Plasma fibronectin concentration associated with various types of canine neoplasia

Fibronectin, a large glycoprotein found in soluble form in plasma and in insoluble form in connective tissue matrices, has been implicated in cell-to-cell and cell-to-substratum interactions, inflammation and tissue repair, phagocytosis, hemostasis, and oncogenic cell transformation. Because fibronectin concentration is diminished or lacking on cell surfaces of many transformed cell lines and because decreased concentration of plasma fibronectin is associated with suboptimal mononuclear phagocyte system function and host defense, plasma fibronectin concentration was evaluated in 119 dogs with various forms of neoplasia. Included were 43 dogs with neoplasia of the skin and soft tissue, 18 with gastrointestinal tract neoplasia, 29 with mammary gland neoplasia, and 29 with various other types of neoplasia. Of the dogs studied, 44 (37%) had evidence of metastatic disease. This group had fibronectin concentration that differed significantly (P less than 0.01) from the plasma fibronectin concentration reference interval. Within this group, 9 dogs (20% of this group) had plasma fibronectin values within the reference interval, 33 (75%) had significantly (P less than 0.01) lower values than the reference interval, and 2 (5%) had significantly (P less than 0.01) higher values than the reference interval. These data suggested that fibronectin concentration determination, when results are abnormal, may be of diagnostic and prognostic interest.     

Binding of curcumin to senile plaques and cerebral amyloid angiopathy in the aged brain of various animals and to neurofibrillary tangles in Alzheimer's brain

The binding of curcumin to senile plaques (SPs) and cerebral amyloid angiopathy (CAA) was examined in the aged brain of various animal species and a human patient with Alzheimer's disease (AD), together with its binding to neurofibrillary tangles (NFTs). Brain sections were immunostained with anti-amyloid β protein 1-42 (Aβ42) and anti-amyloid β protein 1-40 (Aβ40) antibodies. These sections were also stained with alkaline Congo red, periodic acid-methenamine silver (PAM), and curcumin (0.009% curcumin solution) with or without formic acid pretreatment. The sections from the AD brain were also immunostained for anti-paired helical filament-tau (PHF-tau), and were stained with Gallyas silver for NFTs. Some SPs in the AD, monkey, dog, bear, and amyloid precursor protein transgenic mouse (APP Tg-mouse) brains contained congophilic materials, and were intensely positive for curcumin. In addition, curcumin labeled some diffuse SPs negative for Congo red in the AD, monkey, bear, and APP Tg-mouse brains. In all animals, CAA was intensely positive for both Congo red and curcumin. The specific curcumin staining activity was lost by formic acid pretreatment. In the AD brain, NFTs positive for PHF-tau and Gallyas silver were moderately stained with curcumin. These findings indicate that curcumin specifically binds to the aggregated Aβ molecules in various animals, and further to phosphorylated tau protein, probably according to its conformational nature.     

Purification and partial characterization of thyroid hormone binding proteins in canine serum

Thyroxine-binding prealbumin (TBPA) and thyroxine-binding globulin (TBG) were isolated from canine serum and partially characterized. TBPA was isolated by retinol-binding protein (RBP) affinity chromatography and further purified by preparative agarose gel electrophoresis or FPLC ion exchange chromatography. TBG was purified by thyroxine (T₄)-Sepharose chromatography followed by gel filtration on Sephacryl S-300 and preparative electrofocusing in a granulated dextran gel. Molecular weights were estimated by SDS-polyacrylamide gradient gel electrophoresis. Canine TBPA had a tetramer molecular weight of 56,000, an extinction coefficient of 12.8 cm²cg⁻¹, an isoelectric point of 5.26–5.70 and a microheterogeneity pattern similar to that of human TBPA. Partial immunochemical identity with human TBPA was also found. Plasma concentrations of TBPA were measured by rocket immunoelectrophoresis in 43 normal and 35 hypothyroid dogs. Reference levels for TBPA ranged between 205 and 474 mg/l. Hypothyroid dogs had a mean TBPA level of 315.0 mg/l (SD: 91.1 mg/l). TBG had a molecular weight of 75,000 and an isoelectric point of 5.0. No immunochemical identity with human TBG was found. Gel filtration of serum on Sephacryl S-200, identification of T₄-binding proteins with ¹²⁵I-T₄, and protein- and lipoprotein staining of fractions was performed. Thyroxine-binding was found to TBG in the β-globulin region, TBPA in the ₂-region, albumin, and to the high density lipoprotein (HDL₂) in the ₁-region and the very low density lipoprotein (VLDL) in the pre-β region. A corresponding band to the latter protein in serum was masked by TBG and TBPA, and T₄-binding in the ₁-region was not always seen in serum. Many similarities were found between man and dog regarding TBPA, but not TBG. The differences in structure of TBG may in part be responsible for the low serum T₄ levels and rapid T₄ metabolism seen in dogs.     

DNA binding proteins in canine sera. A method for removal of nonspecific DNA binding in the Farr assay

A panel of canine sera, the majority of which were collected from clinically healthy dogs, were investigated for antibodies against double stranded (dsDNA) by the Farr radioimmunoassay technique. Non-specific DNA binding agents interfering with the Farr assay were detected in all sera. Heat inactivation at 60 degrees C or treatment with dextran sulphate was shown to eliminate this kind of unspecific DNA binding while not affecting true antibodies to dsDNA. Canine sera positive in the Farr assay after inactivation at 60 degrees C were positive also in immunofluorescence for anti-nuclear antibody on rat liver sections and for dsDNA with Chrithidia luciliae as antigen preparation. IgG or glycoprotein nature of the non-specific DNA binding could be excluded by means of affinity chromatography on protein A and the lectin lentil.     

Comparison of various canine blood-typing methods

OBJECTIVE: To compare canine blood-typing results determined by use of the card (CARD), gel (GEL), Michigan State University (MSU), and tube (TUBE) tests. SAMPLE POPULATION: Blood samples from 23 healthy dogs. PROCEDURES: Blood samples anticoagulated with EDTA were screened by use of each blood-typing method according to manufacturers' protocols. RESULTS: Strong RBC agglutination reactions were observed with dog erythrocyte antigen (DEA) 1.1 reagents of the CARD and GEL tests as well as MSU test (only after adding Coombs' reagent) in 9 blood samples. By use of the CARD test, RBCs from 4 additional dogs agglutinated weakly; on the basis of MSU test results, these 4 dogs were classified as DEA 1.2 positive. All blood samples agglutinated with the B antigen reagent of the TUBE test. All but 2 blood samples had strong positive reactions with the DEA 4 reagent of the MSU test. All but 3 blood samples reacted with the E antigen reagent of the TUBE test. Three blood samples agglutinated with the DEA 3 reagent of the MSU test and A antigen reagent of the TUBE test. Five blood samples had strong agglutination reactions with the DEA 5 reagent of the MSU test. CONCLUSIONS AND CLINICAL RELEVANCE: Use of the CARD test allows for rapid identification of DEA 1.1 but may produce weak reactions with blood from DEA 1.2-positive dogs. The GEL test is a reliable and rapid clinical laboratory method for identification of DEA 1.1. The MSU test requires Coombs' reagent for identification of DEA 1.1 and 1.2.     

The growth profiles of three types of canine distemper virus on Vero cells expressing canine signaling lymphocyte activation molecule

To know growth profiles of canine distemper virus (CDV) on Vero cells stably expressing canine signaling lymphocyte activation molecule (Vero-DogSLAMtag; Vero-DST cells), the propagation of three strains of CDV was tested in Vero-DST cells in comparison with parental Vero cells. Strain MD77 could grow well in both cell lines, but demonstrated no syncytium formation or indistinguishable rounding cytopathic effects (CPE) in Vero cells. Strains Onderstepoort and KDK-1 also grew well in Vero-DST cells with apparent syncytium CPE, while they grew less or no efficiently, respectively, in Vero cells. All three CDV strains demonstrated the peak titers, in Vero-DST cells before reaching to an extensive CPE and drastic decrease of titers at/after full CPE. Immunohistochemistry revealed that viral antigens of all CDV strains were found exclusively in the syncytia in Vero-DST cells, while in Vero cells, viral antigen was identified in their single cells for strain MD77 but none for other strains. Thus, every strain of CDV could grow well in Vero-DST cells and behaved differently against Vero cells. These results would be of practical value for workers of CDV because 1) In Vero-DST cells, by observation of distinct syncytium CPE, the highest titer or the best growth of virus could be identified; 2) In Vero cells, various CDV strains could be readily classified after propagation in Vero-DST cells.     

The qualitative analysis of canine immunoglobulins and myeloma proteins by immunofixation

The technique of immunofixation was used to locate the immunoglobulin species in canine sera following agarose electrophoresis. The myeloma proteins in two dogs were identified as immunoglobulin G, and a polygonal gammopathy was characterized by this method.     

不同类型抗盐植物整株水平游离脯氨酸的分配

采用田间试验及盆栽试验,对3种不同类型(拒盐、泌盐和稀盐)的抗盐植物植株水平游离脯氨酸的分配特性进行了研究。结果表明,不同类型的植物游离脯氨酸有如下的规律:稀盐植物整株中的游离脯氨酸的含量很低;泌盐植物整株中的游离脯氨酸的含量介于稀盐植物和拒盐植物之间;拒盐植物整株中的游离脯氨酸含量明显高于前2类植物。3种类型的抗盐植物植株中游离脯氨酸分配的共同特点是游离脯氨酸多集中于代谢旺盛的光合器官和生殖器官,在盐逆境下植物优先保护其光合器官和生殖器官。     

The growth of attenuated strains of canine parvovirus, mink enteritis virus, feline panleukopenia virus, and rabies virus on various types of cell cultures

The growth characteristics were studied in the attenuated strains of canine parvovirus CPVA-BN 80/82, mink enteritis virus MEVA-BN 63/82 and feline panleucopenia virus FPVA-BN 110/83 on the stable feline kidney cell line FE, and in the attenuated canine distemper virus CDV-F-BN 10/83 on chicken embryo cell cultures (KEB) and cultures of the stable cell line VERO. When the FE cultures were infected with different parvoviruses in cell suspension at MOI 2-4 TKID50 per cell, the first multiplication of the intracellular virus was recorded 20 hours p. i. In the canine parvovirus, the content of intracellular and extracellular virus continued increasing parallelly until the fourth day; then, from the fourth to the sixth day, the content of extracellular virus still increased whereas that of intracellular virus fell rapidly. In the case of the mink enteritis virus the release of the virus into the culture medium continued parallelly with the production of the cellular virus until the sixth day. In the case of the feline panleucopenia virus the values concerning free virus and virus bound to cells were lower, starting from the second day p. i. When KEB or VERO cultures were infected in cell suspension with the canine distemper virus at MOI about 0.004 per 1 cell, the replicated intracellular virus was first recorded in the KEB cultures five hours after infection but in the VERO cultures only 20 hours after infection, with a timely release of the virus into the culture medium in both kinds of tissue. In the KEB and VERO cultures the highest values of infection titres were recorded on the fourth day p. i., the course of virus multiplication on the cells being parallel with its release into the culture medium.     

Evaluation of ifosfamide for treatment of various canine neoplasms

lfosfamide (3-[2-chloroethyl]-2[(2 chloroethyl)amino]tetrahydro-2H-1,3,2-oxazaphosphorine 2-oxide) is an alkylating agent with a broad spectrum of antitumor activity. The efficacy and toxicity of ifosfamide were evaluated in 72 dogs with spontaneously occurring tumors. Forty dogs (56%) had lymphoma, 31 (43%) had sarcomas, and 1 had a metastatic carcinoma. Five dogs received ifosfamide at dosages <350 mg/m2 IV. Neither toxicity nor response were observed, and the remaining dogs received ifosfamide at 350 mg/m2 (n = 18) and 375 mg/m2 body surface area IV (n = 49). Saline diuresis and the thiol compound mesna were used to prevent urothelial toxicity. Fifty-two dogs had measurable tumors and could be evaluated for response. Complete responses were seen in 1 dog with metastatic leiomyosarcoma of the urinary bladder and in 1 dog with metastatic cutaneous hemangiosarcoma. One dog with lymphoma had a partial response for 112 days. Six dogs with splenic hemangiosarcoma received ifosfamide postsplenectomy and their median survival time was 147 days. The acute dose limiting toxicity was neutropenia 7 days after administration of ifosfamide. The median and mean neutrophil counts 7 days after ifosfamide at 350 mg/m2 were 2,035 cells/microL and 4,773 cells/microL, respectively (n = 12). The median and mean neutrophil counts 7 days after ifosfamide at 375 mg/m2 were 2,500 cells/microL and 3,594 cells/microL, respectively (n = 37). No dog developed clinical or microscopic evidence of hemorrhagic cystitis. Ifosfamide appears safe to use in tumor-bearing dogs, and the evaluation of combination chemotherapy protocols that include ifosfamide should be considered.     

Uropathogenic virulence factor FimH facilitates binding of uteropathogenic Escherichia coli to canine endometrium

Pyometra is a potentially life-threatening condition in bitches and is often caused by Escherichia coli infection. Both pathogenic and non-pathogenic E. coli strains commonly carry the genes for type 1 fimbriae that mediate bacterial adhesion onto host epithelium. To investigate whether the type 1 fimbrial adhesin, FimH, facilitates the binding of uropathogenic E. coli to canine endometrium, the fimH gene was insertionally inactivated in a pathogenic E. coli strain. The ability of E. coli to bind to canine endometrial epithelial cells was determined in vitro using canine uterine biopsies. Binding of the fimH mutant was only 0.3% of that of the wild type. Complementation of the mutation restored the phenotype to that of the parent. This study has developed an in vitro model that allows quantitative and qualitative assessment of bacterial binding to canine endometrium and has demonstrated that the fimH gene plays a role in adherence of pathogenic E. coli to canine endometrium.     

Histochemical differentiation of fiber types in neonatal canine skeletal muscle

Differentiation of fiber types in developing canine skeletal muscle was studied, using morphologic, morphometric, and histochemical techniques. Sample collections were made from 6 muscles from the pectoral and pelvic limbs of 16 healthy pups between 1 day and 12 weeks of age. In newborn pups, 90% to 95% of the fibers in the 6 muscles were classified as undifferentiated or type IIC; the remaining fibers were classified either normal or large-size type I. Large-size type I fibers usually accounted for 2% to 4% of the total population and were considered analogous with the B fiber of Wohlfart. These fibers were larger than all other fiber types and disappeared after pups reached 4 to 5 weeks of age. After 2 to 4 weeks, the number of undifferentiated fibers decreased with the appearance of, and the concomitant numerical increases of, normal size type I and type IIA fibers. The percentages of type I and IIA fibers approached proportions of the adult dog by 12 weeks, at which time a type IIA fiber predominance was present in biceps femoris, lateral head of the gastrocnemius, cranial tibial, and long head of the triceps. Type I fibers predominated in medial head of the triceps and superficial digital flexor after 4 to 5 weeks. The mean fiber diameters of type I and IIA fibers were similar to any given muscle throughout the postnatal development. All fiber types stained uniformly with the oxidative stain nicotinamide adeninedinucleotide-tetrazolium reductase during the first 12 weeks of life, whereas a distinction between type I and II fibers was evident after 3 to 4 weeks with the periodic acid-Schiff stain reaction.     

Histochemical identification of fiber types in canine skeletal muscle.

Proximal and distal skeletal muscles from pectoral and pelvic limbs were histochemically examined in 18 neuromuscular disease-free dogs. On the basis of the human system of classification and nomenclature and results of standard adenosine triphosphatase (ATPase) and glycine-formaldehyde preincubation procedures, the fiber types identified in immature and mature canine skeletal muscles were I, IIA, and IIC.     

硒对犬的营养作用

作者从微量元素硒的主要来源、在犬体内的分布形式、吸收、代谢和排泄,硒对犬的营养功能方面进行比较全面的综述,为种用犬饲养过程中合理利用微量元素硒提供了科学依据。