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Biomedical subjects

J Franz

Publications and source records attributed to J Franz.

At least 19 recordsLinked to original sources

Recombinant fish neurotrophin-6 is a heparin-binding glycoprotein: implications for a role in axonal guidance.

Neurotrophin-6 (NT-6) was identified in the teleost fish Xiphophorus as a new member of the neurotrophin gene family. NT-6 binds specifically the glycosaminoglycan heparin. In this study NT-6 was expressed in a stably transfected mammalian cell line, and in insect cells via a recombinant baculovirus. It was purified to homogeneity and characterized by MS and N-terminal sequencing. NT-6 from both expression systems was proteolytically processed at one of two protease cleavage motifs and was found to be glycosylated. It supported the survival of embryonic chick sensory neurons; half-maximal survival was observed at 100 ng/ml. Furthermore, NT-6 elicited neurite outgrowth in explanted embryonic dorsal root ganglia. Addition of heparin into the medium did not potentiate the activity of NT-6 in survival assays. However, when a sensory ganglion explant was cultured in a collagen gel matrix assay adjacent to a heparin bead coated with NT-6, neurite outgrowth directed towards the bead was observed. This indicated that NT-6 was slowly released from the heparin bead generating a concentration gradient of NT-6 instrumental for axonal guidance in vitro. Thus the interaction of NT-6 with heparin might not be required for the activation of the cellular receptor for NT-6 on responsive cells but rather may serve to control, in vivo, the distribution of NT-6.

Amino Acid Sequence

Spontaneously released substance P and bradykinin from isolated guinea-pig bladder.

OBJECTIVES: To investigate whether the isolated urinary bladder spontaneously releases substance P (SP) or bradykinin (BK), which can act as potent mediators of pain and inflammation of the urinary bladder, and whether peptidase inhibitors enhance peptide release. MATERIALS AND METHODS: Urinary bladder segments (2 x 10 x 0.8-1 mm) were isolated from guinea pigs and studied in vitro; tissue contraction was assessed using force-displacement transducers and the release of peptides by specific enzyme immunoassays. RESULTS: In the absence of any exogenous agonists, the inhibition of neutral endopeptidase and angiotensin-converting enzyme by phosphoramidon and captopril, respectively, increased the frequency and magnitude of spontaneous motility of isolated bladder strips. Phosphoramidon increased the net release of SP-like immunoreactivity (SP-LI) and captopril increased the net release of SP-LI and BK-LI, concomitant with contraction. Peptide-LI was recovered primarily from bladder mucosa and to a lesser degree from detrusor smooth muscle. Similarly, peptidase inhibitors primarily affected the bladder mucosa; phosphoramidon induced a fourfold increase in SP-LI and captopril induced a significant increase of SP-LI and BK-LI from the mucosa. Tissues contracted in response to peptidase inhibitors in the presence of atropine and indomethacin, but contraction was reduced significantly by in vitro capsaicin desensitization or removal of bladder mucosa. BK stimulated SP-LI release from mucosa but not detrusor. SP stimulated increased BK-LI release from mucosa and detrusor. CONCLUSIONS: These findings indicate the basal release of peptide-like immunoreactivity by isolated bladder and further support the concept that peptidases located in the bladder mucosa are important in terminating the effects of endogenous peptides.

Animals

Enhancement of antibody response to bovine herpesvirus 1 with non-specific immunostimulants.

Stimulatory effects of aluminium hydroxide, lipopolysaccharide (LPS), muramyldipeptide (MDP), and empty liposomes on the antigenicity of inactivated bovine herpesvirus 1 were tested in mice. Compared with the standard effect of aluminium hydroxide, stronger antibody responses were observed in mice treated with empty liposomes or LPS alone, or a combination thereof. The strongest antibody response was recorded in mice treated with a combination of inactivated BHV-1, MDP and empty liposomes.

Acetylmuramyl-Alanyl-Isoglutamine

Immunogenicity of infectious bovine rhinotracheitis virus (BHV-1) proteins integrated into ISCOMs or liposomes.

Dynamics and persistence of antibody responses to immunization and reimmunization of rabbits with BHV-1 ISCOMs or BHV-1 liposomes were investigated for 20 weeks and compared with those induced by a commercial vaccine against infectious bovine rhinotracheitis (IBR) or free BHV-1 protein subunits. Comparable levels of antibodies to BHV-1, determined by ELISA, were found throughout the observation period in rabbits treated with BHV-1 ISCOMs or the commercial vaccine. The immunogenicity of BHV-1 liposomes was higher than that of the free virus proteins, but lower than those of BHV-1 ISCOMs or the commercial vaccine. Virus neutralization test confirmed the best immunostimulatory effect of BHV-1 ISCOMs as far as the levels and the persistence of antibodies were concerned. The reactivities of the specific virus protein subunits with anti-BHV-1 antibodies elicited in rabbits immunized with BHV-1 ISCOMs or the commercial vaccine were compared by Western blotting. The patterns of the bands were similar, but some differences were found in their numbers and intensities.

Animals

Comparative between-laboratory trials of the liquid-phase blocking sandwich ELISA for the detection of antibodies to foot-and-mouth disease virus.

Fifty bovine serum samples were tested for the presence or amounts of antibodies to foot-and-mouth disease (FMD) virus serotypes A, O and C by the liquid-phase blocking sandwich ELISA (lpb-ELISA) using reagents prepared by the World Reference Laboratory for Foot-and-Mouth Disease (WRL) in Pirbright, U.K. Twenty of the sera had been collected before extensive vaccination with a commercial inactivated trivalent FMD vaccine was ceased and the remaining thirty originated from animals which had not been vaccinated for more than one year. After the test had been completed, the samples were sent to another two laboratories to be examined by the same assay. Results obtained in the laboratories were compared to assess the degree of agreement in serological tests for FMD in cattle. Antibodies to at least one of the three FMD virus (FMDV) serotypes were demonstrated in 70% of the serum samples. Antibodies to the antigens A5, O1 and C1 were present in 58%, 66% and 58% of the sera, respectively. The overall between-laboratory agreement of the results of the lpb-ELISA for the detection of antibodies against all three serotypes was 96%. Discrepancies in terms of type specificity did not exceed 10%. Most discrepancies were recorded in sera with low antibody titres or optical densities (OD) around the cut-off point. An increase of concentration of C1 antigen in reaction mixture reduced the sensitivity of ELISA and results of screening tests became negative in sera with antibody titres of 1: 90 and lower.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Comparison of the primary and secondary antiphospholipid syndrome: a European Multicenter Study of 114 patients.

PURPOSE: To determine whether the features of the antiphospholipid syndrome (APS) are in any way influenced by the presence or absence of systemic lupus erythematosus (SLE). We followed up patients with 'primary' APS (PAPS) and APS secondary to SLE (APS plus SLE) with the objective of comparing laboratory and clinical events and of determining whether patients with PAPS would have evolution to SLE. PATIENTS AND METHODS: A total of 114 patients from 3 European referral centers were included in this study. Fifty-six had APS plus SLE and 58 had PAPS. Laboratory and clinical data were collected during an average 2-year period. RESULTS: Patients with PAPS and patients with APS plus SLE had similar clinical and laboratory profiles, with the exceptions of autoimmune hemolytic anemia, endocardial valve disease, neutropenia, and low C4 levels, all of which occurred more frequently in patients with APS plus SLE (p values: < 0.05, < 0.005, < 0.01, and < 0.001, respectively). On follow-up, 10 thrombotic episodes occurred in 10 patients, 8 of whom were receiving anticoagulant therapy. No patient with PAPS had either anti-DNA or anti-extractable nuclear antigen antibodies, and these patients had a significantly lower prevalence of antinuclear antibodies (41%) than patients with APS plus SLE (89%). CONCLUSIONS: Patients with APS plus SLE and PAPS have similar clinical profiles, although heart valve disease, hemolytic anemia, low C4 levels, and neutropenia seem to be more common in patients with APS plus SLE. Patients with APS may develop further thrombotic events despite anticoagulation therapy.

Adult

[Antibodies to anticoagulants in rheumatic autoimmune diseases].

The systemic lupus erythematosus (SLE) and the rheumatoid arthritis (RA) as the classic autoimmune diseases exhibit a great number of autoantibodies. Some of them are anticoagulants. Besides inactivating inhibitors against single coagulation factors interfering anticoagulants are known, belonging to the group of anti-phospholipid antibodies and detected as the lupus anticoagulants or anticardiolipin antibodies. Anti-phospholipid antibodies 184 patients with SLE or RA had been checked for. An enzyme immuno assay was used for detection of the anti-cardiolipin antibodies. The relations between occurrence of the anti-cardiolipin antibodies and vascular processes as well as other immunologic parameters had been tested for clinical relevancy.

Adult

Labeling of antibodies with 64Cu using a conjugate containing a macrocyclic amine chelating agent.

The conjugation of a model antibody (rabbit-anti-HSA-IgG) with a functionalized derivative of cyclam (1-(3-aminopropyl)-4-methyl-1,4,8,11-tetraazacyclotetradecane) is reported. Coupling of this derivative to the antibodies was accomplished using commercially available heterobifunctional coupling agents. The conjugation technique produced minimal effects on the antibody binding activity. 64Cu complexes with this cyclam derivative have excellent stability in human serum and aqueous solutions containing 1 mM EDTA. The high stability of these Cu-complexes suggests that this system, or other analogous systems, may be useful for production of stable 67Cu-immunotherapeutic agents.

Copper Radioisotopes

[A new polyvalent vaccine against enteral infections in newborn calves].

The most frequent microbial causative agents of massive diarrheas in new-born calves kept on large cattle farms in the CSSR are rotaviruses, coronaviruses and enterotoxigenic strains of E. coli, manifesting themselves as complicated virus-bacterial infections. An inactivated polyvalent adjuvant vaccine has been developed for the prevention and specific prophylaxis of these enteral infections; the vaccine contains bovine rotavirus, bovine coronavirus and three enterotoxigenic serotypes of E. coli with protective antigen K 99. The rotavirus and coronavirus are propagated on the stable cellular line MDBK and inactivated with 0.2% formalin, the Escherichia strains are submersed in the MINCA culture medium during their cultivation and inactivated with 0.5% formalin. The vaccine was prepared as a blend of the same amounts of rotavirus and coronavirus and of such an amount of bacterin so that 1 ml of the vaccine will contain 10(9) bacteria. One part of oil adjuvant was added to five parts of the virus-bacterial blend and the blend was homogenized in the Ultraturax apparatus. The vaccine is to be used for immunization of pregnant cows and heifers; in these animals it induces the production of specific antibodies to all antigens contained in the vaccine. Its immunogenic effects were checked in 32 calves and 38 cows in the herds with the occurrence of diarrheas caused by both enteropathogenic viruses and enterotoxigenic escherichia. It was demonstrated that the inactivation did not influence in either of the viruses the process of inducing the production of specific antibodies, and the antibody response of the calves and heifers after application of 2 ml of complete inactivated vaccine was equally strong as after application of live vaccine containing only rotavirus and coronavirus. The level of the rotavirus antibodies increased on the average 30 times and 200 times, coronavirus antibodies twice and four times. The antibody response to coronavirus was negatively influenced by the relatively high levels of antibodies before vaccination. The antibody response to antigen K 99 was expressive in all cases.

Animals

[Experimental and natural infection with the enzootic leukosis virus of cattle].

A trial was performed with heifers at the age of six to seven months. The animals were experimentally infected with the lymphocytes of a virus-productive donor. Infection was produced in all the nine cases, as demonstrated by means of the positive syncytial test. As indicated by the results of the trial, the antibodies to the enzootic bovine leucosis virus (BLV) were produced soon after experimental infection. A high sensitivity of the serum-neutralization test and the ELISA method was demonstrated in this connection: by these methods, the antibodies were identified already two to three weeks after experimental infection whereas by the immunodiffusion test they could be detected only after five weeks. Twenty-four animals were exposed to natural contact infection. Within 270 days of the trial, the disease after contact was recorded only in one heifer out of the four that were in close contact with the experimentally infected animals. In this case, as compared with experimental infection, the antibodies were produced much later--after 85 to 93 days. Leucosis was recorded in none of the remaining animals. The reasons why such a favourable result was obtained were the thorough disinfection of the stables after blood collections and the strict observance of the aseptic conditions. The results of experimental infection in three cows were identical with those obtained in young cattle. In the experimentally infected dairy cows, antibodies in milk were determined by the ELISA method. As found, in milk the antibodies to BLV appear two to three weeks later than they do in serum. The ELISA method of BLV antibody detection can be used for the identification of infected animals in herds where enzootic bovine leucosis occurs.

Animals