalian IFN-gamma is applied to chicken splenocytes, it does not yield an equivalent for IFN-gamma/MAF. This suggests that the classification of interferons into types (alpha, beta and gamma), while generally applicable in mammals, may not be applicable in birds. 相似文献
15.
Use of biotinylated antibody for the assay of Hanganutziu-Deicher antibodies and antigens in fluids and tissues from cancer patients 总被引：1，自引：0，他引：1
J K Gathuru H Higashi S Kato O Usuba M Naiki 《The Japanese journal of veterinary research》1989,37(2):71-83
An improved enzyme-linked immunosorbent assay (ELISA) for detection of heterophile Hanganutziu-Deicher (HD) antibodies and antigens, which are frequently detected in sera and/or cancerous tissues from patients with various cancers was developed using biotinylated chicken anti-GM3(NeuGc) antibody and avidin-horseradish peroxidase conjugate. The N-glycolylneuraminyllactosyl-ceramide, GM3(NeuGc) ganglioside was purified from horse erythrocyte membranes. The ELISA procedure required 300 ng GM3(NeuGc) antigen to coat plastic microtiter plates and 190 ng biotinylated antibody per well to give optimum product formation. The technique could detect 6 ng antigen in tissue homogenate as compared to 0.6 ng of the pure compound by inhibition. Chicken anti-GM3(NeuGc) antibody quantitatively inhibited the biotinylated antibody, however, this procedure was not suitable to quantify lower affinity HD antibody in patient sera. Immunostaining specific for HD antigen-positive cells, in tissue sections was by 4 micrograms/ml biotinylated antibody and 200 dilution of Avidin-biotinylated peroxidase complex reagent using pig intestine and lymph node as positive tissues and chicken intestine and lung as negative tissues. 相似文献
16.
Sato T Itou T Sakai T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(5):645-648
The complete coding region sequence of equine muscle-type phosphofructokinase (ePFKM) was obtained from skeletal muscle of a thoroughbred horse. The deduced amino acid sequence of ePFKM showed 97%, 96%, 96%, 96% and 95% identity to canine, human, mouse, rabbit and rat PFKM, respectively. The amino and carboxyl terminal halves of ePFKM presented a structure of tandem repeat, as other mammalian PFKMs. As the amino acid residues constituting various ligand-binding sites were also conserved, it is thought that ePFKM has enzymatic activity similar to PFKM in other mammals. 相似文献
17.
Gordon A Andrews Patricia Sue Chavey Graham Crawford 《Journal of zoo and wildlife medicine》2005,36(4):648-652
Lemurs in captivity progressively accumulate iron deposits in a variety of organs (hemosiderosis) including duodenum, liver, and spleen throughout their lives. When excessive, the toxic effects of intracellular iron on parenchymal cells, particularly the liver, can result in clinical disease and death. The pathogenesis of excessive iron storage in these species has been attributed to dietary factors related to diets commonly fed in captivity. Tissue iron stores can be directly estimated by tissue biopsy and histologic examination, or quantitated by chemical analysis of biopsy tissue, However, expense and risk associated with anesthesia and surgery prevent routine use of tissue biopsy to assess iron status. A noninvasive means of assessing total body iron stores is needed to monitor iron stores in lemurs to determine whether dietary modification is preventing excessive iron deposition, and to monitor potential therapies such as phlebotomy or chelation. Serum ferritin concentration correlates with tissue iron stores in humans, horses, calves, dogs, cats, and pigs. Serum ferritin is considered the best serum analyte to predict total body iron stores in these species and is more reliable than serum iron or total iron binding capacity, both of which may be affected by disorders unrelated to iron adequacy or excess including hypoproteinemia, chronic infection, hemolytic anemia, hypothyroidism, renal disease, and drug administration. We have developed an enzyme-linked immunosorbent assay to measure serum ferritin in lemurs. The assay uses polyclonal rabbit anti-human ferritin antibodies in a sandwich arrangement. Ferritin isolated from liver and spleen of a black and white ruffed lemur (Varecia variegata variegata) was used as a standard. Ferritin standards were linear from 0 to 50 microg/L. Recovery of purified ferritin from lemur serum varied from 95% to 110%. The within-assay variability was 4.5%, and the assay-to-assay variability for three different samples ranged from 10% to 17%. The assay also measures serum ferritin in several other lemur species. 相似文献
18.
Taniguchi K Koida A Mutoh K 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(1):65-70
Although it has been reported that specific proteins are present to take charge in the gustation in the taste buds, there have been only a few reports on the distribution of glycoconjugates binding to glycoproteins on the cellular membranes of the taste cells. In the present study, therefore, binding patters of 24 biotinylated lectins were examined in the three types of lingual papillae in five species of mammals belonging to different orders: cow (artiodactyl), horse (perissodactyl), monkey (primate), dog (carnivore) and mouse (rodent). As the results, lectin binding patterns were different among circumvallate, foliate and fungiform papillae, among the cells of the taste buds, and among animal species. These findings suggest that the different binding patterns of the lectins in the taste papillae and taste bud cells may be involved in different sensitivities of taste among mammalian species. 相似文献
19.
为分离与鸡抗病性密切相关的差异表达基因,并进行功能分析,利用抑制性消减杂交技术,以大骨鸡和海兰褐商品代蛋鸡20周龄时的脾脏组织为试验材料构建消减cDNA文库。文库的插入片段集中在600 bp左右,挑取760个克隆进行PCR筛选获得663个阳性克隆,经过点杂交筛选后选择了531个阳性克隆,从中随机挑取100个阳性克隆进行测序。经过同源性比对归并后得到37个差异表达基因或ESTs序列,其中,32个是已知基因,包括一般抗病性或免疫性能、特异抗病相关基因、细胞信号分子、膜蛋白质、转录因子等差异表达基因;5个为功能尚未确定的基因。并对4个可能影响鸡抗病性能的差异表达基因进行了RT-PCR半定量检测鉴定。该试验结果为进一步研究这些差异表达基因在抗病过程中的重要功能及其调控作用机理奠定了基础。 相似文献
20.
仙居鸡抑制素/活化素β_A亚基成熟区的cDNA克隆及序列分析 总被引：1，自引：0，他引：1
本研究根据发表的来航鸡抑制素 /活化素βA 亚基序列设计引物 ,运用RT PCR技术从仙居鸡卵泡的颗粒细胞总RNA中扩增出抑制素 /活化素βA 亚基成熟区序列 ,并进行了克隆和测序。结果显示 ,所测鸡成熟βA 亚基是由 116个氨基酸 (aa)残基组成的蛋白质 ,共有 9个半胱氨酸残基 ,与发表的鸡及哺乳类相应序列对比 ,其核苷酸序列的同源性分别为 99.2 %和 81%～ 85 .2 % ,其预测氨基酸序列的同源性分别为 10 0 %和 96 .6 %～ 97.4 % ,且所测鸡 βA 亚基成熟区半胱氨酸残基的数目和位置与发表的鸡和哺乳类相同 ,说明该亚基的序列及结构在不同物种间具高度保守性 ,揭示其可能具重要的生理功能 相似文献