Observations on associated histopathology with Aggregata octopiana infection (Protista: Apicomplexa) in Octopus vulgaris

The effect of cimetidine on the praziquantel concentration in the blood of the rockfish Sebastes schlegeli and the consequent effect on the treatment efficacy against Microcotyle sebastis were investigated. Fish were divided into 7 groups and orally administered praziquantel alone (200 and 100 mg kg(-1) body weight [BW]) or in combination with cimetidine (in doses of 200, 100 or 50 mg kg(-1) BW cimetidine with a praziquantel dose of 100 mg kg(-1) BW). The fish in the sixth group were coadministered 50 mg praziquantel and 200 mg cimetidine kg(-1) BW. The fish in the control group were administered only saline. At 24 h post-treatment, the plasma was analyzed for praziquantel by reversed-phase high-performance liquid chromatography (RP-HPLC) using diazepam as the internal standard, and the gills were examined to confirm the effectiveness of each treatment. The praziquantel concentration in plasma of fish administered 100 mg praziquantel + 200 mg cimetidine kg(-1) BW was not significantly different from that of fish treated with 200 mg praziquantel kg(-1) BW and was significantly (p < 0.05) higher (about 2 times) than that of fish administered 100 mg praziquantel kg(-1) BW. The group of fish administered 50 mg praziquantel + 200 mg cimetidine kg(-1) BW showed a similar plasma praziquantel concentration to that in the fish treated with 100 mg praziquantel kg(-1) BW. The treatment efficacies of the groups of fish coadministered 100 mg praziquantel kg(-1) BW and various concentrations of cimetidine (200, 100 and 50 mg kg(-1) BW) were not significantly different from that of the group of fish administered 200 mg praziquantel kg(-1) BW, but were significantly higher than those of the groups of fish fed 100 mg praziquantel kg(-1) BW alone or coadministered 50 mg praziquantel + 200 mg cimetidine kg(-1) BW.     

Purification of murine thymus leukemia antigen (TL). A quantitative assessment of limited proteolysis.

The murine thymus leukemia antigen (TL) has been solubilized from the tumor ASL1 and from an established cell line ASL1W, by papain digestion. When a 15-min digest was chromatographed on Sephadex G-200, two peaks of TL activity were eluted with apparent molecular weights of approximately 58,000 and 31,000. Chromatography of a 30-min digest under the same conditions resulted in elution of a single peak of activity with an apparent molecular weight of 58,000. Additional purification was carried out on the 58,000 molecular weight material by absorption to, and elution from DEAE-cellulose. The combination of gel filtration and ion exchange chromatography resulted in approximately a 150-fold purification.     

Interaction of genes controlling two allotypes in chickens.

This paper presents the results of the investigations of the newly detected antigen of chicken blood serum, called K2. It was established that the K2 antigen which was identified with isoimmune serum was a beta-globulin with the molecular weight over 200 000. The results of the genetic analysis based on sire-dam-offspring combinations seemed to indicate that the antigen under examination was controlled by a gene hypostatic to the gene controlling the previously described K1 allotype.     

Interaction of genes controlling two allotypes in chickens1

This paper presents the results of the investigations of the newly detected antigen of chicken blood serum, called K2. It was established that the K2 antigen which was identified with isoimmune serum was a β-globulin with the molecular weight over 200 000. The results of the genetic analysis based on sire-dam-offspring combinations seemed to indicate that the antigen under examination was controlled by a gene hypostatic to the gene controlling the previously described Kl allotype.     

Hypoglycemic effect of a Lentinus edodes exo-polymer produced from a submerged mycelial culture

The hypoglycemic effect of an exo-polymer produced from a submerged mycelial culture of Lentinus edodes was investigated in streptozotocin-induced diabetic rats. The administration of the exo-polymer (200 mg/kg BW) reduced the plasma glucose level by as much as 21.5%, and increased plasma insulin by 22.1% as compared to the control group. It also lowered the plasma total cholesterol and triglyceride levels by 25.1 and 44.5%, respectively. Gel chromatography of the exo-polymer revealed a single peak which is likely to have been a glycoprotein with a molecular weight of 52 kDa and was found to contain 83.5% carbohydrate and 16.5% protein. The Sugar and amino acid compositions of the exo-polymer were analyzed in detail.     

重组鼠疫菌F1抗原的纯化及其在ELISA检测鼠疫免疫中的应用

从表达rF1抗原的大肠杆菌中以Superdex-200凝胶过滤层析纯化rF1抗原,电泳扫描显示纯化率>90%。SDS-PAGE及琼脂免疫双扩散结果显示纯化的rF1抗原具天然F1抗原的活性。将此纯化的rF1抗原用于间接ELISA分析免疫动物血清中抗F1抗体的水平,并与天然F1抗原相比较,证明rF1抗原优于天然F1抗原分析结果,可用于鼠疫的血清学检测。     

Isolation and partial characterization of a 700 kilodalton human colon carcinoma associated antigen

An antigen of high molecular weight (CA-3) was isolated from the cytosol fraction of GW-39 human colon tumor cells by antibody affinity chromatography. CA-3 was characterized by an acidic pI value of 4.5-4.9, a molecular weight of 700 kilodaltons and a sedimentation coefficient of 13S. It contained all of the commonly occurring amino acids and had an acidic to basic amino acid ratio of 1.4. CA-3 was resistant to dissociation by reducing agents as well as by sodium dodecylsulfate. Quantitation of CA-3 by a radioimmunoassay employing rabbit anti-CA-3 antiserum revealed a marked elevation of CA-3 in the cytosol extracts of human primary colon carcinoma in comparison to normal colon. The molecular properties of CA-3 are compared to those of carcinoembryonic antigen, high molecular weight colon specific antigen CSAp and two other high molecular weight proteins, fibronectin and conglutinin. Colon antigen CA-3 appears to be different from these other molecules in terms of its molecular weight, sedimentation value, isoelectric point and amino acid composition.     

Monoclonal Antibodies Specific for Members of the Genus Endotrypanum

Twenty-six monoclonal antibodies were produced against membrane-enriched preparations of Endotrypanum schaudinni or Endotrypanum sp. promastigotes. Fifteen of these monoclonal antibodies (E1-E15) reacted only with the standard strain of E. schaudinni , M6159. Monoclonal antibodies E16-E26 were considered Endotrypanum specific; no cross reactivity was detected with any other genus of the family Trypanosomatidae (Leishmania, Trypanosoma, Leptomonas. Herpetomonas or Crithidia) by dot-blot radioimmune assay. By indirect immunofluorescence assay, the antigens recognized by Endotrypanum specific monoclonal antibodies appear to be associated with the surface of the parasite. Based on Western blot analysis, 4 antigenic molecules ranging in molecular weight from 24 kD to 160 kD were identified by monoclonal antibodies specific for the strain of E. schaudinni , M6159. Monoclonal antibodies specific for the genus Endotrypanum identified an antigen of molecular weight 48 kD as well as a diffuse component migrating with an apparent molecular weight of 64–200 kD.     

Monoclonal antibodies specific for members of the genus Endotrypanum

Twenty-six monoclonal antibodies were produced against membrane-enriched preparations of Endotrypanum schaudinni or Endotrypanum sp. promastigotes. Fifteen of these monoclonal antibodies (E1-E15) reacted only with the standard strain of E. schaudinni, M6159. Monoclonal antibodies E16-E26 were considered Endotrypanum specific; no cross reactivity was detected with any other genus of the family Trypanosomatidae (Leishmania, Trypanosoma, Leptomonas, Herpetomonas or Crithidia) by dot-blot radioimmune assay. By indirect immunofluorescence assay, the antigens recognized by Endotrypanum specific monoclonal antibodies appear to be associated with the surface of the parasite. Based on Western blot analysis, 4 antigenic molecules ranging in molecular weight from 24 kD to 160 kD were identified by monoclonal antibodies specific for the strain of E. schaudinni, M6159. Monoclonal antibodies specific for the genus Endotrypanum identified an antigen of molecular weight 48 kD as well as a diffuse component migrating with an apparent molecular weight of 64-200 kD.     

Identification, immunoaffinity purification and partial characterization of a human decidua-associated protein

A crude extract of pooled early-pregnancy decidual tissue was enriched for soluble decidual proteins by exhaustive affinity absorption with antibodies to human serum proteins immobilized on Eupergit C. The partly purified extract was used to prepare monoclonal antibodies. A monoclonal antibody was obtained recognizing an antigen present in extract of decidual tissue and not in extract of proliferative endometrium. The monoclonal antibody was used for immunoaffinity purification of the decidua-associated protein. By SDS-PAGE analysis, under reducing conditions it yielded 2 bands at apparent molecular weights of 55,000 and 25,000. Under non-reducing conditions a single protein band at apparent molecular weight of 200,000 was observed. The Mr 200,000 protein was named hDP200 and the Mr 55,000 protein was named hDP55. It is suggested that hDP55 is a subunit of the hDP200. The hDP200 did not react with polyclonal antibodies specific for PP12 and PP14. PP14 has been shown to be immunologically indistinguishable from PEP and alpha 2-PEG. Our data therefore suggest that hDP200 is a novel human decidua-associated protein.     

Characterisation of 24-kD proteins from rat testes using polyclonal sera reactive to human sperm antigens

A group of antigens of 24-kD Mr from rat testes were characterised biochemically. These antigens were part of a larger molecule of approximately 200 kD. On treatment with disulfide bond reducing agent, the 200-kD molecule was reduced to subunits. Immunoreactivity was confined to a doublet of approximately 24 kD and a single band of approximately 50 kD Mr after the reduction. Glycoprotein in nature, this antigen shared immunoreactive epitopes with a 40-kD antigen on human spermatozoa. Antiserum raised in rabbits against the 24-kD antigen from rat testes reacted with antigens on the acrosome of human spermatozoa. Agglutination of sperm could be induced by the antiserum. The carbohydrate residue could be removed by mannosidase digestion. Chemical deglycosylation studies showed a slight decrease in molecular weight. Immunoreactivity was however not completely lost after chemical deglycosylation. Isoelectric focusing of the antigen identified nine isoelectric species. Two relatively minor species showed immunoreactivity. Acrosome-reacted spermatozoa showed loss of antigens from acrosome.     

Antibody response to a solubilized tumor-associated membrane antigen (TAMA) from the murine lewis lung tumor

A tumor-associated membrane antigen (TAMA) has been solubilized from the C57B1/6 murine Lewis lung tumor using the detergent Triton X100. Xenoantiserum prepared in rabbits was used to identify this tumor antigen in Triton extracts by double immunodiffusion in agar. The antigen was partially purified using a combination of DEAE and Sephadex G200 chromatography followed by sucrose gradient isoelectric focusing. Isoelectric focusing also showed the antigen to have a pI at pH 4.7. Sodium dodecylsulfate polyacrylamide gel electrophoresis indicated the antigen to be of 37,000 molecular weight. The 125I protein A antibody-binding assay was used to further characterize the specificity of the antigen using rabbit antisera to the Triton extract. The same assay system also detected a specific antitumor humoral response in syngeneic mice immunized with the Triton extract.     

Brain Atrophy in a Murine Model of Chronic Fatigue Syndrome and Beneficial Effect of Hochu-ekki-to (TJ-41)

Brain-derived neurotrophic factor (BDNF) is associated with the main symptoms of chronic fatigue sydrome (CFS) and neuron apoptosis. Nevertheless, no study has been performed directly to explore the relationship between CFS, BDNF and neuron apoptosis. We induced a CFS model by six injections of killed Brucella abortus antigen in BALB/c mice and treated them with Hochu-ekki-to (TJ-41). Daily running activity, body weight (BW), ratio of cerebral weight to BW (CW/BW) and expression levels of BDNF and Bcl-2 mRNA in the hippocampus were determined. The daily activity and CW/BW decreased significantly in the CFS model. BDNF and Bcl-2 mRNA expression levels in the hippocampus were suppressed in the CFS model and TJ-41 treated mice, while no significant difference was found between them. We improved a murine model to investigate the relationship between CFS and brain dysfunction. In this model, reduced daily activity might have been associated with decreased hippocampal BDNF mRNA expression, hippocampal apoptosis and brain atrophy. TJ-41 increased the daily running activity of the model, which was independent of brain recovery.     

Antigens of Neisseria gonorrhoeae: Characterization by Gel Filtration, Complement Fixation, and Agar-Gel Diffusion of Antigens of Gonococcal Protoplasm

With the use of the agar-gel-diffusion and complement-fixation techniques, it was shown that protoplasm from different gonococcal isolates reacted with sera from some humans with a history of gonorrhea but did not react with "normal" human sera. The reactive antigen(s) could be partially separated from the other antigens by passing the gonococcal protoplasm through Sephadex G-200. The antigen(s) reacting in the gel-diffusion and complement-fixation tests appeared in the same fraction. On the basis of Sephadex gel filtration, the molecular weight of this antigen(s) is probably greater than 200,000.     

Subfragments of the papain solubilized TL antigen.

TL antigen was solubilized from the tumor ASLI (TL. 1,2,3) by papain digestion. The subfragments of 125I-labeled TL were examined by two methods. The first involved immune precipitation followed by electrophoresis on SDS-acrylamide gels. This treatment yielded three bands of molecular weight 39,000 and 19,000, as well as material which migrated with the tracking dye. In the second procedure the papain digested material was partially purified on Sephadex G-200. The active fraction from G-200 was labeled with 125Iodine, mixed with alloantiserum and rechromatographed on G-200. The isolated immune complexes were boiled in SDS and 2-mercaptoethanol, then separated on a SDS-Sephadex G-150 column. Two radioactive peaks were eluted indicating an absence of the 19,000 m.w. component following the latter method of purification.     

Isolation and characterization of alpha-fetoprotein from the mouse hepatoma BW7756.

Purification of alpha-fetoprotein from mouse hepatoma BW7756 extracts was performed using ammonium sulfate precipitations, gel filtration, ion-exchange chromatography and isoelectric focusing. These procedures produced a 5.6% yield of alpha-fetoprotein with 96% purity. Polyacrylamide slab gel electrophoresis, extended agarose electrophoresis and immunoelectrophoresis demonstrated that mouse hepatoma alpha-fetoprotein migrated at pH 8.6 as a rapid alpha1, or postalbumin globulin. Crossed antibody electrophoresis of the agarose zone containing alpha-fetoprotein failed to demonstrate microheterogeneity. Molecular weight analysis of the mouse hepatoma alpha-fetoprotein on a calibrated Sephadex G-200 column yielded a value of 72 000-73 000 for the native protein. Sodium dodecyl sulfate gel electrophoresis subsequently demonstrated a single polypeptide chain with a molecular weight of 72 000. Amino acid analysis showed the alpha-fetoprotein to be an acidic protein dominated by hydrophobic residues. The total carbohydrate content was 5.5%, and 3 mol of sialic acid were detected per mol of alpha-fetoprotein. Although neutral sugars were the principal class present, galactosamine was the most abundant single sugar detected.     

Identification of the core protein carrying the Tn antigen in mouse brain: specific expression on syndecan-3

We isolated glycoproteins carrying the Tn antigen, which was expressed spatiotemporally in the developing mouse brain. The Tn antigen was expressed on two molecular species with a molecular weight from 200 to 350 kDa and 110 to 160 kDa, as judged on SDS-PAGE. Although the two glycoproteins showed different susceptibilities to heparitinase I and solubilities in a salt solution, after treatment with V8 protease they showed the same mobility corresponding to a molecular weight of 90 kDa on SDS-PAGE, suggesting that these two molecules shared a common core protein. Partial N-terminal sequences of the glycoproteins were determined, i.e. AQRXRNENFERPV and ALAAPXAPAMLP, which were identified as the sequences of the N-terminal and central portions of syndecan-3, respectively. Both glycoproteins were reactive to anti-mouse syndecan-3 antibody. These results suggest that one is a soluble syndecan-3 cleaved between mucin-like domain and transmembrane domain, and the other is a membrane-bound syndecan-3 lacking N-terminal glycosaminoglycan attachment sites, and that both glycoproteins have a mucin-like domain characteristic of syndecan-3, in which the Tn antigen may be expressed.     

Apportioning protein requirements for maintenance v. growth for blue-breasted quail (Excalfactoria chinensis) from 7 to 21 days of age

The aim of this study was to investigate protein requirements for the maintenance and growth of blue-breasted quail (Excalfactoria chinensis) from 7 to 21 days of age. A total of 180 quails, 7 days old, were randomly assigned to 36 cages and for 2 weeks were fed diets with a metabolisable energy concentration of 12.13 MJ/kg and a dietary CP concentration of 125, 150, 175, 200, 225 or 250 g/kg. The average BW per cage and the feed intake per cage were recorded daily. The results showed that quails fed 125 g/kg CP could not maintain their BW and had negative feed efficiency. There were linear and quadratic relationships between CP level and response criteria, including BW, weight gain, feed intake, feed efficiency, final body nitrogen mass and body nitrogen accretion (P<0.05). The dietary CP requirements, as calculated using a one-slope quadratic broken-line model, were 211 and 202 g/kg according to weight gain and feed efficiency, respectively. The regression equations, on the basis of metabolic BW, of daily weight gain on daily protein intake according to the model were Y=0.137-2.128(0.113-X) if X<0.113 and Y=0.137 if X>or=0.113 (R2=0.96, P<0.001), which meant that the protein requirement for maintenance was 0.049 times the metabolic BW and that to gain 1 g weight quails needed to ingest an extra 0.47 g protein after the maintenance requirement was satisfied. The regression equations, on the basis of metabolic BW, of daily body nitrogen accretion on daily protein intake according to the model were Y=5.667-76.700(0.119-X) if X<0.119 and Y=5.667 if X>or=0.119 (R2=0.95, P<0.001), which meant that quails had to receive an amount of protein equal to their metabolic BW multiplied by 0.045 to satisfy the requirement for maintenance and then ingest an extra 13 g protein to accrete 1 g body nitrogen. In conclusion, growth or protein accretion rates should be regulated according to dietary CP for specific experimental purposes via apportioning protein requirements for maintenance v. growth.     

Analysis of H-2-linked immune responses involved in resistance to AKR tumor growth

H-2-associated immune response gene(s) govern resistance to growth of a spontaneous AKR lymphoma, BW5147. The antigenic specificities recognized by the anti-BW5147 humoral response have been characterized and include: Thy-1, a T -cell differentiation antigen; gp70, a retroviral envelope protein; and several previously uncharacterized proteins, including a 78 000 molecular mass protein, p78, which is restricted to expression on BW5147 cells and five phosphoproteins with molecular masses of 33 000, 29 000, 23 000, 17 000, and 16 000. Only mice which are able to respond to Thy-1, p78, and the phosphoproteins can survive an inoculation of BW5147. Thus, resistance to BW5147 is complex and involves multiple antigens with possible roles in tumor rejection.Abbreviations used in this paper DMEM Dulbecco's modified Eagle's medium - FCS fetal calf serum - Ir immune response - MuLV murine leukemia virus - NMS normal mouse serum - PBS phosphate-buffered saline - SDS sodium dodecyl sulfate - SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis - TCA trichloroacetic acid     

An antigen common to a wide range of bacteria. I. The isolation of a 'common antigen' from Pseudomonas aeruginosa

In crude water-soluble extracts of Pseudomonas aeruginosa 64 antigens can be demonstrated by crossed immunoelectrophoresis in agarose with polyvalent Pseudomonas-immunoglobulin. One of these antigens cross-reacts with antigens prepared from bacteria of a wide range of taxonomic groups. Monospecific immunoglobulins to this antigen (Common Antigen) were produced by immunization with the appropriate immunocomplex extracted from agarose. Common Antigen was purified by the combination of two fractionation methods: Precipitation of the crude extract with 18% (w/v) sodium sulfate, followed by gel filtration of the supernatant on a Sephadex G-200 column. By this method, 35% of Common Antigen from the crude extract was recovered, more than half of the fractions electrophoretically pure. Electrophoresis of reduced Common Antigen on a dodecyl sodium sulfate polyacrylamide gel revealed two protein bands with apparent molecular weights of 59-62 000 and 62-65 000, respectively. The untreated antigen, however, passed a column of Sephadex G-200 with the void volumen, indicating a substance of high molecular weight (> 4-600 000).